Understanding the Risk of Bat Coronavirus Emergence

RESEARCH Department of Health and Human Services National Institutes of Health Notice of Award Issue Date: 05/27/2014 NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 1 RO 1 Al 110964-01 FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Aleksei President 460 West 34th Street 17th Floor New York, NY 100012317 Award e-mailed to: (b)(6) ----------- Budget Period: 06/01/2014-05/31/2015 Project Period: 06/01/2014 - 05/31/2019 Dear Business Official: The National Institutes of Health hereby awards a grant in the amount of $666,442 (see "Award Calculation" in Section I and ''Terms and Conditions" in Section 111) to ECOHEALTH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the "Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01Al110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STTR} awards. Consult the NIH website http://qrants.nih.gov/qrants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1

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Laura A. Pone Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2

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SECTION I -AWARD DATA -1 R01Al110964-01 Award Calculation (U.S. Dollars) Salaries and Wages Fringe Benefits Supplies Travel Costs Other Costs Consortium/Contractual Cost Federal Direct Costs Federal F&A Costs Approved Budget Federal Share TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS $167,708 $54,168 $21,400 $35,918 $10,000 $227,663 $516,857 $149,585 $666,442 $666,442 $666,442 $666,442 YR THIS AWARD CUMULATIVE TOTALS 1 $666,442 $666,442 2 $630,445 $630,445 3 $611,090 $611,090 4 $597,112 $597,112 5 $581,646 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project Fiscal Information: CFDA Number: EIN: Document Number: PMS Account Type: Fiscal Year: IC CAN 2014 93.855 1311726494A1 RAl110964A P (Subaccount) 2014 2015 Al 8472350 $666.442 $630.445 2016 2017 2018 $611.090 $597,112 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project NIH Administrative Data: PCC: M51 C / OC: 414A / Released: (b)(6) 05/20/2014 Award Processed: 05/08/2014 01 :52:21 PM SECTION 11- PAYMENT/HOTLINE INFORMATION-1R01Al110964-01 For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS-1R01Al110964-01 This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. Page-3

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b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. c. 45 CFR Part 74 or 45 CFR Part 92 as applicable. d. The NIH Grants Policy Statement, including addenda in effect as of the beginning date of the budget period. e. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. (See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain references cited above.) An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the Central Contractor Registration. Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://qrants.nih.gov/grants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01Al110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://qrants.nih.gov/qrants/policy/awardconditions.htm for additional award applicability information. In accordance with P.L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OD-08-033 and the Public Access website: http://publicaccess.nih.gov/. Treatment of Program Income: Additional Costs SECTION IV- Al Special Terms and Conditions -1R01Al110964-01 THIS AWARD CONTAINS GRANT SPECIFIC RESTRICTIONS. THESE RESTRICTIONS MAY ONLY BE LIFTED BY A REVISED NOTICE OF AWARD. RESTRICTION: This award is issued with the knowledge that subjects may be involved within the period of support, but definite plans were not set forth in the application as per 45 CFR 46.118. No human subjects may be involved in any project supported by this award until all requirements for Human Subjects research as identified in the PHS398/SF424 Instructions have been provided to and approved by NIH. RESTRICTION: The present award is being made without a currently valid certification of IRB approval for this project with the following restriction: Only activities that are clearly severable and independent from activities that involve human subjects may be conducted pending the NIAi D's acceptance of the certification of IRB review and approval. No funds may be drawn down from the payment system and no obligations may be made against Federal funds for any research involving human subjects prior to the NIAi D's notification to the grantee that the identified issues have been resolved and this restriction removed. Page-4

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This award includes funds for subcontract/consortium activity with Wuhan Institute of Virology, CHINA and is budgeted as follows: -Yr1 -Yr2 Total Direct Costs F&A Costs @ 8%(MTDC) TOTAL COSTS $123,699 $128,718 $9,896 $10,297 $133,595 $139,015 -Yr 3 $147,335 $11,787 $159,122 -Yr 4 $147,335 $11,787 $159,122 -Yr 5 $147,335 $11,787 $159,122 Consortiums are to be established and administered as described in the NIH Grants Policy Statement. This written agreement with the consortium must address the negotiated arrangements for meeting the scientific, administrative, financial, and reporting requirements for this grant. This award includes funds for subcontract/consortium activity with East China Normal University. CHINA and is budgeted as follows: -Yr 1 Total Direct Costs F&A Costs @ 8%(MTDC) TOTAL COSTS $87,100 $6,968 $94,068 -Yr 2 $67,300 $5,384 $72,684 -Yr 3 $50,108 $4,009 $54,117 -Yr 4 $39,167 $3,133 $42,300 -Yr 5 $14,850 $2,404 $32,454 Consortiums are to be established and administered as described in the NIH Grants Policy Statement. This written agreement with the consortium must address the negotiated arrangements for meeting the scientific, administrative, financial, and reporting requirements for this grant. Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.selectagents.gov/Regulations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm). Research funded under this grant must adhere to the BMBL, including using the BMBL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. Page-5

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If work with a new or additional Agent(s)/Toxin(s) is proposed in the upcoming project period, provide: o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent(s), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted, with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. STAFF CONT ACTS The Grants Management Specialist is responsible for the negotiation, award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Laura A. Pone Email: (b)(6) Phone: (b)(6) Fax: 301-493-0597 Program Official: Erik J. Stemmy Email: (b) (6) Phone: (b)(6) ----- SPREADSHEET SUMMARY GRANT NUMBER: 1R01Al110964-01 INSTITUTION: ECOHEALTH ALLIANCE, INC. Budqet Year 1 Year 2 Year3 Year4 Year 5 Salaries and Waqes $167,708 $167,708 $167,708 $167,708 $167,708 Frinqe Benefits $54,168 $54,168 $54,168 $54,168 $54,168 Suoolies $21,400 $19,250 $7,250 $7,000 $3,500 Travel Costs $35,918 $35,918 $35,918 $35,918 $35,918 Other Costs $10,000 $13,550 $11,050 $9,800 $9,400 Consortium/Contractual Cost $227,663 $211,699 $213,239 $201,422 $191,576 TOTAL FEDERAL DC $516,857 $502,293 $489,333 $476,016 $462,270 TOTAL FEDERAL F&A $149,585 $128,152 $121,757 $121,096 $119,376 TOTAL COST $666,442 $630,445 $611,090 $597,112 $581,646 Facilities and Administrative Year1 Year2 Year 3 Year4 Year 5 Costs F&A Cost Rate 1 44.1% 44.1% 44.1% 44.1% 44.1% F&A Cost Base 1 $339,194 $290,594 $276,094 $274,594 $270,694 F&A Costs 1 $149,585 $128,152 $121,757 $121,096 $119,376 Page-6

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Pl: DASZAK, PETER Title: Understanding the Risk of Bat Coronavirus Emergence Received: 06/05/2013 FOA: PA11-260 Council: 01 /2014 Competition ID: ADOBE-FORMS-B2 FOA Title: RESEARCH PROJECT GRANT (PARENT R01) 1 R01 Al110964-01 IPF: 4415701 Former Number: IRG/SRG: CRFS Subtotal Direct Costs (excludes consortium F&A) Year 1: 499,993 Year 2: 499,469 Year 3: 499,978 Year 4: 499,953 Year 5: 499,974 Senior/Key Personnel: Peter Daszak Zhengli Shi ShuYiZhang Changwen Ke Jonathan Epstein Kevin Olival Parviez Hosseini XingYi Ge Guanjin Zhu Yun-Zhi Zhang Additions for Review Accepted Publication Dual: Accession Number: 3595101 Organization: ECOHEAL TH ALLIANCE, INC. Department: AIDS:N Animals: Y Humans: Y Clinical Trial: N (b) (4) Current HS Code: HESC:N Organization: EcoHealth Alliance, Inc. Wuhan Institute of Virology East China Normal University CDC and Prevention of Guangdong Province EcoHealth Alliance EcoHealth Alliance EcoHealth Alliance Wuhan Institute of Virology Guangdong Entomological Institute Yunnan Center for Disease Control News of manuscripts acceptance Expedited: N New Investigator: N Early Stage Investigator: N Role Category: PO/Pl Co-Investigator Co-Investigator Co-Investigator Co-Investigator Co-Investigator Co-Investigator Co-Investigator Co-Investigator Co-Investigator

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0MB Number: 4040·0001 Expiration Date: 06/30/2011 APPLICATION FOR FEDERAL ASSISTANCE SF 424 (R&R) 3. DATE RECEIVED BY STATE State Application Identifier I I I I 1. • TYPE OF SUBMISSION 4. a. Federal Identifier IGRANT11418218 I D Pre-application D Application IZI Changed/Corrected Application b. Agmy Ro"Uog ldeotm" I I 2. DATE SUBMITTED Applicant Identifier I 06/05/2013 I I I 5. APPLICANT INFORMATION * Organizational DUNS: 10770900660000 I • Legal Name: IEcoHealth Alliance, Inc. I Department: I I Division: I I • Street1: 1460 West 34th Street I Street2: Ii 7th Floor I * City: INew York I County/ Parish: I I • State: I NY: New York I Province: I I • Country: I USA: UNITED STATES I ·ZIP/ Postal Code: ltoOOl-2317 I Person to be contacted on matters involving this application Prefix: lor. I • First Name: !Peter I Middle Name: I I • Last Name: loaszak I Suffix: I I • Phone Number:' (b) (6)] I Fax Number: 1+1. 212. 380. 4465 I Email:( (bf@ I 6. • EMPLOYER IDENTIFICATION (EIN) or (TIN): ls11726494 I 7. * TYPE OF APPLICANT: I M: Nonprofit with 501C3 lRS Status (Other than Institution of Higher Education) I Other (Specify): I I Small Business Organization Type D Women Owned D Socially and Economically Disadvantaged 8. • TYPE OF APPLICATION: If Revision, mark appropriate box(es). IZ! New D Resubmission DA. Increase Award DB. Decrease Award DC. Increase Duration oo. Decrease Duration D Renewal D Continuation Revision DE. Other (specify):! I • Is this application being submitted to other agencies? YesO No~ What other Agencies? I I 9. * NAME OF FEDERAL AGENCY: 10. CATALOG OF FEDERAL DOMESTIC ASSISTANCE NUMBER:! I National Institutes of Health I TITLE: I 11. • DESCRIPTIVE TITLE OF APPLICANT'S PROJECT: !Understanding the Risk of Bat Coronavirus Emergence I 12. PROPOSED PROJECT: '13. CONGRESSIONAL DISTRICT OF APPLICANT • Start Date • Ending Date I 10/01/2013 11 09/30/2018 I INY-010 I 14. PROJECT DIRECTOR/PRINCIPAL INVESTIGATOR CONTACT INFORMATION Prefix: lor. I • First Name: !Peter I Middle Name: I I • Last Name: loaszak I Suffix: I I Position/Title: IP resident I • Organization Name: IEcoHealth Alliance, Inc. I Department:! I Division: I I • Street1: 1460 West 34th Street I Street2: 11 7th Floor I • City: !New York I County/ Parish: I I • State: I NY: New York I Province: I I • Country: I USA: UNITED STABS 1 •ZIP/ Postal Code: !10001-2317 I • Phone Number:~ Cl.>) (6}j I Fax Number: 1+1. 212. 380. 44 65 I • Email: l (b)(6}J I Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-1I-260 Received Date:20l3-06-05Tl8:36:48-04:00

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SF 424 (R&R) APPLICATION FOR FEDERAL ASSISTANCE Page 2 15. ESTIMATED PROJECT FUNDING 16. * IS APPLICATION SUBJECT TO REVIEW BY STATE EXECUTIVE ORDER 12372 PROCESS? a. Total Federal Funds Requested 13,362,338.00 a. YES 0 THIS PREAPPLICATION/APPLICATION WAS MADE AVAILABLE TO THE STATE EXECUTIVE ORDER 12372 PROCESS FOR REVIEW ON: b. Total Non-Federal Funds lo.oo DATE: I I c. Total Federal & Non-Federal Funds 13,362,338.00 b. NO 0 PROGRAM IS NOT COVERED BY E.O. 12372; OR d. Estimated Program Income la. ao ~ PROGRAM HAS NOT BEEN SELECTED BY STATE FOR REVIEW 17. By signing this application, I certify (1) to the statements contained in the list of certifications• and (2) that the statements herein are true, complete and accurate to the best of my knowledge. I also provide the required assurances • and agree to comply with any resulting terms if I accept an award. I am aware that any false, fictitious. or fraudulent statements or claims may subject me to criminal, civil, or administrative penalities. (U.S. Code, Title 18, Section 1001) ~ • 1 agree • The I/st of certifications and assurances, or an Internet site where you may obtain this list, Is contained in the announcement or agency specific Instructions. 18. SFLLL or other Explanatory Documentation I I Add Attachment I Delete Attachment I View Attachment I 19. Authorized Representative Prefix: IMr. I 'First Name: IAleksei I Middle Name: I I • Last Name: lchmur.a I Suffix: I I • Position/Title:IAuthorized Organizational Representacive I • Organization: !EcoHealth Alliance, Inc. I Department: I I Division: I 'Street1: 1460 ~lest 34th Street I Street2: 117th Floor I • City: §ew York I County I Parish: I I * State: I NY: New York I Province: I I • Country: I USA: UNITED STATES I ·ZIP/ Postal Code: 11 o o 01-2317 I • Phone Number:! (b)(, I FaK Number: !+1. 212. 380. 4465 I • Emai1:f (b)(6>) I • Signature of Authorized Representative • Date Signed I Winifred Zubin I I 06/05/2013 I 20. Pre-application I I Add Attachment I Delete Attachment I View Attachment I Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 424 R&R and PHS-398 Specific Table Of Contents SF 424 R&R Face Page-------------------·---- Table of Contents----------------------- Performance Sites------------------------ Research & Related Other Project lnformation----------------- Project Summary/Abstract (Description)•·---------- Public Health Relevance Statement (Narrative attachment---------- Facilities & Other Resources---- Research & Related Senior/Key Person-------------------- Biographical Sketches for each listed Senior/Key Person-------········-··· Research & Related Budget - Year 1-------------------- Research & Related Budget• Year 2------- Research & Related Budget - Year 3-------------·----·------ Research & Related Budget • Year 4---······----·---------- Research & Related Budget - Year 5---------------------- Budget Justification----······-----·····--··· Research & Related Budget - Cumulative Budget--------- Research & Related Budget • Consortium Budget (Subaward 1 ,__ ___________ _ Research & Related Budget - Consortium Budget (Subaward 2)------------------ PHS 398 Specific Cover Page Supplement-------- PHS 398 Specific Research Plan--------------------- Specific Aims---·····----······--····· Research Strategy---------- Human Subjects Sections----------- Protectlon of Human Subjects--------·---- Women &Minorities------ Planned Enrollment Table--------- Children---······-------- Vertebrate Anlmals----------- Select Agent Researcn----------- Bibllography & References Cited Consortium/Contractual----·-------······---··· Letters of Support--- Resource Sharing Plan--------- PHS 398 Checklist-------------------- Table of Contents Page Numbers 3 4 6 7 8 9 12 17 48 51 54 57 60 63 67 68 86 104 106 107 108 120 120 121 122 123 124 127 128 135 136 143 144 Page 3

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Project/Performance Site Location(s) 0MB Number: 4040-0010 Expiration Date: 08/31/2011 □ I am submitting an application as an individual, and not on behalf of a company, state, Project/Performance Site Primary Location local or tribal government. academia. or other type of organization. Organization Name: IEcoHealth Alliance, Inc. DUNS Number: 10770900660000 I • Street1: 14 60 West 34th Street Street2: 11 7th Floor • City: INew York County: * State: INY: New York Province: * Country: lusA: UNITED STATES * ZIP/ Postal Code: ~I l_0_0_0_l_-_2_3_1_7 __________ ~ • Project/ Performance Site Congressional District: JNY-01 O Project/Performance Site location 1 Organization Name: lwuhan Institute D I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. DUNS Number: 15290274740000 I • Street1: lxiao Hong Shan, No. 44 Street2: Jwuchang District • City: Jwuhan • State: of Virology County: Province: IHubei ::==============-----------~ * Country: JcHN: CHINA •ZIP/ Postal Code: ~14_3_0_0_7_1 __________ ~ • Project/ Performance Site Congressional District: ~I ____ ~ Project/Performance Site location a D I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: IEast China Normal University DUNS Number: 14209454 950000 I * Street1: J3663 Zhongshan Beilu Street2: * City: Jshanghai County: • State: Province: Jshanghai • Country: JcHN: CHINA *ZIP/ Postal Code: ._12_0_0_0_6_2 ____________ _, • Project/ Performance Site Congressional District: ,_J ____ ___. Performance Sites Page 4 Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Project/Performance Site Location(s) Project/Performance Site Location 3 D I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: Yunnan Institute of Endemic Diseases Control and Prevention DUNS Number: * Street1: J33 Wenhua Road Street2: * City: Joali • State: Province: J._Y_u_n_n_a_n _____________ ~ * Country: JcHN: CHINA County: •ZIP/ Postal Code: ._I 6_s_o_2_0_1 ____________ __, • Project/ Performance Site Congressional District: .... J ____ __, Project/Performance Site Location a D I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: Center for Disease Control and Prevention of Guangdong DUNS Number: • Street1: J176 Xigang Xilu Street2: * City: JGuangzhou * State: Province: JGuangdong * Country: JcHN: CHINA County: • ZIP/ Postal Code: .... ls_1_0_3_o_o_o ___________ ___, • Project/ Performance Site Congressional District: ,_J ____ __, Additional Location(s) Add Attachment Delete Attachment View Attachment Performance Sites Page 5 Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED Other Project Information 1. • Are Human Subjects Involved? [:g] Yes 1.a If YES to Human Subjects Is the Project Exempt from Federal regulations? D Yes [:g] No If yes, check appropriate exemption number. D 1 D 2 D 3 D 4 D 5 D 6 If no, is the IRS review Pending? 1:8] Yes IRS Approval Date: ~----;::====::'._ ___ ~ Human Subject Assurance Number: 2. * Are Vertebrate Animals Used? [:g] Yes 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? D Yes [:gj No IACUC Approval Date: I 1 o / 15/ 2 o 1 o '------;===-----------, Animal Welfare Assurance Number ~IA_3_4_3_3_-_0_l___ ~ 3. * Is proprietary/privileged information included in the application? D Yes [:gj No 4.a. • Does this project have an actual or potential impact on the environment? D Yes 4.b. If yes, please explain: (8] No ~---------------------------------------------~ 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an environmental assessment (EA) or environmental impact statement (EIS) been performed? D Yes D No 4.d. If yes, please explain: 5. • Is the research performance site designated, or eligible to be designated, as a historic place? 5.a. If yes, please explain: D Yes (8] No ~--------------------------------------------~ 6. * Does this project involve activities outside of the United States or partnerships with international collaborators? [:gj Yes 6.a. If yes, identify countries: China ~--------------------------------------------~ 6.b. Optional Explanation: ~---------------------------------------------~ 7. • Project Summary/Abstract l12 34-NIAID_cov_P ro ject_Summary2 o 13. p~ Add Attachment I Delete Attachment I View Attachment 8. • Project Narrative 11235-NIH_COv_Pro ject_Na rrati ve. pdf Add Attacl1ment j Delete Attachment I View Attachment I 9. Bibliography & References Cited l1236-NIAID_cov_2013_Bibliography .pdf I Add Attachment I Delete Attachment j View Attachment 10. Facilities & Other Resources 11237-FACILITIES_AND_OTHER_RESOURCES. ~ I Add Attachment 11 Delete Attachment I View Attachment 11. Equipment ~---------------~ Add Attachment I Delete Attachment I View Attachment 12. Other Attachments Add Attachments Delete Attachments I View Attachments I D Other Information Page 6 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05TJ8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Project Summary This project will examine the risk of future coronavirus (CoV) emergence from wildlife using in-depth field investigations across the human-wildlife interface in China, molecular characterization of novel CoVs and host receptor binding domain genes, mathematical models of transmission and evolution, and in vitro and in vivo laboratory studies of host range. Zoonotic CoVs are a significant threat to global health, as demonstrated with the emergence of pandemic severe acute respiratory syndrome coronavirus (SARS-CoV) in China in 2002, and the recent and ongoing emergence of Middle East Respiratory Syndrome (MERS-CoV). Bats appear to be the natural reservoir of these viruses, and hundreds of novel bat-Co Vs have been discovered in the last two decades. Bats, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel CoVs. This project aims to understand what factors increase the risk of the next CoV emerging in people by studying CoV diversity in a critical zoonotic reservoir (bats), at sites of high risk for emergence (wildlife markets) in an emerging disease hotspot (China). The three specific aims of this project are to: 1. Assess CoV spillover potential at high risk human-wildlife interfaces in China. This will include quantifying the nature and frequency of contact people have with bats and other wildlife; serological and molecular screening of people working in wet markets and highly exposed to wildlife; screening wild-caught and market sampled bats from 30+ species for CoVs using molecular assays; and genomic characterization and isolation of novel CoVs. 2. Develop predictive models of bat CoV emergence risk and host range. A combined modeling approach will include phylogenetic analyses of host receptors and novel CoV genes (including functional receptor binding domains); a fused ecological and evolutionary model to predict host-range and viral sharing; and mathematical matrix models to examine evolutionary and transmission dynamics. 3. Test predictions of CoV inter-species transmission. Predictive models of host range (i.e. emergence potential) will be tested experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments across a range of cell cultures from different species and humanized mice. Project Description Page 7

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter PROJECT NARRATIVE Most emerging human viruses come from wildlife, and these represent a significant threat to global public health and biosecurity - as demonstrated by the SARS coronavirus pandemic of 2002-03 and an ongoing SARS-like epidemic in the Middle East. This project seeks to understand what factors allow animal Coronaviruses to evolve and jump into the human population by studying virus diversity in a critical group of animals (bats), at sites of high risk for emergence (wildlife markets) in an emerging disease hotspot (China). Public Health Relevance Statement Page 8

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter Daszak, Peter FACILITIES AND OTHER RESOURCES EcoHealth Alliance, New York, USA (Peter Daszak, Jon Epstein, Parviez Hosseini, and Kevin Olival) EcoHealth Alliance is a 40-year old scientific research NGO that specializes in multidisciplinary research on the causes, origins and spread of zoonotic emerging diseases. EcoHealth Alliance scientists have been working on the emergence of Nipah & Hendra virus, SARS CoV, surveillance for zoonotic agents in wildlife, and spatial modeling for over 15 years, and on modeling of infectious disease emergence and spread for over a decade. EcoHealth Alliance is based in New York City with CbH4> square feet of office space including a meeting room and basic laboratory - freezer storage and light microscopy. The scientific staff (25 core scientists, 100+ field staff) is supported by a core ad min staff of 11 which is available for work on this project and is funded through core funds. EcoHealth Alliance is equipped with 35 networked PCs including an NIH ARRA-funded International LifeSize Video Conferencing facility. High-speed video conferencing facilities have been installed with key international collaborators in 2011. EcoHealth Alliance has access to a 24-7 server, server support, and all required software including ArcGIS ArclNFO, Matlab, SPSS, R, Microsoft Office, and Adobe CS5 running on both Apple and Windows Operating Systems. Additionally we have a four-processor, public IP addressed Linux and an eight-processor Mac Pro Server - each with 4TB hard drives, which in combination can be used for intensive computational modeling and database processing by all the grantees. Access to the cloud (Amazon) is provided by core funding to EHA. EcoHealth Alliance is the headquarters of a series of global networks that provide exceptional leverage for the core scientists: 1) The USAID EPT PREDICT consortium. This group conducts human and wildlife surveillance for high-risk pathogens in 24 countries including China. Partners include Dr Ian Lipkin's Center for Infection and Immunity at Columbia University (pathogen discovery), UC Davis, The Smithsonian, GVFI and WCS; 2) The One Health Alliance of South Asia. This is a Rockefeller Foundation funded transboundary disease surveillance program; 3) The EcoHealth Alliance Partners: A global partnership of leading wildlife and global health researchers in tropical and subtropical countries. This gives us unique access to working on-theground in countries where surveillance is difficult, such as China, where our group has proven capacity to export samples from; 4) The Consortium for Conservation Medicine: A unique collaborative institution linking Johns Hopkins Bloomberg School of Public Health, Tufts University School of Veterinary Medicine, The University of Pittsburgh Graduate School of Public Health, The University of Wisconsin-Madison Nelson Institute for Environmental Studies, The USGS National Wildlife Health Center, and EcoHealth Alliance. The CCM provides access to hundreds of high caliber scientists, their facilities, Facilities Page 9

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter Daszak, Peter and their students at 6 leading institutes of public health, veterinary medicine, and environmental science in the USA. East China Normal University, Shanghai, China (Shuyi Zhang and Guangjian Zhu) Dr. Zhang is Dean of the Institutes for Advanced Interdisciplinary Research at East China Normal University. Over Cb) <4J square metres is allocated to his research group at ECNU. The lab is fully capable of carrying out molecular, protein, epidemic disease and evolution research. Experimental equipment includes: Roche 454 (GS FLX Titanium System), Bioinformatics Computer Server, Multi-Channel Neurophysiology Workstation TOT, PCR Amplifier, Real time PCR Amplifier, Electrophoresis, Ultra-low Temperature Freezer, Centrifuge, UV-Visible Spectrophotometer, Two-dimensional Electrophoresis, Vertical Electrophoresis System, Incubator, Clean Bench, and Class 11-Biosafety Cabinet, Hybridization Oven, Water PurificationSystem, and Shaker. Wuhan Institute of Virology, Wuhan, China (Zhengli Shi and Xingyi Ge) The Shi laboratory includes 4 rooms totaling CbH4 , one equipped with two CO2 incubators for tissue culture, one equipped facilities including with high speed centrifijye; 2 -20°C, 3 -80°C freezers, 2 PCR machines, 1 ELISA plate reader, one for molecul~ diagnosis equipped with two biosafety cabinets, and one normal laboratory equipped various small equipment items (mini-centrifuges, gel electrophoresis units, circulating adjustable water baths, and heat blocks). Also available to Dr. Shi's group is a fully equipped biosafety level 3 laboratory, a newly opened BLS-4 laboratory (the first in China) and Institute-supported facility center, which houses full-time staff and equipment for electronic microscopy, ultracentrifugation, confocal microcopy, and sequencing machine. The Wuhan Institute of Virology is China's premier institute for virological research. It consists of three research departments and one center: Department of Molecular Virology, Department of Bio-control, Department of Analytical Biochemistry and Biotechnology, and the Virus Resource and Bioinformation Center of China. It also has the Key Laboratory of Molecular Virology of CAS, the Joint-laboratory of Invertebrate Virology, a HIV Pre-screening Lab and Hubei Engineering and Technology Research Center for Viral Diseases. The institute is further divided into 14 research groups, one of which is run by Dr Zhengli Shi. The supporting system of the institute consists of an Analytical Equipment Center, an Experimental Animal Center, an Editorial Office of "Virologica Sinica"and an Computer Network Center. The virus resource and bioinformation center of China contains the largest virus bank in Asia, curating around 800 viral strains. The Institute collaborates with the World Health Organization (WHO), universities and research institutes in more than 30 counties and regions including EcoHealth Alliance in the USA. There are 14 professors, 36 associated professors, 47 assistant professors conducting research on virology and five of these have been awarded honors in the "Hundred Talents Project". The institute has built a BSL-3 lab and a 600 m2 experimental animal center. In 2013, the first BSL-4 lab in China was opened at this Facilities Page 10

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Daszak, Peter Institute in a purpose-built facility which has been designed with the assistance of the US Centers for Disease Control and the Pasteur Institute. Facilities Page 11

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 0MB Number: 4040-0001 Expiration Date: 06/30/2011 RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE - Project Director/Principal Investigator Prefix: lor. I • First Name:IPeter I Middle Name: I I • Last Name: loaszak I Suffix: I I Position/Title: IP resident I Department: I I Organization Name: IEcoHeal th Alliance, lnc. I Division:! • Street1: 1460 West 34th Street I Street2: 11 7th Floor I • City: INew York I County/ Parish: I I • State: I NY: New York I Province: I I • Country: I USA: UNITED ST1'.TES I •Zip/Postal Code: l10001-2317 I • Phone Number: i: (b) (6)) I Fax Number: 1+1. 212. 380. 44 65 I * E-Mail:t (b)(6J: I Credential. e.g., agency login:[ (b)(©'. I • Project Role: IPD/PI I Other Project Role Category: I I Degree Type: IPh.D. I Degree Year: 11994 I *Attach Biographical Sketch 11244-Peter_Daszak_NIH_bioske~ Add Attachment I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Attachment l View Attachment I PROFILE - Senior/Key Person 1 Prefix:lor. I • First Name:lzhengLi I Middle Name: I I • Last Name: lshi I Suffix:! I Position/Title: Jsenior Scientist I Department: I I Organization Name: Jwuhan Inst it utc of Virology I Division:! • Street1: Jxiao Hong Shan, no. 44 I Street2: l•/\Juchang District I * City: Jwuhan I County/ Parish: I I • State: I I Province: Jttubei I • Country: I CHN: CHINA l •Zip/ Postal Code: J430071 I • Phone Number( (b)(~ I Fax Number: 1+86-27-87198072 I • E-Mail:11 (b) (6}, I Credential, e.g., agency login:I I • Project Role: lea-Investigator I Other Project Role Category: I I Degree Type: !Ph.D. I Degree Year: 12000 I •Attach Biographical Sketch l1245-SHI_Zhengli_Biosketch_2½ Add Attachmen1 I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Allachrnent I View Auacrment I Key Personnel Page 12 Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE • Senior/Key Person 2 Prefix:lor. I • First Name:lshuYi I Middle Name: I I • Last Name: lzhang I Suttix:I I Positionmtle: loean I Department: I I Organization Name: IEast China Normal University I Division:! • Street1: IB319, Science Building 3663 I Street2: !North Zhongshan Road I • City: I shanghai I County/ Parish: I I • State: I I Province: I shanghai I • Country: I CHN: CHINA I •Zip/ Postal Code: 1200062 I • Phone Number:E (b)(~ I Fax Number: I I • E-Mail:t (b) (6)) I Credential, e.g., agency login: I I • Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: ll?h.D. I Degree Year: 11994 I *Attach Biographical Sketch 1124 6-Zhang_Shuyi_Biosketch_K} . Add Allachmenl I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Allachinenl] View Attact>ment I PROFILE - Senior/Key Person 3 Prefix: lor . I • First Name:lchangwen I Middle Name: I I • Last Name: IKe I Suffix: I I Position/Title: lo1recto.r: I Department: I I Organization Name: lcoc and Prevention of Guangdong Province I Division:! * Street1: lxing Gang West Road, no. 176 I Street2: I I • City: !Guangzhou I County/ Parish: I I * State: I I Province: IGuangdong I • Country: I CHN: CHINl\ I •Zip/ Postal Code: ls10300 I • Phone Numberi.. (b) (6)) I Fax Number: I I * E-Mail:t (b)(6)) I Credential, e.g., agency login:I I * Project Role: lco-Invostigator J Other Project Role Category: I I Degree Type: !Ph.D. I Degree Year: 12001 I 'Attach Biographical Sketch l1247-Biosketch_ChangWenKe_C01 Add Attachment I Delete Attachment I View Attachment I Attach Current & Pending Support I I 11 Add Attachment I Delete Attachment I View Atlachment I Key Personnel Page 13 Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE • Senior/Key Person 4 Prefix:lor. I • First Name:IJonathan I Middle Name: IH I • Last Name: !Epstein I Suttix:I I Position(fitle: !Associate Vice President I Department: !conservation Medicine I Organization Name: IEcoHealth Alliance I Division:! • Street1: 1460 W34th Street I Street2: j1 7th Floor I • City: INew York I County/ Parish: I I • State: I NY: New York I Province: I I • Country: I USA: UNITED STATES I •Zip/ Postal Code: 110001-2317 I • Phone Number{ (b)(6)i I Fax Number: I+ 1. 212. 380. 44 65 I • E-Mail:t (b)(ij I Credential, e.g., agency login: I I • Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: lovM I Degree Year: 12002 I *Attach Biographical Sketch I1248-Epstein_BioSketch_NIH -l Add Allachmenl I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Allachinenl] View Attact>ment I PROFILE - Senior/Key Person 5 Prefix: lor . I • First Name:IKevin I Middle Name: I I • Last Name: Ioli val I Suffix: I I Position/Title: lsenior Research scientist I Department: I I Organization Name: IEcoHealth Alliance J Division:! * Street1: 1460 W34th Street I Street2: j1 7th Floor I • City: !New York I County/ Parish: I I * State: I NY: New York I Province: I I • Country: I USA: UNITED STATES I •Zip/ Postal Code: 110001-231 7 I • Phone Number: Ir (b)(~ I Fax Number: 1+1. 212. 380. 44 65 I * E-Mail: I (b) (6)) I Credential, e.g., agency login:I I * Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: !Ph.D. I Degree Year: 12008 I 'Attach Biographical Sketch I1249-0lival_biosketch_NIAID_½ Add Attachment I Delete Attachment I View Attachment I Attach Current & Pending Support I I 11 Add Attachment I Delete Attachment I View Atlachment I Key Personnel Page 14 Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE • Senior/Key Person 6 Prefix:lor. I • First Name:IParviez I Middle Name: I I • Last Name: !Hosseini I Suttix:I I PositionrTitle: lsenior Research Scientist I Department: I I Organization Name: IEcoHealth Al Hance I Division:! • Street1: 1460 W 34th Street I Street2: j1 7th Floor I • City: INew York I County/ Parish: I I • State: I NY: New York I Province: I I • Country: I USA: UNITED STATES I •Zip/ Postal Code: 110001-2317 I • Phone Number{ (b) (6)) I Fax Number:!+ 1. 212. 380. 44 65 I • E-Mail:t (b)(6); I Credential, e.g., agency login: I I • Project Role: lea-Investigator I Other Project Role Category: I I Degree Type: ll?h.D. I Degree Year: 12002 I *Attach Biographical Sketch l12so-HOSSEINI_Biosketch_cov2~ . Add Allachmenl I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Allachinenl] View Attact>ment I PROFILE - Senior/Key Person 7 Prefix: lor . I • First Name:!xingYi I Middle Name: I I • Last Name: IGe I Suffix: I I Position/Title: !Assistant Researcher I Department: !oepan.rnenL of Emerging Viruses I Organization Name: lwuhan Institute of Virology I Division:! * Street1: jxiao Hong Shan, no. 44 I Street2: jwuchang District I • City: jwuhan I County/ Parish: I I * State: I I Province: jHubei I • Country: I CHN: CHINl\ I •Zip/ Postal Code: 1430071 I • Phone Number~ (b) (6)) I Fax Number: I I * E-Mail:t (b)(6)j I Credential, e.g., agency login:I I * Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: !Ph.D. I Degree Year: 12011 I 'Attach Biographical Sketch l12s1-GE_XingYi_Biosketch_201} Add Attachment I Delete Attachment I View Attachment I Attach Current & Pending Support I I 11 Add Attachment I Delete Attachment I View Atlachment I Key Personnel Page 15 Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE • Senior/Key Person 8 Prefix:lor. I • First Name:IGuanjin I Middle Name: I I • Last Name: lzhu I Suttix:I I Position(fitle: !Assistant Researcher I Department: I I Organization Name: lcuangdong Entomological Institute I Division:! • Street1: !Room 1707, Building 622, 3663 Zhongshanbei Rd I Street2: IPutuo District I • City: I shanghai I County/ Parish: I I • State: I I Province: I I • Country: I CHN: CHINA I •Zip/ Postal Code: 1200026 I • Phone Number( (b)(~ I Fax Number: I I • E-Mail:t (b)(6j) I Credential, e.g., agency login: I I • Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: ll?h.D. I Degree Year: 12012 I *Attach Biographical Sketch l12s2-zhu_Gunagjian_BiosketchJ . Add Allachmenl I Delete Attachment I View Attachment I Attach Current & Pending Support I I Add Attachment I Delete Allachinenl] View Attact>ment I PROFILE - Senior/Key Person 9 Prefix: lor . I • First Name:IYun-Zhi I Middle Name: I I • Last Name: lzhang I Suffix: I I Position/Title: laeact of Infectious Disease surveillance I Department: I I Organization Name: !Yunnan Center for Disease Control J Division:! * Street1: 133 Wenhua Road I Street2: I I • City: loali City I County/ Parish: I I * State: I I Province: I Yunnan I • Country: I CHN: CHINl\ I •Zip/ Postal Code: 1671000 I • Phone Number: ( (b)(4 I Fax Number: I I * E-Mail:t (b)(6j) I Credential, e.g., agency login:I I * Project Role: !co-Investigator I Other Project Role Category: I I Degree Type: !Ph.D. I Degree Year: 12010 I 'Attach Biographical Sketch 11253-Biosketch_YunZhiZhang_c} Add Attachment I Delete Attachment I View Attachment I Attach Current & Pending Support I I 11 Add Attachment I Delete Attachment I View Atlachment I Key Personnel Page 16 Tracking Number:GRANT11418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter Program Director/Principal Investigator (Last, First, Middle): Daszak, P. BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors in the order listed on Form Page 2. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME Peter Daszak eRA COMMONS USER NAME (credential, e.g., agency login) (b)(6) POSITION TITLE President & Chief Scientist EDUCATION/TRAINING (Begin with baccalaureate or dher initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MMIYY FIELD OF STUDY (if aoo/icable I Bangor University (UK) BSc. (hons) 07/86 Zoology University of East London (UK) Ph.D 03/93 Infectious Diseases A. Personal Statement The goal of the proposed research is to investigate the ecology, evolutionary biology and transmission dynamics of bat coronaviruses at the human-wildlife interface. Specifically, we will conduct field studies in China to obtain high quality samples from bats, and identify, characterize and isolate known and novel coronaviruses. We will analyze the patterns of coronavirus transmission among bats and other wildlife, and the risk of spillover to humans. I have been working on the dynamics of emerging viral diseases from wildlife for over 15 years, and have the proven scientific vision, leadership and capacity to lead this team and test the hypotheses laid out here. Since working at the CDC Pathology Activity in 1998 during the Nipah virus outbreak, I have specialized in the ecology of viruses emerging from bats. Under my first Nipah virus R01, I developed a multidisciplinary approach combining fieldwork, phylogenetics, virology, and mathematical modeling to isolate and characterize NiV from bats, analyze transmission dynamics, and identify the cause of its emergence. In 2001, I became director of a research program at a New York-based scientific research NGO. This allowed me to expand my research globally, and in 2005, working with current co-investigators Ors Zhang and Zhengli, we were the first team to identify and characterize SARS-like coronaviruses in bats. I have consolidated this work as Pl of: 1) a NIAID R01 to conduct pathogen discovery in bats, and map bat viral diversity; 2) a renewal to my Nipah virus R01 focused on the emergence of NiV in Bangladesh; and 3) a large USAID project (PREDICT) to identify new pathogens in wildlife from emerging disease 'hotspot' regions. The current application builds on this work and leverages my group's unique partnership in China, where we have proven capacity to conduct disease surveillance in humans and wildlife in the markets where SARS emerged, and where we have collaborated at a high level for 12 years. I have a proven record of leading multidisciplinary research teams on emerging viral pathogens from wildlife and have the leadership skills, institutional capacity and network to deliver successful outcomes in the current proposed work. B. Positions and Honors Positions and Em lo ment 1993-8 Senior Faculty Research Scientist, Kingston University 1998 1999-2001 2001- 2001-9 2009- Guest Researcher, Centers for Disease Control and Prevention (CDC) Faculty Research Scientist, University of Georgia Adjunct Faculty, Tufts Univ. Sch. Veterinary Med.; Univ. Georgia; Columbia Univ. Executive Director, Consortium for Conservation Medicine, EcoHealth Alliance, New York President & Chief Scientist. EcoHealth Alliance New York. PHS 398/2590 (Rev. 06/09) Page_ Biographical Sketch Format Page Biosketches Page 17

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Program Director/Principal Investigator (Last, First, Middle): Daszak, P. Other Experience and Professional Membership Keynote speaker Merieux Foundation Conference on Emerging paramyxoviruses, France (2000); UN Millenium Ecosystem Assessment: Lead Author, human infectious diseases (2006); NIH: ad hoc member, ZRG1 IDM-G 90 study section: Virology, Biodefense & Emerg. Diseases (2003-5); Editorial Board, Conservation Biology (Blackwell); Founding Co-Editor EcoHeafth (Springer) (2004-10); NAS - Committee Member, Future Needs in Veterinary Research (2004-5); DIVERSITAS (UNESCO-ICSU): Member of Scientific Committee (2004-11; Treasurer 2007-11 ); NIAID: Steering Committee, workshop on virus-host shifts & emergence of new pathogens (2005); Australian Biosecurity Cooperative Research Center: International Standing Advisory Committee (2005-10); NIH: ad hoc member, ZRG1 IRAP-Q study section (infectious diseases, epidemiology) (2005-7); International EcoHealth Association: Founding board of directors, Treasurer (2006-11); CDC: ad hoc member, ZCD1 SGI, 09PAR07-231, R36 Research Dissertation Awards (2007); European CDC: Keynote speaker, future infectious disease threats (2008); NAS-IOM Committee Member, Global capacity for EID surveillance (2008-9); Scientific Advisory Board, NIAID Center of Excellence, avian influenza (CRISAR), UCLA (2008-9); Reviewer IOM report on Infectious Disease Movements in a Borderless World (2009); NIAID: Steering Committee, workshop on viruses from bats (2009); NAS-IOM Participant, workshop on H1 N 1, Committee on Emerging Microbial Threats (2009); NIH: ZRG1 IRAP-Q Review panel ARRA Challenge grants (2009); Organizing Committee, 1st International One Health Symposium, Australia (2010); Member, Council of Advisors One Health Commission (2010-); Editor-in-Chief, EcoHeafth (2010-); Scientific Advisory Board, Oxford Univ. Clinical Research Unit, Vietnam (2010-); Member of IOM Forum on Microbial Threats 2010-; Steering Committeee, NIAID Workshop on Arboviruses 2011; Organizer IOM Forum on Microbial Threats briefing on MERS-CoV 2013. Honors Meritorious service award, CDC (1999); CSIRO silver medal for collaborative research (2000); Honored by the naming of a new species of centipede, Cryptops daszaki J Nat Hist 2002; 36: 76-106) (2002); ISi Fastbreaking paper (2002); CBS 60 Minutes documentary on Nipah virus research; 6th Annual Lecturer, Medicine & Humanities, Texas A&M (2003); Editor's choice, Science (2006); Zayed International Prize for the Environment (2nd) (2006); Finalist, Director's Pioneer Award (2007); Discovery Channel documentary on Nipah virus research, Bangladesh (2008); Presidential Lecturer, University of Montana (2008) ; Elected member of the Cosmos Club 2012; Honored by the naming of a new species of parasite, lsospora daszaki (Parasitol. Res. 2013; 111: 1463-1466) (2012); Awarded the Hsu-Li Distinguished Lectureship in Epidemiology (2013). C. Peer-reviewed publications (selected from over 190+ ); * = Corresponding author Most relevant to the current a lication 1. Daszak P, Cunningham AA, Hyatt AD (2000). Emerging infectious diseases of wildlife - threats to biodiversity and human health. Science 287: 443-449 2. Li W, Shi Z, Yu M, Ren W, Smith C, Epstein JH, Wang H, Crameri G, Hu Z, Zhang H, Zhang J, McEachern J, Field H, Daszak P, Eaton BT, Zhang S & Wang L-F (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science 310: 676-679. 3. Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, and Daszak P* (Corresponding Author). (2008). Global trends in emerging infectious diseases. Nature 451 :990-993 4. Keesing F, Belden LK, Daszak P, Dobson A, Harvell CD, Holt RD, Hudson P, Jolles A, Jones KE, Mitchell CE, Myers SS, Bogich T & Ostfeld RS. (2010). Impacts of biodiversity on the emergence and transmission of infectious diseases. Nature 468:647-652. 5. Morse SS, Mazet JAK, Woolhouse M, Parrish CR, Carroll D, Karesh WB, Zambrana-Torrelio C, Lipkin WI, Daszak P* (Corresponding Author) (2012). Prediction and prevention of the next pandemic zoonosis. Lancet 380: 1956-1965. 6. Daszak P (2012). Anatomy of a pandemic Lancet 380: 1883-1884. PHS 398/2590 (Rev. 06109) Page_ Biosketches Biographical Sketch Format Page Page 18

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter Program Director/Principal Investigator (Last, First, Middle): Daszak, P. 7. Quan P-L, Firth C, Conte JM, Williams SH, Zambrana-Torrelio C, Anthony SJ, Ellison JA, Gilbert AT, Kuzmin IV, Niezgoda M, Osinubi MOV, Recuenco S, Markotter W, Breiman R, Kalemba L, Malekani J, Lindblade KA, Rostal MK, Ojeda-Flores R, Suzan G, Davis LB, Blau OM, Ogunkoya AB, Castillo DAA, Moran D, Ngam S, Akaibe D, Agwanda B, Briese T, Epstein JH, Daszak P, Rupprecht CE, Holmes EC, Lipkin WI. (2013). Bats are a major natural reservoir for hepaciviruses and pegiviruses. PNAS Published ahead of print April 2013. 8. Anthony SJ, Ojeda-Flores R, Rico-Chavez 0, Navarrete-Macias I, Zambrana-Torrelio CM, Rostal MK, Epstein JH, Tipps T, Liang E, Sanchez-Leon M, Sotomayor-Bonilla J, Avila R, Medellin RA, Goldstein T, Suzan G, Daszak P, Lipkin WI. (2013). Coronaviruses in bats from Mexico Journal of General Virology Online First. 9. Anthony SJ, Epstein JH, Murray KA, Navarrete-Macias I, Zambrana-Torrelio CM, Solovyov A, Ojeda-Flores R, Arrigo NC, Islam A, Ali Khan S, Hosseini P, Bogich TL, Olival KJ, Sanchez-Leon MD, Karesh W, Goldstein T, Luby SP, Morse SS, Mazet JAK, Daszak P*(Co-corresponding Author), Lipkin WI, Estimating viral diversity in Bats. PNAS in review. 10 (b)(6) Additional recent ublications of im ortance to the field from 190+ total 1. Cui J, Han N, Streicker D, Li G, Tang X, Shi Z, Hu Z, Zhao G, Fontanet A, Yi G, Wang L, Jones G, .Field HE, Daszak P* (Corresponding Author) & Zhang, S. (2007) Evolutionary relationships between bat coronaviruses and their hosts. Emerg. Infect. Ois.13: 1526-1533 2. Epstein JH, Prakash V, Smith CS, Daszak P, McLaughlin AB, Meehan G, Field HE, Cunningham AA (2008). Henipavirus infection in fruit bats (Pteropus giganteus), India. Emer, . Infect. Ois.14: 1309-1311. 3. Smith KF, Behrens M, Schloegel LM, Marano N, Burgiel S, Daszak P* (Corresponding Author). (2009). Reducing the risks of the wildlife trade. Science 324:594-595. 4. Epstein J H, Quan PL, Briese T, Street C, Jabado 0, Conlan S, Khan SA, Verdugo D, Hossain MJ, Hutchison SK, Egholm M, Luby SP, Daszak P* (Co-corresponding Author), Lipkin WI. (2010). Identification of GBV-D, a Novel GB-like Flavivirus from Old World Frugivorous Bats (Pteropus giganteus) in Bangladesh. PLoS Patho ens 6 (7): e1000972. 5. Homaira N, Rahman M, Hossain MJ, Epstein JH, Sultana R, Khan MSU, Podder G, Nahar K, Ahmed B, Gurley ES, Daszak P, Lipkin WI, Rollin PE, Comer JA, Ksiazek TG & Luby SP. (2010). Nipah virus outbreak with person-to-person transmission in Thakurgaon, Bangladesh 2007. Epidemiol & Infection 138: 1630-1636. 6. Olival KJ, Islam A, Yu M, Anthony SJ, Epstein JH, Khan SA, Khan SU, Crameri G, Wang L-F, Lipkin WI, Luby SP, Daszak P (2013). Filovirus antibodies in fruit bats, Bangladesh. Erner, . Infect. Dis. 19: 270-273. 7. Sazzad HMS, Hossain MJ, Gurley ES, Ameen KMH, Parveen S, Islam MS, Faruque LI, Podder G, Banu SS, Lo MK, Rollin PE, Rota PA, Daszak P, Rahman M, Luby SP. (2013). Nipah virus infection outbreak with nosocomial and corpse-to-human transmission, Bangladesh. Emerg. Infect. Dis. 19: 210-217. D. Research Support Ongoing Research Support NSF Daszak (Pl) 07/01/10-06/30/15 EcoHealthNet - a Research Coordination Network Funding for student exchange and workshops to fuse veterinary science, ecology and human medical sciences Role: Pl 5R01GM100471 Perrings (Pl) 09/15/11-06/30/15 NIGMS Modeling Anthropogenic Effects in the Spread of Infectious Disease Role: Co-Investigator 1 R56TW009502 Daszak (Pl) NIH Fogarty International Center Comparative Spillover Dynamics of Avian Influenza in Endemic Countries Role: Pl PHS 398/2590 (Rev. 06/09) Page_ Biosketches 09/17 /12-08/31 /14 Biographical Sketch Format Page Page 19

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Program Director/Principal Investigator (Last, First, Middle): Daszak, P. USAID EPT PREDICT Daszak (Pl) 10/01/09- 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Role: Pl on Subcontract 2 R01TW005869 Daszak (Pl) 09/01/08- 08/31/13 NIH Ecology of Infectious Diseases (Fogarty International Center) The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh To conduct mathematical modeling and fieldwork to understand the dynamics of Nipah virus in Bangladesh Role: Pl NSF DEB-1257513 Daszak (Pl) 08/15/12-07/31/13 US-China Ecology and Evolution of Infectious Diseases Collaborative Workshop; Kunming, China - October, 2012 Role: Pl 1 R01AI079231 Daszak (Pl) 09/18/08- 08/31/13 NIAID Non-Biodefense Emerging Infectious Diseases Risk of viral emergence from bats. To model hotspots for bat viral diversity, identify & characterize new bat viruses & understand their pathology Role: Pl HDTRA1-13-C-0029 Preston(PI) 01/11/13-01/10/14 Office of Naval Research, Defense Threat Reduction Agency Global Rapid Identification Tool (GRIT) for Undiagnosed Emerging Infectious Diseases (EID) Events Role: Co-Investigator Com leted Research Su ort NSF BCS 0826779 NSF Human and Social Dynamics Daszak (Pl) 10/01/08- 03/31/12 AOC - HSD - Collaborative Research: Human-related factors affecting emerging infectious diseases To analyze how socio-economic and environmental drivers predict risk of EIDs Role: Pl on lead proposal R01TW005869 - supplemental Daszak (Pl) 09/01/08 - 08/31/11 NIH EID (Fogarty International Center) Supplemental funding: Predicting the risk of global H5N1 spread This project will involve mathematical modeling and fieldwork in Bangladesh and China to understand risk of H5N1 spread. Role: Pl NSF EF-062239 Kilpatrick (Pl) 09/01 /06 - 08/30/11 NSF/NIH: Ecology & Evolution of Infectious Diseases Predicting spatial variation in West Nile virus transmission Study interaction among WNV vector, reservoir host populations across an urban-to-rural gradient. Role: Co-Pl R01 TW05869 Daszak (Pl) 08/01 /02 - 05/31 /07 NIH/Fogarty International Center Anthropogenic change & emerging zoonotic paramyxoviruses To identify the cause of emergence of Nipah and Hendra viruses in Malaysia and Australia. Role: Pl HSD 0525216 Daszak (Pl) 10/15/05- 10/14/06 National Science Foundation: Human and Social Dynamics Collaborative Research: Socio-Economic and Environmental Drivers of Emerging Diseases To analyze patterns of disease emergence globally and produce a broad risk assessment. Role: Pl PHS 398/2590 (Rev. 06109) Biosketches Page_ Biographical Sketch Format Page Page 20

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors inthe order listed on Form Page 2. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME Zhengli Shi eRA COMMONS USER NAME (credential, e.g., agency login} (b)(6) POSITION TITLE Senior scientist EDUCATION/TRAINING (Begin with baccalaureate or dher initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MMIYY FIELD OF STUDY (if ann/icab/e I Department of Biology, Wuhan University, China B.S. 1987 GENETICS Wuhan Institute of Virology, Chinese Academy of M.S. 1990 VIROLOGY Sciences, China University Montpellier II, Montpellier, France Ph.D. 2000 VIROLOGY A. Personal Statement The focus of this project is to understand the risk of coronavirus spillover from bats to people in China, using ecological analyses, fieldwork, receptor binding assays, and modeling approaches. I have worked in lab-based virology for 23 years, specializing in SARS-CoV and SARS-like CoVs since 2002. This includes the discovery of a wide-array of SARS-like coronaviruses in mainland China, including two isolates able to bind to the ACE2 receptor. My lab has established several bat primary cell lines and immortalized cell lines, capacity for pseudovirus generation and SARS-specific binding assays and we have expertise in every laboratory technique in this proposal. I have collaborated with the Pl for over 10 years, and have spent time in laboratories in the USA and Europe. My lab will be responsible for diagnosis, genomics and isolation of coronavirus from wild and domestic animals in Southern China and for analyzing their receptor binding domains. B. Positions and Honors. Positions and Em lo ment 1990-1993 1993-1995 2000- Research assistant. Wuhan Institute of Virology, Chinese Academy of Sciences. China Research scientist, Wuhan Institute of Virology, Chinese Academy of Sciences, China Senior Scientist, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China Other Ex erience and Professional Membershi s 2008- 2001- 2001- 2004- 2004-2009 2010- Honors Member, American Society of Microbiology Member, Chinese Society of Microbilogy Member, Chinese Society of Biochemistry and Molecular Biology Editor board, Chinese Journal of Virology Editor board, Virologica Sinica Associate Editor, Virologica Sinica 1996 Chinese Government Graduate Scholarship, the Ministry of Education, PR China. 2003 Natural Science Award (the second rank) of Hubei province, China. 2004 Outstanding supervisor of graduate student of Hubei province, China. 2005 Visitor scholarship from the Chinese Academy of Sciences. 2006 Outstanding scientist of the Chinese Academy of Sciences. C. Selected peer-reviewed publications (Selected from 82 peer-reviewed) Most relevant to the current ap lication Biographical Sketches for each listed Senior/Key Person 2 Page 21

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 1. Li, W., Shi Z., Yu M., Ren W., Smith C., Epstein H.J., Zhang S., Wang H., Crameri G., Hu Z., Zhang H., Zhang J., Mceachern J., Field H., Daszak P., Eaton T.B. and Wang L. F. (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science, 310(5748), 676-679. 2. Hon, C. C., Lam, T. Y., Shi, Z., Drummond, A. J., Yip, C. W., Zeng, F., Lam, P. Y. and Leung, F. C .. (2008). Evidence of the recombinant origin of a bat severe acute respiratory syndrome (SARS)-like coronavirus and its implications on the direct ancestor of SARS coronavirus. Journal of Virology, 82(4 ), 1819-1826. 3. Yuan, J., Hon,C. C., Li, Y., Wang, D., Xu, G., Zhang, H., Zhou, P., Poon, L. M., Lam, T. T. Leung, F. C. and Shi, Z. (2010). Intra-species Diversity of SARS-Like Coronaviruses (CoVs) in Rhinolophus sinicus and Its Implications on the Origin of SARS-CoVs in human. Journal of General Virology, 91(4),1058-1062. 4. Hou, Y., Peng, C., Yu, M., Li,Y., Han, Z., Wang, L-F., Li, F., Shi, Z. (2010). Bat Angiotensin Converting Enzyme-2 Displays Different Receptor Activity to Severe Acute Respiratory Syndrome Coronavirus Entry. Archives of Virology, 155(10), 1563-1569. 5. Wang, J., Wang, L-F. and Shi, Z. (2008). Construction of a non-infectious SARS coronavirus replicon for application in drug screening and analysis of viral protein function. Biochemical and Biophysical Research Communications, 374(1 ), 138-142. Additional recent publications of importance to the field (in chronological order} 1. Ren, W., Li, W., Yu, M., Hao, P., Zhang, Y., Zhou, P., Zhang, S., Zhao, G., Zhong, Y., Wang, S., Wang, L. F. and Shi, Z. (2006). Full genome sequences of two SARS-like coronaviruses in horseshoe bats and genetic variation analysis. Journal of General Virology, 87(11 ), 3355-3359. 2. Li,Y., Wang, J., Hickey, A. C., Zhang, Y., Li, Y., Wu, Y., Zhang, H., Yuan, J., Han, Z., McEachern, J., Broder, C. C., Wang, L. F. and Shi, Z. (2008). Potential nipah virus infection in Chinese bats. Emerging Infectious Diseases, 14(12),1974-1976. 3. Ren, W., Qu, X., Li, W., Han, Z., Yu, M., Zhang, S., Wang, L. F., Deng, H., Shi, Z. (2008). Difference in receptor usage between SARS coronavirus and SARS-like coronavirus of bat origin. Journal of Virology, 82(4), 1899-1907. 4. Zhou, P., Han, Z., Wang, L.F. and Shi, Z. (2009). lmmunogenicity difference between the SARS coronavirus and the bat SARS-like coronavirus spike (S) proteins. Biochemical and Biophysical Research Communications, 387(2), 326-329. 5. Li, Y., Ge X., Hon C. C., Zhang H., Zhou P., Zhang Y., Wang L. F. and Shi Z. (2010). Prevalence and Genetic Diversity of Adena-Associated Viruses in Bats, China. Journal of General Virology, 91 (10), 2601-2609. 6. Zhang Y., Zhang H., Dong X., Yuan J., Zhang H., Yang X., Zhou Peng., Ge X., Li Y., Wang L-F, and Shi Z (2010). Hantavirus Outbreak Associated with Laboratory Rats in Yunnan, China. Infection, Genetics and Evolution, 10(5): 638-644. 7. Li, Y., Ge X., Zhang H., Zhou P., Zhu Y., Zhang Y., Yuan J., Wang L-F., Shi Z. (2010). Host Range, Prevalence and Genetic Diversity of Adenoviruses in Bats. Journal of Virology, 84(8), 3889-3897. 8. Yu, M., Tachedjian, M., Crameri, G., Shi, Z. and Wang, L.F. (2010). Identification of key amino acid residues required for horseshoe bat angiotensin-1 converting enzyme 2 to function as a receptor for severe acute respiratory syndrome coronavirus. Journal General Virology, 91(7), 1708-1712. 9. Ge, X., Li, Y., Yang, X., Zhang, H., Zhou, P., Zhang, Y. & Shi, Z. (2012). Metagenomic analysis of viruses from bat fecal samples reveals many novel viruses in insectivorous bats in china. Journal of Virology, 86, 4620-4630. 10. Zhou, P., Li, H., Wang, H., Wang, L. F., Shi, Z. (2012). Bat severe acute respiratory syndrome-like coronavirus ORF3b homologues display different interferon antagonist activities. Journal General Virology, 93, 275-281. D. Research Support Ongoing Research Support 30970137 National Natural Science Foundation of China Shi (Pl) 01/01/2010-12/31/2012 Metagenomic analysis of bat intestinal viruses Role: Pl 2011CB504700 National Basic Research program of China Shi (Pl) 01 /01 /2011-12/31 /2015 Biographical Sketches for each listed Senior/Key Person 2 Page 22

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Mechanism of interspecies transmission of zoonotic viruses Role: Pl 81290341 National Natural Science Foundation of China Shi (Pl) Genetic diversity, identification and pathogenesis of bat viruses Role: Pl Completed Research Support 2005CB523004 National Basic Research program of China Interspecies transmission mechanism of zoonotic viruses Role: Pl Shi (Pl) 01/01/2013-12/31 /2017 01/01/2006-12/31/2010 2009ZX10004-109 Key project of infectious diseases Shi (Pl) 01/01/2009-12/31/2010 Rapid and high throughput diagnostic methods for emerging infectious viral pathogens Role: Pl Biographical Sketches for each listed Senior/Key Person 2 Page 23

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME Zhang, Shuyi eRA COMMONS USER NAME (credential, e.g., agency login} (b)(6) POSITION TITLE Dean EDUCATION/TRAINING (Begin with baccalaureate or dher initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MMIYY FIELD OF STUDY (if applicable) Northeast Normal University, China B.Sc 07/87 Biology University of Paris XIII, France D.E.A. 10/90 Ethology University Pierre & Marie Curie, France Ph.D. 12/94 Primatology, Tropical Ecology A. Personal Statement The goal of the current proposal is to work on the ecology and evolutionary biology of a coronaviruses from wildlife, with special emphasis on China. My background is ideally suited to this work because I am originally trained as a wildlife biologist, but have been working on the ecology and evolutionary biology of zoonoses in wildlife for the past decade. My career as a wildlife biologist began with a Ph.D in France on the behavioral ecology of capuchin monkeys (Cebus ape/la) in primary forest of French Guiana. In 1995, I returned to China, working on golden monkeys (Rinopithecus) at the Institute of Zoology, Chinese Academy of Sciences. At the same time, I began to work on the behavioral ecology of bats and rapidly became the leading bat researcher in China, building a large, well-funded group working on the phylogeny, genetics and ecology of bats. In 2003, during the early outbreak of SARS epidemic, I was one of the few Chinese researchers hypothesizing that SARS must have originated from wild birds or mammals, and I became closely involved in the work of the WHO veterinary team investigating potential wildlife reservoirs for SARS. I continued this work with Ors Zhengli Shi, Peter Daszak and Jon Epstein after the WHO team had left, and discovered that bats are the natural reservoir of SARS-like Co Vs. After we published our results in Science in 2005, I continued to work on bat CoVs. bat genetics, molecular biology and immunology. I have worked actively with the EcoHealth Alliance and with the Wuhan Institute of Virology, and am involved in most of the preliminary data that is listed in the current application. I also act as the main, on-the-ground contact for EcoHealth Alliance research in China, and am Country Coordinator for the USAID-EPT PREDICT program. During the past decade, I have demonstrated my capacity to provide access to some of the most sensitive fieldwork sites in China and collaborate with US institutions in this work. This includes: collaborative work at Xinghai Lake, where we successfully isolated the first H5N1 from wild birds; work on hunter-trader cohorts in the wet markets of Guangzhou, where we are collaborating with EcoHealth Alliance to identify novel pathogens spilling over from wildlife to people; and the work we conducted on bat SARS-like CoVs in 20034, which involved the export of samples from wildlife into foreign collaborators labs for sequencing and pathogen discovery. In my current capacity as Dean of a 3- institute collaboration at ECNU, I have unique capability to mobilize resources, and work within my large network of collaborators to facilitate the current project. B. Positions and Honors Positions and Em lo ment 2011- Dean, Institutes for Advanced Interdisciplinary Research, East China Normal University, China 2010- Country Coordinator, USAID-EPT PREDICT 2006-2008 Professor, School of Life Science, East China Normal University, China 1997-2006 Research Professor. Institute of Zoology, Chinese Academy of Sciences, China 1995-1997 Associate Research Professor, Institute of Zoology, Chinese Academy of Sciences, China 1995 Assistant Research Professor, Institute of Zoology, Chinese Academy of Sciences, China Other Ex erience and Professional Membershi s 1997- Chairman of China's Primate Specialist Group, Species Survival Commission, World Biographical Sketches for each listed Senior/Key Person 3 Page 24

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 1999- 2000- Honors 1989 1991 1995 1998 1999 2000 2001 2006 Conservation Union {IUCN-SSC) Secretary General of Bat Specialists Group of China's Mammalogical Society Member of Chinese National Committee for International Union of Biological Sciences Fellowship from the China's Education Ministry for students abroad Fellowship from the French Government for Chinese students Research grant under the "100 Talent Programme" sponsored by the Chinese Academy of Sciences Allowance of the State Department for research and technology Research grant under the "Young Scientist" sponsored by the Chinese Academy of Sciences "Excellent Young Researcher Grant" of the National Natural Science Foundation of China "Young Scientist" award of the Chinese Academy of Sciences Nation Award (class II) for Science and Technology C. Peer-reviewed publications (selected from over 180 peer-reviewed publications) Most relevant to the current a lication 1. He, J.F., Peng, G.W., Min, J., Yu, D.W., Liang, W.J., Zhang, S.Y., Xu, R.H., Zheng, H.Y., Wu, X.W., Xu, J., Fang, L., Zhang, X., Li, H., Yan, X.G., Lu, J.H., Hu, Z.H., Huang, J.C., Wan, Z.Y., Lin, J.Y., Song, H.D., Wang, S.Y., Zhou, X.J., Zhang, G.W., Guo, B.W., Zheng, H.J., Zhang, X.L., Zheng, K., Wang, B.F., Fu, G., Hou, J.L., Wang, X.N., Chen, S.J., Hao, P., Tang, H., Ren, S.X., Zhong, Y., Guo, Z.M., Liu, Q., Miao, Y.G., Kong, X.Y., He, W.Z., Li, Y.X., Chen, Z., Wu, C-1, Zhao, G.P., Chiu, R.W.K., Chim, S.S.C., Tong, Y.K., Chan, P.K.S., Tan, J.S., Lo, Y.M.D. (2004 ). Molecular evolution of the SARS-coronavirus during the course of the SARS epidemic in China. Science, 303, 1666-1669. 2. Li, W.D., Shi, Z.L., Yu, M., Ren, W.Z., Smith, C., Epstein, J., Wang, H.Z., Crameri, G., Hu, Z.H., Zhang, H.J., Zhang, J.H., McEachern, J., Field, H., Daszak, P., Eaton, B.T., Zhang, S.Y., Wang, L.F. (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science, 310, 676-679. 3. Tang, X.C., Zhang, J.X., Zhang, S.Y., Wang, P., Fan, X.H., Li, L.F., Li, G., Dong, B.Q., Liu, W., Cheung, C.L., Xu, K.M., Song, W.J., Vijaykrishna, D., Poon, L.L.M., Peiris, J.S.M., Smith, G.J.D., Chen, H., Guan, Y. (2006). Prevalence and genetic diversity of coronaviruses in bats from China. Journal of Virology, 80, 7481-7490. 4. Ren, W.Z., Qu, X.X., Li, W.D., Han, Z.G., Yu, M., Zhou, P., Zhang, S.Y., Wang, L.F., Deng, H.K., Shi, Z.L. (2008). Difference in receptor usage between SARS coronavirus and SARS-like coronavirus of bat origin. Journal of Virology, 82, 1899-1907. 5. Tang, X.C., Li, G., Vasilakis, N., Zhang, Y., Shi, Z.L., Zhong, Y., Wang, L.F., Zhang, S.Y. (2009). Differential stepwise evolution of SARS Coronavirus functional proteins in different host species. BMC Evolutionary Biology 9, 52, doi:10.1186/1471-2148-9-52. Additional recent ublications of im ortance to the field 1. Wang, L.F., Shi, Z.L., Zhang, S.Y., Field, H., Daszak, P., Eaton, B.T. (2006). Review of Bats and SARS. Emerging and Infectious Disease, 12, 1834 -1840. 2. Li, G., Jones, G., Rossiter, S., Chen, S.F., Parsons, S., Zhang, S.Y. (2006). Phylogenetics of small horseshoe bats from East Asia based on mitochondrial DNA sequence variation. Journal of Mammalogy, 87, 1234-1240. 3. Ren, W.Z., Li, W.D., Yu, M., Hao, P., Zhou, P., Zhang, S.Y., Zhao, G.P., Zhong, Y., Wang, S.Y., Wang, L.F ., Shi, Z.L. (2006). Full-length genome sequences of two SARS-like coronaviruses in 4 horseshoe bats and genetic variation analysis. Journal of General Virology, 87, 3355-3359. 4. Cui, J., Han, N.J., Streicker, D., Li, G., Tang, X.C., Shi, Z.L., Hu, Z.H., Zhao, G.P., Guan, Y., Wang, L.F., Field, H., Jones, G., Daszak, P., Zhang, S.Y. (2007). Evolutionary relationships between bat coronaviruses and their hosts. Emerging and Infectious Disease, 13, 1526-1532. 5. Rossiter, S.J., Benda, P., Dietz, C., Zhang, S.Y., Jones, G. (2007). Rangewide phylogeography in the greater horseshoe bat inferred from microsatellites: implications for population history, taxonomy and conservation. Molecular Ecology, 16, 4699-4714. 6. Cui, J., Counor, D., Shen, D., Sun, G.Y., Deubel, V., Zhang, S.Y. (2008). Detection of Japanese Biographical Sketches for each listed Senior/Key Person 3 Page 25

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter encephalitis virus antibodies in bats, Southern China. American Journal of Tropical Medicine and Hygiene, 78, 1007-1011. 7. Zhang, J.S., Jones, G., Zhang, L.B., Zhu, G.J., Zhang, S.Y. (2010). Recent surveys of bats (Mammalia: Chiroptera) from China II. Pteropodidae. Acta Chiropterologica, 12, 103-116. 8. Liu, Y., Cotton, J.A., Shen. 8., Han. X.Q., Rossiter. S.J., Zhang, S.Y. (2010). Convergent sequence evolution between echolocating bats and dolphins. Current Biology, 20, R53-54. 9. Zhang, L.B., Parson, S., Daszak, P., Wei, L., Zhu, G.J., Zhang, S.Y. (2010). Variation in the abundance of ectoparasite mites in relation to the reproduction status, age, sex and size of flat-headed bats. Journal of Mammalogy, 91, 136-143. 10. Shen, 8., Han, X.Q., Jones, G., Rossiter, S.J., Zhang, S.Y. (2013). Adaptive evolution of Myo6 Gene in Old World Fruit Bats (Family: Pteropodidae). PLOS ONE, 8(4), doi: 10.1371/journal.pone.0062307 D. Research Support Qngoing Research Support CbH4> Morse (Pl) 10/01/09-09/30/14 "===~'"="""~..,.,,.,..-=-:--e-,,-!: PREDICT-Wildlife SMART Surveillance/PREDICT Project to pre-empt at the earlier stages possible, zoonotic diseases that impose significant threat to public health. Role: Collaborator (b)(4) Zhang Pl 01/01/11-12/30/13 CbH4>: Surveillance Emerging Infectious Diseases This ---,--.----,----.....,.,.,----,--.....,.,.-:-=,--,--,---~ project is to conduct surveillance in wildlife in hotspots for new emerging zoonoses. Role: Pl (b)(4) Zhang Pl 01/01/11-12/30/13 CbH4>: Study of the Evolution of SARS Coronavirus This ---------------,--,------ project is to study the evolutionary relationships between bat coronaviruses and their hosts. Role: Pl Completed Research Support (b) (4) Zhang {Pl) ------- : Research on biological characteristics of Bats. Role: Pl 01/01/09-12/30/12 (b) (4) CbH4> Zhang (Pl) 01/01/10-12/30/12 ------- Changjiang Scholars and Innovative Research Team in University in China: Studying and Monitoring Wildlife and Zoonosis in Eastern China This project is to identify new viruses from wildlife in Eastern China, and to examine the pathogenicity and infectiousness for these novel pathogens. Role: Pl Biographical Sketches for each listed Senior/Key Person 3 Page 26

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME POSITION TITLE Ke, ChangWen eRA COMMONS USER NAME (credential, e.g., agency login) Director, Institute of Pathogenic Microbiology Guangdong Center for Disease Control and Prevention EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY (if applicable) West China Medical University M.P.H. 1984 Public Health West China Medical University B.S. 1989 Medicine Sun Yensen University M.D. 2001 Biochemistry and Molecular Biology A. Personal Statement. I have worked in public health and infectious disease research for more than 10 years. As Director of the Institute of Pathogenic Microbiology at Guangdong CDC I have been involved in the study and control of several emerging zoonitic infections, including SARS CoV and most recently, H7N9 avian influenza. Our work under the Guangdong Department of Health and with several Chinese universities and international collaborators has established several syndromic disease surveillance programs and collaborative infectious disease research programs including Chikungunya, enterovirus 71, Avian influenza H7N9 and H5N1, and SARS CoV. Most recently, through partnership with the USAID PREDICT program, we have augmented our lab's ability to identify zoonotic agents in people highly exposed to wildlife such as those working in live animal markets. This ongoing surveillance program has led to the identification of people who have been exposed to animal pathogens, including SARS CoV, and supports the initiative to extend this type of surveillance to other provinces in China. I believe that there is strong evidence that spillover of animal pathogens to people is occurring in China and MERS CoV in the Middle East shows that we should pay more attention to bat coronaviruses. Given the technical expertise and capacity for disease detection at the microbiology lab at Guangdong CDC, I have high confidence that we will be able to contribute to our understanding of coronavirus circulation in human populations and to determine the risk of new CoVs emerging in China. B. Positions and Honors Positions and Em 1989-2000 1994-1996 2003-2004 2004- lo ment Doctor in Charge, Health & Epidemic Prevention Station of Guangdong Province Participant, Department II of Virology, National Institute of Infectious Diseases, Japan Visiting researcher, Virology Department, National Institute of Infectious Diseases.Japan Director, Institute of Microbiology Center for Disease prevention and Control, Guangdong province, China Other Ex erience and Professional Membershi s 2004- Member of National expert committee of Influenza 2006- Member of National Biosafety expert Committee Biographical Sketches for each listed Senior/Key Person 4 Page 27

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter C. Selected Peer-reviewed Publications Most relevant to the current a lication 1. Mo, H., Zeng, G., Ren, X., Li, H., Ke, C.W., Tan, Y., Cai, C., Lai, K., Chen, R., Chan-Yeung, M., Zhong, N. (2006). Longitudinal profile of antibodies against SARS-coronavirus in SARS patients and their clinical significance. Respirology. Jan; 11 (1 ):49-53. 2. Qiaoli, Z.*,Jianfeng, H., De, W., Zijun, W., Xinguang, Z., Haojie, Z., Fan, D., Zhiquan, L., Shiwen, W., Zhenyu, H., Yonghui, Z., Ke, C.W., Yuan D., Liang W., Li D., Chen, P. (2012). Maiden Outbreak of Chikungunya in Dongguan City, Guangdong Province, China: Epidemiological Characteristics. PLOS ONE, 7(8):1-8 3. Wu, 0., Zheng H., Li, H., Monagin, C., Guo, X., Liu, L., Zeng, H., Fang, L., Mo, Y., Zhou, H., Zhang, H., Kou, J., Long, C., Hiromu, Y., & Ke, C.W. (2012). Phylogenetic and molecular characterization of Coxsackievirus A24 variant isolates from a 201 0acute hemorrhagic conjunctivitis outbreak in Guangdong, China Virology Journal, 9.41: 1-9 4. Guan, D., van der Sanden, S., Zeng, H., Li, W., Zheng, H., Ma, C., Su, J., Liu, Z., Guo, X., Zhang, X., Liu, L., Koopmans, M., Ke, C.W.* (2012) Population Dynamic and Genetic Diversity of C4 Strains of Human Enterovirus 71 in Mainland China, 1998-2010. PLOS ONE, 7(9): 1-8 5. Yang, F., He, J.*, Zhong, H., Ke, C.W., Zhang, X., Hong, T., Ni, H., Lin, J. (2012). Temporal Trends of Influenza A (H 1 N 1) Virus Seroprevalence following 2009 Pandemic Wave in Guangdong, China: Three Cross-Sectional Serology Surveys. PLOS ONE, 7(6):1-8 Additional recent ublications of im ortance to the field in chronolo ical order 1. Ke, C.W., Li T.C., Takeda, N. (2005). Positively Charged Amino Acid Residues of VP1 Capsid Protein of Human Polyomavirus BK Influence on the Formation of Virus-like Particles Generated by Recombinant Baculoviruses. Virologica Sinica, 21 (1 )20-23 2. Ke C.W., Zheng, K., Zhang, X., Zhou H.Q., Duan J.H., Lin L.F. (2005). Detection of Dengue virus by realtime polymerase chain reaction with TaqMan MGB probe. Chinese J Zoonosis, 21(8)716-720 3. Yan, J., Ke, C.W., Zheng, H., et al. (2006). Rapid diagnosis and Identification of Human Enteroviruses by sequencing VP4 gene. Chinese Journal of Vaccines and Immunization. 12(6)469-471 4. Zheng, H.Y., Liu L., Guo, X., Ke, C.W. (2006). A Comparative Study of Three lgM ELISA Kits for Measles Detection. Journal of Tropical Medicine, (08) 897-899 5. Ke, C.W., Deng, F. (2007). Surveillance system based on hospital and laboratory network to discover emerging viral diseases Journal Pathogen Biology, 2(1 ): 75-76 6. Ke, C.W., Zou, L.R., Yan, J. (2007). Control strategy for emerging Zoonosis. Chinese J Zoonosis, 23(1 )92- 93. 7. Li, B., Tan, H., Wang, D., et al. (2010). Phenotypic and genotypic characterization of vibrio Cholera 0139 of clinical and aquatic isolation in China. Curr. Microbiol. 8. Ding, X., Jiang, L., Ke, C.W. et al. (2010). Amino acid sequence analysis and identification of mutations under positive selection in Hemagglutinin of 2009 influenza A (H 1 N1) isolates. Virus Genes, 41 :329-340 9. Sun, L.M., Zheng, H.Y., Zheng, X.Z. et al. (2011). An enterovirus 71 epidemic in Guangdong province of China, 2008: Epidemiological, Clinical, and Virogenic manifestations. Jpn. J. lfect. Dis., 64:13-18 10. Su, S., Ning, Z.Y., Zhu, W.J., Jiao, P.R., Ke, C.W., Qi, W.B., Huang, Z., Tian, J., Cao, L., Tan, L.K., Shao, Z.W., Liang, H.B., Huang, W.M., Liao, M., Li, S.J., Zhang, G.H. (2013). Lack of evidence of avian-to-human transmission of avian influenza A (H5N1) virus among veterinarians, Guangdong, China, 2012. Journal of Clinical Virology. 56(4), 365-366. D. Research Support Ongoing Research Support (b)(4) 2012-2015 National Major Projects of Major Infectious Disease Control and Prevention: the Ministry of Science and Technology of the People's Republic of China Biographical Sketches for each listed Senior/Key Person 4 Page 28

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Completed Research Support China-U.S. Collaborative Program on Emerging and Re-Emerging Infectious Diseases Enhanced surveillance on Salmonella in Guangdong province. 30972591 2009-2012 National Natural Science Foundation of China 2010-2011 Epidemiology and molecular mechanism of virulence mutation of dengue viruses in Guangdong World Bank 2005 Establish Laboratory Network for Emergency Response and Surveillance of Infectious Diseases in Guangdong Province and Training. WHO 07.03.01 .AW.01. Epidemiological study on Transmission on Influenza A Virus from Animals to Human WHO grant: 07.02.01 .AW.01. Surveillance on emerging and reemerging infectious diseases pathogen in Guangdong Province Biographical Sketches for each listed Senior/Key Person 4 Page 29

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter BIOGRAPHICAL SKETCH NAME POSITION TITLE Jonathan H. Epstein Associate Vice President & Asia Regional ____________________ _, Coordinator eRA COMMONS USER NAME (crediential, e.g., agency login) I Cb><~ EDUCATION/TRAINING (Begin with baccalaureate or 01her initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE YEAR(s) FIELD OF STUDY (if aoo/icab/e) Brandeis University, MA BA 1996 Biology Tufts University, Sch. Vet. Med., Boston, MA DVM 2002 Wildlife Med., Intl. Med. Tufts University, Sch. Vet. Med., Boston, MA MPH 2002 Epidemiology Tufts University, Sch. Vet. Med., Boston, MA Cert Intl Med 2002 Zoonotic Diseases A. Personal Statement The goal of the proposed research is to investigate the ecology, evolutionary history and transmission dynamics of mammalian coronaviruses at the human-animal interface. Specifically, we will conduct field studies in China to obtain high quality samples from bats and other mammals found in wet market systems and identify and characterize known and novel coronaviruses. We will analyze the patterns of coronavirus transmission among bats and other wildlife, and the risk of spillover to humans. This research will address fundamentally important issues about the diversity of coronaviruses in mammalian hosts and the risk of interspecies transmission and emergence in human populations. My research has focused on the epidemiology and ecology of emerging zoonotic viruses carried by bats (Nipah virus, Ebolavirus, and SARS CoV), and other wildlife, and the drivers that lead to emergence. My work on SARS CoV ecology, in collaboration with coinvestigators Daszak (Pl), Zhang and Shi led to the discovery of several SARS-Like coronaviruses in bats, which appear to be ancestral to SARS-CoV and most recently which utilize the same ACE2 receptor as SARS CoV, suggesting direct spillover to humans is possible. Recently, I led a field team in Saudi Arabia in collaboration with the KSA Minist_ry_Qf Health, to iden![fy the animal origins of the newjy discovered MERS CoV. I continue to be involved in this ongoing investigation along with co-investigators Daszak and Olival. I have also conducted pathogen discovery work in bats, utilizing next generation sequencing technologies, which led to the discovery of a novel flavivirus related to Hepatitis C virus (GBV D). This team brings a high level of expertise in disease ecology, epidemiology, and pathogen discovery, and includes China's leading experts on wildlife zoonoses in partnership with key provincial CDCs. Our team has maintained a highly productive collaboration under several NIH and non-federally funded research projects, generating peer-reviewed papers in high impact journals (including Science, PNAS, and PLoS Pathogens). We have proven through previous work that we can manage logistically challenging projects involving people, wildlife, and animals in the wet markets in China, which gives this proposal a high likelihood of success. Under several federal awards, I have successfully managed the field and molecular investigations of zoonotic viruses in bats in several countries including Saudi Arabia, China, India, Malaysia, Thailand, and Bangladesh, all of which have logistical and political challenges. Using the bat and human samples we have already collected; and new animal samples we propose to collect; we will have the resources available to achieve the aims of this proposal. B. Positions and Honors Positions and Em lo ment 1999 Intern, Brisbane South Public Health Unit & DPI Queensland Animal Research Institute, AUS. 2002 Extern, Division of Viral and Rickettsial Diseases, CDC, Atlanta, GA 2002 Veterinary Intern, Small animal emergency and critical care, Ocean State Vet. Spec., RI 2003- Senior Research Scientist, EcoHealth Alliance, New York, NY. 2003- Adjunct Faculty, Ecology, Columbia Univ., NY & Tufts University Sch. of Vet. Med., MA. 2006- Adjunct Faculty, Mailman School of Public Health, Columbia Univ, NY 2007- Adjunct Asst. Clinical Professor, Public Health & Family Med, Tufts Univ School of Medicine, MA Biographical Sketches for each listed Senior/Key Person 5 Page 30

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 2008- 2008- 2009- 2009- 2009- 2011- 2012- Other Ex 1998- 2003- 2004 2004 2006 2007- 2010- 2008-13. Honors 2002 2002 2002 2004 2006 2007 2012 Postdoctoral fellow, Center for Infection and Immunity, Columbia University, NY, Adjunct Associate Professor, Mt. Sinai School of Medicine Review Editor - EcoHealth Associate Vice President, Conservation Medicine Program EcoHealth Alliance, NY Executive Director, Consortium for Conservation Medicine, EcoHealth Alliance, NY Asia Regional Coordinator, USAID EPT (PREDICT) Admissions committee, Tufts University Masters in Conservation Medicine degree program Board of Directors, International Association of Ecology and Health; Scientific Advisory Board, Lubee Bat Conservancy erience and Professional Membershi s Member: American Veterinary Medicinal Association, American Association of Zoo Vets, Wildlife Disease Association, New York Academy of Sciences, Member, IUCN Veterinary Specialist Group Invited speaker, WHO, Emerging Zoonotic Diseases Working Group meeting Member and Health Advisor, IUCN Bat Specialist Group; Advisory committee, Suffolk Country Board of Public Health; International Assoc. Ecology and Health Member, Delta Omega Public Health Honors Society Leader, Vertebrate Health Task Force, Smithsonian Institution Geological Earth Observatory Program (SIGEO) Scientific Committee Member, DIVERISTAS ecoHEAL TH cross-cutting network (ICSU-UNESCO) (selected) Invited presentations: University of Malaysia, Sarawak - Emerging zoonoses; IOM-NAS Committee on Achieving sustainable global capacity for surveillance and response to emerging infectious diseases; Nipah virus colloquium, University Malaya, Kuala Lumpur, Malaysia, Nipah virus symposium, American Society for Tropical Medicine & Hygiene; International Bat Research Symposium, Prague; American Society of Microbiology, Washington, DC; Australian Animal Health Laboratory (AAHL), Geelong; International Meeting on Emerging Diseases (IMED), Vienna; IOM meeting on MERS CoV and H7N9, Washington DC. First recipient, Certificate of International Veterinary Medicine, Tufts University Sch. Vet. Med. Hills award for excellence in veterinary clinical nutrition Sylvia Mainzer award for outstanding achievement in the field of public health NIH Loan Repayment Award (competitive award for Nipah virus research) Inducted into Delta Omega Honor Society for Public Health (Alpha Rho Chapter - 1st alumni inductee; 1st Inaugural Keynote Speaker) Outstanding Alumnus award, Tufts Cummings School of Veterinary Medicine Young Alumni Achievement Award, Tufts University (selected from all alumni who graduated in past 1 O yrs) D. Selected peer-reviewed publications (from 45). * indicates corresponding author Most relevant to the a lication in chronolo ical order 1. Li, W., Shi, Z., Yu, M., Ren, W., Smith, C., Epstein, J.H., Wang, H., Crameri, G., Hu, Z., Zhang, H., Zhang, J., McEachern, J., Field, H., Daszak, P., Eaton, B.T., Zhang, S. & Wang, L-F. (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science 310: 676-679. 2. Epstein, J.H.*, Quan, P.L., Briese, T., Street, C., Jabado, 0., Conlan, S., Khan, S.A., Verdugo, D., Hossain, M.J., Hutchison, S.K., Egholm, M., Luby, S.P., Daszak, P., & Lipkin, W.I. (2010). Identification of GBV-D, a Novel GB-like Flavivirus from Old World Frugivorous Bats (Pteropus giganteus) in Bangladesh. PLoS Pathogens 6(7): e1000972. doi:10.1371/journal.ppat.1000972. 3. Anthony, S.J, Ojeda-Flores, R., Rico-Chavez, 0., Navarrete-Macias, I., Zambrana-Torrelio, C.M., Rostal, M.K., Epstein, J.H., Tipps, T., Liang, E., Sanchez-Leon, M., Sotomayor-Bonilla, J., Aguirre, A.A., Avila, R., Medellin, R.A., Goldstein, T., Suzan, G., Daszak, P., Lipkin, W.I. (2013). Coronaviruses in bats from Mexico. J. Gen Viral. Published ahead of print January 30, 2013, doi:10.1099/vir.0.049759-0 Biographical Sketches for each listed Senior/Key Person 5 Page 3i

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 4. Wacharapluesadee, S., Sintunawa, C., Kaewpom, T., Khongnomnan, K., Olival, K.J., Epstein, J.H., et al. (2013). Group C betacoronavirus from bat guano 11 fertilizer, Thailand. Emerg Infect Dis. Aug. 12 http://dx.doi.org/10.3201/eid1908.130119 5. Quan, P.L., Firth, C., Conte, J.M., Williams, S.H., Zambrana-Torrelio, C.M., Anthony, S.J., Ellison, J.A., Gilbert, A.T., Kuzmin, I.V., Niezgoda, M., Osinubi, M.O.V., Recuenco, S., Markotter, W., Breiman, R.F., Kalemba, L., Malekani, J., Lindblade, K.A., Rostal, M.K., Ojeda-Flores, R., Suzan, G., D., Lora B., Blau, D.M., Ogunkoya, A.B., Alvarez C., Danilo A., Moran, D., Ngam, S., Akaibe, D., Agwanda, B., Briese, T., Epstein, J.H., Daszak, P., Rupprecht, C.E., Holmes, E.C., & Lipkin, W.I. (2013). Bats are a major natural reservoir for hepaciviruses and pegiviruses. PNAS. doi: 10.1073/pnas.1303037110 Additional recent ublications of im ortance to the field in chronolo ical order 1. Epstein, J.H.*, Field, H.E., Luby, S., Pulliam, J., & Daszak, P. (2006). Nipah Virus: Impact, Origins, and Causes of Emergence. Current Infectious Disease Reports 8: 59-65. 2. Epstein, J.H.*, Rahman, S.A., Zambriski, J.A., Halpin, K., Meehan, G., Jamaluddin, A.A., Hassan, S.S., Field, H.E., Hyatt, A.O., Daszak, P. & HERG. (2006). Feral cats (Felis catus) as possible vectors for Nipah virus. Emerging Infectious Diseases. 12: 1178-1179. 3. Field, H.E., Wang, L.F., Zhang, S., Daszak, P., Smith, C.S., Epstein, J.H., Shi, Z. (2007). Searching for the natural reservoir of the SARS virus. Preventive Veterinary Medicine. 81(1-3): 216-216 Sp. Issue. 4. Epstein, J.H.*, Prakash, V., Smith, C.S., Daszak, P., McLaughlin, A.B., Meehan, G., Field, H.E., and Cunningham, A.A. (2008). Evidence for Henipavirus infection in Indian Pteropus giganteus (Chiroptera; Pteropodidae) fruit bats. Emerging Infectious Diseases 14(8). 1309-11. 5. Epstein, J.H.*, Olival, K.J., Pulliam, J.R.C., Smith, C., Westrum, J., Hughes, T., et al. (2009). Pteropus vampyrus, a hunted migratory species with a multinational home-range and a need for regional management. Journal of Applied Ecology. 46(5):991-1002. 6. Epstein, J.H.*, Price, J.T. (2009). The Significant but Understudied Impact of Pathogen Transmission from Humans to Animals. Mount Sinai Journal of Medicine 76(5):448-55. 7. Homaira, N., Rahman, M., Hossain, M. J., Epstein, J.H., Sultana, R., Khan, M.S.U., Podder, G., Nahar, K., Gurley, E.S., Daszak, P., Lipkin W.I., Rollin, P.E., Comer, J.A., Ksiazek, T.G., Luby, S.P. (2010). Nipah outbreak with person-to-person transmission in Thakurgaon, Bangladesh, 2007. Epidemiology and Infection. 138: 1630-1636. 8. Sohayati, A., Rahman, Hassan, S.S, Olival, K.J., Mohamed, M., Chang, L-Y., Hassan, L., Suri, A.S., Saad, N.M., Shohaimi, S.A., Mamat, Z.C., Epstein, J.H., Field, H.E., Daszak, P., & HERG. (2010). Genetic characterization of Nipah virus isolated from naturally infected Pteropus vampyrus in Malaysia. Emerging Infectious Diseases.16(12).1990-3. 9. Pulliam, J.R., Epstein, J.H., Dushoff, J., Rahman, S.A., Meehan, G., Bunning, M., HERG, Jamaluddin, A.A., Hyatt, A.O., Field, H.E., Dobson, A.P. & Daszak, P. (2011 ). Agricultural intensification, priming for persistence, and the emergence of Nipah virus: a lethal bat-borne zoonoses. Journal of the Royal Society, Interface. Doi:10.1098/rsif.2011.0223 Uournal's most cited article in 2012) 10. Halpin, K., Hyatt, A.O., Fogarty, R., Middleton, D., Bingham, J., Epstein, J.H., Rahman, S.A., Hughes, T., Smith, C., Field, H.E., Daszak, P., & the Henipavirus Ecology Research Group. (2011 ). Pteropid Bats are Confirmed as the Reservoir Hosts of Henipaviruses: A Comprehensive Experimental Study of Virus Transmission. Am J Trap Med Hyg. 85:946-951; doi:10.4269/ajtmh.2011.10-0567 11. Sohayati, R., Hassan, L., Sharifah, S.H., Lazarus, K., Zaini, C.M., Epstein, J.H., Nairn, N.S., Field, H. E., Arshad, S.S., Aziz, J.A., & Daszak, P. (2011 ). Evidence for Nipah virus recrudescence and serological patterns of captive Pteropus vampyrus. Epidemiology and Infection. 139, pp 1570-1579 doi:10.1017/S0950268811000550 12. Daszak, P., Zambrana-Torrelio, C., Bogich, T.L., Fernandez, M., Epstein, J.H., Murray, K.A. & Hamilton, H. (2012). Interdisciplinary approaches to understanding disease emergence: The past, present and future drivers of Nipah virus emergence. PNAS doi:10.1073/pnas.1201243109 Biographical Sketches for each listed Senior/Key Person 5 Page 32

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter D. Research Support Ongoing Research Support Ongoing Research Support USAID Emerging Pandemic Threats: PREDICT Daszak (Pl) 1 0/01 /09 - 09/30/ 14 Modeling hotspots for disease emergence and conducting surveillance in wildlife for new emerging zoonoses. Role: Asia Regional Coordinator: coordinating field and lab activities in Bangladesh, India, Thailand, Malaysia, Indonesia and China; Surveillance Team and Molecular Diagnostic team member. 2 R01TW005869 Daszak (Pl) 09/01/08 - 08/31/14 NIH Ecology of Infectious Diseases (Fogarty International Center) The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh To conduct mathematical modeling and fieldwork to understand the dynamics of Nipah virus in Bangladesh Role: Co-Pl 1 R01AI079231 Daszak (Pl) 09/18/08 - 08/31 /13 NIAID Non-Biodefense Emerging Infectious Diseases Risk of viral emergence from bats. This project is to model hotspots for viral diversity and emergence in bats, to identify new viruses from bats, and to examine the pathogenicity and infectiousness for these novel pathogens. Role: Co-Pl 0955897 NSF Research Coordination Network Daszak (Pl) 07/01/10-06/30/15 EcoHealthNet: Environmental Science and Health Research Network The major goal of this research is to run a series of workshops and student research exchange programs focused on collaborations among the human medical, veterinary, ecological and economic sciences. Role: Co-Pl, Program Director USFWS, F12AP01117 Epstein (Pl). Development of a Great Ape Health Unit in Sabah, Malaysia 09/13/12 - 09/13/14 USFWS, 4500036150 Epstein (Pl) 09/15/12 - 09/14/14 Characterization of Climatic Parameters within Bat Hibernacula, their Influence on Environmental Loads of Geomyces destructans, and Implications for the Migration of White-Nose Syndrome in Bats Completed Research Support 1 K08AI067549 Epstein (Pl) 07 /1 /07 - 07 /30/11 Understanding the Ecology of Nipah Virus in Bangladesh (NIAID) Modeling the dynamics of Nipah virus in Pteropus giganteus and risk of spillover to humans. Role: Pl (collecting Nipah virus epidemiological data from Bats in Bangladesh) Biographical Sketches for each listed Senior/Key Person 5 Page 33

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME Olival, Kevin James eRA COMMONS USER NAME (credential, e.g., agency lo9in} (b)(6) POSITION TITLE Senior Research Scientist EDUCATION(TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY (if applicable) Colorado State University B.S. 05/97 Biology Columbia University MA 10/03 Conservation Biology Ecology and Columbia University PhD 05/08 Evolutionary Biology Molec. American Museum of Nat. History Postdoctoral 08/09 Parasitology NIH Fogarty US Global Health Fellow Postdoctoral 08/11 Intl Emerg. Inf. Dis A. Personal Statement The goal of our proposal is to study the ecology, evolution, and spillover potential of bat coronaviruses. Specifically, we will use a combination of fieldwork, mathematical modeling, and phylogenetic and molecular methods to test several hypotheses related to zoonotic spillover risk and the limits to host range for bat coronaviruses. My research experiences are strongly complementary to these aims. I have been conducting research on bat evolution, ecology, population genetics, and viral discovery for the past 11 years. During my dissertation at Columbia University, I used host population genetics and phylogeography to understand the dynamics of Nipah virus in Southeast Asian fruit bats. As a post-doc at AMNH I discovered several novel malaria parasites in bats, and used molecular systematics to understand the co-evolution and origins of nonhuman Plasmodium. I developed new approaches that combine phylogenetic, ecological and species lifehistory variables to predict viral diversity in bats, and have tested these using data from the literature. As an NIH Global Health Fellow, I expanded our knowledge of Nipah virus ecology in Bangladesh through population genetic analyses of the putative primary reservoir host, Pteropus giganteus, led field investigations on role of non-Pteropus fruit bats in Nipah virus circulation, and discovered a number of novel bat pathogens in these species. I have led field research projects and training workshops to conduct viral discovery throughout Asia, including Malaysia, Bangladesh, India, Vietnam, Cambodia, Thailand, and the Philippines. This included several expeditions to collect Nipah virus samples in Bangladesh and Malaysia, and a three-week long Ebola Reston investigation of bats in the Philippines. Most recently I have led field teams on two 3-week expeditions in Saudi Arabia to identify the animal reservoir of MERS-CoV; this work is ongoing. My latest research is focused on: 1) global viral discovery in bats; 2) integrating phylogenetic and molecular evolution analyses with ecological information to better understand the risk of bat viral spillover. In summary, for the past decade my research has been focused on the evolution and ecology of bats and their associated pathogens, and my current focus of using phylogenetic and evolutionary approaches model viral spillover risk in bats is highly complementary to the aims in this proposal. Biographical Sketches for each listed Senior/Key Person 6 Page 34

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter B. Positions and Honors Positions and Employment 2000-02 Mentor, NSF Undergraduate Mentoring in Environmental Biology (UMEB) for Pacific Islander 2002-08 2003- 2003 2003-08 2005 2005 2005 2006-07 2006-13 2007 2007 2009 2009 2009- 2009- 2009- 2010 2010- 2010- 2011- 2011- 2013- Honors 1993-97 2003 2005-07 2004-07 2008 2013 2013 undergraduates, University of Hawaii Research Collaborator, Consortium for Conservation Medicine, New York Member, Henipavirus Ecology Research Group Lecturer in Disease Ecology, Columbia University Continuing Education course Visiting researcher- bat genetics. Veterinary Research Institute, Malaysia Visiting researcher - bat genetics, Institute for Ecology and Biological Resources, Vietnam Visiting researcher - bat genetics. Pasteur Institute. Cambodia Judge, NY Science and Engineering Fair Mentor, Conservation Genetics High School Internship Program, AMNH, New York Instructor, Columbia University Secondary School Summer Program, New York Steering Committee, Small Matters: Microbes and Their Role in Conservation, New York Symposium Organizer, Bat Hunting and Bushmeat, Phuket, Thailand Symposium Organizer, Bat migration and disease, 1st lnt'I Workshop on Bat Migration, Germany Organizer and Scientific Review Committee, Exploring the Dynamic Relationship Between Health and the Environment, AMNH Spring Symposium, New York Review Editor, EcoHealth Adjunct Research Faculty, Center for Environmental Sustainability, Columbia University, New York. Visiting Research Scientist, American Museum of Natural History, Mammalogy Department. Mentor and Scientific Review Committee, Student Conference on Conservation Science New York Key Personnel and Lead Country Liaison: Thailand, Bangladesh, and Vietnam - USAID PREDICT Lead Field Researcher, FAO-EHA investigation of Ebola Reston reservoirs in Philippines Steering Committee, NSF RCN grant, South-east Asian Bat Conservation Research Group Internship Mentor, NSF RCN grant EcoHealthNet, graduate training in One Health EHA Team lead; MERS-CoV animal reservoir investigations with MoH in Saudi Arabia Colorado State University Distinguished Scholar Award NSF Graduate Student Fellowship, Honorable Mention Bat Conservation International Student Award and Scholarship US Environmental Protection Agency STAR Fellowship Award PhD Dissertation with Distinction, Columbia University Plenary talk on bat virus modeling at 11th Annual ASM Biodefense and EID Research Meeting Invitation to participate in Institute of Medicine panel on novel Coronavirus C. Selected Peer-reviewed Publications (Selected from 25 peer-reviewed publications) Most relevant to the current a lication 1. Turmelle, A. & Oliva I, K.J. (2009). Correlates of viral richness in bats (Order Chiroptera). EcoHealth 6(4 ): 522-39. 2. Rahman, S.A., Hassan, SS, Olival, K.J., Mohamed, M., Chang, L.Y., Hassan, L., Saad, N.M., Shohaimi, S.A., Mamat, Z.C., Nairn, M.S., Epstein, J.H., Suri, A.S., Field, H.E., Daszak, P. & HERG. (2010). Characterization of Nipah virus from Naturally Infected Pteropus vampyrus Bats, Malaysia. Emerging Infectious Diseases 16( 12): 1990-93. 3. Olival, K.J., Epstein, J.H., Wang, L.F., Field, H.E., & Daszak, P. (2012). Are bats unique viral reservoirs? in A. A. Aguirre, R. S. Ostfeld, and P. Daszak, editors. New Directions in Conservation Medicine: Applied Cases of Ecological Health. Oxford University Press, Oxford. pp. 195-212. 4. Levinson, J., Bogich, T.L., Oliva!, K.J., Epstein, J.H., Johnson, C.K., Karesh, W. & Daszak, P. (2013). arneJtio_a survejjlanceJouo.onotLcyirus discoYeJY._Ern_e_ mioaJniectLous Diseases t9(5J H--3-4-Z (b)(4) 5. Biographical Sketches for each listed Senior/Key Person 6

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter Identification of Group C Betacoronavirus from Bat guano fertilizer, Thailand. Emerging Infectious Diseases. Additional recent ublications of im ortance to the field in chronolo ical order 1. Olival, K.J. & Daszak, P. (2005). The ecology of emerging neurotropic viruses. Journal of NeuroVirology 11: 440-45. 2. Pulliam, J.R.C., Field, H.E., Olival, K.J. & HERG. (2005). An alternative explanation of Nipah virus strain variation. Emerging Infectious Diseases 11 (12): 1978-1979. 3. Daszak, P., Plowright, R., Epstein, J.H., Pulliam, J.R.C., Rahman, S.A., Field, H.E., Smith, C.S., Olival, K.J., Luby, S., Halpin, K., Hyatt, A.O., & HERG. (2006). The emergence of Nipah and Hendra virus: pathogen dynamics across a wildlife-livestock-human continuum. In: Disease Ecology: Community structure and pathogen dynamics, In Collinge and Ray, ed. Oxford University Press: Oxford. pp. 188-203. 4. Olival, K.J., Stiner, E.O., & Perkins, S.L. (2007). Detection of Hepatocystis sp. in Southeast Asian Flying Foxes (Pteropodidae) using Microscopic and Molecular Methods. Journal of Parasitology 93(6): 1538- 1540. 5. Epstein, J.H., Olival, K.J., Pulliam, J.R.C., Smith, C.S., Westrum, J., Hughes, T., Dobson, A., Zubaid, A., Rahman, S.A., Basir, M.M., Field, H.E., & Daszak, P. (2009). Management of Pteropus vampyrus, a hunted migratory species with a multinational home-range. Journal of Applied Ecology 46(5): 991-1002. 6. Murdock, C., Olival, K.J. & Perkins, S.L. (2010). Feeding preference of snow-melt mosquitoes (Culicidae: Culiseta and Ochelerotatus) show a link between cervid amplifying hosts for Jamestown Canyon Virus (Bunyaviridae: Orthobunyavirus) and humans. Journal of Medical Entomology 47(2): 226-229 7. Smith, C.S., Epstein, J.H., Breed, A., Plowright, R., Olival, K.J., de Jong, C., Daszak, P. & Field, H.E. (2011 ). Satellite Telemetry and Long-Range Bat Movements. PloS One 6(2): e14696. 8. Bogich, T.L., Olival, K.J., Hosseini, P., Mazet, J., Morse, S., Karesh, W.B., Jones, K.E., Levy, M., Funk, S., Brito, I., Epstein, J.H., Brownstein, J., Joly, D., & Daszak, P. (2012). Using Mathematical Models in a Unified Approach to Predicting the Next Emerging Infectious Disease. New Directions in Conservation Medicine: Applied Cases of Ecological Health. In Aguirre, Ostfeld and Daszak, ed. Oxford University Press. pp. 607-18. 9. Morse, S.F., Olival, K.J., Kosoy, M., Billeter, S.A., Patterson, B.D., Dick, C.W., & Dittmar, K. (2012). Global distribution and genetic diversity of Bartonella in bat flies (Hippoboscoidea, Streblidae, Nycteribiidae). Infection, Genetics and Evolution 12(8): 1717-23. 10. Oliva I, K.J. (2012). Correlates and evolutionary consequences of population genetic structure in bats. In Gunnell and Simmons, ed. Evolutionary History of Bats: Fossils, Molecules, and Morphology. Cambridge University Press, Cambridge. pp. 267-316. 11. Olival, K.J., Islam, A., Yu, M., Anthony, S.J., Epstein, J.H., Khan, S.A., Khan, S.U., Crameri, G., Wang, L.F., Lipkin, W.I., Luby S.P., & Daszak, P. (2013). Ebolavirus Antibodies in Fruit Bats, Bangladesh. Emerging Infectious Diseases 19 ~_: _27_0_-_2_73_. ____________________ .,._..,,.., (b) (4) 12 13 D. Research Support On oin Research Su mort NIH 1 R01AI079231 Daszak (Pl) 09/18/08 - 08/31/13 NIAID Non-Biodefense Emerging Infectious Diseases. "Risk of viral emergence from bats". This project is to model hotspots for viral diversity and emergence in bats, to identify new viruses from bats, and to examine the pathogenicity and infectiousness for these novel pathogens. Role: Key Personnel: lead project implementation, study design, and phylogenetic modeling Biographical Sketches for each listed Senior/Key Person 6 Page 36 (b)(4)

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter USAID EPT PREDICT Daszak (Pl) 10/01/09- 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife for new emerging zoonoses. Role: Key Personnel: Modeling disease risk and managing projects in Asian countries US Geological Survey (USGS) Olival (Co-Pl) 06/18/12-06/17/13 "Genetic Approaches to Defining Taxonomic and conservation Units for the Hawaiian Hoary Bat" Using molecular tools to conserve the endangered Hawaiian Hoary bat. Role: Co-Pl USFWS 4500036150 Epstein (Pl) 09/01/12-12/31/14 Characterization of Climatic Parameters within Bat Hibernacula, their Influence on Environmental Loads of Geomyces destructans, and Implications for the Migration of White-Nose Syndrome in Bats. Role: Co-Pl Completed Research Support NIH 3R01TW005869-06S1 Daszak (Pl) 09/01/09- 8/31/11 NIH Ecology of Infectious Diseases ARRA supplement to "The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh". Examined the ecology of Nipah virus in Bangladesh; population genetic structure of P. giganteus; and the pathogen discovery from a diverse range of bats. Over the course of this award I published >10 papers including 4 in the prestigious journal Emerging Infectious Diseases; presented at >20 national and int'I conferences; and media coverage in the New York Times Science section. Role: Fogarty US Global Health Fellow; lead for ARRA reserach Biographical Sketches for each listed Senior/Key Person 6 Page 37

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the key personnel and other significant contributors in the order listed on Form Page 2. Follow this formal for each person. DO NOT EXCEED FOUR PAGES. NAME POSITION TITLE Parviez R Hosseini Senior Research Fellow ----------------------! eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, and include postdoctoral training.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY (if aoo/icable) Brown University Sc. 8. 12/94 Applied Math - Biology University of California, Santa Barbara Ph.D 06/02 Biological Sciences A. Personal Statement The aims of the proposed research include disease ecology, evolutionary biology, and understanding the transmission dynamics of coronaviruses among wildlife hosts and their spillover to people. The latter includes analyzing patterns of viral prevalence, building and parameterizing mathematical models of pathogen transmission and evolution, and field research on these dynamics. In my career, I have used my training as theoretical ecologist and my broad experience in mathematics, statistics and ecology to focus on analyzing and explaining the process of disease emergence. I have studied disease ecology, with a strong focus on analytical and computation modeling approaches for the past 9 years. This work has involved leading the modeling component of several major research projects across a wide array of disease systems including Mycop/asma gallisepticum in House Finches, Barley and Cereal Yellow Dwarf viruses in California grasslands, Chikungunya virus, Rift Valley fever, and avian influenza. I am now the lead researcher on the modeling component for Influenza and Arbovirus Dynamics at EcoHealth Alliance. My strong interest in the impact of population structure on the emergence of novel pathogens, and my experience in working with computational modeling of emerging diseases give me a perfect background for the current proposed work. I also have considerable experience in working within national and international collaborative groups which will prove invaluable in the current project. B. Positions and Honors Positions and Employment 2002-2005 2005-2009 2009- Post-doctoral Associate, Cornell University, Lab of Ornithology, Ithaca, NY Associate Research Scholar, Princeton University, Dept. of Ecology and Evolutionary Biology. Princeton, NJ Senior Research Fellow, EcoHealth Alliance, NY Professional Activities: 2003 - 2005 Participant, Seasonality and the Population Dynamics of Infectious Diseases, 2004 2004 2006 2007 2008 NCEAS, Santa Barbara, CA Invited Speaker, Ecology of Infectious Disease Meeting, Emory University, Atlanta Invited Speaker, Dept. of Zoology, Oregon State University Invited Speaker, Dept. of Biology, EEOB Seminar Series, University of North Carolina Invited Speaker, Dept. of Ecology and Evolutionary Biology, University of Tennessee Invited Speaker, Dept. of Zoology, Oxford University, UK Biographical Sketches for each listed Senior/Key Person 7 Page 38

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 2008 2009 - present 201 O - present Selected Honors: Invited Speaker, Dept. of Biology, Stanford University, CA Member of modeling team, USAID-EPT PREDICT Review Editor, EcoHealth 2003 NSF RTG/GRT Fellowship on Spatial ecology 2004 Invited to speak at EEID in 2004 and 2011 2004 Member NCEAS group on Recovery plans and de-listing 2005 Member NCEAS group on designing ecological protected areas research 2005 Member NCEAS group on complex population dynamics 2007 PNAS 2007 paper cited by Faculty of 1000 Biology as "Must Read" 2007 PNAS paper listed as Science Editor's choice, 6th April 2007 C. Peer-reviewed publications Most relevant to the current application 1. Hosseini, P.R. (2003). How localized consumption stabilizes predator-prey systems with finite frequency of mixing. American Naturalist 161:567-585. doi:10.1086/368293 2. Hosseini, P.R., Dobson, A. & Dhondt, A.A. (2004). Seasonality and wildlife disease: How seasonal birth, aggregation and variation in immunity affect the dynamics of Mycoplasma gallisepticum in House Finches. Proceedings of the Royal Society of London: Biological Sciences. 271 :2569-2577. doi: 10.1098/rspb.2004.2938 3. Hosseini, P.R. (2006) Pattern Formation and Individual-Based Models: The Importance of Understanding Individual-Based Movement. Ecological Modeling 194: 357-371. doi:10.1016/j.ecolmodel.2005.10.041 4. Seabloom, E.W., Hosseini, P.R., Power, A.G., Borer, E.T. (2009). Causes and implications of co-infection by RNA viruses in natural grasslands. American Naturalist. 173:E79-E98. doi: 10.1086/596529 5. Hosseini, P.R., Sokolow, S.H., Vandegrift, K.J., Kilpatrick, A.M. & Daszak, P. (2010). Predictive power of air travel and socio-economic data for early pandemic spread PLoS One. 5(9):e12763. doi: 10.1371/journal.pone.0012763. Additional recent ublications of im ortance to the field 1. Campbell, S.P., Clark, A., Crampton, L., Guerry, A.O., Hatch, L.T., Hosseini, P.R., Lawler, J.J., O'Connor R.J. (2002). An assessment of monitoring efforts in endangered species recovery plans. Ecological Applications. 12:674-681. doi:10.1890/1051-0761 (2002)012[0674:AAOMEl]2.0.CO;2 2. Kollias, G.V., Sydenstricker, K.V., Kollias, H.W., Ley, D.H., Hosseini, P.R., Connolly, V. & Dhondt, A.A. (2004). Experimental infection of individually caged House Finches with Mycoplasma gallisepticum. J. Wildlife Diseases. 40: 79-86. 3. Dhondt, A.A., Altizer, S., Cooch, E.G., Davis, A.K., Dobson, A., Driscoll, M.J.L., Hartup, B.K., Hawley, D. M., Hochachka, W.M., Hosseini, P.R., Jennelle, C.S., Kollias, G.V., Ley, D.H., Swarthout, E.C.H., Sydenstricker, K.V. (2005). Dynamics of a novel pathogen in an avian host: Mycoplasmal conjunctivitis in house finches. Acta Tropica 94(1 ):77-93. doi:10.1016/j.actatropica.2005.01.009 4. Altizer, S., Dobson, A., Hosseini, P., Hudson, P. Pascual, M., & Rohani, P. (2006). Seasonality and the dynamics of infectious diseases. Ecology Letters 9:467-484. doi:doi: 10.1111/j .1461-0248.2005.00879.x 5. Hosseini, P.R., Dhondt, A.A., & Dobson, A.P. (2006). Spatial Spread of an Emerging Infectious Disease: Conjunctivitis in House Finches - Seasonal Rates and Geographic Barriers, Ecology. 87: 3037-3046. esajournals.org. 6. Borer, E., Hosseini, P.R., Seabloom, E., & Dobson, A.P. (2007). Pathogen-induced reversal of native dominance in a grassland community PNAS. 104:5473-5478 doi:10.1073/pnas.0608573104 7. Ballantyne, F.,Menge, D., Ostling, A., & Hosseini, P.R. (2008). Nutrient recycling affects autotroph and ecosystem stoichiometry, American Naturalist. 171 :511-523. doi: 10.1086/528967 8. Barseghian, D., Altintas, I., Jones, M. B., Crawl, D., Potter, N., Gallagher, J., Cornillon, P., Schildhauer, M., Borer, E.T., Seabloom, E.W. & Hosseini, P.R. (2009). Workflows and extensions to the Kepler scientific Biographical Sketches for each listed Senior/Key Person 7 Page 39

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter workflow system to support environmental sensor data access and analysis. Ecological Informatics. 5(1 ):42-50 doi:10.1016/j.ecoinf.2009.08.008 9. Brandt, A.J., Seabloom, E.W., & Hosseini, P.R. (2009). Phylogeny and provenance affect plant-soil feedbacks in invaded California grasslands. Ecology 90: 1063-1072. 10. Moore, S.M., Borer, E.T., Hosseini, P.R. (2010). Predators indirectly control vector-borne disease: linking predator-prey and host-pathogen models, Journal of the Royal Society Interface. 7:161-176 doi: 10.1098/rsif.2009.0131 D. Research Support Ongoing Research Support NSF EF-1015791 Mitchell (Pl) 07/01/10-6/30/15 National Science Foundation/National Institutes of Health: Ecology of Infectious Diseases program. The community ecology of viral pathogens - Causes and consequences of coinfection in hosts and vectors. To conduct mathematical modeling and fieldwork to understand implications in a wild grass, aphid-vectored disease system. Role: Co-Pl NSF Daszak (Pl) 06/21/10 - 06/20/15 Collaborative research: the community ecology of viral pathogens - causes and consequences of coinfection in hosts and vectors. Role: Co-Pl GHN-A-00-09-00010-00 USAID Emerging Pandemic Threats PREDICT - Wildlife SMART Surveillance Morse (Pl) 10/1 /09-09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Role: Hotspots Modeler National Institutes Of Health Daszak (Pl) Fogarty International Center Comparative Spillover Dynamics of Avian Influenza in Endemic Countries Role: Co-Pl Completed Research Support 09/17/12-08/31/13 NIH 3R01TW005869-07S1 Daszak (Pl) 07/01/10- 06/30/11 Research: The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh, Supplement: Understanding and predicting the spread of H5N1 in Bangladesh, China and Globally, Modeling Research Award. To conduct model development and research to understand the role of wild and domestic poultry and livestock in creating the conditions that allow sustained spillover of human-pathogenic influenza viruses into people. Role: Key Personnel NIH 3R01TW005869-07S2 Daszak (Pl) 07/01/10- 06/30/11 Research: The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh, Supplement: Understanding and predicting the spread of H5N1 in Bangladesh, China and Globally, Field Research Award. To conduct fieldwork to understand the role of wild and domestic poultry and livestock in creating the conditions that allow sustained spillover of human-pathogenic influenza viruses into people. Role: Key Personnel NIH 3R01TW005869-06S4 Daszak (Pl) 07/01/09- 06/30/10 Research: The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh, Supplement: Understanding and predicting the spread of H5N1 in Bangladesh, China and Globally, Modeling Research Biographical Sketches for each listed Senior/Key Person 7 Page 40

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Award. To conduct model development and research to understand the role of wild and domestic poultry and livestock in creating the conditions that allow sustained spillover of human-pathogenic influenza viruses into people. Role: Key Personnel NIH 3R01TW005869-06S3 Daszak (Pl) 07/01/09 - 06/30/10 Research: The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh, Supplement: Understanding and predicting the spread of H5N1 in Bangladesh, China and Globally, Field Research Award. To conduct field work to understand the role of wild and domestic poultry and livestock in creating the conditions that allow sustained spillover of human-pathogenic influenza viruses into people. Role: Key Personnel NSF EID 05-25666 Borer (Pl) 09/01/05 - 8/31/10 Research: Predicting the effects of environmental change and host diversity on the dynamics of insectvectored generalist pathogens. Role: Key Personnel The goal of this project was to assess a community-based strategy for reducing alcohol abuse among older individuals. Role: Pl Biographical Sketches for each listed Senior/Key Person 7 Page 41

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. NAME Ge, Xing Yi Follow this format for each person. DO NOT EXCEED FOUR PAGES. POSITION TITLE Assistant Researcher eRA COMMONS USER NAME (credential, e.g., agency login) EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY (if applicable) Huazhong Agricultural University, Wuhan, China 07/05 Biotechnology Huazhong Agricultural University, Wuhan, China 07/08 Preventive Veterinary Medicine Wuhan Institute of Virology, Chinese Academy of Ph.D 12/11 Biochemistry and Sciences, Wuhan, China Molecular Biology A. Personal Statement Throughout my career, I have received extensive molecular training, including deep sequencing, and collaborated in multiple publications in the field of viral genetic diversity in bats in China. I have investigated the genetic diversity of bat adeno-associated viruses and their virus-host interactions, and isolated 22 novel ssDNA viruses from bat fecal samples using inverse PCR, which were then identified to belong in the Circoviridae family. Additionally, using metagenomic analyses, I participated in the characterization of a totivirus from bat feces in China, which showed its capacity of infecting various insect cell lines, thus having a wide geographical distribution. Our most recent work on SARS-like coronaviruses in bats has shown that there are SARS-like CoVs in bats that use the ACE2 receptor, and therefore could be directly transmissible to humans. The discovery of MERS CoV shows that there are other coronaviruses, most likely from bats, that use different receptors to infect people. For this reason, we should understand the diversity of bat coronaviruses in China and determine whether they can infect people. In the current proposal, which aims to study coronaviruses in China, I will be responsible for the diagnosis, genomics and isolation of coronaviruses and for analyzing their receptor binding domains, in order to understand their viral spillover risk and geographic distribution. We have shown that our lab at Wuhan has the ability to identify and test these viruses for receptor usage, and I am confident that this study will allow us to find many other coronaviruses in nature with zoonotic potential. B. Positions and Honors Positions and Em lo ment 2005-2008 Master's Training, College of Veterinary Medicine, Huazhong Agricultural University, China 2008-2011 Doctoral Training, Wuhan Institute of Virology, Chinese Academy of Sciences, China 2010 Doctoral Training, Unit of Molecular Genetics of RNA Viruses, Department of Virology, Institute Pasteur, Paris, France 2012- Assistant Researcher, Wuhan Institute of Virology, Wuhan, China Honors 2005 2005 2007 2012 Excellent Thesis of Bachelor Degree of Hubei province Innovation Award of Huazhong Agricultural University First Prize of Excellent Graduate student CAS Presidential Scholarship (Excellence Prize) C. Selected Peer-reviewed Publications Biographical Sketches for each listed Senior/Key Person 8 Page 42

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Most relevant to the current a lication 1. Li, Y., Ge X.Y., Hon C.C., Zhang, H., Zhou P., Zhang Y., Wang L.F., Shi Z. (2010). Prevalence and genetic diversity of adeno-associated viruses in bats, China. Journal of General Virology, 91 (10), 2601-9. 2. Ge* X.Y., Rameix-Welti*, M.A., Gault* E., Chase, G., dos Santos Afonso, E., Picard D., Schwemmle, M., Naffakh, N. (2011 ). Influenza Virus Infection Induces the Nuclear Relocalization of the Hsp90 CoChaperone p23 and Inhibits the Glucocorticoid Receptor Response. PLoS One, 6(8), e23368. (*equal contribution) 3. Moisy, D., Jacob, Y., Laoide, B.M., Ge, X.Y., Baudin, F., Naffakh, N., Jestin, J.L. (2012). The HMGB1 protein binds to influenza virus nucleoprotein and promotes viral replication. Journal of Virology, 86(17), 9122-33. 4. Ge, X.Y., Li, Y., Yang X., Zhang H., Zhou P., Zhang Y., & Shi Z. (2012). Metagenomic Analysis of Viruses from the Bat Fecal Samples Reveals Many Novel Viruses in Insectivorous Bats in China. Journal of Virology, 86(8), 4620-30. 5. Wu L., Zhou, P., Ge X.Y., Wang, L.F., Baker M., Shi Z. (2013). Deep RNA sequencing reveals a complex transcriptional landscape of a bat adenovirus. Journal of Virology, 87(1 ), 503-11. Additional recent publications of importance to the field (in chronological order) 1. Li, Y., Ge, X.Y., Zhang, H., Zhou, P., Zhu, Y., Zhang, Y., Yuan, J., Wang, L.F., Shi, Z. (2010). Host range, prevalence, and genetic diversity of adenoviruses in bats. Journal of Virology, 84(8), 3889-97. 2. Zhang, Y., Zhang, H., Dong, X., Yuan, J., Zhang, H., Yang, X., Zhou, P., Ge, X.Y., Li, Y., Wang, L.F., Shi, Z. (2010). Hantavirus outbreak associated with laboratory rats in Yunnan, China. Infection, Genetics and Evolution, 10(5), 638-44. 3. Ge, X.Y., Li, J., Peng, C., Wu, L., Yang, X., Wu, Y., Zhang, Y., Shi, Z. (2011 ). Genetic diversity of novel circular ssDNA viruses in bats in China. Journal of General Virology, 92, 2646-2653. 4. Yang, X., Zhang, Y., Ge, X.Y., Yuan, J., Shi, Z. (2012). A novel totivirus-like virus isolated from bat guano. Archives of Virology, 157(6), 1093-9. D. Research Support Ongoing Research Support Completed Research Support Biographical Sketches for each listed Senior/Key Person 8 Page 43

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter BIOGRAPHICAL SKETCH Provide the following informafion for the Senior/key personnel and other significant contributors. Follow this formal for each person. DO NOT EXCEED FOUR PAGES. NAME Zhu, Gunagjian eRA COMMONS USER NAME (credential, e.g., agency login) xxxx POSITION TlnE Assistant Researcher EDUCATION/TRAINING (Begin with baccalaureate or other Initial professional education, such as nursing. include postdoctoral training and residency /raining if applicable.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY /if applicable) East China Normal University, Shanghai, China B.S. 07/03 Biology Science Hainan Normal University, Haikou, China M.S. 07/03 Ecology East China Normal University, Shanghai, China Ph.D 6/12 Biochemistry and Molecular Biology A. Personal Statement Throughout my graduate studies and work with East China Normal University, I have carried out molecular biology and field ecology research focused on bat genetics and viral diversity. I have co-authored multiple publications in the field of viral genetics and bat ecology under the mentorship of Dr. Shuyi Zhang. I have also worked actively with EcoHealth Alliance on the USAID- EPT PREDICT program as a field team leader for China. For this program I have been responsible for the identification of high-risk interfaces between wildlife and people, where close contact might allow for zoonotic pathogen spillover (e.g. live animal markets). I have also led wildlife surveys which involved bat and rodent capture and sampling for viral discovery. Through this work we have conducted site-selection and wild and domestic animal sampling in Guangxi, Yunnan, Guangdong and Shanghai, and have compiled archived and current samples from birds in Shanghai Chongming Reserve for H7N9 avian influenza analyses. Under the USAID PREDICT program I collected several hundred bat samples which have been tested for coronaviruses (and several other viral families) at the Wuhan Institute of Virology. Under this current proposal, I would be responsible for developing and leading a wildlife team to sample bats, rodents, and other small mammals in the live animal markets of southern China. Through my graduate and professional work I have developed expertise in collecting high-quality, nondestructive samples from wildlife as well as expertise in molecular diagnostics. This combination of experiences allows me to understand the whole process of bringing samples from field to lab with an understanding of how to maximize opportunity for viral detection. I think that the aims of this proposal are important for providing the most current information about viral dynamics in live animal markets in China, particularly in rural areas where wildlife trade still occurs and where there is little data on spillover. I am very enthusiastic about participating in this study and confident that it has the right experts and study plan to succeed. B. Positions and Honors Positions and Em lo ment 2007- Assistant Researcher, Guangdong Entomological Institute, China Other Ex eriences and Professional Membershii:1s Honors 2009 Biology Prize of the 2009 lg Nobel Prize C. Selected Peer-reviewed Publications Most relevant to the current a lication Biographical Sketches for each listed Senior/Key Person 9 Page 44

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 1. Zhu, G., Han, N., Hong, T., Tan, M., Yu, D., Zhang, L. (2008). Echolocation Call, Roost and ND 1 Sequence Analysis of New Record of Nyctalus plancyi ( Chiroptera : Vespertilionidae} on Hainan Island. Zoological Research, 29(4), 447-451. (in Chinese) 2. Zhu, G., Li, D., Ye, J., Hong, T., Zhang, L. (2008). New Record of la io in Hainan Island, its Echolocation Pulses and ND1 Analysis. Chinese Journal of Zoology, 43(5), 69-75. (in Chinese) 3. Sun, Y., Yu, D., Zhu, G., Liu, X., Zhang, S.Y., Chen, J. (2009). Isolation and characterization of 11 microsatellite loci in Scolophi/us kuhlii (Lesser Asiatic Yellow House Bat). Conservation Genetics, 10, 1857-1859. 4. Mao, X., Zhu, G., Zhang, S.Y., Rossiter, S.J. (2010). Pleistocene climatic cycling drives intra-specific diversification in the intermediate horseshoe bat (Rhinolophus affinis) in Southern China. Molecular Ecology, 19(13), 2754-2769. 5. Hua, P., Zhang, L., Zhu G., Jones, G., Zhang, S., Rossiter, S.J. (2011 ). Hierarchical polygyny in multiparous lesser flat-headed bats. Molecular Ecology, 20(17), 3669-3680. Additional recent publications of importance to tt}e field (in chronological oder) 1. Zhu, G, Tang, Z., Liang, B., Zhang, X. (2007). Diet and Roost Site of Cynopterus sphinx in Winter in Haikou. Chinese Journal of Zoology, 42(4 ), 22-27. (in Chinese) 2. Zhang, L., Zhu, G, Jones, G., Zhang, S.Y (2009) Conservation of bats in China: problems and recommendations. ORYX, 43(2), 179-182. 3. Tan, M., Jones, G., Zhu, G., Ye, J., Hong, T., Zhou, S., Zhang, S., Zhang, L. (2009). Fellatio by fruit bats prolongs copulation time. PLoS One, 4(10), e7595. 4. Ma, J., Jones, G., Zhu, G., Metzner, W. (2010). Echolocation behaviours of the Japanese pipistrelle bat Pipistrel/us abramus during foraging flight. Acta Theriologica. 55(4), 315-332. 5. Zhu, G., Chmura, A., Zhang, L. (2011). Morphology, echolocation calls and diet of Scotophilus kuh/ii LC.lli.!,oote.r,a· Vesoe.!!i!.Lo.JlLda~l o.!l.l:lainaJl.Lsland._s_oJJJ!l..C.lli.C!a..Acta ChirooJerolo!'.li_ca . ..li(J) 175.:18.1.- 6. (b) (4] D. Research ;Supporti Ongoing Research Support GHN-A-00-09-00010-00 Morse (Pl) 10/01/09-09/30/14 PREDICT-Wildlife SMART Surveillance/PREDICT Project to pre-empt at the earlier stages possible, zoonotic diseases that impose significant threat to public health. Role: Field Team Leader Completed Research Support Biographical Sketches for each listed Senior/Key Person 9 ~=t#liiiJFffJIH Canment [1]: Other support? Page 45

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FOUR PAGES. NAME POSITION TITLE Zhang, Yun-Zhi Chief Physician, Professor eRA COMMONS USER NAME (credential, e.g., agency login) EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable.) INSTITUTION AND LOCATION DEGREE MM/YY FIELD OF STUDY (if applicable) School of Life Sciences, Yunnan University B.S. 1990 Endemic Diseases Kunming Medical University M.D. 2005 Medicine Wuhan Institute of Virology, Chinese Academy of PhD 2010 Sciences Virology A. Personal Statement My career in public health is focused on virology and surveillance for zoonotic infections, including hantaviruses, henipaviruses and coronaviruses. As head of infectious disease surveillance at Yunnan CDC, I am particularly interested in the risk of new pathogens emerging through the wildlife trade, which Yunnan is on the front line of in China. Through collaborative research with Wuhan Institute of Virology (Zhengli Shi) and EcoHealth Alliance (Peter Daszak, Jon Epstein and Kevin Olival), we have conducted specific surveillance in bats, rodents and primates in Yunnan Province, and on the border with Myanmar, Laos and Vietnam. This has led to our discovery of numerous CoVs in mammals, including bats, and including the recent finding of a bat SL-CoV that uses ACE2. Given my collaboration with this group, and our capacity to do extensive surveillance of wildlife and people in Yunnan, I believe that this project will generate substantial results and help us understand the risk of CoV emergence from wildlife much better in the future. B. Positions and Honors Positions and Em 2003-6 2006-9 2009-Present lo ment Deputy chief physician, Public Health Branch of the Chinese Medical Association Youth Committee Head of infectious disease surveillance, Yunnan Center for Disease Control, Peoples' Republic of China. Head of Infectious disease surveillance, Yunnan Institute of Endemic Disease Control and Prevention, Peoples' Republic of China Other Ex erience and Professional Membershi s 2002-2004 Participant, international Field Epidemiology Training Program (FETP) C. Selected Peer-reviewed Publications Most relevant to the current a lication 1. Li, Y., Wang, J.M., Hickey, A.C., Zhang, Y.Z., Li, Y., Wu, Y., Zhang, H., Yuan, J., Han, Z.G., McEachern, J., Broder, C.C., Wang, L.F. & Shi, Z. (2008). Antibodies to Nipah or Nipah-like viruses in bats, China. Emerging Infectious Diseases, 14(12):1974-1976 Biographical Sketches for each listed Senior/Key Person 1 0 Page 46

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter 2. Zhang, Y.Z., Zhang, H.L., Dong, X.Q., Yuan, J.F., Zhang, H.J., Yang, X.L., Zhou, P., Ge, X.Y., Li, Y., Wang, L.F., Shi, Z.L. (2010). Hantavirus outbreak associated with laboratory rats in Yunnan, China. Infection, Genetics and Evolution, 10(5):638-644 3. Li, Y., Ge, X., Zhang, H., Zhou, P., Zhu, Y., Zhang, Y.Z., Yuan, J., Wang, L.F. & Zhengli, S. (2010). Host range, prevalence, and genetic diversity of adenoviruses in bats. Journal of Virology, 84(8):3889-3897 4. Li, Y., Ge, X., Hon, C.C., Zhang, H., Zhou, P., Zhang, Y.Z., Wu, Y., Wang, L.F. & Shi, Z. (2010). Prevalence and genetic diversity of adeno-associated viruses in bats, China. Journal of General Virology, 91(10):2601-2609 5. Zhang, Y.Z., Yuan, J., Yang, X., Zhou, J., Yang, W., Peng, C., Zhang, H.L., Shi, Z. (2011 ). Novel Hantavirus detected in Yunnan Red-backed Vole Eothenomys miletus, China. Journal of General Virology, 92(3): 1454-1457. Additional recent ublications of im ortance to the field in chronolo ical order 1. Yuan, J.F., Zhang, Y.J., Li, J.l., Zhang, Y.Z., Wang, L.F., Shi, Z.L. (2012). Serological evidence of ebolavirus infection in bats, China. Virology Journal, 9: 236; doi: 10.1186/17 43-422X-9-236 2. Yang, X.L., Zhang, Y.Z., Ge, X.Y., Yuan, J.F., Shi, Z.L. (2012). A novel totivirus-like virus isolated from bat guano. Archives of Virology, 157 (6), 1093-1099, doi: 10.1007/sOO?OS-012-1278-y 3. Ge, X.Y., Li, Y., Yang, XL, Zhang, H.J., Zhou, P., Zhang, Y.Z., Shi, Z.L. (2012). Metagenomic Analysis of Viruses from Bat Fecal Samples Reveals Many Novel Viruses in Insectivorous Bats in China. Journal of Virology. 86(8). 4620-4630, doi. 10.1128/JVl.06671-11 D. Research Support Ongoing Research Su m.QI! Ministry of Science 01/01/2013-12/01/2017 Yunnan region is an important natural reservoir of the virus and the insect vector, pathogen survey Grant No.: 81260437 01/01/2013 -12/01/2016 National Natural Science Foundation of China Yunnan murine viral metagenome important viral epidemic status and related research Grant No.: (b) (4) Yunnan Talented ----young technology leaders Completed Research Support Grant No.: 81060132 National Natural Science Foundation of China, (b) (4) Yunnan novel hantavirus distribution, pathogenicity and receptor research Grant number: (b) (4) Yunnan applied ---- basic research projects Biographical Sketches for each listed Senior/Key Person 10 11/01/ 2012- 11/01/2015 09/01/2011 -12/01/2014 01/01/2011-12/01/2013 01/01 /2011-12/01/2013 Page 47

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter • ORGANIZATIONAL DUNS: ~10_1_10_9_0_0_66_0_0_0_0 ______ ~ • Budget Type: [2J Project 0 Subaward/Consortium Enter name of Organization: !Eco Health Alliance, Inc. Delete Entry I* Start Date: '10101120131 • End Date: !09;30120141 Budget Period 1 A. Senior/Key Person Prefix 1· lor. 2. lor. 3. lor. 4• Dr. 5. • First Name IIPetec IIJonathan IIKevin Parviez 6. ::==~ 7. ::==~ Middle Name • Last Name Suffix • Project Role ioaszak iPD/PI !IH. IIEestoin Jsenior/Key PersonnJ !IJ. lio11va1 isenior/Ke~ Personn~ R. Cal. Acad. Sum. • Requested Base Salary ($) Months Months Months Salary ($) I I I I I c=J~c=Jl IT I c=J~c=11 II 0MB Number: 4040·0001 Expiration Date: 06/30/2011 • Fringe Benefits ($) • Funds Requested ($) (b) (4). (b) (6) IT !I s. =~~==~~~~~~=:~~~~~~~=:=~~~~~~==~~= :=~ -==--==--==--==-~========c=J~c=J~I --~'~' --~~=======: I c=J~c=JI II !I 9. Total Funds requested for al Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel • Number of Personnel ~--------------~ Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical !Research Scientist (tbd) !Program Coordinator • Project Role Add Attachment Total Senior/Key Person 199,498 oo Delete Attachment View Attachment Cal. Acad. Sum. • Requested • Fringe Months Months Months Salary($) Benefits ($) • Funds Requested ($) :====:'·:=' ==:l:=1==::======: :=====: :=======: :==:'::::' ==:l:=1==::=====::::====: :=====: :=====:':=' =~=:::::::====::::::====:::::====::::: II II c=J c=J ~ c=J ~ ~ c=J c=J c=J ~ ~ :===========================::=====:':=' =~':=' =:::::::====::::::====:::::====::::: :============================:::::=::::::'::::' =~1::::1 =:::::::::===:::::::::===::::: ::::===::::: :======================~:==:':=' ==:':='==::=====::::====: :=====: .__ _____________________________ ___,.__ _ _,I._I _ __,l._1 _ __,.___ __, .__ ___ __, .__ ___ __, Total Number Other Personnel RESEARCH & RELATED Budget (A-8) (Funds Re~t3g~fe~fdget - Year 1 Tracking Number:GRANT11418584 Total Other Personnel 1122, 378. oo Total Salary, Wages and Fringe Benefits (A+B) 1221, 876.oo Page48 Funding Opportunity Number:PA-11-260 Received Date:2013-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 1 • ORGANIZATIONAL DUNS: lono<1ooi;wooo I • Budget Type: IZJ Project D Subaward/Consor\lum Enter name of Organization: IEcoHealdt ~.lliar.ce, Inc. Delete Entry I* Start Date: 11 o;n 1 ;;, 01 2 1 •End Date: I ~913012 r 11 Budget Period 1 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment item • Funds Requested ($) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. ::========================================================:::::::=============::::: 11. Total funds requested for all equipment listed in the attached file Total Equipment Additional Equipment: Add Attachment Delete Attachment D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions) 2. Foreign Travel Costs Funds Requested ($) 13,605.00 '32,313.00 Total Travel Cost '3s, 918 . 00 E. Participant/Trainee Support Costs Funds Requested ($) 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other ~ Number of Participants/Trainees Total Participant/Trainee Support Costs ~------~ RESEARCH & RELATED Budget {C·E) (Funds Requested) Detailed Budget - Year 1 Page 49 View Attachment ] Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 1 • ORGANIZATIONAL DUNS: lnn900G&oooo I • Budget Type: ~ Project D Subaward/Consortium Enter name of Organization: IEccHea· th ;,,.''' ancF, ·nc. Delete Entry I Start Date: I· 0 ; 01 , 20 - ~I• End Date: I O<J/ 3012014 I Budget Period F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. !shipping & Communications 9. 10. Funds Requested ($) 121,400 .oo 1227,663.00 110,000.00 Total Other Direct Costs l2s91 063 .oo G. Direct Costs Funds Requested ($) Total Direct Costs (A thru F) ls 16, ss7 .oo H. Indirect Costs Indirect Cost Type Indirect Cost Rate{%) Indirect Cost Base($) 1. IEcoJ-Jeal th Alliance F&A Rate I 14 4 • 10 l2a9 1 195. 00 ~::::::::::=~ 2. l,;cottealth Alliance F&A on 2 subawarl 144 .10 lso, ooo .oo 3. :===============: 4. '-----------------' ._ ___ __, .__ _____ _, • Funds Requested {$) li21, 535.00 122,050.00 Total Indirect Costs h49, 585.oo Cognizant Federal Agency '---------------------------~ (Agency Name, POC Name, and POC Phone Number) I. Total Direct and Indirect Costs Total Direct and Indirect Institutional Costs (G + H) J.Fee Funds Requested ($) 1666, 442. 00 Funds Requested ($) K. * Budget Justification 11239-EHA NIAID COV BUDGETJUSTIFICATIOJ Add Attaci'lment Delete Attachment (Only attach one file.) RESEARCH & RELATED BudQ.et {F·Kl (Funds Requested) "Detailed Budget • Year 1 Page 50 Next Period View Attachment Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Previous Period I RESEARCH & RELATED BUDGET- SECTION A & B, BUDGET PERIOD 2 0MB Number: 4040·0001 Expiration Date: 06/30/2011 • ORGANIZATIONAL DUNS: =10=77=0=9=0=06=6=◊=00=0=============1 • Budget Type: [2J Project 0 Subaward/Consortium Enter name of Organization: !Eco Health Alliance, Inc. Delete Entry I* Start Date: '10101120141 • End Date: lo9;301201sl Budget Period 2 A. Senior/Key Person Prefix 1· lor. 2. lor. 3. lor. 4· Dr. 5. • First Name IIPetec IIJonathan IIKevin Parviez 6. ::==~ 7. ::==~ Middle Name !IH. !IJ. R. • Last Name Suffix • Project Role ioaszak iPD/PI IIEestoin Jsenior/Key PersonnJ lio11va1 isenior/Ke~ Personn~ Cal. Acad. Sum. • Requested • Fringe Base Salary ($) Months Months Months Salary ($) Benefits ($) • Funds Requested ($) I I (b) (4). (b) (6) I I I I I I I lc=]~c=JI II !I I I lc=]~c=11 II !I I s. =~~==~~~~~~=:~~~~~~~=:=~~~~~~==~~= :=~ -==- -=- -=- -=- ~========c=J~c=J~I --~'~' --~~=======: I lc=]~c=]I II !I I 9. Total Funds requested for al Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel • Number of Personnel c=J c=J ~ c=J ~ ~ c=J c=J c=J ~ ~ ~--------------~ • Project Role Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical IResearch Scientist IProqram Coordinator I I I I Total Number Other Personnel RESEARCH & RELATED Budget (A-8) (Funds Re~t3g~fe~fdget - Year 2 Tracking Number:GRANT11418584 Add Attachment Total Senior/Key Person l1os, 265 oo Delete Attachment View Attachment I I I I I I I I Cal. Acad. Sum. • Requested • Fringe Months Months Months Salary($) Benefits ($) • Funds Requested ($) II II 11 Ii I I I II II 11 Ii I I I II II 11 Ii I I I II II 11 11 I I I II II II II I I (bl(•~b)(l II II 11 11 I I I II II 11 11 I I I 11 II 11 11 I I I Total Other Personnel 112 9, 4 68. oo Total Salary, Wages and Fringe Benefits (A+B) lz34 , 733 . 00 Page 51 Funding Opportunity Number:PA-11-260 Received Date:2013-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 2 • ORGANIZATIONAL DUNS: lono<1ooi;wooo I • Budget Type: IZJ Project D Subaward/Consor\lum Enter name of Organization: IEcoHealdt ~.lliar.ce, Inc. Delete Entry I* Start Date: 11 o;n 1 ;;, 01 4 I' End Date: I ~913012 r c; I Budget Period 2 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment item • Funds Requested ($) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. ::========================================================:::::::=============::::: 11. Total funds requested for all equipment listed in the attached file Total Equipment Additional Equipment: Add Attachment Delete Attachment D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions) 2. Foreign Travel Costs Funds Requested ($) 13,605.00 '32,313.00 Total Travel Cost '3s, 918 . 00 E. Participant/Trainee Support Costs Funds Requested ($) 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other ~ Number of Participants/Trainees Total Participant/Trainee Support Costs ~------~ RESEARCH & RELATED Budget {C·E) (Funds Requested) Detailed Budget - Year 2 Page 52 View Attachment ] Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 2 • ORGANIZATIONAL DUNS: lnn900G&oooo I • Budget Type: ~ Project D Subaward/Consortium Enter name of Organization: IEccHea· th ;,,.''' ancF, ·nc. Delete Entry I Start Date: I· 0 ; 01 , 2 0 - 4 1• End Date: I O<J/ 301201 <; I Budget Period 2 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. !shipping and Communications 9. !Local Reimbursement 10. Funds Requested ($) IL9,250.00 12,600.00 1211, 699 .oo ILO,ooo .oo 1950.00 Total Other Direct Costs 12441 499 .oo G. Direct Costs Funds Requested ($) Total Direct Costs (A thru F) Isis, 1so .oo H. Indirect Costs Indirect Cost Type Indirect Cost Rate{%) Indirect Cost Base($) 1. IEcoJ-Jealth Alliance F&A 1144.10 1303 1 450.00 ~::::::::::=~ 2. l,;cottealth Alliance F&A on 2 subawarl 144 .10 lso, ooo .oo 3. :===============: 4. '-----------------' ._ ___ __, .__ _____ _, • Funds Requested {$) li33, 822. 00 122,050.00 Total Indirect Costs Ii 55. 87 2. oo Cognizant Federal Agency '---------------------------~ (Agency Name, POC Name, and POC Phone Number) I. Total Direct and Indirect Costs Total Direct and Indirect Institutional Costs (G + H) J.Fee Funds Requested ($) 1671, 022. 00 Funds Requested ($) K. * Budget Justification 11239-EHA NIAID COV BUDGETJUSTIFICATIOJ Add Attacnment Delete Attachment (Only attach one file.) RESEARCH & RELATED Budg__et {F·Kl (Funds Requested) Tietailed Budget• Year 2 Page 53 Next Period View Attachment Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Previous Period I RESEARCH & RELATED BUDGET- SECTION A & B, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: =10=77=0=9=0=06=6=◊=00=0=============1 • Budget Type: [2J Project 0 Subaward/Consortium Enter name of Organization: !Eco Health Alliance, Inc. Delete Entry I* Start Date: '101011201sl • End Date: !09;30120161 Budget Period 3 A. Senior/Key Person Cal. Acad. Sum. Prefix 1· lor. • First Name Middle Name • Last Name Suffix • Project Role Base Salary ($) Months Months Months IIPetec 2. lor. l!Jonathan 3. lor. 4· Dr. 5. IIKevin ::==~ 6. ::==~ 7. Parviez loaszak !IH. l!Epstoin !IJ. llo11va1 =====:::=::=:::=:::::::::::==::::;:::====; ;::::IPD:::::/::::::P::::::I ====:::::!':=' ===~I ===========:::=:'.::=========::::::::=====::::: Jsenio r /Key PersonnJ I:==========::: !senior/Key PersonnJI =========::~========::====~ :==========::::1 R. ~~c=JI ~~c=i, :=::==::::::::=========:::::::::===========:::===========::::::::=====::::: ~~ -_ -_ -_ -_ -_ -_ -_ -_ -_ -_ -_ -_ -_ -_ -~ ~~ -_ -_ -_ -_ -_ -_ -_ -_ -_ ~~ 8. ~~~I ~~c=11 9. Total Funds requested for al Senior Key Persons in the attached file • Requested Salary($) II II II II 0MB Number: 4040·0001 Expiration Date: 06/30/2011 • Fringe Benefits ($) • Funds Requested ($) (b) (4), (b) (6) 11 11 11 I Total Senior/Key Person 1111, 35 7. oo Additional Senior Key Persons: Add Attachment Delete Attachment View Attachment B. Other Personnel • Number of Personnel ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~--------------~ Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical !Research Scientist !Program Coordinator Total Number Other Personnel • Project Role RESEARCH & RELATED Budget (A-8) (Funds Re~t3g~fe~fdget - Year 3 Tracking Number:GRANT11418584 Cal. Acad. Sum. • Requested • Fringe Months Months Months Salary($) Benefits ($) • Funds Requested ($) II 11 II ll I I II II 11 ll I I II II II ll I I II 11 II ll I I (b) (4), (b) (6) II II 11 11 I I II 11 II ll I I II II II ll I I II II II 11 I I Total Other Personnel 1136, 961. 00 Total Salary, Wages and Fringe Benefits (A+B) 1248, 318.00 Page 54 Funding Opportunity Number:PA-11-260 Received Date:2013-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: lono<1ooi;wooo I • Budget Type: IZJ Project D Subaward/Consor\lum Enter name of Organization: IEcoHealdt ~.lliar.ce, Inc. Delete Entry I * Start Date: I 10 ;n 1 ;;, 01 sl •End Date: I ~ 913012 r "I Budget Period 3 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment item • Funds Requested ($) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. ::========================================================:::::::=============::::: 11. Total funds requested for all equipment listed in the attached file Total Equipment Additional Equipment: Add Attachment Delete Attachment D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions) 2. Foreign Travel Costs Funds Requested ($) 13,605.00 '32,313.00 Total Travel Cost '3s, 918 . 00 E. Participant/Trainee Support Costs Funds Requested ($) 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other ~ Number of Participants/Trainees Total Participant/Trainee Support Costs ~------~ RESEARCH & RELATED Budget {C·E) (Funds Requested) Detailed Budget - Year 3 Page 55 View Attachment ] Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: lnn900G&oooo I • Budget Type: ~ Project D Subaward/Consortium Enter name of Organization: IEccHea· th ;,,.''' ancF, ·nc. Delete Entry I Start Date: I· 0 ; 01 , 2o sl• End Date: I O<J/ 30 ; 2 oH I Budget Period 3 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. !shipping and Communications 9. !Local Reimbursement 10. Funds Requested ($) 11 ( 250. 00 12,600.00 l2u, 238 .oo 11' 500. 00 1950.00 Total Other Direct Costs 12311 538 .oo G. Direct Costs Funds Requested ($) Total Direct Costs (A thru F) Isis, 774 .oo H. Indirect Costs Indirect Cost Type Indirect Cost Rate{%) Indirect Cost Base($) 1. IEcoJ-Jeal th Alliance F&A Rate I 14 4 • 10 1302, 53 6. 00 ~ IEcoHealth Alliance F,A Rate on 2 subl4t.10 150,ooo.oo 3. ::===========:::::::::=I====: ::=======: 4. L..I ___ _, '--------' • Funds Requested {$) li33, 418.00 122,050.00 Total Indirect Costs Ii ss, 4 68. oo Cognizant Federal Agency ~-------------------------~ (Agency Name, POC Name, and POC Phone Number) I. Total Direct and Indirect Costs Total Direct and Indirect Institutional Costs (G + H) J.Fee Funds Requested ($) 1671, 242. 00 Funds Requested ($) K. * Budget Justification 11239-EHA NIAID COV BUDGETJUSTIFICATIOJ Add Attacnment Delete Attachment (Only attach one file.) RESEARCH & RELATED Budg__et {F·Kl (Funds Requested) Tietailed Budget• Year 3 Page 56 Next Period View Attachment Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Previous Period I RESEARCH & RELATED BUDGET- SECTION A & B, BUDGET PERIOD 4 0MB Number: 4040·0001 Expiration Date: 06/30/2011 • ORGANIZATIONAL DUNS: =10=77=0=9=0=06=6=◊=00=0=============1 • Budget Type: [2J Project 0 Subaward/Consortium Enter name of Organization: !Eco Health Alliance, Inc. Delete Entry I* Start Date: '1010112ol6I • End Date: !09;30120171 Budget Period 4 A. Senior/Key Person Prefix 1· lor. 2. lor. 3. lor. 4· Dr. 5. • First Name IIPetec IIJonathan IIKevin Parviez 6. ::==~ 7. ::==~ Middle Name !IH. !IJ. R. • Last Name Suffix • Project Role ioaszak iPD/PI IIEestoin Jsenior/Key PersonnJ lio11va1 isenior/Ke~ Prsonn~ Cal. Acad. Sum. • Requested • Fringe Base Salary ($) Months Months Months Salary ($) Benefits ($) • Funds Requested ($) I I (b) (4), (b) (6) I I I I I c=J~c=Jl IT IT I c=J~c=11 II !I s. =~~==~~~~~~=:~~~~~~~=:=~~~~~~==~~= :=~ -==--==--==--==-~========c=J~c=J~I --~'~' --~~=======: I c=J~c=JI II !I 9. Total Funds requested for al Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel • Number of Personnel ~--------------~ Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical !Research Scientist !Program Coordinator • Project Role Add Attachment Total Senior/Key Person 1116, 925 oo Delete Attachment View Attachment Cal. Acad. Sum. • Requested • Fringe Months Months Months Salary($) Benefits ($) • Funds Requested ($) :====:'·:=' ==:l:=1==::======: :=====: :=======: :==:'::::' ==:l:=1==::=====::::====: :=====: :=====:':=' =~=:::::::====::::::====::::: :====::::: II II c=J c=J ~ c=J ~ ~ c=J c=J c=J ~ ~ :======================::::::::~=::::1:::1 ==:1::::1 =:::::1~1 =====:1:::::1 =====: :===~ :======================~:=====:l::::1=~11 I :='=====::===~ :::=======================::=~ll~==:11 I ~'====::::====: '---------------------------~~-_,11, _ ___,II I ~I---~~---~ Total Number Other Personnel RESEARCH & RELATED Budget (A-8) (Funds Re~t3g~fe~fdget - Year 4 Tracking Number:GRANT11418584 Total Other Personnel 1143, 808. oo Total Salary, Wages and Fringe Benefits (A+B) lz60 , 733 . 00 Page 57 Funding Opportunity Number:PA-11-260 Received Date:2013-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET· SECTION C, D, & E, BUDGET PERIOD • ORGANIZATIONAL DUNS: lono<1ooi;wooo I • Budget Type: IZJ Project D Subaward/Consor\lum Enter name of Organization: IEcoHealdt ~.lliar.ce, Inc. Delete Entry I * Start Date: I 10 ;n 1 ;;, 01 6 1' End Date: I ~ 913012 r 7 1 Budget Period 4 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment item • Funds Requested ($) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. ::========================================================:::::::=============::::: 11. Total funds requested for all equipment listed in the attached file Total Equipment Additional Equipment: Add Attachment Delete Attachment D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions) 2. Foreign Travel Costs Funds Requested ($) 13,605.00 '32,313.00 Total Travel Cost '3s, 918 . 00 E. Participant/Trainee Support Costs Funds Requested ($) 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other ~ Number of Participants/Trainees Total Participant/Trainee Support Costs ~------~ RESEARCH & RELATED Budget {C·E) (Funds Requested) Detailed Budget - Year 4 Page 58 View Attachment ] Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 4 • ORGANIZATIONAL DUNS: lnn900G&oooo I • Budget Type: ~ Project D Subaward/Consortium Enter name of Organization: IEccHea· th ;,,.''' ancF, ·nc. Delete Entry I Start Date: I· 0 ; 01 , 2o f I• End Date: I O<J/ 301201 7 I Budget Period 4 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. !shipping & Communications 9. !Local Reimbursement 10. Funds Requested ($) 11 ( 000. 00 12,600.00 1201, 422 .00 16,250.00 1950.00 Total Other Direct Costs bs, 222 .oo G. Direct Costs Funds Requested ($) Total Direct Costs (A thru F) ls14, a13 .oo H. Indirect Costs Indirect Cost Type Indirect Cost Rate{%) Indirect Cost Base($) 1. IEcoJ-Jealth Alliance F&A 1144.10 1313 1 452.00 ~::::::::::=~ 2. l,;cottealth Alliance F&A on 2 subawarl 144 .10 150, ooo .oo 3. :===============: 4. '-----------------' ._ ___ __, .__ _____ _, • Funds Requested {$) l13a, 232. oo 122,050.00 Total Indirect Costs ii Go, 222. oo Cognizant Federal Agency '---------------------------~ (Agency Name, POC Name, and POC Phone Number) I. Total Direct and Indirect Costs Total Direct and Indirect Institutional Costs (G + H) J.Fee Funds Requested ($) 1675, 155. 00 Funds Requested ($) K. * Budget Justification 11239-EHA NIAID COV BUDGETJUSTIFICATIOJ Add Attacnment Delete Attachment (Only attach one file.) RESEARCH & RELATED Budg__et {F·Kl (Funds Requested) Tietailed Budget• Year 4 Page 59 Next Period View Attachment Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Previous Period I RESEARCH & RELATED BUDGET- SECTION A & B, BUDGET PERIOD 5 0MB Number: 4040·0001 Expiration Date: 06/30/2011 • ORGANIZATIONAL DUNS: =10=77=0=9=0=06=6=◊=00=0=============1 • Budget Type: [2J Project 0 Subaward/Consortium Enter name of Organization: !Eco Health Alliance, Inc. Delete Entry I* Start Date: '10101120171 • End Date: !09;301201sl Budget Period 5 A. Senior/Key Person Prefix • First Name Middle Name • Last Name Suffix • Project Role 1- lor. IIPetec loaszak IPD/PI :::::=::====::::::::=====~:==========:::==::::: ;::::===========! 2. :=====:::::l:=IJ=o=na=t=h=a=n====::::!l:=H=. ==========::::ll:=E:'.::ps=t=o=i=n======:::::=====::::: lsenio r /Key PersonnJ 3. IIKevin !IJ. llo11va1 !senior/Key PersonnJ ~~ 4. Parviez R. :::::==::::::::======:::!:=:======:::!!:::::::::::::======::::::====:::! :::=:::::::::::::::::::::::::::::::=:::::::::::::::::::::::::::::::::::! 5. 6. ~~ 7. ~~ Cal. Acad. Sum. • Requested • Fringe Base Salary ($) Months Months Months Salary ($) Benefits ($) • Funds Requested ($) I I (b) (4), (b) (6) I I I I I c=J~c=Jl IT IT I c=J~c=11 II !I s. =~~==~~~~~~=:~~~~~~~=:=~~~~~~==~~= :=:~ -==--==--==--==-~========c=J~c=J~I --~'~' --~~=======: I c=J~c=JI II !I 9. Total Funds requested for al Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel • Number of Personnel ~--------------~ Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical !Research Scientist !Program Coordinator • Project Role Add Attachment Total Senior/Key Person 112 2 ,· 772. oo Delete Attachment View Attachment Cal. Acad. Sum. • Requested • Fringe Months Months Months Salary($) Benefits ($) • Funds Requested ($) :====:I·:=' ==:1:=1 ===::======: :=====: :=======: ::====:'::=' ==:1::::1 ==:::!:=======::::====: :=======: :=====:1::::1 =~1::::1 =::::::::::===:::::::::===::::: :===::::: II 11 c=J c=J ~ c=J ~ ~ c=J c=J c=J ~ ~ :======================::::::::~=::::1:::1=~1::::1 =::::::l~I =====:1:=I =====: :===~ :======================~:=====:l::::1=~11 I :='=====::===~ :==========================::==:::::u~==::::11 I ~'======::=====: '---------------------------~~ _ _,U, _ ___,11 I ~I---~~---~ Total Number Other Personnel RESEARCH & RELATED Budget (A-8) (Funds Re~t3g~fe~fdget - Year 5 Tracking Number:GRANT11418584 Total Other Personnel 11 so, 99 9. oo Total Salary, Wages and Fringe Benefits (A+B) 1273 , 77l. 00 Page 60 Funding Opportunity Number:PA-11-260 Received Date:2013-06-0ST18:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 5 • ORGANIZATIONAL DUNS: lono<1ooi;wooo I • Budget Type: IZJ Project D Subaward/Consor\lum Enter name of Organization: IEcoHealdt ~.lliar.ce, Inc. Delete Entry I* Start Date: 11 o;n 1 ;;, 01 71 •End Date: I ~913012 r 8 I Budget Period 5 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment item • Funds Requested ($) 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. ::========================================================:::::::=============::::: 11. Total funds requested for all equipment listed in the attached file Total Equipment Additional Equipment: Add Attachment Delete Attachment D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico and U.S. Possessions) 2. Foreign Travel Costs Funds Requested ($) 13,605.00 '32,313.00 Total Travel Cost '3s, 918 . 00 E. Participant/Trainee Support Costs Funds Requested ($) 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other ~ Number of Participants/Trainees Total Participant/Trainee Support Costs ~------~ RESEARCH & RELATED Budget {C·E) (Funds Requested) Detailed Budget - Year 5 Page 61 View Attachment ] Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET - SECTION F-K, BUDGET PERIOD 5 • ORGANIZATIONAL DUNS: lnn900G&oooo I • Budget Type: ~ Project D Subaward/Consortium Enter name of Organization: IEccHea· th ;,,.''' ancF, ·nc. Delete Entry I Start Date: I· 0 ; 01 , 2o ,I• End Date: I O<J/ 3012018 I Budget Period 5 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. !shipping & Communications 9. !Local Reimbursement 10. Funds Requested ($) 13,500.00 12,600.00 1191, 576 .00 16,250.00 lsso. oo Total Other Direct Costs 1204 1 476 .oo G. Direct Costs Funds Requested ($) Total Direct Costs (A thru F) ls14, 16s .oo H. Indirect Costs Indirect Cost Type Indirect Cost Rate{%) Indirect Cost Base($) 1. IEcoJ-Jeal th Alliance F&A I 14 4 • 10 1322, 588. 00 ~::::::::::=~ 2. l,;cottealth Alliance F&A on 2 subawarl 144 .10 150, ooo .oo 3. :===============: 4. '-----------------' ._ ___ __, .__ _____ _, • Funds Requested {$) li42, 262. 00 122,050.00 Total Indirect Costs Ii 64. 312. oo Cognizant Federal Agency '---------------------------~ (Agency Name, POC Name, and POC Phone Number) I. Total Direct and Indirect Costs Total Direct and Indirect Institutional Costs (G + H) J.Fee Funds Requested ($) 1678, 477. 00 Funds Requested ($) K. * Budget Justification 11239-EHA NIAID COV BUDGETJUSTIFICATIOJ Add Attacnment Delete Attachment (Only attach one file.) RESEARCH & RELATED Budg__et {F·Kl (Funds Requested) Tietailed Budget• Year 5 Page 62 View Attachment Tracking Number:GRANTl1418584 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter ECOHEALTH ALLIANCE BUDGET JUSTIFICATION A. Key personnel: The PD/Pl, Dr. Peter Daszak, will commit CbH4),(b)(6) per year in each year of this budget. He will be primarily responsible for overseeing the project, general management, communication and collaboration with subawardees, as well as contributing to data analysis and manuscript writing. Senior/Key Personnel, Dr. Epstein, will commit CbH4),(b)(6) per year. Dr. Epstein will lead the design and implementation of the bat epidemiology fieldwork; supervise field teams, lab data analyses as well as conduct field training. He will participate in regular conference calls, help with data analysis, and draft manuscripts. Senior/Key Personnel, Dr. Olival, will commit )(4), (b)( per year. Dr. Olival will direct the bat population genetics work, assist with data analyses, and manuscript writing. He will also advise on the modeling and provide training for field teams. Senior/Key Personnel, Dr. Hosseini, will commit (b)(4),(b)(6) per year. Dr. Hosseini will perform spatial analyses and data mapping in collaboration with of Dr. Epstein and Dr. Olival. B. Other personnel: A research scientist will be hired at 12 months time per year to provide direct assistance and oversight of field activities in China; maintain equipment and logistics; and coordinate animal and human sample shipment to the labs in China and in the US. This person will be based at EcoHealth Alliance, but will spend significant time in the field. Mr. Aleksei Chmura Cb) <4>, Cb) (6) per year) in Y1 -Y5 will fulfill program assistance duties as well as conduct field research in China. Mr Chmura will coordinate regular calls, reports, maintain EcoHealth Alliance budget and financial reporting, draft subcontracts, and setup project database advise field activities and assist with statistical analysis. Once we secure IRBs for human sampling in Y1, we will hire three medical officers from China provincial CDCs as consultants to work in Guangxi, Hunan, and Fujian during Y2- Y5. These medical officers will be responsible for all I RB approved human sampling as well as maintaining cold chain for storage and shipping of samples. For all EcoHealth Alliance personnel that will have salary covered by this grant, we have included the EcoHealth Alliance 5% per annum increase in salary. C. Fringe benefits. Fringe benefits are calculated for EcoHealth Alliance's federally approved rate of 30% of base salary. D. Consultant: Once all permits are in place in Y2-5, EcoHealth Alliance will contract three technician-consultants trained in phlebotomy - one in each province: Guangxi, Hunan, and Fujian. The technicians will conduct interviews as part of the human wildlife contact survey as well as collect blood samples from volunteers in animal markets. These will be given daily rate of $67 for 5 months work per year (3 technicians x 5 Budget Justification Page 63

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter months x 20 days per month x $67 = $20,000 per year) as well as funds to cover shipping and maintaining cold-chain ($333 x 3 months x 3 provinces = $3,000 per year) from provincial areas to Wuhan Institute of Virology. We also will support the technician's allowable room/transportation/food costs expected to average monthly at food ($62), room ($100), and transportation ($100): $262 x 3 technicians x 5 months = $3,923 per year. E. Equipment: N/A F. Supplies: For Y1-Y5, we request annual support for tubes. syringes (5k); computer. phone, GPS (8k); lab reagents and buffer (10k); shipping (10k); PPE (10k); bat catching equipment (1 Ok); food/accomodation for field team (10k); dry shippers. liquid Nitrogen (8k) G. Travel Domestic travel: $4,400 is requested for years 1-5, comprising $2,200 each for the Pl (Dr. Daszak) and Senior/Key Personnel (Dr. Epstein) for travel to collaborating labs, to group meetings, and domestic scientific conferences to present results of our work. International travel: $33,000 is requested p.a. in Years 1-5. This will support 4 RT flights p.a. from New York to Shanghai or Guangzhou for the field veterinarian; 3 for the Senior/Key Personnel and 1 for the Pl (Daszak) @ $2500 ea; Food and accommodation at $8,500 p.a. for Senior/Key Personnel and the field team in China including field activities. Field vehicle rental & driver hire $500/wk x 9 wks. H. Participant support costs: We are requesting consortium/contractual support for our two partners: East China Normal University and Wuhan Institute of Virology. We are requesting $34,560 per year for Y1-Y5 for East China Normal University and for Wuhan Institute of Virology $93,960 in Y1 and $81,000 in Y2-Y5. These amounts are justified as follows: EAST CHINA NORMAL UNIVERSITY EcoHealth Alliance Budget Justification, H. Participant Support Costs (ctd} a) Senior Personnel: Dr. Shuyi Zhang (b)(4).(b)(6) per year in Y1-Y5 Dr. Zhang will oversee the field sample collection and coordination of sample transfer to Wuhan or US partners. He will not request any salary from this grant. His salary will be covered by his instiutional discretionary funds. b) Other personnel: A full time field biologist, Dr. GuanJian Zhu (b) (4), (b) (6) per year), will implement field site visits. sample collection. and sample shipment to Wuhan or to Co-Pis. A full time field technician, Mr. Junpeng Zhang (b)(4),(b)(6) per year), will assist with sample collection, handling, and transport from field to lab as well as with sample shipping from East China Normal University to Wuhan or US partners. Both Mr. Zhang and Dr. Zhu will work full-time for in Y1-Y5. Budget Justification Page 64

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter c) Fringe Benefits: Fringe benefits are provided at ECNU rate of 5% to Dr. Zhu and Mr. Zhang in Y1-Y5. d) Equipment: N/A e) Supplies: $560 are allocated per year in Y1-Y5 to support Field Biologist and Field Technician costs for telephone internet, and GPS: Phone= 15x2= 30; Internet= 5x2 = 10, GPS (batteries)= 3.5x2 = 10; total per month= $47. f) Travel: Dr. Zhang will provide travel costs for Field Biologist and Field Technician from discretionary funding. We request $2,000 per year to support Dr. Zhang and Dr. Zhu to travel to US for Co-Investigator meetings in either Boston or New York. Support will provide room and board for the two at per diem rate of $250 for 4 days (= 250x4x2 = $2,000). Dr. Zhang is already supported for travel funds of his own in those years. g) Participant support costs: N/A h) Other direct costs: NIA i) Indirect Costs. All administrative costs are charged directly. WUHAN INSTITUTE OF VIROLOGY a) Key personnel. Dr. Zhengli Shi, Senior Virologist. CbH4).~ per year in Y1-Y5. Dr. Shi will oversee the coronavirus screening for all samples collected in China. She will work with the Pl, Co-Investigators, and Senior/Key Personnel to analyze data and write manuscripts. She will also coordinate data and material sharing with the coInvestigators. Dr. Shi will not take salary on this grant and is funded by discretionary sources at her Institute. b) Other personnel: Mr. Jialu Li, Lab technician. (b)(4).(b)(6) p.a. in Y1-Y5. The lab technician will test all bat and other animal samples collected in China for coronaviruses and will conduct molecular characterization and phylogenetic analyses of new coronavirus strains identified as well as catalog and ship samples and maintain a sample database. EcoHealth Alliance Budget Justification, H. Participant Support Costs (ctd) c) Fringe benefits: Wuhan Institute of Virology benefit rate of 5% is applied to salary for Mr. Jialu Li in Y1-Y5. d) Equipment: The subcontractor will purchase one ultracold -80°C freezer for dedicated sample storage for this project. $12,960. Budget Justification Page 65

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter e) Supplies: Annual costs for Laboratory reagents (20k); shipping to US (twice per year x 2.5k = 5k); testing costs (30k); expendable equipment costs (5k) are requested in Y1-Y5. f) Travel: Round-trip airfare to Boston/NYC for Co-Investigator (Dr. Shi) for attending PD/Pl meetings once-per year in Y1-Y5: $2,600. Dr. Shi has discretionary funds to supplement travel expenditures. We also request $2,400 to cover in-country transportation costs of supplies and samples. g) Participant support costs: N/A h) Other direct costs: We request annual support costs for Telephone ($180), Printing ($144 ), Conference Calls ($504 ), and local shipping charges ($172) for Dr. Shi's laboratory and Mr. Li in Y1-Y5. i) Indirect Costs. All administrative costs are charged directly. H. Other direct costs: We request $71,000 in year one for sample collection materials including bat catching equipment, syringes, tubes, and reagents, and 2 liquid nitrogen dry shippers at $3,000each. We also request $10,000 for shipping supplies from NY to China in yr 1, then $5,000 p.a. for shipping in years 2-5. We also request $4,000 in years 3 and 4 for lab reagents to complete population genetics tests. Other expenses for publishing and communications including video conferencing facilities will be covered by EcoHealth Alliance. I. Indirect Costs We are requesting the EcoHealth Alliance federally-approved indirect cost rate of 30.0% on all applicable direct costs. Indirect is taken only on the first $25,000 for each consortium/contractual agreement. As there are 2 (one to Wuhan lnsititute of Virology and the other to East China Normal University), a total of $15,000 ($7,500x2) is taken as indirect on consortium/contractual agreements only in Y1 and included as part of direct cost calculations. In Y2-Y5 no indirect is taken on consortium/contractual agreements. Budget Justification Page 66

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other 2 10.Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Cumulative Budget Tracking Number:GRANTl1418584 Totals($) 118, 02s. oo 1161,565.00 lss, 400. oo 110,400.00 11,045,598.00 140,000.00 13,400.00 lsss, 817. oo 1683, 614 .00 11,239,431.00 1179, 590. 00 11,157,798.00 12,576,819.00 Ins, 519.oo 13,362,338.00 Page 67 Funding Opportunity Number:PA-11-260 Received Date:20 l3-06-05T!8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 1 * ORGANIZATIONAL DUNS: 5290274740000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology * Start Date: 10-01-2013 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Zhengli Xingyi • Last Name Shi Ge Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Laboratory Technician 100 Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 1 Suffix * End Date: 09-30-2014 • Project Role Base Salary ($) Co-Investigator Senior Research Technician Mime Type: Budget Period: 1 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 68 • Funds Requested ($) (b) (4). (b) (6) [(b) (4). (b~ * Funds Requested ($) (b) (4). (b) (6) (b) (4). (b) (6) 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 1 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2013 • End Date: 09-30-2014 Budget Period: 1 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 69 • Funds Requested ($) Funds Requested ($) 2,060.00 2,060.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 1 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2013 F. Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations • End Date: 09-30-2014 Budget Period: 1 Funds Requested ($) 95,737.00 Total Other Direct Costs 95,737.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. Wuhan Institute of Virology F&A Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate (%) 8.00 Total Direct Costs (A thru F) 123,699.00 Indirect Cost Base ($) • Funds Requested ($) 123,699.00 9,896.00 Total Indirect Costs 9,896.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + H) 133,595.00 IJ. Foe F,nds Roq,osted ($) I K. * Budget Justification File Name: 1242-WIV NIAID GOV BUDGET JUSTIFICATION .pdt (Only attach one file.) RESEARCH & RELATED Budget {F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Mime Type: application/pd! Page 70 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 2 * ORGANIZATIONAL DUNS: 5290274740000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology * Start Date: 10-01-2014 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Zhengli Xingyi • Last Name Shi Ge Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Laboratory Technician Total Number Other Personnel , ... & HE:LAI t:U Budget {A·l:3) (1-unds Requested) Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 1 Suffix * End Date: 09-30-2015 • Project Role Co-Investigator Senior Research Technician Base Salary ($) Mime Type: I Budget Period: 2 Cal. Acad. Sum. • Fringe • Funds Requested ($) Months Months Months 'Requested Salarv (Sl Benefits!}),_ ____ ~~~~ (b) (4). (b) (6) Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits I Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 71 [(b) (4). (b~ * Funds Requested ($) (b) (4). (b) (~ (b) (4). (b~ (1 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 2 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2014 • End Date: 09-30-2015 Budget Period: 2 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 72 • Funds Requested ($) Funds Requested ($) 2,060.00 2,060.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 2 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2014 F. Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations G. Direct Costs H. Indirect Costs Indirect Cost Type 1. Wuhan Institute of Virology F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) • End Date: 09-30-2015 Indirect Cost Rate (%) 8.00 Budget Period: 2 Total Other Direct Costs Funds Requested ($) 100,756.00 100,756.00 Funds Requested ($) Total Direct Costs (A thru F) 128,718.00 Indirect Cost Base ($) • Funds Requested ($) 128,718.00 10,297.00 Total Indirect Costs 10,297.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + H) 139,015.00 IJ. Foe F,nds Roq,osted ($) I K. * Budget Justification File Name: 1242-WIV NIAID GOV BUDGET JUSTIFICATION .pdt (Only attach one file.) RESEARCH & RELATED Budget {F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Mime Type: application/pd! Page 73 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 3 * ORGANIZATIONAL DUNS: 5290274740000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology * Start Date: 10-01-2015 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Zhengli Xingyi • Last Name Shi Ge Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Laboratory Technician Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 1 Suffix * End Date: 09-30-2016 • Project Role Base Salary ($) Co-Investigator Senior Research Technnician Mime Type: Budget Period: 3 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($) Benefits ($) I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 74 • Funds Requested ($) (b) (4), (b) (6) [(b) (4), (b~ * Funds Requested ($) (b) (4), (b) (6) (b) (4), (b) (6) 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2015 • End Date: 09-30-2016 Budget Period: 3 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 75 • Funds Requested ($) Funds Requested ($) 2,060.00 2,060.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2015 F. Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations G. Direct Costs H. Indirect Costs Indirect Cost Type 1. Wuhan institute of Virology F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) • End Date: 09-30-2016 Indirect Cost Rate (%) 8.00 Budget Period: 3 Total Other Direct Costs Funds Requested ($) 119,373.00 119,373.00 Funds Requested ($) Total Direct Costs (A thru F) 147,335.00 Indirect Cost Base ($) • Funds Requested ($) 147,335.00 11,787.00 Total Indirect Costs 11,787.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + H) 159,122.00 IJ. Foe F,nds Roq,osted ($) I K. * Budget Justification File Name: 1242-WIV NIAID GOV BUDGET JUSTIFICATION .pdt (Only attach one file.) RESEARCH & RELATED Budget {F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Mime Type: application/pd! Page 76 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 4 * ORGANIZATIONAL DUNS: 5290274740000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology * Start Date: 10-01-2016 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Zhengli Xingyi • Last Name Shi Ge Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Laboratory Technician Total Number Other Personnel , ... & HE:LAI t:U Budget {A·l:3) (1-unds Requested) Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 1 Suffix * End Date: 09-30-2017 • Project Role Base Salary ($) Co-Investigator Senior Research Technician Mime Type: Budget Period: 4 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) I * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 77 • Funds Requested ($) (b) (4). (b) (6) * Funds Requested ($) (b) (4). (b) (~ 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 4 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2016 • End Date: 09-30-2017 Budget Period: 4 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 78 • Funds Requested ($) Funds Requested ($) 2,060.00 2,060.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 4 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2016 F. Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations G. Direct Costs H. Indirect Costs Indirect Cost Type 1. Wuhan Institute of Virology F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) • End Date: 09-30-2017 Indirect Cost Rate (%) 8.00 Budget Period: 4 Total Other Direct Costs Funds Requested ($) 119,373.00 119,373.00 Funds Requested ($) Total Direct Costs (A thru F) 147,335.00 Indirect Cost Base ($) • Funds Requested ($) 147,335.00 11,787.00 Total Indirect Costs 11,787.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + H) 159,122.00 IJ. Foe F,nds Roq,osted ($) I K. * Budget Justification File Name: 1242-WIV NIAID GOV BUDGET JUSTIFICATION .pdt (Only attach one file.) RESEARCH & RELATED Budget {F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Mime Type: application/pd! Page 79 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 5 * ORGANIZATIONAL DUNS: 5290274740000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology * Start Date: 10-01-2017 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Zhengli Xingyi • Last Name Shi Ge Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Laboratory Technician Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 1 Suffix * End Date: 09-30-2018 • Project Role Base Salary ($) Co-Investigator Senior Research Technician Mime Type: Budget Period: 5 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 80 • Funds Requested ($) (b) (4), (b) (6) 1(b> (4), Cb><~ * Funds Requested ($) (b) (4), (b) (6) (b)(4), ~ 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 5 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2017 • End Date: 09-30-2018 Budget Period: 5 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 81 • Funds Requested ($) Funds Requested ($) 2,060.00 2,060.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 5 • ORGANIZATIONAL DUNS: 5290274740000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology • Start Date: 10-01-2017 F. Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations G. Direct Costs H. Indirect Costs Indirect Cost Type 1. Wuhan Institute of Virology F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) • End Date: 09-30-2018 Indirect Cost Rate (%) 8.00 Budget Period: 5 Total Other Direct Costs Funds Requested ($) 119,373.00 119,373.00 Funds Requested ($) Total Direct Costs (A thru F) 147,335.00 Indirect Cost Base ($) • Funds Requested ($) 147,355.00 11,787.00 Total Indirect Costs 11,787.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + H) 159,122.00 IJ. Foe F,nds Roq,osted ($) I K. * Budget Justification File Name: 1242-WIV NIAID GOV BUDGET JUSTIFICATION .pdt (Only attach one file.) RESEARCH & RELATED Budget {F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 1 Mime Type: application/pd! Page 82 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Tracking Number: GRANT11418584 Subaward 1 Totals($) 5 10,300.00 554,612.00 79,325.00 50,185.00 129,510.00 10,300.00 554,612.00 694,422.00 55,554.00 749,976.00 Page 83 0MB Number: 4040-0001 Expiration Date: 0613012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter WUHAN INSTITUTE OF VIROLOGY BUDGET JUSTIFICATION, SUBAWARD A. Senior/Key Personnel: Co-Investigator, Dr. Zhengli Shi, a Senior Research Scientist at the Chinese Academy of Science's Wuhan Institute of Virology will commit CbH4),(b)(6) per yearCbH4>.CbH6) to this project to refine study protocols, coordinate research, oversee implementation of all activities, analyze data, lead regular meetings with other PD/Pl and Other Senior/Key Personnel as well as draft papers. Dr. Xingyi Ge, Senior Research Technician, will commit CbH4>, (b)(6) per year to perform all laboratory work and directly supervise the laboratory technician. B. Other Personnel One laboratory technician will commit CbH4), (b)(6) per year CbH4), (b)(6) each to this project to perform all required laboratory assays and maintenance as well as participate in selected meetings, perform research for papers, and assist Dr. Shi in performing the work under this award. All Wuhan Institute of Virology salaries include the US "overhead" or "fringe", so this is not calculated separately. C. Equipment No equipment over $5,000 will be purchased. D. Travel We are requesting $2,060 per year for all years for Senior/Key Personnel Dr. Shi and Dr. Ge to travel to Shanghai to visit partner laboratory at East China Normal Univeristy (ECNU) and meet with the PD/Pl as well as with collaborators on this proposal: these include EcoHealth Alliance, East China Normal University, Yunnan CDC, Shanghai CDC, and Guangdong CDC. Travel is calculated at one round trip airfare from Wuhan to Shanghai ($300), three-night hotel in Shanghai ($150 per night), and four days per diem (at $70 per day) F. Other Direct Costs We are requesting support for laboratory experiments and related costs. RNA Extractions We will be running RNA Extractions for 1,000 bats per year (three samples per bat: oral, anal, and blood) in each year of the project. This will cost $13,922 per year (QIAamp ViralRNA Mini Kit with Axygen Pipette Tips and Filter Tubes at $4.64 per sample). Extracted RNA per animal will be pooled. RT-PCR Costs for 1-Step RT-PCR assays for Coronavirus conducted on 1,000 samples per year for each year of the project total $7,123 and are detailed as follows: Superscript 111 one step kit ($5.18 per sample); Platinum Tag DNA Polymerase ($0.57 per sample); nuclease-free water ($0.16 per sample); and Axygen Pipette Tips and Filter Tubes ($1.21 per sample). DNA Sequencing In each year of the project, DNA Sequencing will be performed on 3,200 samples at a cost of $2.91 per reaction. We request a total of $9,325 per year in each year. Subaward 1 Budget Justification Attachment Page 84

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Laboratory Supplies We request support for in vitro infection experiments using pseudoviruses carrying the spike proteins (wild type or mutants) or live viruses in cell lines of different origins, binding affinity assays between the spike proteins (wild type or mutants) and different cellular receptor molecules, and humanized mouse experiments. In Year 1, $65,367 is requested: Lipofectamine2000 transfection reagent at a cost of $2,428; cell lines from bats and other mammals including primates and humans at a cost of $971; in vitro infection experiments require GIBCO Fetal Bovine Serum ($3,562), GIBCO antibiotic antimycotic ($563), GIBCO medium ($2,914) as well as $19,426 for Corning Cell culture; receptor-mutant pseudovirus binding assays require Luciferase assay system E1500 ($858), pseudovirus package ($3,885), and sequencing ($22,664); $8,094 is required for protein expression from the binding affinity assays. In Year 2, $70,385 is requested: Lipofectamine2000 transfection reagent at a cost of $2,428; cell lines from bats and other mammals including primates and humans at a cost of $971 - sufficient cell lines will be established by the end of Year 2, so this cost requirement will discontinue in Years 3-5; increased number of in vitro infection experiments require slightly more funding for GIBCO Fetal Bovine Serum ($4,047) as well as GIBCO antibiotic antimycotic ($563), GIBCO medium ($2,914) as well as $19,426 for Corning Cell culture; receptor-mutant pseudovirus binding assays require Luciferase assay system E1500 ($858), pseudovirus package requirements will approximately double from Y1 ($6,799), and sequencing ($22,664); $9,713 is required for protein expression from the increased Year 2 number of binding affinity assays at a slightly higher cost than year one as well. In Years 3, 4 and 5, $89,002 is requested per year: Lipofectamine2000 transfection reagent at a cost of $2,428 per year; increased number of in vitro infection experiments require slightly more funding for GIBCO Fetal Bovine Serum ($5,828 per year) as well as GIBCO antibiotic antimycotic ($563 per year), GIBCO medium ($2,914 per year) as well as $19,426 per year for Corning Cell culture; receptor-mutant pseudovirus binding assays require Luciferase assay system E1500 ($858 per year), pseudovirus package requirements will be $6,799 per year, sequencing ($22,664 per year) and gene synthesis ($12,915 per year) will also be required; $9,713 per year is required for protein expression from binding affinity assays; in only years 3, 4, and 5 humanized mouse in vivo experimental animals will be raised at an annual cost of $4,857 per year. H. Indirect Costs We are requesting an extremely indirect cost of 8% on all direct costs. Subaward 1 Budget Justification Attachment Page 85

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 1 * ORGANIZATIONAL DUNS: 4209454950000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2013 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Shuyi Guangjian • Last Name Zhang Zhu Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel 0 Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 2 Suffix * End Date: 09-30-2014 • Project Role Base Salary ($) Co-Investigator Research Technician Mime Type: Budget Period: 1 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B} Page 86 • Funds Requested ($) (b) (4). (b) (6) * Funds Requested ($) 25,000.00 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 1 • ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University • Start Date: 10-01-2013 • End Date: 09-30-2014 Budget Period: 1 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 87 • Funds Requested ($) Funds Requested ($) 2,700.00 2,700.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 1 * ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project e Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2013 • End Date: 09-30-2014 Budget Period: 1 F. Other Direct Costs Funds Requested ($) 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Fieldwork Support Costs 59,400.00 Total Other Direct Costs 59,400.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. East China Normal University F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate(%) 8.00 Total Direct Costs (A thru F) 87,100.00 Indirect Cost Base ($) • Funds Requested ($) 87,100.00 6,968.00 Total Indirect Costs 6,968.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + HJ 94,068.00 IJ. Fee Funds Reqoested ($) I K. * Budget Justification File Name: 1243-ECNU NIAID COV BUDGET JUSTIFICATION.pd! (Only attach one lile.) RESEARCH & RELATED Budget (F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Mime Type: application/pdf Page 88 0MB Number: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 2 * ORGANIZATIONAL DUNS: 4209454950000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2014 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Shuyi Guangjian • Last Name Zhang Zhu Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel 0 Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 2 Suffix * End Date: 09-30-2015 • Project Role Base Salary ($) Co-Investigator Research Technician Mime Type: Budget Period: 2 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B} Page 89 • Funds Requested ($) (b) (4). (b) (6) I (b) (4). (b) <~ * Funds Requested ($) (b) (4). (b) (6) 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 2 • ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University • Start Date: 10-01-2014 • End Date: 09-30-2015 Budget Period: 2 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 90 • Funds Requested ($) Funds Requested ($) 2,700.00 2,700.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 2 * ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project e Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2014 • End Date: 09-30-2015 Budget Period: 2 F. Other Direct Costs Funds Requested ($) 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Field Work Support Costs 39,600.00 Total Other Direct Costs 39,600.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. East China Normal University F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate(%) 8.00 Total Direct Costs (A thru F) 67,300.00 Indirect Cost Base ($) • Funds Requested ($) 67,300.00 5,384.00 Total Indirect Costs 5,384.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + HJ 72,684.00 IJ. Fee Funds Reqoested ($) I K. * Budget Justification File Name: 1243-ECNU NIAID COV BUDGET JUSTIFICATION.pd! (Only attach one lile.) RESEARCH & RELATED Budget (F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Mime Type: application/pdf Page 91 0MB Number: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 3 * ORGANIZATIONAL DUNS: 4209454950000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2015 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Shuyi Guangjian • Last Name Zhang Zhu Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel 0 Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 2 Suffix * End Date: 09-30-2016 • Project Role Base Salary ($) Co-Investigator Research Technician Mime Type: Budget Period: 3 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B} Page 92 • Funds Requested ($) (b) (4). (b) (6) I (b) <4). (b) c4 * Funds Requested ($) (b) (4). (b) (oj 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 3 • ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University • Start Date: 10-01-2015 • End Date: 09-30-2016 Budget Period: 3 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 93 • Funds Requested ($) Funds Requested ($) 2,700.00 2,700.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 3 * ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project e Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2015 • End Date: 09-30-2016 Budget Period: 3 F. Other Direct Costs Funds Requested ($) 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Field Work Support Costs 29,700.00 Total Other Direct Costs 29,700.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. East China Normal University F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate(%) 8.00 Total Direct Costs (A thru F) 50,108.00 Indirect Cost Base ($) • Funds Requested ($) 50,108.00 4,009.00 Total Indirect Costs 4,009.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + HJ 54,117.00 IJ. Fee Funds Reqoested ($) I K. * Budget Justification File Name: 1243-ECNU NIAID COV BUDGET JUSTIFICATION.pd! (Only attach one lile.) RESEARCH & RELATED Budget (F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Mime Type: application/pdf Page94 0MB Number: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 4 * ORGANIZATIONAL DUNS: 4209454950000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2016 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Shuyi Guangjian • Last Name Zhang Zhu Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel 0 Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 2 Suffix * End Date: 09-30-2017 • Project Role Base Salary ($) Co-Investigator Research Technician Mime Type: Budget Period: 4 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B} Page 95 • Funds Requested ($) (b) (4). (b) (6) l(b) (4). (b) <~ * Funds Requested ($) (b) (4). (b) (6) 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 4 • ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University • Start Date: 10-01-2016 • End Date: 09-30-2017 Budget Period: 4 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 96 • Funds Requested ($) Funds Requested ($) 2,700.00 2,700.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 4 * ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project e Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2016 • End Date: 09-30-2017 Budget Period: 4 F. Other Direct Costs Funds Requested ($) 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Field Work Support Costs 19,800.00 Total Other Direct Costs 19,800.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. East China Normal University F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate(%) 8.00 Total Direct Costs (A thru F) 39,167.00 Indirect Cost Base ($) • Funds Requested ($) 39,167.00 3,133.00 Total Indirect Costs 3,133.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + HJ 42,300.00 IJ. Fee Funds Reqoested ($) I K. * Budget Justification File Name: 1243-ECNU NIAID COV BUDGET JUSTIFICATION.pd! (Only attach one lile.) RESEARCH & RELATED Budget (F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Mime Type: application/pdf Page97 0MB Number: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter RESEARCH & RELATED BUDGET - SECTION A & B, BUDGET PERIOD 5 * ORGANIZATIONAL DUNS: 4209454950000 * Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2017 A. Senior/Key Person 1. 2. Prefix Dr. Dr. • First Name Middle Name Shuyi Guangjian • Last Name Zhang Zhu Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: 8. Other Personnel * Number of Personnel 0 Post Doctoral Associates Graduate Students Undergraduate Students Secretarial/Clerical Total Number Other Personnel Trac~ing Numbor: CRANT11418S84 File Name: * Project Role Subaward 2 Suffix * End Date: 09-30-2018 • Project Role Base Salary ($) Co-Investigator Reasearch Technician Mime Type: Budget Period: 5 Cal. Acad. Sum. 'Requested • Fringe Months Months Months Salary($! Benefits (!). I Total Senior/Key Person Cal. Acad. Sum. * Requested Months Months Months Salary ($) * Fringe Benefits Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B} Page 98 • Funds Requested ($) (b) (4), (b) (6) I (b) (4), (b) (6) * Funds Requested ($) (b) (4), (b) (6) 0MB Numbei: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTION C, D, & E, BUDGET PERIOD 5 • ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project • Subaward/Consortium Enter name of Organization: East China Normal University • Start Date: 10-01-2017 • End Date: 09-30-2018 Budget Period: 5 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Addltlonal Equipment: File Name: D. Travel 1. Domestic Travel Costs (Incl.Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2.Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget (C-E} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Total Equipment Mime Type: Total Travel Cost Total Participant/Trainee Support Costs Page 99 • Funds Requested ($) Funds Requested ($) 2,700.00 2,700.00 Funds Requested ($) 0MB Number: 4040-0001 Expiration Date: 06/3012011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- SECTIONS F-K, BUDGET PERIOD 5 * ORGANIZATIONAL DUNS: 4209454950000 • Budget Type: 0 Project e Subaward/Consortium Enter name of Organization: East China Normal University * Start Date: 10-01-2017 • End Date: 09-30-2018 Budget Period: 5 F. Other Direct Costs Funds Requested ($) 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Field Work Support Costs 14,850.00 Total Other Direct Costs 14,850.00 G. Direct Costs Funds Requested ($) H. Indirect Costs Indirect Cost Type 1. East China Normal University F&A Rate Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Indirect Cost Rate(%) 8.00 Total Direct Costs (A thru F) 30,050.00 Indirect Cost Base ($) • Funds Requested ($) 30,050.00 2,404.00 Total Indirect Costs 2,404.00 I. Total Direct and Indirect Costs Funds Requested ($) Total Direct and Indirect Institutional Costs (G + HJ 32,454.00 IJ. Fee Funds Reqoested ($) I K. * Budget Justification File Name: 1243-ECNU NIAID COV BUDGET JUSTIFICATION.pd! (Only attach one lile.) RESEARCH & RELATED Budget (F-K} (Funds Requested) Tracking Number: GRANT11418584 Subaward 2 Mime Type: application/pdf Page 100 0MB Number: 4040-0001 Expiration Date: 06/30/2011

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH & RELATED BUDGET- Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1 . Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Tracking Number: GRANT11418584 Subaward 2 Totals($) 13,500.00 163,350.00 96,875.00 96,875.00 13,500.00 163,350.00 273,725.00 21,898.00 295,623.00 Page 101 0MB Number: 4040-0001 Expiration Date: 0613012011

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter EAST CHINA NORMAL UNIVERSITY BUDGET JUSTIFICATION, SUBAWARD A. Senior/Key Personnel: Co-Investigator Dr. Shu-Yi Zhang will commit CbH4>, (b)(6) per year to this project to refine field work design, coordinate research, oversee implementation of all activities, analyze data, lead regular meetings with other PD/Pl and Other Senior/Key Personnel as well as draft papers. To keep costs low, Dr. Zhang will not take any salary. Research Technician, Dr. Zhu Guangjian, will commit CbH4),(b)(6) in Years 1 and 2; 34 weeks or CbH4>, (b)(6) in Year 3; H4>, CbH6J inYear 4 and only CbH4>, CbH6J in Year 5 to directly coordinate field work, liaise with local CDCs, ensure shipment of samples to Wuhan Institute of Virology, and participate in some meetings as well as assist with drafting papers. B. Other Personnel There are no additional personnel. All East China Normal University salaries include the US "overhead" or "fringe", so this is not calculated separately. C. Equipment No equipment over $5,000 will be purchased. D. Travel We are requesting $2,700 per year for all five years of this award to cover 3-per-year round-trip flights each from Shanghai, China, to Yunnan, Guangdong, and Guangxi for Dr. Zhu Guangjian to meet with collaborating institutions, train field teams, and ensure sample collection, storage, and shipments. Each flight is estimated at $300. F. Other Direct Costs Fieldwork Support Costs In year 1, we are requesting $59,400 to support 12-months of fieldwork costs. This we estimate as follows: $7,200 for driver ($600 per month) and car rental ($600 per month); $2,700 for gas ($450 per month); $10,800 to support a field team of three ($600 per month); and $9,000 for meals and lodging at a rate of $50 per day ($1,500 per month). In year 2, we are requesting $39,600 to support 8-months of fieldwork costs. This we estimate as follows: $7,200 for driver ($600 per month) and car rental ($600 per month); $2,700 for gas ($450 per month); $10,800 to support a field team of three ($600 per month); and $9,000 for meals and lodging at a rate of $50 per day ($1,500 per month). In year 3, we are requesting $29,700 to support 24-weeks or 6-months of fieldwork costs. This we estimate as follows: $7,200 for driver ($600 per month) and car rental ($600 per month); $2,700 for gas ($450 per month); $10,800 to support a field team of three ($600 per month); and $9,000 for meals and lodging at a rate of $50 per day ($1,500 per month). In year 4, we are requesting $19,800 to support 16-weeks or 4-months of fieldwork costs. This we estimate as follows: $7,200 for driver ($600 per month) and car rental ($600 per month); $2,700 for gas ($450 per month); $10,800 to support a field team of three ($600 per month); and $9,000 for meals and lodging at a rate of $50 per day ($1,500 per month). Subaward 2 Budget Justification Attachment Page 102

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter In year 5, we are requesting $14,850 to support 12-weeks or 3-months of fieldwork costs. This we estimate as follows: $7,200 for driver ($600 per month) and car rental ($600 per month); $2,700 for gas ($450 per month); $10,800 to support a field team of three ($600 per month); and $9,000 for meals and lodging at a rate of $50 per day ($1,500 per month). H. Indirect Costs We are requesting an extremely indirect cost of 8% on all direct costs. Subaward 2 Budget Justification Attachment Page 103

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter PHS 398 Cover Page Supplement 0MB Number: 0925-0001 1. Project Director/ Principal Investigator (PD/Pl) Prefix: lor. I • First Name: !Peter I Middle Name: I I • Last Name: loaszak I Suffix: I I 2. Human Subjects Clinical Trial? {8] No 0Yes • Agency-Defined Phase Ill Clinical Trial? No 0Yes 3. Applicant Organization Contact Person to be contacted on matters involving this application Prefix: IDr. I ' First Name: !Peter I Middle Name: I • Last Name: loaszak I Suffix: I I • Phone Number: ( (b)(6}) I Fax Number: 1+1. 212. 380. 4 465 I Email: I (l>f(6Ji I • Title: le resident I • Street1: 1460 West 34th Street I Street2: 117th Floor I • City: INew York I County/Parish: I I • State: I NY: Nev. York I Province: I I • Country: lusA: UNITED STATES I ·Zip/ Postal Code: !10001-2317 I Clinical Trial & HESC Page 104 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05Tl8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter PHS 398 Cover Page Supplement 4. Human Embryonic Stem Cells • Does the proposed project involve human embryonic stem cells? 0Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://stemcells.nih.gov/research/registry/. Or, if a specific stem cell line cannot be referenced at this time, please check the box indicating that one from the registry will be used: Cell Line(s): D Specific stem cell line cannot be referenced at this time. One from the registry will be used. Clinical Trial & HESC Page 105 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05TJ8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 0MB Number: 0925-0001 PHS 398 Research Plan 1. Application Type: From SF 424 (R&R) Cover Page. The response provided on that page, regarding the type of application being submitted, is repeated for your reference, as you attach the appropriate sections of the Research Plan. *Type of Application: 1:8:J New D Resubmission Renewal D Continuation D Revision 2. Research Plan Attachments: Please attach applicable sections of the research plan, below. 1. Introduction to Application I I , Add Attachment I [Delete Attachment! View Attachment [ (for RESUBMISSION or REVISION only) 2. Specific Aims 11240-NIH_COv_Specific_Aims.~ Add Attachment I Delete Attachment I View Attachment [ 3. *Research Strategy 11241-Understanding_the_riskL Add Attachment I Delete Attachment I View Artachmem I 4. Inclusion Enrollment Report I I Add Attachment I Delete Attachment J View Attachment [ 5. Progress Report Publication List I I Add Attachment I Delete Attachment I View Attachment I Human Subjects Sections 6. Protection of Human Subjects I 12 5 4-NIH_COv_P rotection_Huml Add Attachment I Delete Attachment I View Attachment I 7. Inclusion of Women and Minorities 11255-NIH_COv_Inclusion_of_WI Add Attachment I ~ Delete Attachment I View Attachment] 8. Targeted/Planned Enrollment Table 11256-China_CoV_Planned_enrol Add Attachment I Delete Attachment I View Attachment [ 9. Inclusion of Children 112 5 7-NIH_COv _Incl us ion_of_C~ Add Attachment I Delete Attachment I View Attacl1men1 [ Other Research Plan Sections 10. Vertebrate Animals l12s8-NIH_COv_Vertebrate_Ani~ Add Attachment I Delete Attachment! View Attachment [ 11 . Select Agent Research l12s9-NIH_cov_Select_Agent_RI , Add Attachment I Delete Attachment [ , View Attachment [ 12. Multiple PD/Pl Leadership Plan I I 1, Add Attachment I Delete Attachment [ .. View Attachment I 13. Consortium/Contractual Arrangements l1260-NIH_cov_Consortium_Cont, Add Attachment [ Delete Attachment [ View Attachment [ 14. Letters of Support I 12 61-NIAID_cov _2013_Al1Supp~ Add Attachment [ Delete Attachment [ View Attachment I 15. Resource Sharing Plan(s) 112 62-NI H_COv_Resource_Shar i~ Add Attachment [ Delete Attachment [ View Attachment I 16. Appendix Add Attachments I Remove Attachments [ View Attachments [ List of Research Plan Attachments Page 106 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05TJ8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter SPECIFIC AIMS: Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-Co Vs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel Co Vs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2) how this process is affected by CoV evolutionary potential. and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses· screen eo le who have hi h occu ational ex osure to bats and other wildlife· and examine the enetics and rece tor bindin properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-Co Vs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological op ortunit for contact betweens ecies so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Genbank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecolo ical and h lo enetic traits to ex lain atterns of viral sharin betweens ecies. We will test for positive selection in market vs. wild-sampled viruses, and use data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and Co Vs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. Specific Aims Page 107

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESEARCH STRATEGY A. SIGNIFICANCE: General: Severe Acute Respiratory Syndrome, like many other emerging human pathogens ( 1). originated in a wildlife reservoir host, initially thought to be terrestrial mammals (2), and later shown by our group to be bats (3). Bats harbor the most closely-related viruses to SARS-CoV, and are traded widely for food in the wildlife markets of China (4). The diversity of bat-CoVs is very high, and some studies even suggest that the Coronaviridae originated within bats (3, 5-9). Recently a novel CoV emerged in the Middle East (MERS-CoV) ( 10) and available data (including from our group) suggest that MERS-CoV also has bat origins ( 11-13). Given that hunting and eating of bats continues across Asia, future spillover of bat-CoVs is likely. Yet salient questions remain: How diverse are bat-Co Vs? Can the conditions in these markets enhance bat-Co V evolution and spillover of bat-Co Vs? The proposed work addresses these issues and examines viral diversity in a critical zoonotic reservoir (bats), at sites of high risk for emergence (wildlife markets) in an EID hotspot (China) ( 14). SARS and bat-CoVs: Coronaviruses are found in a wide range of animal species ( 15). Before the SARS epidemic, only two human coronaviruses (HCoVs) had been characterized (HCoV-229E and HCoV-OC43) ( 16, 17). Since then three more human coronaviruses (HCoV-NL63, HCoV HKU-1, and MERS-CoV), in addition to SARS-CoV, have been identified in individuals with respiratory infections (16, 18, 19). One of these, HCoV-NL63, is thought to be zoonotic and of bat origin (6). Our group recently identified a CoV from bats in Bangladesh closely-related and likely ancestral to HCoV-OC43 (20) and is currently characterizing CoVs from bats in Saudi Arabia. The animal origins of SARS-CoV were first suspected due to the association among index cases and the trade in wildlife for food (21). Initially, civets and other mammals consumed in restaurants in southern China were implicated (2), however these species did not exhibit the high seroprevalence and low viral (PCR) prevalence expected from a natural wildlife reservoir of a zoonotic virus (21). In 2004, our group discovered SARS-llke (SL) CoVs In free-living wild bat species In China and demonstrated that human SARS-CoV nestled phylogenetically within this group (4). However, SARS-CoV uses the angiotensin-converting enzyme 2 (ACE2) receptor to gain entry to human cells (22), and bat SL-CoVs appeared unable to bind to ACE2. A large number of novel Alpha- and Betacoronaviruses have since been discovered in Old and New World bats, but few have been isolated ( 8, 11-13, 23-21). In 2012, we isolated and characterized two bat SL-CoVs from Rhinolophus sinicus from Yunnan Province, China that use the ACE2 receptor and are closely related to SARS-CoV (Fig. 1) (28). We found a seasonal shedding pattern for this SL-CoV, with peak prevalence of 30-50%. Bats from this population are hunted for human consumption, posing two crucial questions: 1) What is the risk of these CoVs emerging in humans? 2) Will the conditions that exist in live animal markets in Asia promote further emergence of bat-Co Vs in human populations? Figure 1. Phylogenetic tree of receptor binding domain sequences of SARS-CoVs (Red), bat SARS-like CoVs discovered by our group in the last 2 years (Blue), and bat SL-CoVs that we published previously in our paper proposing a bat-origin for SARS in 2005 (Green) (3). In 2012, we isolated two novel SL-CoVs (SL-CoV-SHC014 and 3367, blue arrows) and have shown for the first time that a bat SL-CoV use the ACE2 receptor which SARS-CoV uses to infect human cells. Unpublished data from Ge et al. (in review) (28). Evolution, host-virus co-phylogeny and risk of CoV emergence: There is wide variation in the propensity of viruses for cross-species transmission, within and among viral genera and families (29). Coronaviruses undergo genetic recombination by a genomic template-switching mechanism and generate point mutations at a rate similar to that of other RNA viruses, perhaps explaining their capacity for host switching and zoonotic transmission ( 15, 30). This capacity is heightened by the ecology of host species, opportunity for contact, characteristics of the pathogen, and evolutionary (phylogenetic) relationships between hosts (31-33). Bats (Order Chiroptera) are the second most diverse group of mammals (~1,200 species) with a wide range ecological and life-history traits that affect their ability to share viruses (34, 35) and may explain variation in viral diversity (36, 37). Phylogenetic relationships may determine limits to viral binding at receptor sites and to cross-species transmission (31, 33), and these factors could be used to predict the risk of spillover (see Specific Aim 2). Apart from our own work (see Section C2b, Fig. 7), bat and CoV co-evolutionary patterns haven't been rigorously examined. Recent work suggests that most bat-CoV clades correspond to specific bat species or genera (38, 39), with little evidence of bat-CoV spillover among species roosting together in the same cave (40). There is also evidence for geographically distributed, but related, bat taxa sharing related CoV strains (8, 38). In contrast, other studies of wild-caught bats did not find strict co-evolutionary congruence in bat-Co Vs for host Research Strategy Page 108

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter species, genera or families ( 41-43). Thus, the same CoV strains may circulate in different bat genera ( 41), and multiple diverse CoV lineages can be found in the same bat species and even individuals ( 7, 40, 44, 45). This, and density of some bat species populations, suggests that viral recombination may be possible in these hosts (6). Forced contact in wildlife markets could also facilitate recombination, and may explain divergent Gammacoronavirus strains ancestral to those in birds, in two mammals species in Southern Chinese wetmarkets ( 46). In this proposal, we will look for generalizable patterns among bat species and the CoV genotypes they harbor, and use this to examine how phylogeny and contact affect CoV spillover risk. Host-CoV interactions: an evolutionary approach: The interaction between CoV receptor binding domains (RBDs) and host receptors, e.g. ACE2 for SARS-CoV; dipeptidyl peptidase 4 (OPP4) for MERS-CoV; carcinoembryonic antigen-related cell adhesion molecules (CEACAM) for mouse hepatitis virus; and aminopeptidase N (APN) for hCoV-229E, is critical to understanding limits to host species range (47-52). Bats have highly diverse ACE2 receptors at a nucleotide and especially protein level (Fig. 2). This is in contrast to other viral receptors in bats, e.g. Ephrin-B2 receptors for henipaviruses (53, 54), and DPP4 for MERS-CoV appear to be highly conserved (51). Several different genera of bats (e.g. Myotis, Rhinolophus, and Rousettus) have receptors that support viral mediated entry by the SARS-CoV Spike protein (52, 55). A B Figure 2. Mammal ACE2 phylogeny using: A) nucleotide data (~2400bp) of ACE2 gene; B) translated protein sequences of same ACE2 genes. All mammal species with available data, including primates (purple), lagomorphs (pink); carnivores (blue); ungulates (green), rodents (brown), and bats (red). Bats are monophyletic and species group with expected taxonomic relationships using nucleotide sequence data (A); but they are paraphyletic a when analyzing protein-level differences (B). This shows functional ACE2 diversity may differ from nucleotide data, and a need to better characterize receptor diversity in a wider range of hosts. While our preliminary results suggest interesting patterns in bats (Fig. 2), the limited number of bat ACE2 sequences precludes ,.- --- .,... ..... _.... robust comparison of co-phylogenetic patterns. In this study, we propose to sample dozens of species more than 5 bat families in China, and compare sequence with bats we've sampled globally. This will allow us to build a testable, phylogenetically informed models to examine the extent of, and limits to, batCoV host-range; and will allow us to analyze other receptors like DPP4 for MERS-CoV. Modeling risk of human infection: The use of mathematical, computational models of viral dynamics has become a standard tool to understand risk of pathogen emergence and spread ( 56-60). However, models that characterize the risk of wildlife-to-human infection require data on contact among populations ( 61), evolutionary constraints of pathogens (29, 62), and diversity of novel pathogens (63). Because these datasets are usually unavailable, mathematical models can often be theoretical, and of reduced value in predicting risk of pathogen spillover and spread. Building on our group's experience in modeling disease emergence (64-67), we will develop a mathematical model that explicitly describes the transmission dynamics and evolutionary dynamics of CoVs in wildlife markets and in bat caves. These models will be parameterized with data we have already collected, and new data from this study, to predict whether novel CoV strains we discover are likely to emerge. Tests of host range in vitro: Receptor usage in different animals is a primary determinant of viral host range. However, while the receptor and receptor binding domains (RBDs) of human-infecting CoVs have been studied intensively, bat-CoVs have not (22, 47). In this study, we will determine the RBD of bat-CoVs, develop pseudovirus assays ( 68), and work with a humanized mouse model expressing ACE2 receptor. This provides a way to experimentally test hypotheses on the host-range of novel coronaviruses, even from sequence data. However, despite a plethora of novel CoVs in the recent literature (38, 39, 44, 45), there has been little work towards this goal. Furthermore, the recent discovery of MERS-CoV, which uses DPP4, and the use of other receptors for other CoVs (69) suggest that this work will be highly significant for other CoVs. B. INNOVATION: This project is an innovative fusion of virology, ecology, and mathematical modeling. The analysis of CoV genetic diversity in bats and other mammals in southern China, combined with characterization of and co-phylogenetic analysis with CoV functional genes (e.g. ACE2, receptor of SARS-CoV and DPP4 for MERS-CoV) has not yet been attempted, and will allow us to better understand the patterns of host-switching. Previous studies using molecular clock analysis have found that the bat SARS-like-CoV to civet/human SARS-CoV divergence Research Strategy Page 109

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter ranged from 7-17 years (mean 4.9) before the 2003 outbreak (7, 70). We will use a novel phylogenetic and mathematical modeling approach to examine how dynamics of contact and pathogen transmission among hosts in markets drives viral evolution and emergence. We will determine how many years it takes for a coronavirus to evolve an Ro > 1 and therefore have epidemic potential using a modeling framework that combines evolutionary changes and multi-host dynamics (Specific Aim 3), expanding on published approaches (71-73). We will then simulate coronavirus emergence under different market conditions to identify most likely scenarios that can inform strategies to prevent future outbreaks. Finally, we will use pseudovirus binding assays, in vitro infections and humanized mice expressing ACE2 to test our analyses on the novel viruses we have, and will, identify. We will use our multidisciplinary approach to examine fundamental questions on how the wildlife trade, wetmarkets and other target interfaces promote the emergence of coronaviruses, and assess the risk of future spillover of Co Vs from bats or other mammals and humans. In particular, despite 10 years since the emergence of SARS and the discovery of 60+ novel bat-Co Vs, three significant issues remain unanswered: 1) What are the natural limits to CoV host range, and can this be predicted by the host-receptor-virus relationship; 2) Are the conditions in wildlife markets sufficient to allow enough interspecies transmission that coronaviruses can evolve the ability to infect new hosts, including humans, either by accumulation of point mutations, or by recombination events; or 3) Is the expansion of the wildlife trade bringing expanded diversity of CoVs into the enhanced human-animal interface present in wet markets? C.APPROACH ~1: Specific Aim 1. Assessment of CoV spillover potential at high risk human-wildlife interfaces: C1a) General strategy and supporting studies: SARS-CoV emerged in live animal markets in Guangdong, with unrelated spillover events in at least five of seven municipalities, suggesting widespread introduction into wildlife markets within this city (21). We propose to characterize the species composition of bats and small mammals in wildlife markets where there is a high degree of contact between animals and people. We will identify additional high risk interfaces that may occur in southern China such as guano collection, which we have recently identified as a potential CoV exposure risk in Thailand ( 12). We will interview people at high-risk interfaces and who are enrolled in acute respiratory or influenza-like illness surveillance programs conducted by our colleagues at CDCs in Shanghai, Guangdong, Yunnan, and Guangxi. These data will be used to parameterize the contact process (z) in our mathematical model of CoV emergence (see Aim 3). We will assess 1) whether market conditions provide enhanced capacity (increased evolutionary opportunity) for bat-CoVs to evolve the ability to infect other hosts, either via repeated inter-species transmission, positive selection or recombination events; and 2) whether the intake of wildlife from Southeast Asia by China introduces a greater diversity of hosts and a correspondingly diverse group of CoVs (increased ecological opportunity). We will conduct CoV pathogen discovery in samples from humans and wildlife at these sites and examine their receptor binding domains to identify their ability to bind to ACE2, DPP4, or CEACAM receptors in humans. We will compare CoV diversity in China with that in wildlife across Southeast Asia (from our current work on other funded programs, and published data) that may potentially enter China's wildlife trade. Data from this aim will be used to assess the likelihood of inter-species bat-CoV transmission (see also Specific Aim 2). Working in high-contact human-wildlife interfaces can be challenging. However, we have already collected significant preliminary data to accomplish Aim 1. We have located and surveyed wildlife markets in Yunnan, Guangdong, GuangXi and Fujian provinces, and have identified populations that hunt and consume bats in Yunnan province. We have begun to characterize the species composition of free-ranging bat populations and have collected samples from over 1000 bat individuals (28 spp.) from 35 localities in over 15 (two-thirds of all} Chinese Provinces. We will also utilize archived wild bat, rodent, and civet samples collected by our team in Malaysia, Thailand and Indonesia on another large federally-funded project to provide samples of species regularly imported into China (section C1b) (21, 74). Research Strategy Page 110

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Wildlife Markets: Ten years following the SARS-CoV outbreaks, there is little information available on the current diversity of bats and other mammals available in the wet markets in southern China. One study found that 91 species of vertebrates, including 40 mammal species, were being traded in Guangxi, China ( 75). Further, little data is available on the origin of wild animals brought into the market system. In some cases, animals may be locally collected, while in other cases animals may be imported from Southeast Asia, including adjacent Vietnam (74-76)- factors which will affect the diversity of CoVs. Captive and free-ranging rodents are found in markets and may be an additional host for Co Vs ( 77). We have worked with Yunnan Institute of Endemic Diseases Control and Prevention since June 2012 (see Letters of Support). We have conducted initial surveillance in Nujiang, Baoshan Denong and Xishuangbanna prefectures and Ruili, which is a major wildlife trade gateway between Myanmar and China (Fig. 3). We have collected 187 small mammals from markets in Yunnan and tested them for coronaviruses using a 1-step PCR assay ( 78), finding 2/21 shrews (Crocidura attentuata) are CoV-positive. Figure 3: Map of wildlife trade routes from Southeast Asia into China. Modified from ( 79). Other animal samples available for this project: To date, our group has collected more than 90,000 high quality specimens from 15,000 animals representing key wildlife reservoirs for zoonoses such as bats, rodents and primates under our USAID-EPT PREDICT project. Clinical samples include blood, throat swabs, feces and urogenital swabs and represent animals from 10 different countries including Bangladesh, India, Malaysia, Thailand, Indonesia, China, Brazil, Bolivia, Colombia, Peru, and Mexico. 50,000 of these samples originate from Asia, and are currently being screened for novel coronaviruses (See Section C2a, Fig. 6). We have also collected more than 500 bat specimens representing seven species from the Kingdom of Saudi Arabia in collaboration with Saudi Arabia's Ministry of Health and Columbia University. Nearly 20,000 of our samples come from bats, and will be used to analyze CoV diversity along with novel CoVs we identify. Identifying novel Co Vs in wild bats in China: We have already conducted significant CoV surveillance in China for bats, other wildlife and humans. For this, we use pan-coronavirus PCR protocols based on conserved RNA-dependent RNA polymerase (RdRp) motifs A and C to screen samples at Wuhan Institute of Virology (80). Besides a large number of SL-CoVs, we have detected several novel bat-CoVs including strains closely related to CoV HKU4/5, CoV 1A &1 B, CoV HKU 2, 6, & 8. For the first time, we have also isolated and characterized a bat-CoV from China that uses ACE2 receptors (see Section C3a preliminary data) (28). In all. we have identified sequences from 268 novel bat-CoVs (140 from China alone) from bat species collected in Bangladesh, Thailand, Mexico, Brazil and China (See Section C2a, Fig. 6). We have an additional 5,000+ clinical samples from free-ranging bats and rodents from Guangdong province, from an ongoing study which are being screened for viral pathogens, including CoVs at Guangdong Entomological Institute. Survey of people highly exposed to wildlife in Guangdong, China: We have worked with Guangdong CDC since 2008, under a currently active IRB protocol, to interview and sample people working in live animal markets, hunters and restaurant workers with a high level of exposure to animals. We have interviewed volunteer participants about the nature and frequency of animal interactions; collected biological samples (blood, feces, sputum}, and trained participants to collect animal blood samples (dried blood spots on filter paper) from animals they butchered or hunted. We enrolled 1300 participants across 12 sites within Guangdong Province (Fig. 4). ·=:_°""'·1 Figure 4: Sites of current human sample collection by Guangdong CDC for zoonotic pathogen surveillance in Guangdong Province, Southern China. Each star represents a large wildlife market where we have enrolled market and restaurant workers (total= 1,300) for our zoonotic pathogen spillover study. Seventeen people had lgG antibodies to SARS-CoV and a follow-up study is underway. Samples have been tested for antibodies to animal pathogens, including SARS-CoV. Of the 1300 serum samples screened using a SARS-CoV ELISA. 17 were positive for lgG antibodies to SARS-CoV. These patients were not acutely ill at the time of sample collection, and this finding suggests one of three possibilities: 1) that SARS-CoV is still circulating in Guangdong markets; 2) that these people may have been exposed during the time of the 2002-3 outbreak; or 3) that the ELISA used is cross-reacting to another CoV. Review of their history of wildlife exposure is currently underway. In Shanghai, the Shanghai Municipal Center for Disease Control and Prevention (see Letters of Support) currently conducts surveillance on people with influenza-like illness in rural communities surrounding Shanghai. We will develop a similar study of people in Research Strategy Page 111

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter these communities who have exposure to wildlife. We will review and re-screen archived blood samples at Guangdong CDC for other bat coronaviruses once we determine candidates that could likely infect humans. to see whether there is exposure to Co Vs other than SARS. We will re-screen these samples with specific serological assays based on bat-Co Vs that will help differentiate between SARS-CoV lgG and other bat-CoV lgG to see whether there is exposure to CoVs other than SARS (3, 81). We will expand our survey to Guangxi, Fujian, Shanghai and Yunnan provinces to survey regions where SARS-CoV was not reported, but where wildlife trade, hunting, and bat guano collection is common. C1 b) Market characterization, wildlife sampling and human surveys: We have conducted surveillance at the wildlife markets of Guangdong where early cases of SARS-CoV were identified. From 2011-2013 we interviewed and sampled animal vendors, hunters and restaurant workers who butcher wildlife (See Section C1a, Fig. 4). For this proposed study, we will identify 10 markets in Guangxi, Yunnan, and Fujian Provinces (Fig. 5). We will characterize the physical size, number of vendors, diversity and abundance of mammalian species in each market. A questionnaire will be developed based on the one we used in Guangdong, to collect data on the nature and frequency of animal exposure of people who work in markets or hunt wildlife. We will conduct interviews to determine which bat species are sold, typical numbers, and source locations. We will collect information about recent acute respiratory illness and include those who have had undiagnosed acute respiratory symptoms within 3 months of the survey. We will then screen volunteers from this cohort for bat-CoV antibodies using existing and newly developed assays. We will compare exposure rates between people who are highly exposed to wildlife and a control group from the same regions. ·, -·r· ;w? - ~~.-- Figure 5: Proposed sampling sites in Southern China (Guangdong, Guangxi, and Fujian Provinces) for the current study. Arrows indicate wildlife trade routes. Letters indicate wild animal markets in Guangdong (A-R), Guangxi (S-W), Hunan (X) and Fujian (Y). ';:rt--...~ • GtNA In Shanghai, where wildlife markets are less common than southern provinces, we will L interview voluntary participants under surveillance by Shanghai CDC for influenza-like ----- illness. We will compare CoV exposure rates in people with acute respiratory illness to a control group from the same region (see letter of support). Wildlife sampling: We will locate wild bat populations used to supply local markets in Yunnan, Guangdong, Guangxi, and Fujian. We will sample a minimum of 30 individuals from 30 different bat species representing but not limited to the following families: Rhinolophidae, Hipposideridae, Vespertillionidae, Mollossidae, and Pteropodidae, all of which are known to carry alpha- or betacoronaviruses and are consumed by people ( 4, 7, 82). Bat SL-CoV PCR prevalence is 10%-38% ( 4, 24). Given 10% prevalence in bat populations, sampling 30 individuals would ensure a CoV detection probability of 95%. In all wildlife markets, we will opportunistically sample a variety of insectivorous and frugivorous bats, and other mammals if available, taking fresh feces or rectal swabs, saliva (oropharyngeal swab), and blood. A small number of bats will be sacrificed as vouchers and to collect intestinal tissue for CoV receptor analyses if required. We will use cyt-b to identify host species. Human exposure to Co Vs study: Expanding on our work in Guangdong, we will develop a voluntary study of animal vendors and hunters in Guangxi, Yunnan, and Fujian provinces in cooperation with local Bureaus of Public Health and CDCs. We will develop a survey to identify people with high exposure to wildlife, particularly bats, and will recruit volunteers, collect blood, sputum, and stool sample from each enrolled participant. We will screen sera for antibodies to SARS-CoV, other alpha & beta coronaviruses including MERS-CoV, and novel bat-CoVs. We will screen stool from CoV seropositive participants for CoV nucleic acid. We will also develop specific bat-CoV serological assays and share these with our Chinese collaborators. In each province in southern China we will aim to include 10 markets and survey 20 vendors per market; 20 additional wildlife hunters per province (220 case subjects); 400 control subjects from the general population near the markets in each province (total of 620 people per province). For Shanghai, we will enroll 200 acute respiratory illness cases and 400 non-respiratory controls (600 total), The total number of human subjects will be 2460. The study will be conducted in Guangxi, Yunnan, Fujian and Shanghai provinces (see Section E, Human Subjects). C1 c) Data analysis: Human sera and stool samples will be tested at provincial CDC labs (see letters of support) and animal samples will be screened at the Wuhan Institute of Virology (Co-I, Shi). Serum or plasma samples will be tested for CoV antibodies using ELISAs specific for SARS-CoV and bat SL-CoVs that we have developed ( 4, 68, 83). Fecal and saliva samples will be tested for CoV viral nucleic acid using a series of pancoronavirus PCR assays that target a region in the RdRp that is highly conserved among coronaviruses and for which we have a positive control, developed by our group under another federally-funded contract ( 13, 23, Research Strategy Page 112

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 84). The RdRp gene will be sequenced from all positive PCR samples and used to build co-phylogenetic trees (see Specific Aim 2). We will also test these pathogens for recombination events in markets vs. wild sampled Co Vs after viral strains are characterized. Data from Aim 1 will be used to parameterize mathematical models of viral transmission (Specific Aim 3) in markets to estimate relative risk of emergence depending on different diversity of mammals, contact rates, size of markets, and evidence for human exposure to bat-Co Vs. C1d) Potential Pitfalls and Solutions: We may find lower than expected levels of wildlife diversity in markets in Southern China. If this occurs, we have access to tens of thousands of wildlife samples from over 20 countries globally from work on a current NIAID R01 (Daszak Pl) to assess diversity of viral pathogens in bats in Asia and Latin America, a large multi-year contract from USAID (Emerging Pandemic Threats: PREDICT program, Daszak Pl) to conduct surveillance and pathogen discovery in wildlife in Asia and Latin America and two Nipah virus R01 s. We have already discovered >250 novel Co Vs from bats in these countries (Section C2a) including >100 from China. A second setback would be that access to markets becomes restricted due to political sensitivities. We are working closely with long-term local collaborators at ECNU and the Institute of Virology, Wuhan, both of which institutions are well respected nationally. The Institute of Virology is the National Center of Excellence for viral pathogens, and has Federal authority for viral research. Furthermore, we have shown through our work with Guangdong CDC that we can conduct long-term collaborations in these sites. Finally, we have selected a large number of wildlife market sites, so the closing of one will not affect all sampling activities. C2: S~ecific Aim 2. Rece~tor evolution, host range and redictive modeling of bat-CoV emergence risk'.: C2a) General strategy and supporting studies: Can we use information on Co V sequence, host sequence and behavioral traits and population dynamics at critical human-wildlife interfaces to predict which Co Vs are most likely to emerge?To answer this, we will use data from our characterization of bat-CoVs, host range, receptor genes, serological data, and from field-collected data to build and parameterize three related models. First, using phylogenetic reconciliation we will map the co-phylogenetic patterns of bats and their CoVs using neutral and functional markers (RBDs and host receptor genes). We will compare free-ranging and market-sampled species assemblages and test the related hypothesis that wildlife markets disrupt 'natural' bat - CoV host associations and increase recombination and/or accelerated evolution to facilitate emergence. Second, we will construct generalized linear models that encompass phylogenetic information to test the two related hypothesis that spillover potential and host-range of bat-CoVs is limited by: 1) opportunity for contact; or 2) phylogenetic relatedness of host species and their receptor genes. Third, we will use mathematical matrix modeling to investigate bat-CoV transmission and evolutionary dynamics, and test the potential of novel CoVs to infect humans, bats, and other market animals. This model will be informed by serological data, market surveys, and receptor binding data from bat cell line and humanized mouse inoculation studies. Phylogenetic studies of known and novel bat-Co Vs: Phylogenetic methods can be used to identify recent host shifts and spillover events of Co Vs, often these events are due to anthropogenic changes to host ecology, e.g. Rhinolophus spp. and human/civet SARS-CoV in the wildlife trade ( 4, 7). It has been proposed that repeated passage between civets and humans in wet markets facilitated SARS-CoV evolution towards greater human and civet ACE2 receptor affinity (85), and accelerated evolution and positive selection in CoVs was detected after host spillover (86). It is not known if bat-CoVs follow predictable patterns of co-phylogeny between host and virus; many studies found unique CoV strains circulating in different bat lineages, but also multiple - CoV strains have been identified in the same bat species and individuals ( 7, 40, 44, 45). --=-• Figure 6 (above): Phylogenetic tree (RdRp) of selected bat-CoVs from Genbank, including as "-----'-=._'----- - • .....,,_ subset of the 268 novel bat-CoVs discovered by our group through our USAID-EPT PREDICT ...... • .. ,~ pathogen discovery work in China, Brazil, Bangladesh and Mexico. Wildlife trade and market dynamics may promote the cross-species transmission of distinct bat-CoV strains and facilitate viral recombination within these hosts ( 46); the extent of this will depend on the role of host phylogeny vs. contact in limiting bat-CoV spillover. Using our extensive database of bat and other wild animal Co Vs that we have characterized, isolated, or are available on Genbank, we will examine these constraints for known and novel Co Vs we identify. Over the past four years. we have conducted large surveys of bat pathogens globally. including the discovery of sequences from 268 novel bat-CoVs (including 140 from China) (Fig. 6). Research Strategy Page 113

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Figure 7 (left}: Host-pathogen co-phylogeny of bat-CoVs from China (43). Bat genera: R, ::-:~:., Rhinolophus; Mm Miniopterus; Mr, Myotis; P, Pipistrellus; V= viral sequence, B= bat sequence. This E _ figure suggests rhinolophid CoVs may have a greater ability to jump hosts. Warrants further · =: - ~ investigation using functional genetic markers and data from more species. -... ., C2b) Co-phylogenetic analysis of bat-CoVs: We will use coronavirus and host sequence data generated in this project, from archived samples that we collected from bats just after the SARS outbreak, and previously published CoV strains from a diverse range of host species to quantify co-evolutionary patterns and host range in bat-CoVs. Combined analyses of host and viral phylogenies will allow us better understand if host phylogeny (and receptor gene similiarity) can predict CoV host switching and whether or not market systems have disrupted the "natural" patterns of CoV association (e.g. Fig. 7, from (43)). We will reconstruct phylogenetic relationships of CoVs using a combination of the HEL, N, RdRp, and S genes, as each has a different evolutionary rate and will allow us to test patterns of cophylogeny at different taxonomic scales. We will reconstruct host species relationships from tissue collected in our study using both neutral (mitochondrial and nuclear, e.g. cytB and RAG2} and functional (e.g. ACE2 CoV receptor) host genetic markers. Multiple alignments will be performed MAFFT (87), and phylogenies estimated using maximum likelihood (88) and Bayesian inference (89) for each viral and host gene, and concatenated virus datasets when no viral recombination is detected. We will test for statistical significance using ParaFit implemented in CopyCat (90) and AxPcoords (91), and visualize these using TreeMap v2.0213 (92). These methods will allow us to identify which particular host-virus associations contribute most to the observed patterns. We will partition our dataset by collection localities and higher-level taxonomic groups to test co-phylogenetic significance at multiple spatial and taxonomic scales. To test the null hypothesis that there is no pattern of co-evolution we will perform permutations to randomized hosts-virus associations and then measure congruence relative to the host tree. By comparing the patterns of host-CoV co-phylogeny in natural bat communities (cave sites) vs. wet markets, we will be able to identify anomalies that may likely signal recent spillover events. To test for genetic recombination in market vs. wild-collected bat-CoVs, we will use sliding window analysis (7) and RDP3 v3.44 software (93). We will use previous methods to test for positive selection and identify specific virus residues under selective pressure ( 94). Quantifying CoV strain sharing between host species: We will use viral sequence data from RdRp and S genes to delimit unique CoV "species" or "genotypes" at different taxonomic and sampling levels. We will test for non-random patterns of association of viral community assemblages between species (95-97) (98). This will involve calculating Jaccard's index of similarity (J) for the viral assemblages between pairs of species and testing for deviations from that expected by random chance using Monte Carlo randomizations ( 99). Deviation from the null model will be calculated as the difference between the mean J observed (Jobs) in the data and the mean J expected, such that Jc1ev= Jobs - Jnufl• Positive values of Jdev will thus indicate that CoV community assemblages between host species are more similar than would be expected by random chance, while negative values would indicate greater dissimilarity in the viral assemblages than would be expected by chance. C2c) Predictive model of CoV host-range and diversity: We will develop a predictive model of host-range for bat-CoVs using data of bat distribution in natural caves and the markets. geographic ranges, ecological and behavioral characteristics of host species from our field studies and the literature. host and viral phylogenies. and associations of host species to particular CoV strains/clades. We will include phylogenetic distance between bat species and other mammal hosts from various neutral and receptor genes generated in this study. We will use CoV similarity indices (Jaccard, above} as our response variables in multiple regression models, i.e. generalized linear models (GLMs) and phylogenetic generalized linear mixed models (PGLMMs) with relevant bat ecological, phylogenetic, morphological. behavioral, and life history traits as our predictor variables, to assess the relative contribution of host phylogeny, viral traits, or species-specific ecological traits in explaining CoV diversification and sharing. We will calculate indices of host specificity that account for host phylogeny ( 100, 101), to further test hypotheses of whether bat-Co Vs are more likely shared between host ecological groups or among species with similar life-history traits vs. relatedness. All statistical analyses will be conducted in R with relevant packages for community ecology and species diversity (vegan, fossil), and phylogenetic modeling (ade4, ape). Extension of this model beyond China will allow us to map a global spatial and phylogenetic risk gradient for CoV emergence based on host species traits, mammalian phylogeny (including functional CoV receptor genes), and relatedness of CoVs. Further, we can use the results from our logistic regressions to identify gaps in surveillance, where bat species are found to share a lower than expected number of CoV strains given a threshold level of contact and relatedness with other host species. We will test our predictions of host range from the analytical model for bat-Co Vs using synthetic reconstruction of bat-Co Vs and in vitro studies of ortholog Research Strategy Page 114

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter receptor binding with different mammalian cell lines (Aim 3). Specifically, we will evaluate the ability of novel bat-CoVs to recognize and bind to selected receptors (ACE2, CEACAM, APN, receptor for alpha-CoV, or DPP4/CD26, receptor of MERS-CoV) reconstructed from divergent bat taxa. We envision an iterative process over the first few years of this grant whereby initial data are generated from known host-CoV associations. results from the model will be tested experimentally. and then data from experimental studies will be used to refine the models and better inform field sampling in China and globally. Analyses of literature database: We have built a database of virus-host associations for 131 bat species and all 50 unique ICTV recognized bat viruses. We used a logistic GLM regression approach with host and virus variables, and found that host phylogeny (i.e. phylogenetic distance to other bat host species) was a strong predictor of observed virus sharing across bat species (trend with phylogeny only shown in Fig 8). We will adapt this approach by using host genetic distance of functional receptor genes instead of neutral markers, and CoV data collected from our standardized survey efforts. Fig 8. Scatterplot showing a decrease in the number of shared viruses with decreasing phylogenetic relatedness among bat species. Dataset includes all bat species pairs with >3 shared viruses for -200 bat-virus associations form the literature (Olival, unpublished). Pairwise phylogenetic distance from maximum likelihood tree using cytochrome B mtDNA data. C2d) Modeling the dynamics of CoV spillover risk: A key question in EID research is the role of viral evolution in enabling pathogen emergence. While some EID ·--~-·""""" pathogens cause epidemic or pandemic disease because they readily transmit among humans (Ro.Human> 1, e.g., HIV, A/H1N1pdm), or only spillover directly from animals (Ro.Human= 0, e.g. West Nile Virus). Others, including MERS-CoV, may spillover regularly to humans, and even cause small clusters of human-to-human transmission, but have not yet caused a major epidemic or pandemic (1 > Ro.Human > 0, e.g., Nipah virus, monkeypox, Influenza H5N1 ). A looming issue is the likelihood of such a pathogen evolving to become a major epidemic or pandemic (i.e., Ro.Human> 1 ). Divergence times between ancestral bat-CoVs and hCoVs can vary widely and provide a timeline of past spillover events, e.g. 560+ years between hCoV-NL63 and its progenitor alpha-CoV ( 6) and ~20 years between bat SARS-like Co Vs and human or civet SARS-CoV ( 7, 70). The limits on SARS emergence are still unclear: Were the bat SL-CoVs unable initially to bind to human receptors, or was it necessary for a precursor CoV to evolve and adapt to humans for SARS-CoV to emerge? Were civets or similar non-bat, non-human hosts a critical intermediate evolutionary step in the transition from bats to humans, or were they incidentally infected along with humans simply by virtue of similar receptors? To examine the timeline for different emergence pathways, we have built a model framework (below) to represent the wildlife market environment and include viral ecology and evolution. We will use a matrix framework (72, 102) to determine how the pathogen is transmitted among different host species and between locations. We have already built the framework of this model (below), and have listed the data that we will collect in the current study to parameterize it (Table 1, below). To incorporate strain variation and evolution, we will adapt the approach of Antia et al. ( 71) by integrating a branching process approach to our matrix framework. We will use these techniques to develop "What If" scenarios that predict how different strains of CoV would emerge, and potentially evolve, in different market systems within Asia and elsewhere (e.g. scenarios with different host diversity and different levels of host-host and host-human contact within markets). To examine strain evolution, we will model n possible strains, where strain 1 is the initial variant, and strain n is the variant that has a human R0 >1, with n-2variants in between, which may each have their own R0 that depends on the host community using 'Who-Acquires-Infection-From-Whom' (WAIFW) matrix framework (below). Following Antia et al. (71), we assume the mutation rate,µ, is the same for all variants, that only single mutations can occur, and we ignore back-mutation. However, we will reconsider these assumptions if changes in these can alter the expected outcomes of the mathematical results. We also assume that the total number of secondary infections generated by an individual with variant i is Poisson distributed with mean R/>. A proportion µ of the variants will have mutated or recombined into type i+ 1, while the proportion (1- µ) remains the same, as type /. We will separate out the cases of mutation and recombination by placing different restrictions on the changes that could occur in the strains as they move from type i to type i+ 1. These assumptions result in the probability generating functions: - (i) ' (i) fi(s1,s2, ... ,s,,,)-e)q>(-(1-µ)R 0 (J-s,))exp(-µ,R 0 (l-si+1)) for i<m ( 5) otherwise fm(s1,-~2, ... ,sm) = exp ( - R~"')(l -s 111)) Research Strategy Page 115

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Through this branching process approach we can gain insight into the limitations and possibilities that stochastic processes may impose on the evolution of strain diversity in both limited diversity settings (e.g., only bats and humans), and highly diverse environments (e.g., markets with other hosts such as civets and bamboo rats). We can also adapt this methodology to compare mutation, which we expect to take small incremental movements in a fitness landscape that may have low fitness valleys between a wild-host adapted strain and a human or other host adapted strain, and recombination which may be able to take larger leaps across a given fitness landscape. Using this framework we can vary the R0(i) depending on the fitness of the mutants in various hosts, and the host diversity and abundance, simulating the complex fitness landscapes of real CoV systems. We can calculate the number of secondary hosts infected as Ro= z<frr, where -r is the duration of infectiousness, and z is the rate of contact. Our receptor binding studies and predictive GLM models of host range will be used to inform ¢, the joint probability that a susceptible host becomes infected when exposed. We will model our system both mathematically from a simple R0 perspective for insight, as well as using a spatial stochastic-birth-death simulation implementation to understand the implications of multiple scales of variation, including mutation and recombination and the implications for stochasticity for CoV emergence. To do this we will expand our basic equation, Ro= z(J-r, into a matrix formulation to incorporate the multiple hosts within this system. Each strain and spatial location (e.g., market), can be represented by a different matrix. Thus we have: x,. - . . . q,k - . . . Tk - . . . _ [Xl-1,k . . l _ [r/>1,1,k . . l _ [7"J,l,k . . l ( 1 ) · · X,,J,k · · 'Pi,j,k · · r;,j,k which we can use to define a 'WAIFW' nk matrix (73, 103) of which the eigenvalue gives us an estimate of Ro for the whole system, for a given strain and location. The 'WAIFW' matrix is: Ok= [ . . . ;\l,L,,r/>·1,l,k7°1,I,!: '. l (2) . ' X,.J,!-efii,J.k1'i,J,!s Critically, this enables us to analyze certain 'what-if' scenarios. For example, we can examine the role of civets in emergence by assuming that the strain which initially infected civets had to evolve in order to then infect humans. This would give us two strains in a single location, each with its own R0. Alternatively, we can assume that all three SARS-CoV host species (bats, civets, humans) were in the same market place, and a single CoV strain. In this case we would have a single matrix, with all three species, and values in every cell of the matrix. By keeping the separate pieces of the R0 = z~equation, in the matrix form, we can examine potential public health control measures (e.g. quarantine, culling or separating species into different market locations) (104), which might also vary depending on the nature of receptor binding and strain evolution. To account for assumptions, we will investigate the implications of mixing in a stochastic environment. We have already built a stochastic-birth death, discrete event simulation of the spread of EIOs for Avian Influenza in multi-species markets and farms. We will adapt these simulations for strain and receptor diversity interactions with multiple species of CoV hosts. This suite of modeling approaches will allow us to integrate our ecological and molecular approaches to understanding the potential pandemic emergence threat posed by the whole suite of bat-CoVs. Table 1: Data Needs tor Model: Parameter Description Sources 't Human. 't Bat, Duration of infectiousness, Humans, Bats, Humans (57, 105-108), Bats ( 7), other 't Other'.' other spp. species ( 108, 109) <!>Human->Human Joint probability an infected Human can (57, 105-107) transmit to susceptible Human <l>sat->Human, <!>other->Human Joint probability an infected host can *( 109) <l>sat->Other, <l>sat->Human transmit to susceptible; can use receptor <l>other->Other binding in host species for parameterization <!>? As above Generally assume O or <l>i.i = <l>J,i * Xi,J Contact rates Market Surveys, using map overlap for non-market areas. µ Mutation rate Literature I:. Recombination rate Literature NHuman Nbat Nother Population density of bats, humans, other Market surveys, census & transect data. * Use knowledge of receptor bindings to appropriately upscale or downscale relative to human-to-human case of SARS and laboratory studies on other animals. We will run sensitivity analyses for these parameters. Research Strategy Page 116

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter We will assume that twice the estimate for SARS Ro.Human rounded up to the max of the 95% Cl to give 5 or 10 represents a near maximum, and O forms a lower boundary. We will assume -r is constant regardless of species and again do sensitivity analysis using SARS-CoV values. We test the following hypotheses: 1) That recombination can either substantially boost (HA: !lPr > 0) or mutation have the same effect (HA: !lPr > 0) on the probability of CoV spillover into humans, or that only recombination and mutation together provide a substantial boost to spillover probability (HA: !lPr > O); 2) That known (e.g. civets) or unknown intermediate animal hosts or no intermediate hosts are necessary for CoV spillover to humans; 3) That high diversity of intermediate hosts either increases or decreases the probability of CoV spillover into humans. We will use our modeling framework to examine the potential CoV spillover in different markets, using the market data from Specific Aim 1, evolutionary characteristics of the CoVs from Aim 2, and specifically-acquired data to parameterize the model. Table 1 (above) lists parameters in the model, and gives available sources for data. Previous experience of modeling disease emergence: Our group has used mathematical models to test hypotheses on zoonotic disease emergence for over 15 years. We use computational models that are tailored for the specific pathogen type or combination of hosts involved, and parameterize these with extremely detailed datasets specific for the emergence event. We then run simulations to test hypotheses on the spillover of viruses and the emergence of zoonoses. For Nipah virus (NiV), another bat-borne zoonosis, we obtained data from pig production facilities in Malaysia ( 110, 111), from experimental infection of bats and in vitro under BSL-4 conditions for viral transmission parameters ( 112, 113). We used this approach to demonstrate the cause of NiV emergence ( 111). We have successfully used similar approaches to demonstrate viable causal mechanisms for the emergence of Hendra virus ( 114), Avian influenza ( 115-117) and West Nile virus ( 118-120). C2e) Potential pitfalls and solutions: The diversity of coronaviruses that we identify may be inadequate for robust co-phylogenetic analysis. We have already shown proof of concept in preliminary data through USAID and NIAID funded projects that we have detected new coronaviruses in most bat species examined; there has been a large amount of research from several groups showing a broad diversity of coronaviruses; previous studies from us and other groups have provided evidence of a diversity of coronaviruses associated with bats and there is high likelihood that we will identify more. In China specifically, 23% of bat samples we have screened were positive for CoVs, thus we do not anticipate a lack of diverse CoVs (28). For modeling studies, not all necessary parameters may be easily obtained. We will use information from the SARS-CoV outbreak, where we have detailed data from the WHO investigations on serology and viral isolation from market wildlife; and from our recent and current work in Guangdong province; and an ongoing study on avian influenza in Shanghai and Guangdong markets (Co-I Zhang). Lastly, for parameters that we cannot actually estimate, we may be able to posit reasonable limits. For example we can constrain the probability of spillover: it must be greater than 0, since SARS did in fact spillover ( 106), but it is very unlikely that this probability is higher than the within species transmission probability. If the rate of transmission within a host species is unestimatable, we can use data from other diseases in similar species, such as bat rabies. Thus we can readily perform a sensitivity analysis for unknown parameters within a range that is biologically plausible, using sensible constraints. ~3: Specific Aim 3. Testing predictions on CoV inter-species transmissio": How can we test predictive strategies to understand which viruses have the capacity to 'iump hosts'? To answer this, we will analyze the interspecies infection or transmission of CoVs we have identified, particularly the SARS-like CoVs and CoV HUK4/5 that is closely related to MERS-CoV (hCoV-EMC) from Saudi Arabia. Our main approach will be: 1) in vitro infection experiments using pseudoviruses carrying the spike proteins (wild type or mutants) or live viruses in cell lines of different origins; 2) binding affinity assays between the spike proteins (wild type or mutants) and different cellular receptor molecules; and 3) humanized mouse experiments if viruses are identified of significant human infection potential (see Ralph Barie, Letter of Support). C3a} General strategy and supporting studies: We will sequence the spike (or other receptor binding/fusion) protein genes from all bat-CoVs we identify, creating mutants of these to identify how significantly each would need to evolve to use ACE2 or CD26/DPP4 (receptor for MERS). We will then use receptor-mutant pseudovirus binding assays, in vitro studies with a wide range of cell lines from bats, other mammals including primates and human cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated (see Ralph Barie, Letter of Support). These tests will provide direct public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. Research Strategy Page 117

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Experience working with receptor mutants & pseudovirus binding assays: We have established a stable pseudovirus assay for SARS-CoV and SARS-like CoV and tested the infectivity of these spike proteins in cells expressing ACE2 from human, civet and bats (52, 68). We have demonstrated that several bat species are susceptible to the SARS-CoV and that some SARS-like CoV strains can use human ACE2 for cellular entry (52). Minor mutations in S proteins or ACE2s greatly affected the receptor binding and finally abolish the pseudovirus entry ( 68, 121) (Fig. 9). Recently, we have discovered a number of alpha and beta CoVs including HKU4/5 ( 122)(Ge et al., Co-infection of alphacoronaviruses in one bat community, in China. unpublished results). The established approaches will enable us to analyze the receptor usage of these CoV S proteins and understand the host range ... -." and potential interspecies transmission ability of these novel Co Vs and finally I ll UI ~ predict the potential spillover probability of these viruses to humans or other I_' hosts at a molecular level. -- - Figure 9: Top panel: HIV pseudovirus carrying spike proteins from human SARS-CoV (BJ01-S) and bat SARS-like CoV (SHC014-S, 3367-S and Rp-S). Bottom panel: lnfectivity assay with the above pseudoviruses in HeLa cell lines expressing ACE2 from human, civet and bat. In vitro cell lines & Humanized mouse model: We have developed primary cell lines and transformed cell lines from 9 bat species using kidney, spleen, heart, brain and intestine. We have used these for virus isolation, infection assays and receptor molecule gene cloning. We also have a large number of cell lines from humans and animals that we will use for virus infectivity assays. We have obtained a letter of support from Dr Ralph Barie, who is keen to collaborate with us initially to infect his humanized mouse model with our bat SL-CoV that uses ACE2, and subsequently to use other Co Vs we identify (see Dr Ralph Barie, Letter of Support). C3b) Receptor-mutant pseudovirus binding assays: We will amplify ACE2, DPP4 or other receptor genes of human and bats and clone them into eukaryotic expression vector pc0NA3.1 to construct cells expressing these molecules. We will amplify full length spike genes (S) of bat-CoVs detected from different bat species. The full length S gene, particularly RBDs, will be codon optimized, then cloned into eukaryotic expression vector pcDNA3.1 ( 68, 123). For packaging pseudovirus, S-expressing plasmids ( or empty vector control) and pH IV-Luc (pNL4.3.Luc.R E· -Luc) bone plasmid will be co-transfected into 4 x 106 293T cells using calcium phosphate transfection system (Promega), after 8 hours, replacing the medium with fresh medium, and supernatants will be harvested at 48 hours post transfection and separated from cell debris by centrifugation at 3,000g, then by passing through a 0.45µm filter (Millipore). The filtered supernatants will be stored at -80°C in aliquots until the use. We will use prepared pseudoviruses bearing different S proteins to infect human and bat ACE2 or DPP4 receptor expressing cells (in Hela cell model), 24 hours post infection, receptor usage by different S proteins will be determined by measuring luciferase activities. We will also induce site mutations in S proteins using site-directed mutation method, then do receptor-mutant pseudovirus binding assays. Pseudovirus infectivity on different human cell lines (A549, 293T, Caco, Huh 7, and etc), primary and immortalized bat cell lines (listed below) and other mammalian cell line (mouse, pig, hamster, monkey, and ect) will be also determined by luciferase assay. The results will provide information whether bat-CoVs could use known bat and human ACE2. DPP4 or other known CoV receptors to enter cells. and allow us to determine critical receptor binding sites. viral host range. and to better predict the capacity of our Co Vs to infect people. C3c) In vitro studies: We will isolate bat-Co Vs using Vero E6 cell (susceptible SARS-CoV and MERS-CoV) and primary or transformed bat cell lines that we have developed from Myotis davidii, Rhinolophus sinicus, Myotis chinensis, Rousettus leschenaultia and other bats of China ( 124, 125). CoV PCR-positive bat samples (in 200 µI buffer) will be 3,000-12,000 rpm gradient centrifuged, and supernatant will be diluted at 1 :10 in DMEM medium, then added to cells, incubated at 37°C for 1 h, the inoculum removed and replaced by fresh DMEM medium with 2% fetal calf serum, and cells checked daily for cytopathic effect (CPE). Double dose triple antibiotics (penicillin 200 IU/ml, streptomycin 0.2 mg/ml, amphotericin 0.5 µg/ml-Gibco) will be included in all culture media. Three blind passages will be carried out for each sample and the culture supernatant and cell pellet examined for presence of virus by RT-PCR using primers targeting the RdRp or S gene after each passage (28, 126). Live bat-CoVs will be sequenced to confirm viral receptor and by comparing viral infection in ACE2 or DPP4 expression cells and virus infectivity and replication on different human cell lines (A549, 293T, Caco, Huh?, and etc), bat cells and others (mouse, pig, hamster, monkey) using plaque assay, real time-PCR, and Immunological Fluorescence Assay (IFA). These in vitro assays will be used to test viral host species range and transmission possibility of bat-CoVs to human and other mammal, as predicted by our GLM and matrix models. Research Strategy Page 118

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter C3d) Humanized mouse in vivo infection experiments: To evaluate pathogenicity of bat-CoVs we will perform in vivo infection experiments in humanized mice modified to carry human ACE2 or DPP4 gene in the Wuhan Institute of Virology BSL-3 animal facility. We will passage isolated bat-CoVs in permissive cells twice, administer a specific inoculum (e.g. 1x106 TCID50) to intranasally or intraperitoneally. Mouse body temperature will be monitored with implanted microchips (LifeChip Bio-thermo, Destron Fearing), and mice will be weighed and observed for clinical signs of illness daily. Dead or moribund mice will be euthanized, organs harvested and sectioned. Live animals will be euthanized at three weeks post-inoculation and organs harvested. We will test for neutralizing antibodies against bat-Co Vs on days 10, 15 and 21 pi. We will collect nasal washes, oral swabs, and rectal swabs, and urine every two days and quantify virus using qRT-PCR. We will conduct routine histology, immunohistochemistry, qRT-PCR, and virus isolation on tissues. This work will provide information about viral pathogenicity, tissue tropism, transmission route, and infection symptom. C3e) Binding affinity assay: The recombinant S proteins and receptor molecules (e.g. ACE2 or DPP4) will be expressed in insect cells or eukaryotic cells. Octet RED platform (ForteBio, Menlo Park, CA)) will be used to perform binding affinity kinetics experiments. Streptavidin-coated sensor tips from Fortebio will be used to capture biotinylated S protein onto the surface of the sensor. After reaching baseline, sensors will be moved to the association step containing indicated concentrations of wild or mutant receptor molecules diluted with kinetics buffer for 30 min and then dissociated for 30 min at 25°C. Binding affinity will be determined by collecting the dissociation constants KO, Kon (association-rate), and Koff (dissociation-rate) determined by fitting binding chromatogram data with the Octet®User Software. C3f) Potential pitfalls and solutions: Through our targeted sampling in China, we may only identify a small portion of the huge diversity of bat-CoVs in bat populations. To resolve this, we plan to expand our sampling locations to include samples from across SE Asia and improve our detection methods targeting more virus sequences. We will also synthesize the S genes based on the published data for viruses we do not obtain. Virus isolation may be a big challenge for this specific aim. In our previous work, we have isolated a number of novel bat viruses including adenovirus, reovirus and SARS-like CoV and have refined and optimized our methods for virus isolation. We will also attempt to construct additional bat cell lines which are lacking interferon response or over expressing the receptor molecules and more susceptible for virus infection to increase isolation success. D. TIMELINE & MANAGEMENT PLAN: Marketldentlfica~~~\haracterirntion 2013 2014 2015 2016 2017 2018 This project will take 5 years to complete. The AolmalSamplln uisl1lon 1,5 initial phase will involve filing the IRB application, Lab Testln of s 1RBApp11ca11on 1.5 identifying sampling sites, and conducting animal HumanSamplln 2.25 sampling and testing. Mid-project efforts will Lob Testin of Human 5.amples J .5 Lab Data Analysis and Modeling 4,0 involve initial human sampling, analyses of lab results and production of models. The final phase will involve testing human and wildlife samples and analyses and modeling to maximize results. Project Management: Funds will be managed via subcontracts originating with EcoHealth Alliance, which is an A 133 (low risk)-audited 501 (c) 3 organization specializing in international research on emerging diseases. Pl Daszak will oversee all aspects of the project management. He is an experienced manager, with over 15 years of federally-funded research experience. Prof. Shi, based at the Wuhan Institute of Virology, will oversee all laboratory testing and analyses. Prof. Shuyi Zhang will manage field sampling work. EcoHealth Alliance staff will manage all modeling and analytical approaches (Aims 1 & 2). Communication will be via monthly video-conferences using EHA's NIH ARRA-funded video-conference facility. Travel budget has been requested to enable regular face-to-face meetings for all key staff. Research Strategy Page 119

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter PROTECTION OF HUMAN SUBJECTS: HUMAN SUBJECTS RESEARCH 1. Risk to subjects This project is a study of human exposure to animal coronaviruses in southern China. Subjects will be enrolled on a voluntary basis and a single interview and sample collection will be conducted. Informed consent will be obtained. People found to be infected with an animal coronavirus will be followed up after 6 months with a secondary interview and collection of biological specimens to determine whether infection is persistent and exposure is ongoing. Primary subjects will be male or female adults who are highly exposed to wildlife through hunting, butchering, or general handling in the context of live animal markets or restaurants that prepare and serve wild animals. The study population will be selected in Shanghai, Yunnan, Fujian, and Guangxi provinces, China, and will be open to people of all ethnicities that fit the subject criteria. We will target human subjects, comprising 220 subjects (market workers and hunters) and 400 controls from the general population in Yunnan, Fujian, and Guangxi provinces plus 600 subjects in Shanghai (total enrolled: 2460). The market types are defined in Specific Aim 1, Human exposure to CoVs. There are no data to suggest an ethnic bias for coronavirus exposure or infection, therefore subjects will be enrolled based on exposure criteria, though subjects will not be excluded based on ethnicity or gender. We will endeavor to have an equal number of men and women, if the composition of animal vendors in markets allows. Sources of Materials: Samples to be collected and screened for coronaviruses include blood, saliva and stool samples. 1 0 ml of blood will be collected from each subject. Subjects will also be asked to provide saliva and stool in sterile containers. An initial sample collection and interview will be performed by trained medical personnel from the local CDC under the provincial Public Health Bureau. Sample collection will be done once in years 2-4 of the study. Samples will be screened for coronaviruses using PCR and an ELISA at the appropriate CDC microbiology lab or at the Wuhan Institute of Virology. Samples that test positive for coronavirus or antibodies to coronavirus will be followed up after 6 months with a secondary interview designed to determine the current level of exposure to wild animals, and whether exposure at the current level was consistent between the first and subsequent interview. Repeated clinical samples will also be collected and tested for coronaviruses. In all instances, volunteers will be given a medical exam and informed of their test results. Potential risks: The potential risks to study participants resulting from study participation are minimal. The volume of blood being collected is within normal safety limits. The interview questions will be designed to assess exposure risk, and may ask personal questions, but surveys will be done in private and anonymized to protect privacy. There may be some stress to subjects who are informed that they have been exposed to an animal virus, but counseling will be available and options for medical care will be included in the discussion. 2. Adequacy of protection against risks Recruitment and informed consent: Prospective study participants will be identified by the research team at each site in partnership with CDC personnel. The team will be thoroughly trained on communicating the research objectives and will be able to address any questions that potential subjects may have. Both written and oral descriptions of the study will be provided in Chinese (in Mandarin or via an interpreter in local dialect if necessary) as part of the informed consent process. Contact details of the collaborators at local CDCs and the study Pl will be provided to all subjects, and CDC personnel on the research team will be available on site to answer questions from the study subjects. Test results will be communicated to each subject and counseling offered to minimize stress. 3. Potential benefits to Subjects and Others There are potential benefits to the study subjects including receiving a physical exam/health check from a medical officer and the potential benefit of identifying an occupational health hazard. At the conclusion of the study, we will deliver an educational workshop for high risk individuals (open to study subjects and non-study subjects) describing the health benefits of using PPE and hand-washing during animal handling activities throughout the day. 4. The importance of knowledge to be gained. There are valuable potential benefits to the general public from the knowledge to be gained by this study, as it may identify sources of zoonotic coronaviruses in the market system or which are commonly hunted. Avoidance of these animals or extra care when handling them may substantially reduce the risk of CoV (and other zoonotic pathogen) transmission. Protection of Human Subjects Page 120

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter INCLUSION OF WOMEN AND MINORITIES: Inclusion of Women: This proposal will enroll men and women as study subjects. Depending on local gender composition of animal vendors, we will make every effort to have men and women equally represented in this study. Inclusion of Minorities: Subjects will be enrolled in this study without regard to ethnicity. Occupational exposure to wildlife in a market, hunting, or butchering context will be the primary criteria for identifying subjects. Women &Minorities Page 121

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Program Director/Principal Investigator (Last, First, Middle): Daszak, Peter Targeted/Planned Enrollment Table This report format should NOT be used for data collection from study participants. Study Title: The ecology of bat coronaviruses and the risk of future coronavirus emergence. Total Planned Enrollment: 2460** TARGETED/PLANNED ENROLLMENT: Number of Subjects Ethnic Category Females Hispanic or Latino ,0 Not Hispanic or Latino 1,230 Ethnic Category: Total of All Subjects* 1,230 Racial Categories American Indian/Alaska Native 0 Asian 1,230 Native Hawaiian or Other Pacific Islander 0 Black or African American 0 White 0 Racial Categories: Total of All Subjects* 1,230 • The "Ethnic Category: Total of All Subjects" must be equal to the "Racial Categories: Total of All Subjects.'' (** all study subjects will be enrolled at foreign sites in China) Males 0 1,230 1,230 0 1,230 0 0 0 1,230 Total 0 2,460 2,460 0 2,460 0 0 0 2,460 PHS 398/2590 (Rev. 06/09) Page_ Targeted/Planned Enrollment Table Format Page Planned Enrollment Table Page 122

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter INCLUSION OF CHILDREN: Inclusion of Children: Children will not be included in this study. Children do not normally work in wildlife markets, and are not normally involved in the wildlife trade in China. Children Page 123

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter VERTEBRATE ANIMALS: 1. Detailed description of animal use. All work with vertebrate animals will be conducted in China. Capture and sampling techniques for all wild animals described in this study have been previously approved by UC Davis IACUC (Mazet and Epstein; UC Davis 15898; current). Experimental work using humanized mice will be conducted at the Center for Animal Experiment Biosafety 3 lab of Wuhan University at the School of Medicine in Wuhan, China. The Center is AMLAC accredited and has both an Institutional Biosafety Committee and an Institutional Animal Care and Use Committee. Animals will be housed in a BSL-3 facility and will be under the care of a full-time veterinarian. We will submit our protocols for IACUC approval should this proposal be funded. Conditions for animal use are described below. Note: The majority of wild animals captured and sampled will be done using non-destructive, techniques. In a small number of instances (~ 2 bats per species), where intestine and lung tissue is required to establish cell lines, animals will be humanely euthanized and a necropsy performed according to accepted protocols (see euthanasia section) Bat capture. Free-ranging bats will be captured using either a mist net or harp trap. The net system is manned by two people during the entire capture period, and bats are removed from the net as soon as they become entangled to minimize stress and prevent injury. In the Co-Pl's (Dr. Epstein) experience, a maximum of 20-30 bats can be safely held and processed by a team of three people per trapping period. Duration of trapping will depend on the capture rate. Bats are placed into a pillowcase or small cloth bag and hung from a branch or post until samples are collected. Bats are held for a maximum of six hours. Wild rodent capture. Free-ranging rodents will be captured through pit traps and box traps; captive rodents, including resident free-ranging wild rats/rodents in markets, will be manually captured or captured through traps. Traps will be checked a minimum of once daily in the morning. If adverse weather (extreme heat, rain) is expected or researchers are working in areas where predation is common, traps will be checked more frequently, and closed during the adverse weather. Handling of rodents will involve morphometric measurements. Captive and wild rodent sampling procedures (including anesthesia if necessary), will involve manual restraint, venipuncture, mucosal swabs, fecal, urine, and external parasite collection. Following capture, small animals will be restrained with a fine mesh bag to minimize entanglement, taking precautions to ensure the animals are not traumatized by the hoop of the net or through net removal. Larger rodents will be restrained for sampling in specialized squeeze-cages, allowing adjustments appropriate to the size of the animal. Squeeze-cages consist of a wooden frame with a plasticized wire bottom and a Plexiglas shield used to press the animal, while ensuring visible communication between the field veterinarian and the animal. Once squeezed, a rod is inserted to keep the plastic shield in place. The box is then inverted, allowing sampling to be conducted through the open wire bottom and abdomen of the animal when the animal is safely immobilized. Anesthesia for small rodents will be conducted using plastic tubes, with the animals transferred directly from the traps to the tubes containing a cotton swab soaked in ether, isoflurane, or methoxyflurane for anesthetic induction. For larger rodents, chemical restraint and anesthesia (ketamine alone, or ketamine combined with xylazine) will be applied either through the squeeze cages by syringe if applicable. Laboratory mice. Lab mice will be sourced commercially by the Wuhan Center for Animal Experiment at Wuhan University. Sample Collection. Bats will be manually restrained during sampling. Bats: Depending on the species and size of bat, swabs will be taken from the oropharynx, urogenital tract, and rectum. Fresh feces will be collected if available, in which case a rectal swab will not be collected. Blood will be collected from fruit bats either from the cephalic vein or from the radial artery or vein using a 25 gauge needle and 1 cc syringe. Blood will be collected from bats weighing less than 1 00g according to published techniques ( 126). Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Vertebrate Animals Page 124

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Rodents: Rodents will be anesthetized prior to sampling. Once anesthetized a small blood sample will be collected using a capillary tube placed into the retro-orbital sinus. Only trained technicians will perform retro-orbital bleeding and it will only be performed on anesthetized rodents. Femoral or jugular venipuncture may be used for larger rodents (e.g. rats). In all rodents, blood volumes of no more than 1 % of body weight will be withdrawn. (example 0.2 ml blood from a 20 gram rodent). Civets and other small mammals: Anesthesia will be used to restrain small free ranging mammals according to published protocols. Animals will be monitored continuously while recovering from anesthesia. Animals that are sampled in the marketplace, and that may potentially be consumed, will not be anesthetized. Manual restraint will be used and blood will be drawn from the femoral artery or saphenous vein. Laboratory Mice. Humanized mice will be bred at the University of Wuhan. Mice will be inoculated with a specific dose (e.g. 1x106 TCID50) of virus through different routes (intranasally and intraperitoneally). Mouse body temperature will be monitored with implanted temperature sensing microchips (LifeChip Bio-thermo, Destron Fearing), and mice will be weighed daily. Animals will be observed daily for clinical signs of illness. Moribund mice will be euthanized, according to AVMA recommendations. Live animals will be euthanized at three weeks post-inoculation and organs harvested. We will collect sera on days 10, 15 and 21 to test for neutralizing antibodies against bat CoVs. We will collect nasal washes, oral swabs, and rectal swabs, and urine every two days. These are minimally invasive procedures, and will be performed by experienced lab technicians under the supervision of a full-time veterinarian. 2. Justify use of animals, choice of species, numbers to be used. Species and number used in study: The purpose of this study is to conduct multi-regional surveillance in large populations of animals to detect coronaviruses that may pose a risk to the health of both humans and animals. The experimental work is designed to understand the ability of bat coronaviruses to bind to human receptors. Because we don't have prevalence estimates for novel strains of coronaviruses, we assume a conservative estimate of 10% prevalence. SARS-like coronaviruses have been found in between 10% and 38% of bats studied ( 4, 25). A 10% in wild populations of bats would require a sample of 30 individuals per species in order to ensure detection of an infected individual with 95% confidence. Wild bats: We will sample 30 individuals from 30 different species in each province in China (2 per species euthanized for organ tissue); representing but not limited to the following families: Rhinolophidae, Hipposideridae, Vespertillionidae, Mollossidae, and Pteropodidae, all of which are present in Southern China and potentially in the wildlife markets. Bats in wet markets: We will opportunistically sample a wide variety of insectivorous and frugivorous bats according to what is present in markets. In addition to bats, we will sample civets, raccoon dogs, rats, bandicoots, bamboo rats, and other rodents present in the markets that may act as intermediate hosts. Numbers of animals sampled from markets will be limited to animal availability. In every situation, sampling of wildlife will be conducted in the most humane manner while minimizing the impacts on individual animals and their wild populations. In cases where feces are collected for testing, non-invasive techniques will be used. In all instances, the fewest number of animals will be sampled that will provide valid information and statistical inference for the pathogen and disease of interest and every effort will be made to minimize stress and discomfort for the animal. A small number of bats (maximum 2 per species) representing each of the species in this study may be euthanized in order to collect lung and intestinal tissue required for characterizing coronavirus receptors. Voucher specimens may also be collected at the discretion of the team leader for the accurate identification of species using molecular methodology. Humanized mice for experimental infection for Specific Aim 3: In order to understand whether bat coronaviruses that utilize receptors found in people have the potential to infect people, we will use Swiss albino mice (standard breed at Wuhan University) that have been genetically modified to have human receptors. We'll infect them with cultured bat coronaviruses and determine which organs become infected and whether these mice are capable of shedding infectious virus. Humanized mice will be genetically modified to carry human ACE2 or DPP4 gene will be used to evaluate pathogenesis of CoVs. We cannot anticipate exactly how many viruses we will find that are candidates for experimental models, however we estimate that we will use Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Vertebrate Animals Page 125

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter four adult mice (2 male, 2 female) per virus and that we will identify approximately 20 viruses that will be used for mouse infection experiments. This will require a total of 80 mice over the study period. 3. Provide information on veterinary care. For wild caught animals, there is no specific veterinary care that is appropriate, nor will clinical veterinary facilities be available. Animals that are injured during the capture or sampling process will be assessed by an experienced team leader, and if the animal is determined to be unlikely to survive if released, it shall be euthanized humanely (see euthanasia section). Animals will be released within hours of capture. In the markets, animals will be sampled using manual restraint or anesthesia. Animals will be returned to vendors after sampling, or, if wild caught in the markets (e.g. rodents), they will be released in the area outside the marketplace. Laboratory mice will be housed in the BSL-3 small animal facility Center for Animal Experiment at Wuhan University. Experimental animals will be regularly monitored by experienced staff and a supervising veterinarian. The animal facility operates 24 hours a day and has full-time veterinarians on staff. All animals will be provided with food and water ad libitum and will otherwise receive standard care. 4. Procedures for ensuring animal comfort, lack of distress, pain, or injury: Animals will not be held longer than 6 hours. Co-Pis, Ors. Epstein and Olival have extensive experience in capture, anesthesia, and sampling wildlife, including bats. In our experience, bats and rodents tolerate the described procedure well. Mist nets will be attended continuously during capture periods, and bats will be extracted from the net as soon as they become entangled. This will minimize stress and injury from entanglement. Bats will be placed individually in cotton bags and hung from tree branches while awaiting processing and during recovery. The bags are sufficiently porous as to allow for ventilation and are designed for bat capture. The enclosed environment seems to calm the bats, as they do not struggle once inside, but they hang quietly. Animals will be monitored by a veterinarian or experienced field team member during all stages of capture, processing, and release. Animals will be kept in a cool place while in the pillowcases. Rodent traps will be set overnight and all traps will be checked in the morning while it still cool outside. Rodents will be kept in a cool, shaded environment during sampling and will be released within 10 hours of capture. The procedures used in this experiment (blood draw, nasal, oral, and rectal swabs) are minimally invasive, however, mice that show signs of morbidity post-infection will be examined and euthanized according to AVMA standards (see below). 5. Euthanasia: In the event of injury to an animal that results in pain and suffering, and reasonable veterinary care is unavailable, the animal will be euthanized by a veterinarian or trained field team member using ketamine injected intramuscularly 37 .5mg/kg and sodium pentobarbital injected intravenously at a dose of 1.0ml per 5kg injected intravenously. This protocol is in accordance with the AVMA euthanasia report (2007). Any animal that is euthanized using a chemical agent will be disposed such that it will not be permitted to enter the food supply either through markets or hunting. Vertebrate Animals Page 126

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter SELECT AGENT RESEARCH/BIOHAZARDS. No select agent research as of 5/25/12 SARS-CoV caused outbreaks with significant case fatality rates, and there are no vaccines available for this agent. SARS-CoV is classified as a BSL-3 agent. The work proposed in this application will involve two aspects: field work and laboratory work. Fieldwork involves the highest risk of exposure to SARS or other Co Vs, while working in caves with high bat density overhead and the potential for fecal dust to be inhaled. There is also some risk of exposure to pathogens or physical injury while handling bats, civets, rodents or other animals, their blood samples or their excreta. The Co-Pl is a veterinarian with extensive experience working with wildlife species and high-biosecurity pathogens (Nipah virus, ebolavirus, SARS), and great care will be taken in the field to limit the risk of accidental exposure to known or unknown animal pathogens. We have strict procedures for handling bats and working with samples from them as they are secured in the field and transported to the lab. Field team members handling animals will be trained to utilize personal protective equipment and practice proper environmental disinfection techniques. This includes wearing coveralls or dedicated clothing, nitrile gloves, eye protection, and a P95 or P100 respirator. All field clothing and equipment will be disinfected using Virkon disinfectant. All biological waste from field surveys will be disposed of in the appropriate container (sharps box or an autoclave bag) and will be autoclaved at local hospitals or university labs. All personnel will be vaccinated against rabies and have a neutralizing antibody titer, in accordance with WHO and CDC recommendations. Field teams will carry rabies boosters in the field and will receive a booster in the event of a potential rabies exposure. Field safety protocol: Our procedures to deal with bites, needle-sticks etc. are as follows: The wound is washed thoroughly with soap and water to clean away dirt and debris, then vigorously scrubbed with a sterile gauze bandage and benzalkonium chloride for 5 minutes. If bleeding, pressure is applied with a sterile bandage for until bleeding has stopped. If the wound continues to bleed, medical attention at the nearest hospital is sought. The bat from which the bite or exposure originated is identified, and the samples collected from it labeled on the data sheet that these were involved in an exposure. Our procedures require that the person potentially exposed reports to a major hospital within 24 hours to have wound examined and receive a rabies booster (as per WHO/CDC protocols). The laboratory work is lower risk, as samples placed in lysis buffer will be non-infectious. Samples placed in viral transport medium and frozen will be stored at ultra-low temperatures (-86C) until viral isolation is required. Serum will be heat inactivated (56C for 30 min) prior to testing. Lab biosafety: Wuhan Institute of Virology and the Wuhan University Center for Animal Experiment BSL-3 lab have an Internal Biosafety Committee and are accredited BSL-2 and BSL 3 laboratories. All experimental work using infectious material will be conducted under appropriate biosafety standards. Disposal of hazardous materials will be conducted according to the institutional biosafety regulations. Select Agent Research Page 127

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Bibliography & References Cited 1. L. H. Taylor, S. M. Latham, M. E. J. Woolhouse, Risk factors for human disease emergence. Philosophical Transactions of The Royal Society 8-Biological Sciences 356, 983 (2001 ). 2. Y. Guan, B. J. Zheng, Y. Q. He, X. L. Liu, Z. X. Zhuang, C. L. Cheung, S. W. Luo, P.H. Li, L. J. Zhang, Y. J. Guan, K. M. Butt, K. L. Wong, K. W. Chan, W. Lim, K. F. Shortridge, K. Y. Yuen, J. S. M. Peiris, L. L. M. Poon, Isolation and characterization of viruses related to the SARS coronavirus from animals in Southern China. Science 302, 276 (2003). 3. W. Li, Z. Shi, M. Yu, W. Ren, C. Smith, J. H. Epstein, H. Wang, G. Crameri, Z. Hu, H. Zhang, J. Zhang, J. McEachern, H. Field, P. Daszak, B. T. Eaton, S. Zhang, L.-F. Wang, Bats are natural reservoirs of SARS-like coronaviruses. Science 310, 676 (2005 Oct 28 (Epub 2005 Sep, 2005). 4. W. D. Li, Z. L. Shi, M. Yu, W. Z. Ren, C. Smith, J. H. Epstein, H. Z. Wang, G. Crameri, Z. H. Hu, H.J. Zhang, J. H. Zhang, J. McEachern, H. Field, P. Daszak, B. T. Eaton, S. Y. Zhang, L. F. Wang, Bats are natural reservoirs of SARS-like coronaviruses. Science 310, 676 (Oct, 2005). 5. J. F. Drexler, V. M. Corman, T. Wegner, A. F. Tateno, R. M. Zerbinati, F. Glaza-Rausch, A. Seebens, M.A. Muller, C. Drosten, Amplification of Emerging Viruses in a Bat Colony. Emerging Infectious Diseases 17, 449 (Mar, 2011 ). 6. J. Huynh, S. Li, B. Yount, A. Smith, L. Sturges, J. C. Olsen, J. Nagel, J. B. Johnson, S. Agnihothram, J. E. Gates, M. B. Frieman, R. S. Barie, E. F. Donaldson, Evidence Supporting a Zoonotic Origin of Human Coronavirus Strain NL63. Journal of Virology 86, 12816 (Dec, 2012). 7. S. K. P. Lau, K. S. M. Li, Y. Huang, C. T. Shek, H. Tse, M. Wang, G. K. Y. Choi, H. F. Xu, C. S. F. Lam, R. T. Guo, K. H. Chan, B. J. Zheng, P. C. Y. Woo, K. Y. Yuen, Ecoepidemiology and Complete Genome Comparison of Different Strains of Severe Acute Respiratory Syndrome-Related Rhinolophus Bat Coronavirus in China Reveal Bats as a Reservoir for Acute, Self-Limiting Infection That Allows Recombination Events. Journal of Virology 84, 2808 (Mar, 2010). 8. P. L. Quan, C. Firth, C. Street, J. A. Henriquez, A. Petrosov, A. Tashmukhamedova, S. K. Hutchison, M. Egholm, M. 0. V. Osinubi, M. Niezgoda, A. B. Ogunkoya, T. Briese, C. E. Rupprecht, W. I. Lipkin, Identification of a Severe Acute Respiratory Syndrome Coronavirus-Like Virus in a Leaf-Nosed Bat in Nigeria. Mbio 1, (Sep-Oct, 2010). 9. S. Tong, C. Conrardy, S. Ruane, I. V. Kuzmin, X. Guo, Y. Tao, M. Niezgoda, L. Haynes, B. Agwanda, R. F. Breiman, L. J. Anderson, C. E. Rupprecht, Detection of novel SARS-like and other coronaviruses in bats from Kenya. Emerg Infect Dis 15, 482 (Mar, 2009). 10. M. Tahir, R. Gajraj, M. Bardhan, H. Mohammed, L. Dyke, P. Charlemagne, R. Alves, D. Kirrage, D. Killalea, K. James, M. Kemp, H. Duggal, R. Carr, M. Afza, N. Aigbogun, B. Sibal, R. Harrell, 0. Edeghere, K. Neal, S. Ibbotson, N. Wickramasinghe, N. Sherwood, B. Oppenheim, L. Hopton, H. Osman, E. Smit, S. Atabani, J. Workman, S. Wilson, C. Overton-Lewis, M. Logan, R. Mccann, M. Petrovic, V. Bothra, W. Welfare, B. lsalska, J. Barker, A. Ashworth, I. Fedor, C. Seng, D. Kumar, B. McCloskey, J. Nguyen-Van-Tam, P. Cosford, A. Bermingham, J. Ellis, M. Galiano, A. Lackenby, R. Myers, R. Gopal, M. Zambon, R. Pebody, L. Thomas, N. Boddington, H.K. Green, H. Zhao, I. Kennedy, I. Abubakar, J. Jones, N. Phin, M. Catchpole, J.M. Watson, H.P. A. U. K. N. Hlth Protection Agcy, Evidence of person-to-person transmission within a family cluster of novel coronavirus infections, United Kingdom, February 2013. Eurosurveillance 18, 4 (Mar, 2013). 11. A. Annan, H.J. Baldwin, V. M. Corman, S. M. Klose, M. Owusu, E. E. Nkrumah, E. K. Badu, P. Anti, 0. Agbenyega, B. Meyer, S. Oppong, Y. A. Sarkodie, E. K. V. Kalka, P.H. C. Lina, E. V. Godlevska, C. Reusken, A. Seebens, F. Glaza-Rausch, P. Vallo, M. Tschapka, C. Drosten, J. F. Drexler, Human Betacoronavirus 2c EMC/2012-related Viruses in Bats, Ghana and Europe. Emerging infectious diseases 19,456 (2013-Mar, 2013). 12. S. Wacharapluesadee, C. Sintunawa, T. Kaewpom, K. Khongnomnan, K. J. Olival, J. H. Epstein, A. Rodpan, P. Sangsri, N. lntarut, A. Chindamporn, K. Suksawa, T. Hemachudha, Identification of Group C Betacoronavirus from Bat guano fertilizer, Thailand. Emerging Infectious Diseases [Internet], (2013). 13. S. Anthony, R. Ojeda-Flores, 0. Rico-Chavez, I. Navarrete-Macias, C. Zambrana-Torrelio, M. K. Rostal, J. H. Epstein, T. Tipps, E. Liang, M. Sanchez-Leon, J. Sotomayor-Bonilla, A. A. Aguirre, R. Avila, R. A. Medellfn, T. Goldstein, G. Suzan, P. Daszak, W. I. Lipkin, Coronaviruses in bats from Mexico. Journal of General Virology 94, (2013). References Cited Page 128

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter 89. 90. 91. 92. 93. 94. 95. 96. 97. 98. 99. 100. 101. 102. 103. 104. 105. 106. 107. 108. 109. 110. 111. 112. 113. J. P. Huelsenbeck, F. Ronquist, MrBayes: Bayesian inferences of phylogeny. Bioinformatics 17, 754 (2001 ). J. P. Meier-Kolthoff, A. F. Auch, D. H. Huson, M. Goker, Copycat: cophylogenetic analysis tool. Bioinformatics 23, 898 (2007). A. Stamatakis, A. F. Auch, J. Meier-Kolthoff, M. Goker, AxPcoords & parallel AxParafit: statistical CDphylogenetic analyses on thousands of taxa. Bmc Bioinformatics 8, (Oct. 2007). M.A. Charleston, R. D. M. Page. (2002). D. P. Martin. P. Lemey, M. Lott, V. Moulton, D. Posada, P. Lefeuvre, RDP3: a flexible and fast computer program for analyzing recombination. Bioinformatics 26, 2462 (October 1, 2010, 2010). A. Demogines, M. Farzan, S. L. Sawyer, Evidence for ACE2-Utilizing Coronaviruses (CoVs) Related to Severe Acute Respiratory Syndrome CoV in Bats. Journal of Virology 86, 6350 (Jun, 2012). J. Diamond, M. Gilpin, Examination of the "null'' model of connor and simberloff for species cooccurrences on Islands. Oecologia 52, 64 (1982101101, 1982). E. F. Connor, D. Simberloff, Species Number and Compositional Similarity of the Galapagos Flora and Avifauna. Ecological Monographs 48, 219 (1978). D. M. Raup, R. E. Crick, Measurement of Fauna! Similarity in Paleontology. Journal of Paleontology 53, 1213 (1979). A. E. Magurran, Measuring biological diversity. (Blackwell Publishing, Malden, MA, 2004). N. J. Gotelli, NULL MODEL ANALYSIS OF SPECIES CO-OCCURRENCE PATTERNS. Ecology 81, 2606 (2000/09/01, 2000). R. Poulin, D. Mouillot, Parasite specialization from a phylogenetic perspective: a new index of host specificity. Parasitology 126, 473 (May, 2003). R. Poulin, Decay of similarity with host phylogenetic distance in parasite faunas. Parasitology 137, 733 (Apr, 2010). 0. Diekmann, J. A. Heesterbeek, J. A. Metz, On the definition and the computation of the basic reproduction ratio RO in models for infectious diseases in heterogeneous populations. J Math Biol 28, 365 (Jan 1, 1990). A. Dobson, J. Foufopoulos, Emerging infectious pathogens of wildlife. Philosophical Transactions of the Royal Society of London Series 8-Biological Sciences 356, 1001 (Jul 29, 2001 ). C. Fraser, S. Riley, R. Anderson, N. Ferguson, Factors that make an infectious disease outbreak controllable. P Natl Acad Sci Usa 101, 6146 (Jan 1, 2004). G. Chowen, C. Castillo-Chavez, P. Fenimore, C. Kribs-Zaleta, L. Arriola, J. Hyman, Model parameters and outbreak control for SARS. Emerg Infect Dis 10, 1258 (Jan 1, 2004). G. Chowen, P. Fenimore, M. Castillo-Garsow, C. Castillo-Chavez, SARS outbreaks in Ontario, Hong Kong and Singapore: the role of diagnosis and isolation as a control mechanism. Journal of Theoretical Biology 224, 1 (Jan 1, 2003). M. Lipsitch, T. Cohen, B. Cooper, J.M. Robins, S. Ma, L. James, G. Gopalakrishna, S. K. Chew, C. C. Tan, M. H. Samore, D. Fisman, M. Murray, Transmission dynamics and control of severe acute respiratory syndrome. Science 300, 1966 (Jun 20, 2003). N. Nagata, N. Iwata-Yoshikawa, F. Taguchi, Studies of severe acute respiratory syndrome coronavirus pathology in human cases and animal models. Vet Pathol 47, 881 (Sep, 2010). B. E. Martina, B. L. Haagmans, T. Kuiken, R. A. Fouchier, G. F. Rimmelzwaan, G. Van Amerongen, J. S. Peiris, W. Lim, A. D. Osterhaus, Virology: SARS virus infection of cats and ferrets. Nature 425, 915 (Oct 30, 2003). P. Daszak, Plowright R, Epstein JH, Pulliam J, Abdul Rahman S, Field HE, Smith CS, Olival KJ, Luby S, Halpin K, Hyatt AD, & (HERG), in Disease Ecology: Community structure and pathogen dynamics., R. S. Collinge S Ed. (Oxford University Press Oxford 2006) ... e,._. 1_8_6_-_2_0_1. _________ ,...__,.., (b) (4) R. Fogarty, K. Halpin, A. D. Hyatt, P. Daszak, 8. A. Mungall, Henipavirus susceptibility to environmental variables. Virus Research 132, 140 (Mar, 2008). K. Halpin, A. D. Hyatt, R. Fogarty, D. Middleton, J. Bingham, J. H. Epstein, S. A. Rahman, T. Hughes, C. Smith, H. E. Field, P. Daszak, HERG, Pteropodid bats are confirmed as the reservoir hosts of References Cited Page 133

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Principal Investigator/Program Director (Last, first, middle}: Daszak, Peter 114. 115. 116. 117. 118. 119. 120. 121. 122. 123. 124. 125. 126. 127. henipaviruses: A comprehensive experimental study of virus transmission. American Journal of Tropical Hygiene and Medicine, (2011 ). R. K. Plowright, P. Foley, H. E. Field, A. P. Dobson, J.E. Foley, P. Eby, P. Daszak, Urban habituation, ecological connectivity and epidemic dampening: The emergence of Hendra virus from flying foxes (Pteropus species). Proceedings of the Royal Society 8-Biological Sciences 278, 3703 (2011 ). P. Hosseini, S. H. Sokolow, K. J. Vandegrift, A. M. Kilpatrick, P. Daszak, Predictive Power of Air Travel and Socio-Economic Data for Early Pandemic Spread. PLoS One 5, (Sep, 2010). P. R. Hosseini, P. Daszak, paper presented at the Eight Annual Scientific Conference of Chittagong Veterinary and Animal Sciences University: Networking for Promoting Change Towards One World One Health Chittaa~>naJ B~lade_.,s__,h0 ,,_ ...,2.,_0....,1_0_..._. _____________________ ...,._..,.,. (b) (4) A. M. Kilpatrick, L. D. Kramer, S. R. Campbell, E. 0. Alleyne, A. P. Dobson, P. Daszak, West Nile virus risk assessment and the bridge vector paradigm. Emerging Infectious Diseases 11, 425 (Mar, 2005). A. M. Kilpatrick, P. Daszak, M. J. Jones, P. P. Marra, L. D. Kramer, Host heterogeneity dominates West Nile virus transmission. Proceedings of the Royal Society 8-Biological Sciences 273, 2327 (Sep, 2006). A. M. Kilpatrick, L. D. Kramer, M. J. Jones, P. P. Marra, P. Daszak, West Nile virus epidemics in North America are driven by shifts in mosquito feeding behavior. PLoS. Biol. 4, 606 (Apr, 2006). W. H. Li, C. S. Zhang, J. H. Sui, J. H. Kuhn, M. J. Moore, S. W. Luo, S. K. Wong, I. C. Huang, K. M. Xu, N. Vasilieva, A. Murakami, Y. Q. He, W. A. Marasco, Y. Guan, H. Y. Choe, M. Farzan, Receptor and viral determinants of SARS-coronavirus adaptation to human ACE2. Embo Journal 24, 1634 (Apr 20, 2005. (b) (4) S. M. Poutanen, D. E. Low, B. Henry, S. Finkelstein, D. Rose, K. Green, R. Tellier, R. Draker, D. Adachi, M. Ayers, A. K. Chan, D. M. Skowronski, I. Salit, A. E. Simor, A. S. Slutsky, P. W. Doyle, M. Krajden, M. Petric, R. C. Brunham, A. J. McGeer, N. M. L. Canada, C. S. A. Respiratory, Identification of severe acute respiratory syndrome in Canada. New England Journal of Medicine 348, 1995 (May 15, 2003). L. J. Wu, P. Zhou, X. Y. Ge, L. F. Wang, M. L. Baker, Z. L. Shi, Deep RNA Sequencing Reveals Complex Transcriptional Landscape of a Bat Adenovirus. Journal of Virology 87, 503 (Jan, 2013). Y. Li, X. Y. Ge, H.J. Zhang, P. Zhou, Y. Zhu, Y. Z. Zhang, J. F. Yuan, L. F. Wang, Z. L. Shi, Host Range, Prevalence, and Genetic Diversity of Adenoviruses in Bats. Journal of Virology 84, 3889 (Apr, 2010). Y. Li, X. Ge, H. Zhang, P. Zhou, Y. Zhu, Y. Zhang, J. Yuan, L. F. Wang, Z. Shi, Host range, prevalence, and genetic diversity of adenoviruses in bats. J Virol 84, 3889 (Apr, 2010). C. S. Smith, C. E. de Jong, H. E. Field, Sampling small quantities of blood from microbats. Acta Chiropterologica 12, 255 (2010). References Cited Page 134

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter CONSORTIUMICONTRACTUAL ARRANGEMENTS: Consortium!Contractual Arrangements This project is a multi-institutional collaboration led by EcoHealth Alliance, New York (Oaszak, Pl), which will subcontract funds to two institutions: the East China Normal Univerisity (Dr S. Zhang) and the Wuhan Institute of Virology (Dr. Z. Shi), which are both foreign institutions. Dr. Daszak has over 15 years previous experience managing collaborative projects including two R01 son Nipah virus ecology that involved 5 separate foreign institutions, a 5-year NSF/NIH Ecology of Infectious Disease award on West Nile virus which involved multiple subcontractees, an R01 on bat viral discovery that involves multiple international contracts, and a multi-million dollar p.a. contract from USAID that involves 12 international partners. The applicant organization (EcoHealth Alliance) is justified in taking the lead on this project because it specializes in understanding the ecological, and virological processes underlying zoonotic disease emergence. Dr Daszak has conducted significant preliminary work on this issue including 10-years of research on the ecological and related factors of the emergence of SARS and 11-years of work in China. The subcontractees will work on specific issues and areas in which they have proven expertise. These areas are: human and animal field sampling (East China Normal University, Dr. Zhang) and viral discovery, pathogenesis as well as sample storage and shipping (Wuhan Institute of Virology, Dr. Shi). Dr Daszak has launched and co-directed a joint institute in China with Dr Zhang, and has been involved in contractual arrangements with ECNU for 8 years. Ors Shi, Zhang, and Daszak have collaborated together since 2002 and have been involved in running joint conferences, and shipping samples into and out of China. Consortium/Contractual Page 135

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter ,,.), ..t ~ 1P ~ ~ -ffi ~ #: J <f> ,(!)• ~-'J i8 Shanghai Municipal Center for Disease Control and Prevention ~---~------~------------------------ M!.J.il:.: 'f IEJ:.~rj, 'f J.iiffJ3t 1380 -It, 200336 Add:1380 Zhongshan Rd.(w) Shanghai, 200336 P.R.China te.it-(Tel): +86-21-62758710 #-Ji.(Fax): +86-21-62756323 J#Uil:.(Website): http://www.scdc.sh.cn Dr. Peter Daszak President EcoHealth Alliance 460 W 34th St. 17th Floor New York, NY 10001 USA Dear Dr. Daszak, I am writing in response to a request for collaboration on an upcoming NIAID funded ROl entitled "Understanding the risk of bat coronavirus emergence." The Shanghai CDC has a high interest in working with EcoHealth Alliance and its scientists in identifying and preventing the transmission of bat coronaviruses to human populations. The Shanghai CDC recognizes the mutual benefits to be gained through research cooperation and a successful partnership with EcoHealth Alliance in the field of identification and prevention of zoonotic disease transmission. It is vital to not only identify the diseases themselves, but also identify high-risk human populations and the actions that put them at risk for infection along with evaluating approaches to intervention and disease management. Understanding and preventing exposure and transmission of zoonotic diseases from wildlife to humans remains a high priority for prevention of pandemics. In our discussion with EcoHealth Alliance, we have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to the outbreak of epidemic diseases, particularly those of animal origin. To assist in this study, we will provide participating laboratories in China with human epidemiological information, both new and archived, to support research in bat coronaviruses. We at the Shanghai CDC look forward to our collaboration with the EcoHealth Alliance team and working further on this worthwhile study. Sincerely ~ /4-JL Fan Wu, M.D. Director General Shanghai Municipal Center for Disease Control and Prevention Letters of Support Page 136

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter WUHAN INSTITUTE OF VIROLOGY The CHINESE ACADEMY OF SCIENCES Address: Xiaohongshan 44, Wuchang, Wuhan 430071, Hubei, P.R. Chfna Tel: -+86-27-87198117 Fax: +86-27-87198072 http://www.whiov.ac;.cn May 23, 2013 To whom it may concern: On behalf our Institute, I am very pleased to express my strong support for Dr. Zhengli Shi for applying for the R01 entitled "Understanding the Risk of Bat Coronavirus Emergence• under the project managed by Peter Daszak, president of EcoHealth Alliance. Dr. Shi has extensive expertise in viral pathogen discovery. Since 2004, Dr. Shi's laboratory has discovered a variety of genetically diverse bat viruses including bat SARS-like coronavirus, bat adenovirus, and adeno-associated viruses. She has established a worldwide collaborative-group of leading experts on viral pathogens and ecology covering identification of emerging viruses, epidemiology on bat-borne viruses including Hendra and Nipah virus and SARScoronavirus. Her work with Dr. Peter Daszak led to the discovery of bat SARS-fike coronavirus in 2005. Our Institute would provide all necessary support to Dr. Shi for accomplish the project if it is approved. C Xia Wuh Letters of Support Virology Page 137

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter UNGt SCHOOL OF PUBLIC HEALTH DEPARTMENT OF EPIDEMIOLOGY Dr. Peter Daszak President Eco Health Alliance 460 W 34"' St. 17"' Floor NewYork, NY 10001 USA Dear Dr. Daszak, 5/31/2013 I am writing in response to a request for collaboration on an upcoming NIAID R01 grant entitled "Understanding the risk of bat coronavirus emergence." I agree that studies are definitely needed to identify the key risk factors and develop strategies that prevent the transmission of bat coronaviruses to human populations. Understanding and preventing exposure and transmission of zoonotic diseases from wildlife to humans remains a high priority for prevention of pandemics. Our laboratory has developed a variety of animal models for understanding human coronavirus pathogenesis in vivo. We have developed transgenic mouse models in the C57BL/6 mice, expressing hACE2 in ciliated cells from the FOXJ1 promoter. Unlike other epithelial cell promoters (e.g., K18, hACE2 expression from FOXJ1 should be specific to the airway epithelium. FOXJ1 (hepatocyte nuclear factor-3/forkhead homologue 4; HFH-4) is a member of the forkhead/wnged helix family of transcription factors whose expression is tightly restricted to cells possessing motile cilia or flagella. Inoculation of these mice with wild type SARS-CoV resulted in lethal respiratory tract infections characterized by high virus titers (>108 PFU/day 4), hemorrhage, severe pneumonia and acute respiratory distress syndrome between days 2-7 post infection (Fig 1). We also have aged and immunosenescent models that are highly vulnerable to synthetically reconstructed strains of SARS-CoV from early in the epidemic. This letter states my Vvillingness to collaborate with your group to evaluate the in vivo pathogenesis of interesting bat and anirrel SARS-like coronaviruses. It vvas a pleasure talking with you the other day. I believe your proposal asks fundamentally important questions in the evolution of new Letters of Support Ace2 Transgenic Weight Loss 110 60 6 Days Post Infection Figure 1. FOXJ1 ACE2 Transgenic Mice are Highly Susceptible to wildtype SARS-CoV Infection. Two different lines were infected with Wildtype SARS-virus and clinical disease evaluated at different times postinfection. human coronaviruses from bats, contributes Page 138

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter dramatically to our understanding of coronavirus variation in natural populations, and provides key insights into the ecology of new emerging infectious diseases. Let me know if I can be of any additional assistance. Sincerely, Ralph S. Barie, Professor Department of Epidemiology Department of Microbiology and Immunology Ph: 00~~-~= Email: (b) (6) -------- Letters of Support Page 139

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Wi YUNNAN INSTITUTE OF ENDEMIC DISEASES CONTROL AND PREVENTION (YIEDC) Dr. Peter Daszak President EcoHealth Alliance 460 W 34th St. 17th Floor New York, NY 10001 USA Dear Dr. Daszak, I am writing in response to a request for collaboration on an upcoming NIAID funded R01 entitled "Understanding the risk of bat corona virus emergence." The Yunnan Institute of Endemic Diseases Control and Prevention (EDC) has a high interest in working with EcoHealth Alliance and its scientists in identifying and preventing the transmission of bat corona viruses to human populations. The Yunnan EDC recognizes the mutual benefits to be gained through research cooperation and a successful partnership with Eco Health Alliance, and long term colleague ZhengLi Shi, in the field of identification and prevention ofzoonotic disease transmission. It is vital to not only identify the diseases themselves, but also identify high-risk human populations and the actions that put them at risk for infection along with evaluating approaches to intervention and disease management. Understanding and preventing exposure and transmission of zoo no tic diseases from wildlife to humans remains a high priority for prevention of pandemics. In our discussion with Eco Health Alliance, we have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to the outbreak of epidemic diseases, particularly those of animal origin. To assist in this study, we will provide participating laboratories in China with human samples, both new and archived, and support research in bat coronaviruses. We at the Yunnan EDC look forward to our collaboration with the EcoHealth Alliance team and working further on this worthwhile study. Sincerely, 7 A/4-m Zhang Yunzhir~li/ It" Yunnan Institute of Endemic Diseases Control and Prevention Tel: (b)(6) E-mail: (b)( Add. 33wenhua Rd., Dali City, Yunnan. PR.China i:1315: (Tel) :0872-2125196 /~Ii: (Fax) :0872-2125437 !ll!!!Jiijj, (PO Box) :671000 Letters of Support Page 140

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter Dr. Peter Daszak President Eco Health Alliance 460 W 34 th St. 17 1h Floor New York, NY 10001 USA Dear Dr. Daszak. I am writing in response to a request for collaboration on an upcoming NIAID funded ROl entitled" Understanding the risk of bat coronavirus emergence." The Guangdong CDC has a high interest in working with EcoHealth Alliance and its scientists in identifying and preventing the transmission of bat coronaviruses to human populations. The Guangdong CDC recognizes the mutual benefits to be gained through research cooperation and a successful partnership with EcoHealth Alliance in the field of identification and prevention ofzoonotic disease transmission. This partnership will continue a successful five year relationship between the Guangdong CDC and Eco Health Alliance. It is vital to not only identify the diseases themselves, but also identify high-risk human populations and the actions that put them at risk for infection along with evaluating approaches to intervention and disease management. Understanding and preventing exposure and transmission of zoonotic diseases from wildlife to humans remains a high priority for prevention of pandemics. In our discussion with EcoHealth Alliance, we have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to the outbreak of epidemic diseases, particularly those of animal origin. To assist in this study, we will provide participating laboratories in China with human samples, both new and archived, and support research in bat coronaviruses. We at the Guangdong CDC look forward to our collaboration with the Eco Health Alliance team and working further on this worthwhile study. 7;:_·'~~ Ke Changwen \d,I: ()1111,i:111 H .... .i. lla~lii Tto\\11. rall) 11 Jli,1ri..i. L11a11;.:Ll11111. (;11.111;.:1lu11;.:. c:l1i1111.:'i 11-t.10 1~ i,5-: ( T,·I l : 020-310:'i I CHIO (1:: !\: ( V.1, i : 0::!0-:I I 051 :'i02 Letters of Support Page 141

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter -- ,_ ff It ,E.. ,t M-"f '1 ~ * ~ ·~'1, ~~ 6]f 'ft,~ Institutes for Advanced Interdisciplinary Research , ECNU Dr. Peter Daszak President Eco Health Alliance 460 W 341h St. 17th Floor New York, NY 10001 USA Dear Dr. Daszak, 25 May 2013 As Dean of Institutes for Advanced Interdisciplinary Research, I am delighted at the prospect of our continued collaboration on the NIAID funded ROl "Understanding the Risk of Bat Coronavirus Emergence." Since 2005, our organizations have collaborated via our School of Life Science. We have a joint-MOU as well. I have enjoyed our close working relationship with EcoHealth Alliance especially on issues related to emerging infectious diseases and health. Our collaborations include past and current research projects in Guangzhou, Guangxi, Yunnan, Hainan, and Shanghai as well as capacity building, training, and over 20 joint publications including Science papers, which have led to increased understanding of the ecology of disease dynamics and garnered invaluable data towards predicting and preventing zoonotic disease emergence. My field and laboratory teams based in Beijing, Shanghai, Guangxi, and Guangzhou are ideally positioned to conduct both research and surveillance as we work towards reducing the risk of zoonosis in China. In our discussion with EcoHealth Alliance, I have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to the outbreak of epidemic diseases - particularly those of animal origin. I look forward to our continued collaboration and the results of this exciting and timely project. Sincerely, Dr. Zhang Shu-Yi Dean of Institutes for Advanced Interdisciplinary Research East China Normal University B319, Science Building, 3663, North Zhongshan Road, Shanghai 200062 China (b)(6) Letters of Support Page 142

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter RESOURCE SHARING PLAN: Data Sharin Plan: Sequence data will be made publicly available via GenBank, and shared when requested by other scientists, as soon as a publication is in press. Viral isolates will remain at the Wuhan Institute of Virology initially. Isolates, reagents and any other products, should they be developed, will be made available to other NIH-funded researchers via applicable Wuhan Institute of Virology and EcoHealth Alliance Material Transfer Agreements and/or licensing agreements. Sharin Model Or anisms: We do not anticipate the development of any model organisms from this study. Should any be developed, they will be made available to other NIH-funded researchers via applicable Wuhan Institute of Virology and EcoHealth Alliance Material Transfer Agreements and/or licensing agreements. Genome Wide Association Studies (GWAS): N/A Resource Sharing Plan Page 143

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter PHS 398 Checklist 0MB Number: 0925-0001 1. Application Type: From SF 424 (R&R) Cover Page. The responses provided on the R&R cover page are repeated here for your reference, as you answer the questions that are specific to the PHS398. • Type of Application: IZJNew D Resubmission ORenewal D Continuation D Revision Federal Identifier: I GRANTl 1418 218 I 2. Change of Investigator / Change of Institution Questions D Change of principal investigator/ program director Name of former principal investigator/ program director: Prefix: I • First Name: I Middle Name: I • Last Name: I Suffix: I D Change of Grantee Institution • Name of former institution: I I 3. Inventions and Patents (For renewal applications only) • Inventions and Patents: YesD No 0 If the answer is "Yes" then please answer the following: • Previously Reported: Yes D No 0 Checklist Page 144 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05TJ8:36:48-04:00

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Principal Investigator/Program Director (Last, first, middle): Daszak, Peter 4. * Program Income Is program income anticipated during the periods for which the grant support is requested? Oves If you checked "yes" above (indicating that program income is anticipated), then use the format below to reflect the amount and source(s). Otherwise, leave this section blank. *Budget Period *Anticipated Amount ($) •source(s) D I I D I I CJ I I D I I D I I 5. * Disclosure Permission Statement If this application does not result in an award, is the Government permitted to disclose the title of your proposed project, and the name, address, telephone number and e-mail address of the official signing for the applicant organization, to organizations that may be interested in contacting you for further information (e.g., possible collaborations, investment)? IZ]Yes Checklist Page 145 Tracking Number:GRANTI 1418584 Funding Opportunity Number:PA-11-260 Received Date:2013-06-05TJ8:36:48-04:00

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RESEARCH Department of Health and Human Services National Institutes of Health Notice of Award Federal Award Date: 06/10/2015 NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 5R01Al110964-02 FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Aleksei Chmura President 460 West 34th Street 17th Floor New York, NY 100012317 Award e-mailed to: (b)(6) ----------- Period Of Performance: Budget Period: 06/01/2015 -05/31/2016 Project Period: 06/01/2014 - 05/31/2019 Dear Business Official: The National Institutes of Health hereby awards a grant in the amount of $630,445 (see "Award Calculation" in Section I and 'Terms and Conditions" in Section 111) to ECOHEALTH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the "Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01Al110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STTR} awards. Consult the NIH website http://qrants.nih.gov/qrants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1

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Laura A. Pone Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2

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SECTION I -AWARD DATA- 5R01Al110964-02 Award Calculation (U.S. Dollars) Federal Direct Costs Federal F&A Costs Approved Budget Total Amount of Federal Funds Obligated (Federal Share) TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS $502,293 $128,152 $630,445 $630,445 $630,445 $630,445 YR THIS AWARD CUMULATIVE TOTALS 2 $630,445 $630,445 3 $611,090 $611,090 4 $597,112 $597,112 5 $581,646 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project Fiscal Information: CFDAName: CFDA Number: EIN: Document Number: PMS Account Type: Fiscal Year: IC CAN Al 8472350 Allergy, Immunology and Transplantation Research 93.855 1311726494A1 RAl110964A P (Subaccount) 2015 2015 $630,445 2016 2017 $611,090 $597,112 2018 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project NIH Administrative Data: PCC: M51C / OC: 414E / Released: (b)(6)L 06/09/2015 Award Processed: 03/23/2015 01 :36:12 PM SECTION 11- PAYMENT/HOTLINE INFORMATION- 5R01Al110964-02 For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://qrants.nih.gov/grants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS - 5R01Al110964-02 This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. c. 45 CFR Part 75. d. National Policy Requirements and all other requirements described in the NIH Grants Policy Statement, including addenda in effect as of the beginning date of the budget period. e. Federal Award Performance Goals: As required by the periodic report in the RPPR or in the final progress report when applicable. f. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. (See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain Page-3

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references cited above.) Research and Development (R&D): All awards issued by the National Institutes of Health (NIH) meet the definition of "Research and Development" at 45 CFR Part§ 75.2. As such, auditees should identify NIH awards as part of the R&D cluster on the Schedule of Expenditures of Federal Awards (SEFA). The auditor should test NIH awards for compliance as instructed in Part V, Clusters of Programs. NIH recognizes that some awards may have another classification for purposes of indirect costs. The auditor is not required to report the disconnect (i.e., the award is classified as R&D for Federal Audit Requirement purposes but non-research for indirect cost rate purposes), unless the auditee is charging indirect costs at a rate other than the rate(s) specified in the award document(s). An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the Central Contractor Registration. Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://grants.nih.gov/qrants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01Al110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://grants.nih.gov/grants/policy/awardconditions.htm for additional award applicability information. In accordance with P .L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OO-08-033 and the Public Access website: http://publicaccess.nih.gov/. Treatment of Program Income: Additional Costs SECTION IV - Al Special Terms and Conditions - 5R01Al110964-02 This Notice of Award (NoA) includes funds for consortium activity with Wuhan Institute of Virology- CHINA awarded in the Total Costs amount of $139,015 ($128,718 Direct Costs+ $10,297 F&A Costs). Future year commitments are as follows: Year 3 Total Costs: $159,122 Year4 Total Costs: $159,122 Year 5 Total Costs: $159,122 This Notice of Award (NoA) includes funds for consortium activity with East China Normal University - CHINA awarded in the Total Costs amount of $72,684 ($67,300 Direct Costs+ $5,384 F&A Costs). Future year commitments are as follows: Page-4

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Year 3 Total Costs: $54,117 Year 4 Total Costs: $42,300 Year 5 Total Costs: $32,454 Consortiums are to be established and administered as described in the NIH Grants Policy Statement (NIH GPS). The referenced section of the NIH Grants Policy Statement is available at http:1/grants. nih.gov/grants/policy/nihgps_2013/nihgps_ ch 15.htm# _ Toc271265264. The written agreement with the consortium must address the negotiated arrangements for meeting the scientific, administrative, financial and reporting requirements for this grant. No foreign performance site may be added to this project without prior approval of the National Institute of Allergy and Infectious Diseases. Although a specific amount has been awarded for each consortium, the grantee retains standard rebudgeting authorities. Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.selectagents.gov/Requlations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm). Research funded under this grant must adhere to the BMBL, including using the BMBL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. If work with a new or additional Agent(s)/Toxin(s) is proposed in the upcoming project period, provide: o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent(s), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted, with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. Page-5

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STAFF CONT ACTS The Grants Management Specialist is responsible for the negotiation, award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Laura A. Pone Email: (b)(6) Phone: (b)(6) Fax: 301-493-0597 Program Official: Erik J. Stemmy Email: (b)(6) Phone: (b)(6) ----- SPREADSHEET SUMMARY GRANT NUMBER: 5R01Al110964-02 INSTITUTION: ECOHEALTH ALLIANCE, INC. Facilities and Administrative Costs Year2 Year3 Year4 Year5 F&A Cost Rate 1 44.1% 44.1% 44.1% 44.1% F&A Cost Base 1 $290,594 $276,094 $274,594 $270,694 F&A Costs 1 $128,152 $121,757 $121,096 $119,376 Page-6

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RPPR FINAL A. COVER PAGE Project Title: Understanding the Risk of Bat Coronavirus Emergence Grant Number: 5R01Al110964-02 Project/Grant Period: 06/01/2014- 05/31/2019 Reporting Period: 06/01/2014 - 05/31/2015 Requested Budget Period: 06/01/2015 - 05/31/2016 Report Term Frequency: Annual Date Submitted: 05/01/2015 Program Director/Principal Investigator Information: Recipient Organization: PETER DASZAK , PHD BS ECOHEAL TH ALLIANCE, INC. ECOHEAL TH ALLIANCE, INC. Phone number: I (b) (6)1 460 W 34TH ST Email:! (b) (6)] 17TH FLOOR NEW YORK, NY 100012320 DUNS: 077090066 EIN: 1311726494A1 RECIPIENT ID: 07-049-7012 Change of Contact PD/Pl: No Administrative Official: Signing Official: ALEKSEI CHMURA ALEKSEI CHMURA 460 W 34th St., 17th Floor 460 W 34th St., 17th Floor New York, NY 10001 New York, NY 10001 Phone number: (b)(6)~ Phone number: (b)(6)~ Email:1 (b)(6) Email:1 (b)(6) Human Subjects: Yes Vertebrate Animals: Yes HS Exempt: No Exemption Number: Phase Ill Clinical Trial: hESC:No Inventions/Patents: No RPPR Page 1

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RPPR FINAL B. ACCOMPLISHMENTS 8.1 WHAT ARE THE MAJOR GOALS OF THE PROJECT? Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-CoVs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel CoVs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2} how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses; screen people who have high occupational exposure to bats and other wildlife; and examine the genetics and receptor binding properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-CoVs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological opportunity for contact between species so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Gen bank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecological and phylogenetic traits to explain patterns of viral sharing between species. We will test for positive selection in market vs. wild-sampled viruses, and use data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. 8.1.a Have the major goals changed since the initial competing award or previous report? No B.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? File uploaded: Accomplishments.pdf B.3 COMPETITIVE REVISIONS/ADMINISTRATIVE SUPPLEMENTS For this reporting period, is there one or more Revision/Supplement associated with this award for which reporting is required? No B.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? File uploaded: Professional Development.pdf RPPR Page2

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RPPR B.5 HOW HAVE THE RESULTS BEEN DISSEMINATED TO COMMUNITIES OF INTEREST? 1) Conference and University lectures FINAL • Pl Daszak, and Co-investigators Olival and Shi gave >10 invited University lectures that included specific discussion of the current project and results. 2) Agency and other USG briefings • NRC, 2015: Invited speaker, IOM Forum on public health preparedness, lnteragency meeting on Medical Countermeasures. Pl Daszak specifically reported on the findings from Year 1 of this project and the risk of SARS-like viruses causing future pandemics • World Health Summit, Berlin 2014: Pl Daszak was an invited panelist at a session on pandemic risk, and specifically reported the results and aims of this project • International bat virus conference, Colorado, 2014: Pl Daszak and Co-investigator Olival presented results from this study • National Academies, Division of Earth & Life Studies, Spring Advisory Committee Meeting, DC. Pl Daszak presented results from this study as part of an invited talk. • Consortium of Universities for Global Health Conf., Washington DC, 2014. Pl Daszak presented data from this study in a session on disease ecology 3) Public outreach • Pl Daszak reported on this project at an EcoHealth Alliance meeting hosted by the Cosmos Club, 2014 B.6 WHAT DO YOU PLAN TO DO DURING THE NEXT REPORTING PERIOD TO ACCOMPLISH THE GOALS? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. Early in Year 2 of the study, ii is anticipated that all of the qualitative research (i.e, 5-7 focus groups and -100 ethnographic interviews) will be completed, transcribed and translated. It is anticipated that a total of approximately 100 ethnographic interviews and five to seven focus groups will be conducted in targeted areas with known bat populations in Yunnan, Guangxi, Guangdong and Fujian over the next few months. At least one of the focus groups and an estimated 35-40% of the interviews and surveys will be conducted with women. Subjects are enrolled in this study without regard to ethnicity. Preliminary analyses will be conducted and will focus on the factors least understood, but crucial to the development of a behavioral risk survey that captures relevant behaviors and practices. Factors include specific human-animal interactions, experiences of unusual illness in both humans and animals, and an assessment of the context within which these activities occur. Because of the unique dataset and the expected richness of the data, additional research questions will be developed and explored using grounded theory, as well as more recently developed methods such as narrative analysis and case oriented understanding. Results from preliminary analyses will contribute to the development of the behavioral risk survey. A behavioral survey sampling frame and recruitment materials are currently being developed. After pilot testing the behavioral survey, we will begin concurrent biologic specimen collection from bats, other wildlife and humans to compare circulating CoV strains in the bat population with serological exposure in human populations. The behavioral risk survey will facilitate the identification of explicit behavioral risks and practices that are found among study participants seropositive for SARS-like corona virus. These findings will be used to develop better risk mitigation policies and targeted intervention strategies. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. Future steps to optimize the model of role of species diversity in CoV emergence risk will include: 1. Parameterizing with actual data on species diversity and abundance of animals from Southern China markets. 2. Parameterizing with species-specific data on CoV prevalence and strain variation in different bat species from field surveillance, e.g. if Rhinolopus spp. represent the highest risk for SARS-related CoV emergence, these species will be given a higher weight. 3. Incorporation of CoV lineage specific probabilities for inter-host spillover based on receptor binding data. We will also conduct further modeling activities, including: 1. Comparative cophylogenetic analyses of bat host and CoV RdRp and Spike gene phylogenies, to assess patterns of evolutionary congruence and frequency of cross-species transmission. a. Using previously published data from literature and Genbank b. Using sequence data from our S. China surveillance 2. Calculate CoV divergence times using Spike RBD sequences for S. China. 3. Construct initial generalized linear mixed model to predict CoV diversity using S. China data and bat host-specific trait data. Update model regularly with new data from CoV screening in different bat species. Specific Aim 3: Testing predictions of CoV inter-species transmission. The following experiments will be undertaken in Year 2: 1. Animal infection experiment with SARS-like CoV Option 1. Virus infection through ACE2 humanized mouse. Human ACE2 promotor (9-10 kb) and ACE2 will be inserted into a expressing vector and sent to a commercial company to generate transgenic mice. The stably expressed human ACE2 mice will be used for virus infection. Option 2. Virus infection through SARS-CoV susceptible animals such as ferrets. All above animal infection experiment will be performed under the containment of BSL3. 2. Continued surveillances of SARS-like CoVs in Yunnan and Guangdong provinces and isolation of novel virus strains. 3. Surveillance of infection in human populations by SARS-like CoVs. This work will be performed at two locations, one each in Yunnan and Guangdong provinces. PCR and ELISA will be used, respectively, for detection of viral replicase gene and antibody against the viral RPPR Page3

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RPPR FINAL I nucleocapsid protein. RPPR Page4

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B.2 (Accomplishments.pdt) 8.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? Daszak, Peter, Pl Year 1 Report for Understanding the Risk of Bat Coronavirus Emergence Award Number: 1R01Al110964-01 82: What was accomplished under these goals? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. In the first year of this R01, we have: 1) Designed a behavioral risk study using an iterative approach that begins with rapid and focused qualitative research at or near biological surveillance sites in China where bats have previously been captured, sampled and found to contain novel CoVs. The study design includes: 1) structured observation and mapping of public spaces, 2) focus groups and 3) ethnographic interviews. The primary enrollment criteria are related to occupational exposure to bats and residence near bats. This research is conducted with two groups of individuals: those involved in the bat value chain (from hunter through market to consumer) and those highly exposed to bats (e.g., cave dwellers). The qualitative data will be used to inform a behavioral risk survey, as well as to contextualize findings from behavioral surveillance analyses. 2) Conducted observational research and mapping in: Yunnan: In and around Xiang Yun village (two clinics and one wildlife restaurant); in and around the remote Lu Feng village (1 wildlife farm, 1 wildlife butcher and 1 wildlife restaurant) and at the An Ning communicable disease hospital complex; Guangxi: In and around LiPu, (two markets, 3 wildlife farms, 1 wildlife restaurant); and Guangdong: Guangzhou wildlife market, Foshon wildlife market (this market is where the first cases of SARS were traced back to in 2003). 3) Secured local IRB approval in November 2014 from Wuhan University School of Public Health, Hubei Province, to conduct qualitative research, to administer behavioral surveys and to collect biological data including blood (no more than 550ml), sputum, and stool samples from humans. We secured US IRB approval through Hummingbird IRB (2014- 23 approval letter sent to NIH) in November 2014 for qualitative, quantitative and biological specimen data collection. 4) Drafted protocols, guides, and training modules for Observational Research, Focus Groups, and Ethnographic Interviews and pilot tested these. The Observational Guide and Ethnographic Interview materials were pilot tested in live animal markets in Queens, New York City. Consistent with the original proposal, we have trained interviewers and identified key informants. Key informants include community health workers from three different administrative level CDCs, Barefoot Doctors, public health clinicians, local wildlife farmers and wildlife restaurant owners, as well as market vendors and workers. Ethnographic and Focus Group Interviews to be conducted pending NIH approval of IRB approval letter. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. 1) Collation and preliminary analysis of published bat Coronavirus data to optimized specimen collection and taxonomic targets for surveillance. RPPR Page5

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B.2 (Accomplishments.pelf) RPPR Daszak, Peter, Pl Over the last decade a large number of bat viral discovery studies have been published globally (including a large number focused on CoVs). In year 1, we conducted the first ever systematic analysis of these data. We collated literature from over 100 viral discovery studies in bats, to examine patterns of host range and known viral diversity in different bat taxa (Young and Oliva!, In Review). We found that Coronavirus diversity has been most thoroughly characterized in a few bat families, including the Vespertillionidae and 5 other families, but several bat taxa remain under-represented in global virus surveillance efforts (Fig 1 ). Identification of these surveillance gaps allows us to better target our field surveillance towards bat taxa where CoV diversity is largely unknown (blue and light colored cells, Fig 1 ). These analyses were completed at various taxonomic levels, including by bat subfamily and genera (Family level analysis only shown). 1 n 25 ~ Pb:lmovlridoo Cif00Yiridae 20 PaMYirldae 1$ Fllowtdae Retrovlfldae 10 Reo,iridae Papillomavlrldae 1--= OrllW:>my>a>vlrldae Callcivitrdao Togwlridae 8<Jny,Mridae HcpednQ\'iridae lflavirid.ae FlPoeyomavlrldaft Rhabdovlrldae Corona\/iridae AstroVtrldae H.,_v,ridae Ade<,Ovlridae Paramyxoviridoo f ! !I' I ! i I I ~ ~ ~ I i 5: 0' ~- '8 i i i i I a 3 I "- i .. l gI C " f 5: .. 8 f f ~ is: = .. .. " .. i Figure 1. Heat map of viral richness by bat host and viral family, clustered by similarity in viral richness across host and viral families. To maximize our chances of discovering CoVs, we need to define the number of specimens required for our bat surveillance work and the bat taxonomic groups on which to focus our surveillance. We used generalized linear mixed models (GLMM) and applied this to a subset of our collated data for CoVs alone. We found that sample type screened (feces), collection methods, and the number of specimens tested best explains the probability of finding an individual CoV positive sample. We will now use these Page6

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B.2 (Accomplishments.pelf) Daszak, Peter, Pl approaches to increase the likelihood of getting positive samples in our fieldwork in China. 2) Preliminary 'What-if' Model: Role of species diversity in CoV emergence risk. RPPR We built a mathematical model to analyze different scenarios of CoV spillover. We began with an assessment of how the diversity of wildlife (and other factors) in wet markets may affect the probability of CoV zoonotic spillover. We modeled evolution of CoVs within wildlife in a market following the initial introduction of a novel virus in one specific host. We assume this initial virus is a single genotype that does not yet have a great enough rate of spread to create an epidemic, but has a rate of spread close to this threshold. When this virus infects a new host, a new genotype is generated, based on random drift from the infecting genotype. We use Neutral Theory of Species Diversity to specify the species distribution in the market, for a given total number of species and total abundance of animals. We assume 500 animals in the market, and alter the species diversity from 3 to over 40. These numbers are easily attained in a small to medium market in Southern China (and in year 2 we will groundtruth these assumptions) As the number of species present in a market increases from 3 to 20, the percent of simulations where zoonotic spillover occurred from any of the animals into humans increases (Fig 2). However, the risk remains fairly level if wildlife biodiversity increases above that level. The probability of epidemic failure is inverse to the probability of a zoonotic spillover taking hold and decreases with increasing species diversity (Fig 2). Therefore our null model shows that reducing the diversity of species in live animal markets could reduce the risk of zoonotic spillover, including of potentially pandemic CoVs. - Zoonotic Spillover - Epidemic Failure 100 "' 80 C 0 -~ 'S 60 E ui 0 c 40 CD e &. 20 0 0 10 20 30 40 Number of Species in Market Figure 2. 'What-if' scenario model based on the Neutral Theory of Species Diversity to examine the role of wildlife species diversity for CoV spillover in markets. Page 7

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B.2 (Accomplishments.pdf) Daszak, Peter, Pl S ecific Aim 3: Testing predictions of CoV inter-species transmission. 1) Bat Corona virus Surveillance in 2014 We collected 1555 anal swab samples, 1357 fecal samples, 461 blood samples, 469 serum samples and 24 tissue samples from> 14 bat genera in 5 provinces and in Laos (Table 1). Table 1 Bat Samples collected for CoV surveillance in 2014 Anal Oral Fecal Blood Serum tissue Jan. 2014 Mengla, Yunnan 164 -- -- -- -- -- Mar. 2014 Beihai, Guangxi 30 -- -- -- -- -- Apirl 2014 Shenzhen 77 -- -- -- -- -- Ruyuan, 167 -- -- -- -- -- Guangdong Chuxiong, Yunnan 52 52 103 -- 8 16 May 2014 Jinning, Yunnan -- -- 131 -- -- -- Mojiang, Yunnan 25 25 103 -- -- 3 May-Sep. Xianning, Hubei 583 2014 -- -- -- -- -- Jun. 2014 Guangdong 77 -- -- -- -- -- Jul. 2014 Hainan 460 -- -- -- -- -- Aug. 2014 Yichang, Hubei -- -- 114 -- -- -- Guilin,Guangxi 121 122 -- 122 122 -- Sep. 2014 Guangdong 335 337 -- 335 335 -- Jul.--Sep. Mojiang, Yunan 96 2014 -- -- -- -- -- Oct. 2014 Jinning, Yunan 13 13 6 3 3 4 RPPR Page 8

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B.2 (Accomplishments.pdf) RPPR Daszak, Peter, Pl Mojiang, Yunan 34 34 100 1 1 1 Laos 121 Total 1555 583 1357 461 469 24 CoV was detected in 14% (336/2329) samples (Table 2). Diverse alphacoronaviruses were identified, including isolates closely related to Bat CoV 1 A, 1 B, HKU2, HKU6, HKU7, HKU8 and HKU10. Groups of novel alphacoronaviruses were discovered in a variety of bat species (Fig 3). Novel SARS-like coronaviruses were detected in Rhinolo hus bats collected in different re ions of Guan don rovince. Diverse novel betacoronaviruses related to HKU5 were detected in Pipistrellus bats and la io in Guangdong and in Asel/iscus stoliczkanus in Mengla, Yunnan. Novel coronaviruses related to HKU9 were found in Cynopterus sphinx and Rousettus leschenaulti in Mengla (Fig 3A). In addition, sequences significantly divergent to other CoV were obtained from three samples of /a io and Hipposideros bats. Page9

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B.2 (Accomplishments.pelf) A Daszak, Peter, Pl 55 MLI-10055/Astlliscu sto6ClUJlu.s (2) .------......,.""'l lll,1401,()5-51/Roumtulmhmanltl (1) :'.'.ll.140056,'Asdllicus uolk:zltaans (2) 67 l!LU00M-ZJIRhillolopbat a1liD.4 '------.Alpha-CoVXw1G Spaiu.2007 HQIS-W55 --------< :ardiodean..b.t-CoV Keny~'KY43.2006 HQ 728-180 74'---------1 OWfflUS-bat-C-0V-HKU!O 1Q9&92"1 100 ipposidetO!-bat-CoV-HKUIO JQ9S9'.166 .---------l;·otiub,·idii-CoVisobt~ XYMd70 KF569991 -----Alpha-CoV-Psp,X.Spain.1007 HQl8~060 BtC-O\',AS19l2005 DQ64SS'36 at-CoV-IIKlJ6 DQ24922-I !\fi.,140031-31/:'.'.fiDJopmus schrtibenji \ll.UOOlJ...151\finiopttnu tcbrrib.nii ai.COV-1B DQ66633S at-C-OV-IA Et:4:!0138 ,--___.!l!2["--fl:at-CoV-HKUS DQ2492:S ~ll..1400!81 Miwopttrlll ,cbnibmii (3) !Il.140030/ lfiru•ptmu scbrttl>trsii LOO '-1L1~003J-~?/11miopttm scllrtibtrsii 68 Bat..COV-HKU1 DQ249"..26 at-CoV-HKU2 ££203067 JOO ~Il,140001/Rbinolopbu, affiais 100 ULH0026-lS/Rh.inolophus :dlin:is Beu-CoV-E.isaM.'Spaiu.,2007 HQIS-4062 eta-CoV -Epteiicu;, 13RS3S4 _ 26·1tdy101l KB U399 lil,140062-MIAsdlimu stoliakanus ln..140066/A,eDlscus stoliczkanus '-----&t:.C-OV-HKU5 EF065509 ~------+<a-CoV-HKl.4 EF065505 ,._ _____________ _,,uman-CoV-HKl:1 AYSS~OOI 99 78 0.1 RPPR lll.1-10137-51.JCynopm-ussplwu ~Il.U0J37-5.4/Cynopm,u lJLU0136/Cynopterus sphinx (4) :I.Il,l40l05-561Rousett1u ltscbtnaulti (-0 Bat-C-OV-HKU9 EF06551~ 100 at-SARS-CoV-R.fl DQ412042 at-SARS.COV-HKU3 DQ022305 Ba-SARS-Co\'.Rp3 DQ0716l5 7 sSHCOl~ KCSS 1005 3367 KCSSI006 96 ARS-C-OV,QZ02 AY390556 ARS-Co \. -SZ3 A Y304486 62 ARS-C-0 V.BJOI A Y27S48S Page 10

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B.2 (Accomplishments.pelf) B .----, 0.05 RPPR 5 l~lppo,ldrrounni\ter M/Spain'2001 HQ 184062 &13-CoV-tptesicus/13RS384_26/lialyt2012 KF312399 '-LH0460 t;J l'lplSll~llus plpisirtUus ~84-36/Ph>l>lr•llu• l!ll!l<trellu< (7) NI.139551Myot1• pOos u 1 XLI JOJ551P!J!illnllas PJRislrtllus N Ll JS84-371PiP-istreUus PlPJ.tlmJuj; 9~ NLIJ893-531J'iplsUcUus1>lpblreUus BltCo V(BtCo V,A4 3 712005) DQ64 &820 Sf.1314'7 26/f'lpl-<il"NIIU<iabnunu, llll!t-Co V-HKU5 EF065,5P9 DQ648~~ p. Human-CoV-HKUl AY834 Bat-CoV-HKU9 EF065514 Bal-SARS-CoV-HKUJ D@ill05 1.NLl-10-100/klunoiOpbus:lDkU.S Bat-SARS-CoV-Rfl D0412042 L\3973-61/Rhlnolop••JS slnlCUl 9 l,.U0391/RhlOOklphUU1Dl(tlS . t,SARS.cC<>.V-. NLHOJ52/Rhlnol (3) NL140494-l9lfllp er RsSHCOIJ KCSSIOO; 61 Rs3367 KC&81006 SARS-CoV-SZ3 AY304486 9 SARS•CoV•GZ02 A \"390556 SARS-CoV-BJOl AY278488 NLJ JOJJ5-l llR•ioolopbus sioicus (2) :,,;"Lt-&OJ88/Rhinolopliutisioicul (3) Bat-Co\'-RKU2 EF203067 l:-1,14035Q-J6/Rhlnq)opb•• pus111u, Ol!J1-f1/Rblnohll)htn pul.OJII" marrot1s(7) I Ill r--'""l___).!'~~~~~;;s 8 .xLI J0.&60-1 "/ riol\l rr1hn pjp.JilrtJtus NLI J0j99/Hlpposldff'os arm~•· (2) NL1389J SI/Pipburh$,pp. (8) :-.'LJ40A61/Hyps"l!0S•••ll (~lit) (2) XLI 40480/IIJppesldtros aro,Jjier Alpba-CoV N.laslGISpain.12007 HQ 184055 ll'------t llous<ltua-bat-CoV-HKUlO J0989h71 9 Hioposidaos-b:IL•Co\1-HKU 10 J0989266 Myotis-davidii,CoV isolatcXYMd70 Kf56999 I 11 abramut 2007 IIQI 84060 8836 $ ~...;....~~--"--''-""'2) 124 ti ~IS NL13946-S.l/l\l ·otls l!illt! s ~I 13845 !Mo fo IO NLL.llOJJ-;51 Hlppo,i<lcro, jll'l>lll :',LIJI015-161 Hlru10>1<1rro annlg<r Daszak, Peter, Pl Page 11

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B.2 (Accomplishments.pelf) RPPR C 81 0.1 Daszak, Peter, Pl 14J2J 7/:\·lyotis pllosus BY1405S2-Sl/Nlyotisborsr ... ldii 141312/:\llnlopcerus spp. (21) I U !62-!\9/:\llnlopttnn spp. B:tt-CoV-lB DQ666.3l8 H1355/Rhinoloplws sioicllS HD131585r.\flnlopieru.sspp. ◄) Hl-4J7-3-I/Pipts,reHusabramo, (2) Bal-CoV-lA .EU420138 ~------Ba1-CoV-HKU7 DQ249226 51 UJ262-56/lllniopterus spp. Bat-CoV-HKUS DQ249228 J41286r.lllniop1en1s s1>p. (6) '--------------L14 t :3!>2/lllpposlderos pontona (3) 100 Rousenus-ba1-CoV-1iKUJO JQ989271 Hipposidero .. bat,CoV-HKUlO JQ98926<! 1Il>l31599/Rlllnolophus spp. (9) 1 "16301 Hlpposideros pomona Bat-CoV-8.KU2 EF'203067 '----- Hl2500lyotis pil0$11$ '-----::-::-i .141378/Rbinolopbus pusillus 14HJ!'/Rhinolopbus p11Sillus ~----------Mvolib-da,idii-COV isolate XY.Md70 ..KE-'69991 BYH05J~l0,Plpls1rtlus ftbrnn>bS -141228-8/M)·otis pllosus ~-------AJpha,CoV-P.sp•K.Spain/2007 HQ184060 BtCoV,A81912005 ~8836 U:013.1580-29 .:\Jyotis pflosus (S) HDUlS81-~l)'OliS pllosus (12) J. 131<51Hq)posldu·O$ powooa Ba1-CoV-HXU6 DQ249224 BYl-40S3~1/Pipisb'tllosabnmus (10) BY140~65-621Plplst rtllus ObNlmUS Bat-CoV-HKU5 EF065509 BatCoV(BtCoV 1Al2061200.S) DQ6-4S802 141 Hl/Pipistrellusspp. BatCoY(BtCoV A437/2005) DQ648820 HH311Pipistrellus abramus Beta-CoV-EJsaf~Spain.2001 HQ184062 B•ta-CoV-Eptesii:w l3RS384_26111aly12012 KF312399 Bat-CoV-8KU4 EF065505 GZ131660/TyloinycterK pac.hypus Human-CoV-m...'vl AYSS400l ~-------_._,.>U-JBV1 05Sl/1\lrotUbo1•sflddU ~-----""'-I BYH056~661Plpistn,Uus abrllllUS (-4) '-------- Bat-COV-l:[KU9 Ef065514 100 76 HDl3158l-56/ 61 HD131601-2/Rbinolopbus spp. (2) ~---< Bat.SARS.CoV..Rfl DQ412042 RD131S80-27 Bat-SARS--CoV-HKU3 DQ022305 141352/Rbinolopbussinicus (9) 141437-38 141577(3) 141600/Rbinolophus pusillus (4) B•t.-SARS-CoV-Rp3 OQ07 I 615 9 RsSHC014 KC881005 7A Rsl367 KC881006 94 SARS-CoV-OZ02 AY390556 62 ~~~~~~~~ii :;;~;::: Figure 3: Phylogenetic analysis of partial RdRp gene of CoV. Co Vs identified in this study are in bold and named by the sample numbers. Sequence amplified from samples co-infected with two CoV strains are indicated in red. (A) CoVs detected in Mengla, Yunnan. (B) CoVs detected in Ruyuan, Guangdong. (C) CoVs detected in other regions in Guangdong. Page 12

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B.2 (Accomplishments.pelf) RPPR Daszak, Peter, Pl 2) Complete S gene sequencing and recombination analysis of novel SARSlike CoV We amplified the full-length S gene of the novel SL-CoV detected in a Rhinolophus sinicus colony in Yunnan Province. In addition to our previously reported Rs3367 and RsSHC014, we now have 24 new full-length S gene sequences from 22 samples. Phylogenetic analysis showed that these SL-CoV are diverse, and identified two strains of novel SL-CoV more closely related to SARS-CoV than Rs3367 (Fig 4A). Our new strains named Rs4841 and Rs487 4 share the highest homology to SARS-CoV than any other known SL-CoV, including those we published previously in Nature. These viruses are highly similar to SARS-CoV in receptor-binding domain (RBD) sequence but also in N-terminal domain (NTD) (Figure 48). Analysis of the complete S protein shows > 97% amino acid identify to that of SARS-CoV isolates. 100 100 too too 1(1) Human SARS-CoV BIOi Human SARS-CoV GZ02 Cint SARS.CoV SZ3 R.4841 100 '---------BatSL-CoV LYRall ~l Rs4092 ~------------ 1 -°'--rRs4075 Rs4122 ~--------B1tSL-CoVRfl 100 ~---Bat SL-CoV HKU3 Bat SL-CoV Rp3 '-----Bat SL.CoV Rml BatSL-CoV Rs672 Rs410S '------:::too:-"l..l.Rs4249 Rs4255 Rs40Sl 100 Rs40S7-2 ·• Rs4096 Figure 4A Phylogenetic analysis of novel SLCoVs discovered in Year 1 of this project (Bold}, based on amino acid sequences of complete S gene. Page 13

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B.2 (Accomplishments.pelf) RPPR .'IAl\lt'"CnV 5;:0:; !lA.l\.!J•CoV ltJOL !IAllta-eov 't'o.-2 111•••-e•V a:e, tu'IGU l\<!f-42)1 ... ...,,,., A.•13Z7 1t,08HCO.l• f\o>39:P SL•C',iV R,,p) IJt.•CoV HKlll ,. •• 2.1 ft.-'IZ3~ "--'~=, l\-42SS ••4081 in..-eov l'l.•i72 81rC'oV ~ :,r, ... e,v f\tt 111. .. 11015 IJL-cov :.v•a.U. SA.R.9-CoV ~;:Q::? I QA. ........ '-'OY ..... o.l- $A,fli$•COV fo~2 J u~eov tZ3 lh'l•O P.•4231 a..33,n ,.,1327 "•8HCOU ,..,," :n.-,:ov Rp) ai.-eov ttKt.tl A.t0'1Z'l'T 1'.s'l<l3S l't•"•24 ... 111:u,r; A.1tt08l 81,•CoV fl.sS'1Z •~· e .. v l'\ml ir,.-e,;v p,f1 lt.4015 -'Lo•COY .lll'lh.J..I. IJAl\e•C'oV C:t:02 a>....._..eov S.JO.l 8Al\8--C'oV rod '""..-cov Oi:3 A•48U ll•4::?31 ,._,33'7 I\ .. .,~:., "-••tto:OJ.'1 ~...,,,, •L>•<eov ..,.,, 81,--CoV HKtJl f\.11>'12'1., ...... tc.):) ltaf52' .... z.ss kn08J. 8t,,-,,CoV f\e1.1% :tt,,-,,C'oV l'Jnl IJJ.•C:oV Rtt ...... o,s 8AlltJ·C'oV &~OZ 8AA8·C'oV &J'Ol. .... ,. •• CoV l'o.,~ "'""-••c:-ov s;::) lt.48U -•.c:,.a M33n il-•1321 ~UUtCOL4 lbll,9 81.-C'o\l 11.p) 8L·C'o\l KKIJl ~-•t•'l' lb423.S k•6S2E ,..12~s "-'"4n1u ii,.c,v ~d.,i :,1.,-c,v -t ,o.-~ .. v t<n ,. .. ,0;5 81"-CoV t,'(11,ell ~""'-•·eov •zuz v.Re-.CoV WO.l $.'-1:t••cov tor2 -'IA.RS-CoV :IZ3 lbt8fl "-•U:31 A.:J36"' A-s'1~27 ,._dutcO.l◄ A.•33,9 8lrCoV fl.p) •r..-e .. v Ml("' llle424'J ,. •• 2)5 ... ,2,~ A•'IOh S'- C.,(IV ><•¥'): !llr-Cov """-l 8t.-CoV fl.Cl l\ootQ;,) 3~c::,v 1,-o;,.11 8AA8•CoV &a02 13..,P<a•CoV WOt, IS.u\!S•CoV l'oi,2 -""M-eov sa, lls48U -'•'1%31 ... s~,., 111•7327 AsfU,CO.l• " .. ,:,o,. 81,,-CoV R.pJ :~;~:~ t{l(-,J "-••23.S fli,6526 l\•4-',)~ 111•4081 IJl,-CoV R•072 8t,,-CoV ,._1 s1,-cov 11.tl 111.,. 1:1-,.5 SL-C:oV LYll.•J l \'t''i:or~!I 1'1?1T0~!1 lT'.t.'1'8)1' \'lf'l>Sll~UI! l1',t,.'J'838 !'.7':!:;s 4T~V81:N'• J."li.V8tl'i ,1.•r~v;o....,. ri..>,i,t.A. ,~ ;~~~ ... ... ... ... ... ... ... .. , .. , ~· ... ... "' ... ... ... ... ... ... ... ... vo~-ooc-.1: PUOEIKt>Cffl? P'O-Pp.KPC't'f't> YIIIP~1'1C'~V VlllPC4QDCMV oo---------- ~········· ·----------- t-·•-·-···- , ........... . !----------- l" seOiKeC'l't'e Daszak, Peter, Pl ,,. ,,. ,,. ,,. ,,. ,a ,eo , .. ... ,eo ,., ,., ,., ,., ,., ,., ,., ,., ,., ,., ,,. ,., HO HO HO HO HO HO Hl '" '" ,u uo m t?'l' ... l96 ... .,. ••• . .. .. , ... '" Page 14

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B.2 (Accomplishments.pdf) Daszak, Peter, Pl RPPR Figure 4B Alignment of amino acid sequences of S1 (aa1-680) of SARS-CoV and bat SL-CoVs. We performed recombination analysis and detected potential recombination events in S genes of multiple SL-CoV strains suggesting that that the region around nt1000 in RBD is a recombination hotspot. In addition, a novel SL-CoV strain (Rs4075) with an NTD sequence distinct from all other SL-CoVs was identified (Figure 4). The results suggest that the high genetic diversity of SL-CoV in this colony is related to the frequent recombination. 3) Virus isolation and characterization Isolation on Vero E6 cells was conducted on all CoV PCR-positive samples using an optimized protocol. Repoducible CPE was observed for Rs4841 (the strain closely related to SARS-CoV in both the RBD and NTD region of the S protein). Purified virions displayed typical coronavirus morphology under electron microscopy, and this novel isolate was named SL-CoV-WIV16. We conducted virus infectivity studies (using Hela cells expressing or not expressing ACE2 from humans, civets or Chinese horseshoe bats) to determine whether SL-CoVWIV16 can use ACE2 as a cellular entry receptor (Figure 5). We found that WIV16 is able to use ACE2 of different origins as an entry receptor. HelabACE2 HelacACE2 HelahACE2 Hela DAPI fITC Cy3 Merged . -, - - - - 4 - - - - - - - -tflµm . ' Page 15

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B.2 (Accomplishments.pdf) Daszak, Peter, Pl Figure 5. Analysis of receptor usage of SL-WIV16 determined by immunofluorescence assay. Determination of virus infectivity in Hela cells without the expression of ACE2. b, bat; c, civet; h, human. Nuclei are stained with DAPI. The columns (from left to right) show staining of nuclei (blue), ACE2 expression (green), virus replication (red) and merged triple-stained images. To assess its cross-species transmission potential, we conducted infectivity assays in cell lines from a range of species. Our results (Figure 6) show that SL-CoV-WIV16 can grow in human alveolar basal epithelial (A549), pig kidney-15 (PK15), Rhinolophus sinicus kidney (RSKT), Macaca mulatta Kidney cell lines (MK2) and human lung carcinoma (NCI-H292), but not in human cervix (Hela), Syrian golden hamster kidney (BHK21 ), Myotis davidii kidney (BK), Myotis davidii intestine (MDI), Rousettus leschenaulti kidney (RLK), Rhinolophus sinicus brain (RSBT), Rhinolophus sinicus heart DAPI C 3 (RSHT), Rhinolophus sinicus Lung (RSLuT), Rhinolophus A549 sinicus intestine (RSI) or Pteropus alecto kidney (PaKi) cell lines. LLC-MK2 PSKT PK15 H292 VeroE6 RPPR Figure 6 Cell infection with SLCoV WIV16 determined by immunofluorescence assay with antibody against SARS-like coronavirus nucleocapsid protein. The columns (from left to right) show staining of nuclei (blue), virus replication (red) and merged double-stained images. Page 16

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B.4 (Professional Development.pelt) 8.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? Daszak, Peter, Pl Accomplishments for Understanding the Risk of Bat Coronavirus Emergence Grant Number 5R01Al110964 84: Opportunities for Training and Professional Development In year 1 of this work, we trained undergraduate interns from Columbia University in modeling approaches to understand bat risk of harboring zoonotic CoVs. In the behavioral risk work, we used standardized training materials for all three qualitative behavioral risk data collection methodologies have been created. Materials were used to train six people in New York City and 12 people in Yunnan, China, of which 11 were from three different administrative levels of local government Centers for Disease Control (CDC). The trainees include the Chinese EcoHealth Alliance Field Coordinator and Yunnan Provincial CDC personnel: six researchers from Xiangyun County CDC (4 women, 2 men), two from Yunnan Institute for Endemic Diseases (Yunnan Provincial CDC; 2 men), and three from Lu Feng County CDC (3 men). RPPR Page 17

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RPPR FINAL C. PRODUCTS C.1 PUBLICATIONS Are there publications or manuscripts accepted for publication in a journal or other publication (e.g., book, one-time publication, monograph) during the reporting period resulting directly from this award? Yes Publications Reported for this Reporting Period ·i• ~:..,,,._...,_.,._;,,.,• ,----------------------------- N/A: Not Journal Olival KJ, Weekley CC, Daszak P. Bats and Viruses. Wang L editor. New York: John Wiley & Sons, Inc.; 2015. What we know and need to know Non-Compliant (b)(4)- PMC Journal - In process (b) (4) C.2 WEBSITE(S) OR OTHER INTERNET SITE(S) NOTHING TO REPORT C.3 TECHNOLOGIES OR TECHNIQUES NOTHING TO REPORT C.4 INVENTIONS. PATENT APPLICATIONS. AND/OR LICENSES Have inventions, patent applications and/or licenses resulted from the award during the reporting period? No C.5 OTHER PRODUCTS AND RESOURCE SHARING C.5.a Other products NOTHING TO REPORT C.5.b Resource sharing NOTHING TO REPORT RPPR Page 18

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RPPR FINAL D. PARTICIPANTS D.1 WHAT INDIVIDUALS HAVE WORKED ON THE PROJECT? .;,.-'>--,cv~.~•"·•-~!,,:.ot --;;:,:-~;4-w,,;f:'. )"""/•?,< ~:---::-::11-<·~., '"'"-,;'v:-;-.f-'. • 1rr~ ~i'Y.Jr;, 1:,;,_,, '.-:,..~.-: ,>,,:.-,:,-,.,, .. ~;,·11~· ~i~~,~ (.;>.,,l ~--,.,., 1~· ,!,..,_-,_,J 'I:'· .• ,,, ~~$• .. ~;\ r~";-·.~ • 1~1••->'.i< Plr.<- ,~::,;, (b) (4) (b) (6) I (b) (6)) y DASZAK, PETER y KE, CHANGWE N y ZHANG, YUNZHI y ZHU, GUANGJIA N (b) (6)] y SHI, ZHENGLI I (b)(6): N CHMURA, ALEKSEIA (b) (6)] y OLIVAL, KEVIN J I (b)(~ y HOSSEINI, PARVIEZ RANA (b) (6)] y ZHANG, SHUYI y GE, XINGYI (b)(6): y EPSTEIN, JONATHAN H Glossary of acronyms: S/K - Senior/Key DOB - Date of Birth Cal - Person Months (Calendar) RPPR (b)(, (b)(4 (b)(, (b)(~ (b)(~ I (b) (6)] BS,PHD PHD (b) (6)j PHO (b)(6): PHD (b)(~ PhD (b)(6): BS (b) (6)] PHD (b)(~ BS,PHD (b)(6): PHO PHD (b)(6): MPH,DV M,BA,PH D PD/Pl Co- CDC and Investigator Preventio n of Guangdo ng Province Co- Yunnan Investigator Institute of Endemic Diseases Control & Preventio n Co- East Investigator China Normal Universit y Co- Wuhan Investigator Institute of Virology NonStudent Research Assistant CoInvestigator CoInvestigator Co- East Investigator China Normal Universit y Co- Wuhan Investigator Institute of Virology CoInvestigator Foreign Org - Foreign Organization Affiliation SS - Supplement Support RE - Reentry Supplement DI - Diversity Supplement Page 19 NA CHINA NA CHINA NA CHINA NA CHINA NA NA NA NA CHINA NA CHINA NA NA

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RPPR FINAL I Aca - Person Months (Academic) OT- Other Sum - Person Months (Summer) NA - Not Applicable I D.2 PERSONNEL UPDATES D.2.a Level of Effort Will there be, in the next budget period, either (1) a reduction of 25% or more in the level of effort from what was approved by the agency for the PD/Pl(s) or other senior/key personnel designated in the Notice of Award, or (2) a reduction in the level of effort below the minimum amount of effort required by the Notice of Award? No D.2.b New Senior/Key Personnel Are there, or will there be, new senior/key personnel? No D.2.c Changes in Other Support Has there been a change in the active other support of senior/key personnel since the last reporting period? No D.2.d New Other Significant Contributors Are there, or will there be, new other significant contributors? No D.2.e Multi-Pl (MPI) Leadership Plan Will there be a change in the MPI Leadership Plan for the next budget period? No RPPR Page 20

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RPPR E. IMPACT E.1 WHAT IS THE IMPACT ON THE DEVELOPMENT OF HUMAN RESOURCES? Not Applicable FINAL E.2 WHAT IS THE IMPACT ON PHYSICAL, INSTITUTIONAL, OR INFORMATION RESOURCES THAT FORM INFRASTRUCTURE? NOTHING TO REPORT E.3 WHAT IS THE IMPACT ON TECHNOLOGY TRANSFER? Not Applicable E.4 WHAT DOLLAR AMOUNT OF THE AWARD'S BUDGET IS BEING SPENT IN FOREIGN COUNTRY(IES)? RPPR Page 21

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RPPR FINAL F.CHANGES F.1 CHANGES IN APPROACH AND REASONS FOR CHANGE Not Applicable F.2 ACTUAL OR ANTICIPATED CHALLENGES OR DELAYS AND ACTIONS OR PLANS TO RESOLVE THEM NOTHING TO REPORT F.3 SIGNIFICANT CHANGES TO HUMAN SUBJECTS, VERTEBRATE ANIMALS, BIOHAZARDS, AND/OR SELECT AGENTS F.3.a Human Subjects No Change F.3.b Vertebrate Animals No Change F.3.c Biohazards No Change F.3.d Select Agents No Change RPPR Page 22

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RPPR FINAL G. SPECIAL REPORTING REQUIREMENTS G.1 SPECIAL NOTICE OF AWARD TERMS AND FUNDING OPPORTUNITIES ANNOUNCEMENT REPORTING REQUIREMENTS NOTHING TO REPORT G.2 RESPONSIBLE CONDUCT OF RESEARCH Not Applicable G.3 MENTOR'S REPORT OR SPONSOR COMMENTS Not Applicable G.4 HUMAN SUBJECTS G.4.a Does the project involve human subjects? Yes Is the research exempt from Federal regulations? No Does this project involve a clinical trial? No G.4.b Inclusion Enrollment Data Report Attached: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 G.4.c ClinicalTrials.gov Does this project include one or more applicable clinical trials that must be registered in ClinicalTrials.gov under FDAAA? No G.5 HUMAN SUBJECTS EDUCATION REQUIREMENT Are there personnel on this project who are newly involved in the design or conduct of human subjects research? Yes As reported by Dr. Peter Daszak (Pl) to NIH in May 2014, all of the following senior/key/other personnel were enrolled in and passed the Human Subjects Research Course provided by the Collaborative Institutional Training Initiative (CITI Program) at the University of Miami (http://citiprogram.org ). The CITI Program is a leading provider of research education content with web based training materials serving millions of learners at academic institutions, government agencies, and commercial organizations in the U.S. and around the world. Peter Daszak, Pl Zhengli Shi, Co-Investigator Shuyi Zhang, Co-Investigator Changwen Ke, Co-Investigator Jonathan Epstein, Co-Investigator Kevin Oliva!, Co-Investigator Parviez Hosseini, Co-Investigator Xingyi Ge, Co-Investigator Guangjian Zhu, Co-Investigator Yunzhi Zhang, Co-Investigator Aleksei Chmura, Program Coordinator G.6 HUMAN EMBRYONIC STEM CELLS (HESCS) RPPR Page 23

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RPPR FINAL Does this project Involve human embryonic stem cells (only hESC lines listed as approved In the NIH Registry may be used In NIH funded research)? No G.7VERTEBRATE ANIMALS Does this project involve vertebrate animals? Yes G.8 PROJECT/PERFORMANCE SITES ;!_,_, ::,.;_;.,.,: .. '.t:.,:,.:~_"j r~~<'=· 1·~!~ ... ~_\c-<;.:;~;::,~ 1·1~ ''• ' '\; . -------·------------------- Primary: EcoHealth 077090066 NY-010 460 West 34th Street Alliance, Inc. 17th Floor New York NY 100012317 Wuhan Institute of Virology 529027474 Xiao Hong Shan, No. 44 Wuchang District Wuhan East China Normal University G.9 FOREIGN COMPONENT 420945495 Organization Name: East China Normal University Country: CHINA Description of Foreign Component: Institution of Co-Investigators Dr. Shuyi Zhang and Dr. Guangjian Zhu Organization Name: Wuhan Institute of Virology Country: CHINA Description of Foreign Component: 3663 Zhongshan Beilu Shanghai Primary Laboratory and Institute of Co-Investigators Dr. Zhengli Shi and Dr. Xingyi Ge Organization Name: Yunnan Institute of Endemic Diseases Control and Prevention Country: CHINA Description of Foreign Component: Institution of Co-Investigator Dr. Yunzhi Zhang Organization Name: Center for Disease Control and Prevention of Guangdong Country: CHINA Description of Foreign Component: Institution of Co-Investigator Dr. Changwen Ke G.10 ESTIMATED UNOBLIGATED BALANCE G.1 0.a Is it anticipated that an estimated unobligated balance (including prior year carryover) will be greater than 25% of the current year's total approved budget? No G.11 PROGRAM INCOME Is program Income anticipated during the next budget period? No RPPR Page24

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RPPR G.12 F&A COSTS Is there a change in perfonnance sites that will affect F&A costs? No RPPR FINAL Page 25

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RPPR Inclusion Enrollment Report Inclusion Data Record (IDR) #: 166195 Study Title: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 Foreign/Domestic: Foreign Planned Enrollment Report Planned Enrollment Total: 2,460 FINAL NOTE: Planned enrollment data exists in the previous format; the PD/Pl did not enter the planned enrollment information in the modified format and was not required to do so. Only the total can be provided. Cumulative Enrollment Report NOTE: No cumulative inclusion enrollment data exists in the previous inclusion format or modified format. Although prompted to do so, the PD/Pl did not enter information in the modified format. No data can be provided. RPPR Page 26

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Notice of Award RESEARCH Federal Award Date: 05/05/2017 Department of Health and Human Services National Institutes of Health NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 5R01Al110964-03 REVISED FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Aleksei Chmura President 460 West 34th Street 17th Floor New York, NY 100012317 Award e-mailed to: (b)(6) ----------- Period Of Performance: Budget Period: 06/01/2016 - 05/31/2017 Project Period: 06/01/2014 - 05/31/2019 Dear Business Official: The National Institutes of Health hereby revises this award (see "Award Calculation" in Section I and "Terms and Conditions" in Section Ill) to ECOHEALTH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the "Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01 Al 110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STIR) awards. Consult the NIH website http://qrants.nih.gov/grants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1 ID)

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Philip E. Smith Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2

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SECTION I -AWARD DATA- 5R01Al110964-03 REVISED Award Calculation (U.S. Dollars) Salaries and Wages Fringe Benefits Personnel Costs (Subtotal) Materials & Supplies Travel Other SubawardsfConsortium/Contractual Costs Federal Direct Costs Federal F&A Costs Approved Budget Total Amount of Federal Funds Obligated (Federal Share) TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS YR THIS AWARD CUMULATIVE TOTALS 3 $611,090 4 $597,112 5 $581,646 $167,708 $54,168 $221,876 $7,250 $35,918 $11,050 $213,239 $489,333 $121,757 $611,090 $611,090 $611,090 $0 $611,090 $597,112 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project Fiscal Information: CFDAName: CFDA Number: EIN: Document Number: PMS Account Type: Fiscal Year: IC CAN Al 8472350 Allergy and Infectious Diseases Research 93.855 1311726494A1 RAl110964A P (Subaccount) 2016 2016 $611,090 2017 $597,112 2018 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project NIH Administrative Data: PCC: M51C / OC: 414E / Released (b)(6) 05/05/2017 Award Processed: 05/05/2017 07:00:56 PM SECTION II- PAYMENTfHOTLINE INFORMATION- 5R01Al110964-03 REVISED For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS- 5R01Al110964-03 REVISED This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. c. 45 CFR Part 75. d. National Policy Requirements and all other requirements described in the NIH Grants Page-3

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Policy Statement, including addenda in effect as of the beginning date of the budget period. e. Federal Award Performance Goals: As required by the periodic report in the RPPR or in the final progress report when applicable. f. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. (See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain references cited above.) Research and Development (R&D): All awards issued by the National Institutes of Health (NIH) meet the definition of "Research and Development" at 45 CFR Part§ 75.2. As such, auditees should identify NIH awards as part of the R&D cluster on the Schedule of Expenditures of Federal Awards (SEFA). The auditor should test NIH awards for compliance as instructed in Part V, Clusters of Programs. NIH recognizes that some awards may have another classification for purposes of indirect costs. The auditor is not required to report the disconnect (i.e., the award is classified as R&D for Federal Audit Requirement purposes but non-research for indirect cost rate purposes), unless the auditee is charging indirect costs at a rate other than the rate(s) specified in the award document(s). An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the System for Award Management (SAM). Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://grants.nih.gov/grants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01Al110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://grants.nih.gov/grants/policy/awardconditions.htm for additional award applicability information. In accordance with P.L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OD-08-033 and the Public Access website: http://publicaccess.nih.gov/. In accordance with the regulatory requirements provided at 45 CFR 75.113 and Appendix XII to 45 CFR Part 75, recipients that have currently active Federal grants, cooperative agreements, and procurement contracts with cumulative total value greater than $10,000,000 must report and maintain information in the System for Award Management (SAM) about civil, criminal, and administrative proceedings in connection with the award or performance of a Federal award that reached final disposition within the most recent five-year period. The recipient must also make semiannual disclosures regarding such proceedings. Proceedings information will be made publicly available in the designated integrity and performance system (currently the Federal Awardee Performance and Integrity Information System (FAPIIS)). Full reporting requirements and procedures are found in Appendix XII to 45 CFR Part 75. This term does not apply to NIH fellowships. Treatment of Program Income: Additional Costs Page-4

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SECTION IV- Al Special Terms and Conditions - 5R01Al110964-03 REVISED The Research Performance Progress Report (RPPR), Section G.9 (Foreign component), includes reporting requirements for all research performed outside of the United States. Research conducted at the following site(s) must be reported in your RPPR: San Pya Clinic, BURMA lnstitut Pasteur du Cambodge, CAMBODIA Primate Research Center at Bogor Agricultural University, INDONESIA Conservation Medicine, Ltd, MALAYSIA King Chulalongkorn Memorial Hospital, THAILAND Hanoi Agricultural University, VIETNAM REVISED AWARD: This Notice of Award is revised to provide approval for collaboration with the Wuhan University School of Public Health (CHINA) in accordance with the request submitted by Aleksei Chmura, Ecohealth Alliance, Inc. on October 6, 2016. Supersedes previous Notice of Award dated 7/26/2016. REVISED AWARD: This Notice of Award is revised to provide approval for collaboration with the Wuhan University School of Public Health (CHINA) in accordance with the request submitted by Aleksei Chmura, Ecohealth Alliance, Inc. on October 6, 2016. Supersedes previous Notice of Award dated 7/26/2016. No funds are provided and no funds can be used to support gain-of-function research covered under the October 17, 2014 White House Announcement (NIH Guide Notice NOT-OD-15-011). Per the letter dated July 7, 2016 to Mr. Aleksei Chmura at EcoHealth Alliance, should any of the MERS-like or SARS-like chimeras generated under this grant show evidence of enhanced virus growth greater than 1 log over the parental backbone strain you must stop all experiments with these viruses and provide the NIAID Program Officer and Grants Management Specialist, and Wuhan Institute of Virology Institutional Biosafety Committee with the relevant data and information related to these unanticipated outcomes. This Notice of Award (NoA) includes funds for consortium activity with: • Wuhan Institute of Virology - CHINA awarded in the Total Costs amount of $159,122 ($147,335 Direct Costs+ $11,787 F&A Costs). Future year commitments are as follows: Year 4 Total Costs: $159,122 and Year 5 Total Costs: $159,122 • East China Normal University - CHINA awarded in the Total Costs amount of $54,117 ($50,108 Direct Costs+ $4,009 F&A Costs). Future year commitments are as follows: Year 4 Total Costs: $42,300 and Year 5 Total Costs: $32,454 Consortiums are to be established and administered as described in the NIH Grants Policy Statement (NIH GPS). The referenced section of the NIH Grants Policy Statement is available at http://qrants.nih.gov/qrants/policy/nihqps 2013/nihqps ch15.htm# Toc271265264. The written agreement with the consortium must address the negotiated arrangements for meeting the scientific, administrative, financial and reporting requirements for this grant. No foreign performance site may be added to this project without prior approval of the National Institute of Allergy and Infectious Diseases. Page-5

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Although a specific amount has been awarded for each consortium, the grantee retains standard rebudgeting authorities. ***** This award may include collaborations with and/or between foreign organizations. Please be advised that short term travel visa expenses are an allowable expense on this grant, if justified as critical and necessary for the conduct of the project. ***** Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.setectagents.gov/Regulations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbf5toc.htm). Research funded under this grant must adhere to the BMBL, including using the BM BL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. If work with a new or additional Agent(s)/Toxin(s) is proposed in the upcoming project period, provide: STAFF CONT ACTS o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent(s), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted, with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. The Grants Management Specialist is responsible for the negotiation, award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These Page-6

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individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Jenny L. Greer Email: (b)(6) Phone: (b)(6) Fax: 301-493-0597 Program Official: Erik J. Stemmy Email: (b) (6) Phone: ____ (b)(6) _,, SPREADSHEET SUMMARY GRANT NUMBER: 5R01Al110964-03 REVISED INSTITUTION: ECOHEALTH ALLIANCE, INC. Budget Year3 Year4 Years Salaries and Wages $167,708 $167,708 $167,708 Fringe Benefits $54,168 $54,168 $54,168 Personnel Costs (Subtotal) $221,876 $221,876 $221,876 Materials & Suoolies $7,250 $7,000 $3,500 Travel $35,918 $35,918 $35,918 Other $11,050 $9,800 $9,400 Subawards/Consortium/Contractual Costs $213,239 $201,422 $191,576 TOTAL FEDERAL DC $489,333 $476,016 $462,270 TOTAL FEDERAL F&A $121,757 $121,096 $119,376 TOTAL COST $611,090 $597,112 $581,646 Facilities and Administrative Costs Year 3 Year4 Years F&A Cost Rate 1 44.1% 44.1% 44.1% F&A Cost Base 1 $276,094 $274,594 $270,694 F&A Costs 1 $121,757 $121,096 $119,376 Page-?

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RPPR FINAL A. COVER PAGE Project Title: Understanding the Risk of Bat Coronavirus Emergence Grant Number: 5R01Al110964-03 Project/Grant Period: 06/01/2014- 05/31/2019 Reporting Period: 06/01/2015 - 05/31/2016 Requested Budget Period: 06/01/2016 - 05/31/2017 Report Term Frequency: Annual Date Submitted: 05/13/2016 Program Director/Principal Investigator Information: Recipient Organization: PETER DASZAK , BS PHD ECOHEAL TH ALLIANCE, INC. ECOHEAL TH ALLIANCE, INC. Phone number: I (b) (6)1 460 W 34TH ST Email:! (b) (6)] 17TH FLOOR NEW YORK, NY 100012320 DUNS: 077090066 EIN: 1311726494A1 RECIPIENT ID: Change of Contact PD/Pl: N/A Administrative Official: Signing Official: ALEKSEI CHMURA ALEKSEI CHMURA 460 W 34th St., 17th Floor 460 W 34th St., 17th Floor New York, NY 10001 New York, NY 10001 Phone number: (b)(6)~ Phone number: (b)(6)~ Email:1 (b)(6) Email:1 (b)(6) Human Subjects: Yes Vertebrate Animals: Yes HS Exempt: No Exemption Number: Phase Ill Clinical Trial: hESC:No Inventions/Patents: No RPPR Page 1

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RPPR FINAL B. ACCOMPLISHMENTS 8.1 WHAT ARE THE MAJOR GOALS OF THE PROJECT? Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-CoVs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel CoVs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2} how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses; screen people who have high occupational exposure to bats and other wildlife; and examine the genetics and receptor binding properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-CoVs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological opportunity for contact between species so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Gen bank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecological and phylogenetic traits to explain patterns of viral sharing between species. We will test for positive selection in market vs. wild-sampled viruses, and use data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. 8.1.a Have the major goals changed since the initial competing award or previous report? No B.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? File uploaded: Year 2 NIAID CoV Report Final.pdf B.3 COMPETITIVE REVISIONS/ADMINISTRATIVE SUPPLEMENTS For this reporting period, is there one or more Revision/Supplement associated with this award for which reporting is required? No B.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? File uploaded: Year 2 NIAID CoV Report Professional Development.pdf RPPR Page2

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RPPR FINAL B.5 HOW HAVE THE RESULTS BEEN DISSEMINATED TO COMMUNITIES OF INTEREST? 1) Conference and University lectures: Pl Daszak, and Co-investigators Shi, Epstein, Oliva I, Ge, and Zhang gave >100 invited University and Conference lectures including Forum on Microbial Threats (National Academies of Science), Symposium at Ecole du Val-de-Grace in Paris, Leadership Roundtable at Concordia University Montreal, 1st annual Global Pandemic Policy Summit at Texas A&M Univ., Intl. Conf. of the Wildlife Disease Association in Australia, Intl. Conf. of Conservation Biol in Montpellier France, Michigan State University, Duke University, WDA, ISID conference, Zoological Society of London Symposium, Future Earth meeting, North American Bat Research Symposium, and others that included specific discussion of the current project and results. 2) Agency and other briefings: Pl Daszak and Research Technician Dr. Guangjian Zhu introduced this project to potential collaborators within the following agencies: Forestry Dept of Peoples' Republic of China, FAO, TNC, TRAFFIC, China CDC, and TA Foundation in Beijing China in meetings (2015) and also at presentations at the first Wildlife and Public Health Workshop in China (2016) co-hosted by EcoHealth Alliance, the State Forestry Administration of China, and China CDC. 3) Public outreach: Pl Daszak presented this work to members of the NIH, NSF, DoD, IUCN, EPA, and the general public, at an EcoHealth Alliance meeting hosted by the Cosmos Club, Washington D.C. (2015); Pl Daszak and Co-investigator Zhu reported on this project at a Wildlife Trade and Public Health Seminar, Beijing (2016); Pl Daszak introduced this project in a lecture on Pandemics at a New York Academy of Science Panel (2016); Co-Pl Y-Z Zhang presented project and results-to-date to department heads and senior researchers at Infectious Disease Departments of four Yunnan Hospitals (2015) B.6 WHAT DO YOU PLAN TO DO DURING THE NEXT REPORTING PERIOD TO ACCOMPLISH THE GOALS? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. - Given the reduced amount of wildlife in the local markets within Southern China, and the continued expansion of the Chinese wildlife trade within SE Asia, we would like to conduct short field trips to assess markets, identify wildlife in them, and sample species of bats and other high-risk hosts in countries that neighbor China (Myanmar, Vietnam, Cambodia, Lao PDR) and others that supply wildlife to the international trade to China (Thailand, Malaysia, Indonesia. EcoHealth Alliance has other activities in these countries which would provide leverage to reduce costs of fieldwork, and samples would be tested in Wuhan, China. - Following the successful collection of ethnographic interviews and focus groups in Year 2, we will be analyzing the qualitative data collection from Years 1 and 2. - Finalize and conduct survey collection tool for a network study of wildlife farmers using a questionnaire to characterize and map the wildlife value chain. - After the success of our pilot studies in Year 2, we will continue targeted (at individuals with high risk of exposure to bats), integrated behavioral and biological survey work in Yunnan and expand to Guangxi and Guangdong provinces. - We will commence our anonymized, surveillance data collection from acutely ill hospital in-patients who satisfy syndromic eligibility criteria; have complete medical records; non-normative laboratory confirmed diagnostic results; and suspected acute viral infection. Eligibility criteria are: (a) suspected acute viral infection; (b) fever> 38'C, and (c) presenting symptoms of at least one of the following: •Encephalitis of unknown origin •Hemorrhagic fever of unknown origin •Respiratory disease olnfluenza-like illness (Ill) oSevere Acute Respiratory like Illness (SARI) •Rash •Diarrhea Some patients with particular infections such as with HIV, HCV, and HBV, may be excluded from the study on that basis. Hospital surveillance has the advantage of monitoring an acutely ill population. Anonymized, passive hospital surveillance allows for data collection and viral testing from all eligible hospital patients thereby limiting population sample bias and increasing the likelihood of identifying positive cases. The strengths of this approach are enormous: an unbiased patient population; prospectively collected, anonymized patient data; a low resource effort with a high efficiency design; and impactful research potential for both case series and case control studies. We have already secured approval from the Institutional Review Boards of the Wuhan School of Public Health and Hummingbird IRB. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. Future steps to optimize the model of role of species diversity in CoV emergence risk will include: - Test and implement our respondent-driven survey to collect specific data on the diversity, abundance, and turnover of species along the wildlife trade network in south China. - Model viral mixing across the full range parameters found along the wildlife trade network to identify the trade nodes with highest mixing potential. This will include a network analysis of market facility/site connectivity including wild harvest sites, wildlife farming operations, transit holding facilities, and small and large wildlife markets. - Phylogeographic study of bat-CoV to better understand the geographic distribution and evolution of bat-CoV genetic diversity in south RPPR Page3

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RPPR FINAL China. - Phylogeographic study of bat host (Rhinolophus) species to assess the connectivity of bat populations and infer their historical movements and demographic history to improve our understanding of CoV transmission among bat populations in southern China. Preliminary sequences data has been generated and will be completed and analyzed. - Cophylogenetic analyses of bat host and CoV phylogenies to assess frequency of cross-species transmission. Comparison of Alphaand Beta-CoV cophylogenetic patterns building on Year 2 analyses using published sequences and also including Spike gene and additional sequences obtained in Year 2. - Test and implement our respondent-driven survey to assess diversity, abundance, and turnover of species along the wildlife trade network. - Examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes; - Parameterize mathematical models that predict CoV evolutionary and transmission dynamics - Continued surveillances of SARS-like CoVs and lineage C betacoronaviruses (MERS-related CoVs) in Southern China; - Full-length genome sequencing and evolution analysis of SARS-like coronaviruses identified from different bat species and different geographical locations across China; - Full-length genome sequencing and evolution analysis of Lineage C betacoronaviruses identified from different bat species and different geographical locations across China; - Full-length genome sequencing and evolution analysis of HKU9-related and HKU10-related bat coronaviruses in China; Specific Aim 3: Testing predictions of CoV inter-species transmission. The following experiments will be undertaken in Year 2: - Humanized mice with human ACE2 receptors will be infected with WIV1 and the two rescued chimeric SARS-like coronaviruses to determine the tissue tropism and pathogenicity of bat SL-CoV - Isolation of novel bat coronaviruses. Live virus or pseudovirus will be used to infect cells of different origin or expressing different receptor molecules. Spillover potential for each isolated virus will be assessed. - An infectious clone of full-length MERS-CoV will be constructed using reverse genetic method. Using the S sequence of different MERS-related viruses identified from Chinese bats, the chimeric viruses with S gene of bat MERS-related coronaviruses and backbone of the infectious clone of MERS-CoV will be constructed to study the receptor usage and infectivity of bat MERS-related coronavirus. - Surveillance of infection in human populations by SARS-like CoVs. This work will be performed at locations in Yunnan, Guangxi, and Guangdong provinces, in previously identified areas with human populations of high risk of exposure to bats. PCR and ELISA will be used, respectively, for detection of viral replicase gene and antibodies against the viral nucleocapsid protein. RPPR Page4

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B.2 (Year 2 NIAID CoV Report Final.pdf) 8.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? 1R01Al110964 Year 2 Report Pl: Daszak, Peter Year 1 Report: Understanding the Risk of Bat Coronavirus Emergence Award Number: 1R01Alll0964-02 ************* Section 8: Accomplishments B.1 What are the Major Goals of the Project Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome {MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-CoVs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel Co Vs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2) how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses; screen people who have high occupational exposure to bats and other wildlife; and examine the genetics and receptor binding properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-CoVs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological opportunity for contact between species so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Genbank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecological and phylogenetic traits to explain patterns of viral sharing between species. We will test for positive selection in market vs. wild-sampled viruses, and use RPPR Page5

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. B.la Have the major goals changed since the initial competing award or previous report? No. B.2 What was accomplished under these goals? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife inter/aces In year 2, we continued and expanded the qualitative research begun at the end of Year 1. In addition, a community based integrated biological behavioral surveillance system was developed and pilot tested to identify specific animal exposure risk factors associated with biological evidence of exposure to SARSlike CoV (i.e., seropositive status). QUALITATIVE RESEARCH Targeted, in-depth ethnographic interviews were conducted with 47 individuals (18 women; 29 men) in rural Southern China where wildlife trade routes have been documented. Yunnan, Guangxi and Guangdong provinces were specifically selected for study because they have large wildlife populations, a diversity of wildlife species and numerous live animal markets. Individuals who were 18 years of age or older and who were able to provide informed consent were eligible to participate. Twenty-three (49%} in-depth interviews were conducted in Yunnan province at nine different sites, 24 (51%) in Guangxi province at six different sites. In addition, one focus group was conducted in Guangxi. The study was approved by the Institutional Review Boards of the Wuhan School of Public Health and Hummingbird IRB. Recruitment sites in each province included forested areas or preserves, wildlife farms, hunting areas, wildlife restaurants, live animal markets, caves where people dwell or collect guano and residential areas/farms near known bat caves or roosts. Participants were recruited primarily through local contacts developed as part of wildlife conservation and health research conducted by team members over the past decade. Contacts including wildlife conservationists and researchers, local government health outreach workers and wildlife farmers facilitated introductions and provided referrals. To achieve a sample with sufficient representation of categories of interest, participants were recruited using RPPR Page6

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter purposive sampling, which provides minimum quotas in terms of sex, age and wildlife exposure setting (e.g., live animal market, forest preserve). The five core themes that guided the in-depth discussions are: 1) human-animal contact, 2) unusual illness experience and response, 3) socioeconomics and daily living, 4) biosafety and 5) human environments and movement/travel. An ethnographic interview guide was developed with examples of questions that could be asked for each theme. In addition, field based participant-observation was ongoing throughout the study and involved observing and talking informally with people in their own natural setting. Field notes were maintained of these ongoing observations and discussions. Table 1: Species Observed in Wetmarkets in Guangdong Province from 2015 - 2016 Interviews were conducted between March Genus species Prionailurus bengalensis Nyctereutes procyonoides Sus scrota Lepus sinensis Arctonyx co/loris Hystrix brachyura Marmota sp. Common Name Leopard Cat Raccoon Dog Wild Boar Chinese Hare Hog Badger Porcupine Marmot and June 2105 by 10 trained interviewers, none of whom had social science training. Interviewers conducted between one and 22 interviews; three interviewers conducted two thirds of all interviewers. Interviews lasted between 20 and 60 minutes, and were taperecorded and transcribed verbatim before they were translated into English. All participants received cooking oil valued at US$10 in appreciation of their time. Rhizomes sinensis Erinaceus sp. Mustela putorius Muridoe Myocastor coypus Vulpes sp. Mustela sibirica Pagumo larvata Fe/is catus Con is lupus familioris Cervinae Ovis aries Capra sp. Rotus norvegicus Bamboo Rat Hedgehog Ferrets Rat (species unknown) Nutria Fox Siberian weasel Masked Palm Civet Domestic Cat Domestic Dog Sam bar Deer Sheep Domestic Goat Common Rat The data are currently being coded and an analytic database is being constructed. Initial insights include observations by a number of participants, especially those who are older, that there has been a decrease in wildlife in the surrounding environment. This decrease is attributed to many factors including infrastructure development. The government has invested resources to build new roads and renovate local infrastructure with the intention of increasing tourism. This has reduced forested area. Observations by research staff in live animal markets in Guangzhou found wildlife to be plentiful (see Table 1), although no bats were seen for sale during the observation period. In contrast, wildlife was not found in live animal markets at the sites we visited in either Yunnan or Guangxi. This is a change from previous research visits to the same or similar communities, when bats, rodents and wild boar could be found. Locals in Yunnan and Guangxi attribute the change to conservation law enforcement. The success of conservation enforcement may have moved hunting and trapping underground and made the capture of local wildlife less economically feasible than other income generating activities. RPPR Page 7

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Preliminary analyses are underway. Three specific studies in support of Specific Aim 1 are being developed: the changing wildlife trade in Southern China, the economics of wildlife farming, and zoonotic disease risks resulting from a rapidly changing wildlife trade. INTEGRATED BIOLOGICAL BEHAVIORAL SURVEILLANCE PILOT STUDY Currently, mechanisms of zoonotic viral spillover are unknown. In order to evaluate potential risk factors, it is necessary to measure both exposure and outcome data. Therefore, a behavioral risk survey was developed that assessed both animal exposure and experiences of unusual illness both during lifetime and in the past 12 months. In addition, participants were requested to provide serum to test for previous exposure to SARS-like CoV. The integrated surveillance was pilot tested in October 2015 among residents living near bat caves or roosts where SARS-like-CoV has been previously detected in the bat population in Jinning County, Yunnan. Please view the full survey here: https://www.dropbox.com/s/sv62neywuvl027r/Questionnaire%20Complete.docx?dl=O 4% Occupation 9% \ ■ Farmer ■ Student Of 218 participants, 139 (64%) were women and 79 (36%) were men, with a mean age of 48 (range: 12-80). Most reported being farmers (87%, and see chart to left); a majority were long term residents (97%). Animal exposures in the past year were extensive, including ■ Other general (e.g., buying live animals at markets (61%]) and intimate (e.g., being scratched or bitten [9%], slaughter (38%]). In fact, two-thirds of participants reported handling recently killed animal parts and 2 out of 5 reported slaughtering animals. Only 20 (9%) participants reported known exposure to bats. Standardized syndromic case definitions informed questions concerning unusual illness experience (e.g. severe acute respiratory infections [SARI], influenza-like illness [ILi]). Lifetime, 12 month and unusual illness experience in family for the past 12 months were assessed for all participants. In the past year, SARI was reported by 4 (2%) respondents and for 4 additional family members. Table 2 provides data for all unusual illness experience assessed. None of the participants were found to be seropositive for SARSlike CoV. Table 2. Unusual Illness Experience . I . . I . ' I ' . ' I , . . Severe Acute Respiratory Infections (SARI) 15 (6,9%) 4 (1.8%) 4 (1.8%) Influenza Like Illness (ILi) 54 (24.8%) 16 (7.3%) 26 (11.9%) Encephalitis 19 (8.7% 4 (1.8%) 3 (1.4%) Hemorrhagic Fever 0 (0.0%) 0 (0.0%) 0 (0.0%) Fever with Diarrhea /Vomiting 12 (S.S.%) 2 (0.9%) 3 (1.4%) Fever with Rash 2 (0.9%) 2 (0.9%) 3 (1.4%) RPPR Page 8

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Although the sample size was small, animal exposures among those who reported unusual illness experiences in the past 12 months were evaluated. Of the four respondents who reported SARI symptoms, 75% reported: raising animals, animals in the home, preparing recently killed animals and buying live animals; 50% reported slaughter. Among the 16 respondents who reported ILi symptoms, 12 (75%) reported handling/preparing recently killed animals, 11 (69%) Handling live animals or having animals in the home, 10 (63%) reported slaughtering/killing animals or buying live animals at wet market, 9 (56%) raised live animals, 7 (44%) reported a pet, and 1 (6%) reported animal feces near food or eating animal touched or damaged food, hunting, or eating raw/undercooked animal products. Finally, among the four respondents who reported encephalitis symptoms, 3 (75%) reported hunting, handling or raising animals, 2 (50%) reported animals in the home, 1 (25%) reported having animals as pets, slaughtering/killing animals, or having bought live animals at wet market. Respondents were asked about the source of their unusual illnesses. None reported any kind of animal exposure as a potential source of infection and most stated they had no idea how they had become infected. However, when asked about potential behavior changes made at live animal markets in the last 12 months, participants reported a great deal of change. In particular, respondents reported buying live animals less often (38%), only buying farmed wildlife (54%) or buying meat at the supermarket (23%). (See Table 3). Table 3: Behavior Change at Wet Market in the last 12 months Behavior N (%) The results of this pilot study conducted with a largely female farmer population found high Wear a mask Wear gloves Wash hands Sometimes shop for meat at supermarket Buy live animals less often Buy only farmed wildlife 4 5 80 30 so 71 (3.0) (3.8) (60.6) (22.7) (37.9) (53.8) levels of unusual illness, as well as high levels of exposure to animals. There was a notable lack of knowledge of animals' ability to transmit infection. Despite this lack of knowledge, there may be a sense of unease about animal exposures, given the fairly dramatic behavior changes reported at live animal markets. The finding of a reduction in wildlife purchase may be due to sensitivity to the legality of wildlife trade, biasing respondents towards not admitting purchasing wildlife. Although, there were no participants seropositive for SARS-like CoV, serological data may add support to the findings from self-reported syndromic surveillance, once serological assays No are optimized. longer buy wildlife at wet 39 (29.5) market In preparation for full implementation of the integrated biological behavioral surveillance, the survey has been programmed as an application for use on either a mobile device or computer. Electronic data collection will facilitate survey implementation in the field and quality control of the data being collected. Four field team leads were trained on behavioral survey data collection, data collection technologies (the tablet application) and analysis. Nucleic acid test results of human biological samples Testing High-Risk Human Populations for Coronavirus Infection RPPR Page9

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Surveillance of CoV infections in human populations by SARS-like CoVs was significantly expanded in Year 2, including both custom-built ELISA serology (an assay developed by the Wuhan Institute of Virology to test antibodies against the N protein of SL-CoV) and PCR detection of viral RNA. Serological test for SL-CoV antibodies in human samples from }inning, Yunnan Province In order to assess past exposure to bat CoVs, 223 human sera samples were collected in villages in proximity to the bat habitat from which two SL-CoVs with potential for interspecies infection, WIVl and WIV16, were discovered in our previous research. An ELISA developed by the Wuhan Institute of Virology was used to test antibodies against the N protein of SL-CoV. A number of human specimens generated high OD values and neutralization test to WIVl and WIV16 was then performed. These findings are encouraging; however, no neutralization antibodies were detected. In Year 3, we will continue to validate and optimize these ELISA assays and other serological tests to obtain data on past CoV exposure. PCR test for CoV Nucleic Acid in human samples from several Provinces We tested 405 individual human samples for CoV RNA to identify evidence of active infection in human populations and to obtain sequence data on strain variation. Individual samples (4 each) were pooled prior to nucleic acid extraction then tested using PCR. When a group tested positive, we then conducted the confirmation test in the individual samples. One single sample (14XN611) from someone who had identified as having had a fever and suffered both a cough and headache in the past 7-days was then identified to be positive for HCoV-HKUl. The low number of PCR detections in human specimens is not unexpected, and will be improved in Year 3-5 by better targeting syndromic individuals for specimen collection and continuing to optimize PCR assays. Refined serological assays (above) will provide sufficient data to assess past exposure to specific CoV lineages, and optimizing of PCR detections will allow for more CoV positive human sequences moving forward. Bat CoV PCR detection and sequencing from live-sampled bat populations We collected 1,714 anal swab samples, 677 fecal samples, 53 blood samples, and 38 serum samples from 15 bat genera in Guangdong, Yunnan, Sichuan, Hubei, Hunan, Guizhou, Guangxi provinces (Table 4). Table 4 Bat Samples collected for CoV surveillance in 2015 Sample date Sample location Anal Fecal Blood Serum Mar. 2015 Huidong, 69 -- -- -- Guangdong Jun. 2015 Guangdong 495 -- 12 -- Apr. 2015 Menglun, Yunnan 51 -- -- -- May 2015 Jinning, Yunnan -- 193 -- -- May. 2015 Mojiang, Yunnan 93 -- -- -- Oct. 2015 Jinning, Yunnan 30 -- -- -- RPPR Page 10

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Jingna, Yunnan 15 15 13 13 Dec, 2015 Miaoxin, Yunnan 42 28 25 Jul, 2015 Zigong, Sichuan 128 -- -- -- Aug,2015 Hubei 332 Sep,2015 Xianning, Hubei 95 Aug,2015 Jishou, Hunnan 204 Aug-Sep, 2015 Tongren, Guizhou 438 Dec,2015 Longzhou, Guangxi 191 Total 1714 677 53 38 We tested 2,256 samples for CoV RNA and 280 tested positive. The total positive rate is 12.4% (Table 5). Diverse alphacoronaviruses related to Bat CoV lA, lB, HKU2, HKU6, HKU7, HKU8 and HKUlO were identified; SARS-like coronaviruses were detected in Rhinolophus bats in both Yunnan and Guangdong (Fig 1). Novel lineage B betacoronaviruses more distantly related to SARS-CoV than other SL-CoVs were detected in Vespertilo superans in Sichuan. HKU4-related coronaviruses were found in Tynolycteris pachypus in Guangdong and Guangxi while HKU5-related coronaviruses were found to be highly prevalent in Vespertilio superans in Zigong, Sichuan (41 bats out of 128 tested positive). RPPR Page 11

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Table 5 Test result of bat CoV surveillance in 2015 - 12% positive {280/2,256) Yunnan Guangdong Hubei Sichuan Guangxi Guizhou Hunan Total Bat species No. positive/No. tested Rhino/ophus spp. 47/98 12/103 16/225 8/63 83/489 Hipposideros spp. 0/35 0/51 26/152 0/131 0/91 26/460 la io 0/3 0/3 Pipistrellus spp. 1/1 0/19 0/2 0/4 1/26 Miniopterus spp. 6/7 34/83 2/6 42/96 Eonycteris spp. 0/3 0/3 Vespertilio superans 41/128 41/128 Myotis spp. 1/38 0/70 0/35 1/143 Taphozous spp. 0/25 0/1 0/26 Tynolycteris pachypus 8/25 27/191 35/216 Scotophilus kuhlii 1/1 1/1 Eptesicus f uscus 0/1 0/1 Tadrida spp. 0/5 0/5 Barbastella 0/1 0/1 Nyc/atus velutiaus 0/10 0/10 Fecal samples 28/468 22/180 50/648 Sub-total 82/637 56/326 48/332 41/128 27/191 18/438 8/204 280/2256 RPPR Page 12

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report A B r--1 0.1 0...-.-on>n•IINo- I 8 DQ666J1R (;0Vl-8(11l.M1n1VJ)CCIU> ahrab<r11 CoVH6WM1111opccrus pu.all11> ( 10) CoV1-8621/M,n,or<<N< >ol-.-c,bcto, • 10) 9$ CoV1-1i6S0-2S ~i1n10p1tn1s ~hml><r$i __ _._.., B.tf..,..on,vrru.1ol,\ Et:UOIJII CuV 1-WJ MJJ.11up1.._ .. pu;,llu. ( ll • .._ ____ &HOfOIUiv1ru.,-IIKU1 0()249216 CoVZ•N>H M1111opcm,, 11<htt1l><r11 B.il•<"""1.lv11u_..lll<l/8 [)()!49".28 Cu\'2-!Ui6.S.l1Mtruopt<ru• pu>ll.tu. ,--------<~ llippu~tdcr<b-'b.tH;:orona,IJ'U),o-ltKUIO JQ989266 Rct1«11u .. 1>o1-a,mnavirus-HKUIO J()<.189271 -----c·oVl,X1~0 ~«,1c,phll11> "llhh 1-----t\•V2-k717 '-1)\tllll rldt..1ll l'------ 1 8..ll-cCllOn.mru,-lll{t:6 DQ249~~ Myo1is d3-.1d11 c:orona,uu, oolatc XYl'\1d';O RllRp KJ-5"99'11 ~---("o\. 2--.:7,),t, Rh,n,,l,>phu~ puutlu, Co\..:! )il;"S'"' RJ11nolnrtua .1f11111 8.11--t-oron,nm,,-.IIKl Z LI :o_lC)fl .._ ____ 13J1-roron.-.rn .. 11KUS,I Pr06S509 ,.__ _________ Hunl•n•cutClftl\ll'IIS>IIKUI AY88400 I .____. 0 I llal·COrooa,uus-HKU,-2 1.tO!lSS14 B.i1-S/\RS-tunlll,1>1n,"'HKt J DQOZ:!.i.05 CnV1•~361 Rlunol11phm 'lllnu:u> 8Jl•SARS-cor0n11\10J.i,ok.fl OQ412041 rnv2-~,.,. tuuootophu., pw;,Uu, CuV2-~S;lk Rhm<llopb111 an,cus <S) 8:11-SAR$,"Of'u.1lo, 1rua.-Rpl DQ0? 1615 W367 KC3ll1006 R.,SIICOl4 K("88100S II SAR.~-C'<>IONl\;ru,.BJOI AY271'4N8 6 SARS-co1ono"ru..,GZ02 AY390SS6 SARS•coro,-irus-SZ3 AYJ~416 B1l•Cormi;ivm1,-I B [)()li663J o\ :. 196$: M101<1114cru., ,pp (51 ( 'uV.?·CJl'l.1 P11l1strl.'llu,11:1.1H ...1tdhl$ Ba1-e<llona, lnL~-1AEl'420 I Jg cov;. ·q66 Rhinoloohu,.,oo ( 5, 8a1-cornna, iru<-Hl<U7 1)0149226 I C'tlV2°71Hh \l1n1op1cr1L,Spp 9 c:uV2-~0ij' :1 Ba1-0or11nuvinis-HKU8 DQ249228 ,-----8:ll•Cl)r0113Vl'IS•HKU6 00249224 .-----coV2-80061HioPOGidero•Soo ,---~11-0V2-7960-6tl Rhinoloobusson (IO> o, V2•K I ti5 Ncthicus ,1ohakanus .._ __ -i ,Hinonsicltros-h.11-cororuvirus-111< UI O J(.)989266 IO Rouscnus-b:11-coronavirus-Hl<UI 0JQ989l71 '----Mvotuda\ldncoronavims ,..,latcXYMd KF569991 lb1-coro1Ul\ U'U>•HJ<Ul E1203067 ----1-{.'11\2-7~61l-~fillth1nolooh11~ !OD 45 \ ( o\ 1-7119'--\i Rh1nolophu••PP ('o\2-7'.11-1.,.1 Rhh•1l~phu•<pp ~![::::::::_8;;.3l•COIOIIOl'IIUS•HKU4• I EF065S05 &1-toron.1,· uu.-llKUS-1 cl'06SS()') -------11u111an-i:011>11.11 lrn,-I IKUI AY88400 I 1331-coron.ovlrtL<-lll-.l'0-2 H'06S~ 14 'o\2-7.V6-U Rh1nolooht1~hOO ((, I Co\ z. 79.31-L--2I !Uumlophusspp ll•1-SARS-curonaviru,1-IIKU3-1 DQOll30S llai-SARS-coro,~,, inis-Rfl DOl 12042 lla1•SARS-r111011n, ,ru .. Rpl 1)()(17161$ Co \11-9400/ Rhmolophus s1nku1 c SI 3367 KC881006 RsSHCO 14 KC88 I 005 s SARS-coronu\'irus-BJOI /\Y278488 6 'ARS-corona\'inl~-OZ02AY390SS6 S~orona,irus-SZJ AY304486 Pl: Daszak, Peter Fig 1 Phylogenetic analysis of partial RdRp gene of CoV (440-nt partial sequence). Co Vs identified in 2015 are named by the sample numbers. Sequence amplified from samples co-infected with two CoV strains are indicated in red. (A) CoVs detected in Guangdong. (B) CoVs detected in Yunnan. RPPR Page 13

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Cophvloqenetic analysis of CoV host switching We completed preliminary cophylogenetic analysis of bat host - CoV sequences using data published in the literature and available on Genbank. Two figures from these analyses are highlighted below (Figs 2 and 3) and these methods are currently being extended using partial RdRp CoV and bat mitochondrial DNA sequences from a large number of bat specimens found CoV positive in Year 2 (Table 5, above). ---- - -------------- .:- _. Figure 2: Tanglegram depicting the pattern of infection of bats (and outlier mammalian hosts) by Co Vs. The CoV tree was reconstructed from DNA sequences available in GenBank (partial RdRp gene} using Bayesian inference (MrBayes). The topology of host tree was reconstructed using the mammal and bat phylogenies available in Asher & Helgen (2010} and Agnarsson et al. (2011), using methods our group has previously applied to bat parasite cophylogenetic analyses (Lei and Oliva! 2014). Both ParaFit (ParaFitGlobal = 64957.61, p-value = 0.001) and PACo (m2 = 366.44, p-value = 0.013) provided evidence for significant global congruence between the two topologies, and evidence for coevolution. Lines connecting taxa indicate host-CoV associations. Red lines indicate significant host-CoV associations as indicated by ParaFit (p ~ 0.05, 999 permutations). CNJ I I ..... ~:. I LI 0,,.,,. =.,., I .,,. - -= ,_,.. .. ""' I ...... CN.!'Wf ~ I ---- --! Q\t_ly I T~'M• ~ ~-=- :::.;;:= - QN <H - - -I "' I ......... ........... "'-"'~ ftn¥01.U - ,:,,,,,_,w ·-- /4 <N4 ............ <N,..k ""'"" --c.N.c..iw ,_ ""'-" &~ L~ Figure 3: Reconstruction of one of 3 potentially optimal solutions of reconciled host-CoV trees recovered from a Jane analysis. Black and blue lines represent the host and CoV trees, respectively. For each solution, the number of co-speciation events inferred by Jane was always significantly greater than expected by chance. Jane inferred 4 co-speciation events (hollow colored circles), 1 duplication (solid RPPR Page 14

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter colored circle), 14 host switches (solid colored circle with arrow), 0 loss and 0 failure to diverge. Our findings demonstrate co-speciation alone is not sufficient to explain the observed co-phylogenetic pattern and several host switches can be specifically identified. This is the case even if a significant global signal of co-speciation has been detected. This work highlights, the need for these types of detailed cophylgoenetic analyses to best explain the evolutionary history and host-switching of bat-CoVs. References cited for the above analysis: Agnarsson, I., Zambrana-Torrelio, C.M., Flores-Saldana, N.P. & May-Collado, L.J. (2011) A time-calibrated species-level phylogeny of bats (Chiroptera, Mammalia). PLOS Currents, 3:RRN1212. Asher, R.J. & Helgen, K.M. (2010) Nomenclature and placental mammal phylogeny. BMC Evolutionary Biology, 10, 1-9. Lei BR, Oliva! KJ (2014) Contrasting Patterns in Mammal-Bacteria Coevolution: Bartone/la and Leptospira in Bats and Rodents. PLoS Neg/ Trap Dis 8(3): e2738. Market Characterization Model Parameterization Our ongoing observational research and mapping of farms and markets suggests that rapid changes in the market and regulatory environment are changing the nature and location of the wildlife market trade. The nexus of the wildlife trade and the potential hotspots of interspecies viral mixing is now in many cases in animal storage facilities and transport between high-volume customers. To define realistic parameters for intermixing wildlife species in areas of high potential mixing, we have developed a preliminary survey and sampling protocol to assess these values as animals move along the value chain - through these storage facilities - using respondent-driven questionnaires to follow and sample along the wildlife trade network and reveal hidden nodes and sites of intermixing of species. We have expanded our intermixing modeling framework to incorporate the variations along this value chain, where the diversity, abundance, residence time, and contact rates between species change as animals move through the trade network. Specific Aim 3: Testing predictions of CoV inter-species transmission. In Year 2, we continued surveillance for novel SARS-like CoVs from bats in Yunnan and Guangdong provinces and obtained full genome sequence for 11 CoV isolates. Full genome analysis of these CoV isolates was completed, including phylogenetic and recombination analyses. Importantly, recombination analysis of the full-length SL-CoV genome sequences from a single bat population revealed that frequent recombination events among different SL-CoV strains occur. Several SL-CoVs that are more genetically similar to SARS-CoV (2003) than any previously discovered were also identified from bat populations in Yunnan province. Full genome analysis suggests that an epicenter of SL-CoV occurs in rhinolophid bats and provides more insight into the evolutionary origin of SARS-CoV. Full-length genome sequencing of SL-Co Vs identified from a single bat colony To date, including preliminary data submitted for this R0l that we are now analyzing under the current funding, we have conducted 5-years of surveillance of SL-CoV in a single bat colony in Yunnan Province (from 2011 to 2015), leading to the discovery of diverse novel SL-Co Vs. Based on genotyping of these SLCoVs by the region corresponding to the receptor-binding domain (RBD) of SARS-CoVs, 11 isolates were selected and full-length genome sequencing was performed in Year 2. These SL-CoVs, including four others isolated previously from this colony, Rs3367, RsSHC014, WIVl and WIV16, are highly diversified in the S gene, but share similar sequence identity to SARS-CoV in ORFlab (Fig 4). Genomic phylogenetic analysis showed that the SL-CoVs detected in this colony are more closely RPPR Page 15

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter related to SARS-CoVs from other geographic regions, especially three isolates, WIV16, Rs4874 and Rs4231 (Fig 5). Notably, among the 15 SL-CoVs, two isolates, Rs4084 from Rhinolophus sinicus and Rf4092 from Rhinolophus ferrumequinum, are highly similar to SARS-CoV in the ORF8 region (Fig 5). Rf4092 possessed a single ORF8 of the same length (369bp) as that in civet SARS-CoV SZ3, and the sequence showed only 10 nucleotide substitution (Fig 6). The ORF8 sequence of Rs4084 is highly similar to that of Rf 4092, however in the region corresponding to the 29-bp deletion acquired in human SARS CoVs (e.g Tor2), a shorter deletion of only 5-bp is present, resulting in two overlapping ORF8s, ORF8a and ORF8b. The position of start codon and stop codon of the two ORFs were consistent with those in human strains (Fig 6). 0.9 o.es 0.8· 0.75 07 0.65· 05 04 0.3 -Rs4081 -Rs4255 -Rs4247 -Rs4237 -Rs6526 -Rf4092 -Rs4231 -\l\4V16 -Rs4874 -Rs7327 -Rs9401 R$3367 -\l\4V1 -RSSHC014 Rs4084 Query sequence: SARS-Co V SZ3 0 2,000 4/JOO 6/JOO 0JJJ0 10/JOO 12/JOO H,000 16,000 1~.000 ,rJJRO 22/RO 24,000 2fillf$J 28,0W 30/JOJ - ~w 1 SOO bp, 9:~ XlO bp, OepSt1'): On, Kirue (2..pwameter), TA 2.0 Fig 4. Simplot analysis of the 15 SL-Co Vs identified from a single bat colony in Yunnan. SARS-CoV 523 is used as query sequence. RPPR Page 16

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter , .. .. .. ,,. Human SARS Co V Tor2 • llumanSAR CoV BJOI .. Human SARS Co V GZ02 Civet SARS CoV SZ3 Civet SARS CoV PC4-227 Rs4874 WI 16 Rs42.l1 Rs7327 Rs9401 Rs3367 WI I Rs BC014 Rs4084 Rf4092 Rs/YN2013 Rs408] Rs42SS Rs672 As6526 Rs4247 Rs4237 RfNNLF31C l._--1 RftY LF 34C Rp3 Rs/GX2013 ,----LYRall C.D 2011 .....----RKU3-1 ,----Rrnl '---Rs/HuB2-0 I 3 .---- Rp/SAXlOI I Rf/JL20I2 Rfl Rf/SAX2013 Rf/Jle820I3 ,..__ _______ BM48-3 I .__ _________________ IJKU9-I Fig 5. Phylogenetic analysis of full-length genome sequences of SL-CoVs and SARS-CoVs. Isolates identified in the single investigated bat colony in Yunnan in in bold. ORFSla> ORFSb or Tor2 > ORn, oCTor2 < .... I.JOl .... .,, .. , 1.'N: .u.~TttTCJ.~'""1'--C.1,C"TT,ot.i.-'MTC'Tc-'rATCe1.C1'-Tc<"a"TATOCAC~T ... ~,. .... c..t.c«<.T<l'NeA'T't:1'A1t.T.1.u.eci 1.TC nc.u.c1.~etE::._ ...._ ... _ ..... -.............. Eco--TAeuc-. ,-,c 1:ao : I.TO UCTTtTCATTGTTIT-•G~r.1-T"TKTCl.t.wt.LGT•TGClTJCliCMTGTlH&.•-CAIXCiCTGT«ATCTUT.U.1CC AT!; ~UOJ.TCC -----·-------- GO--TACAJ.CA nG uo : 1.TG Cnt;TCATTGTTTT• .. ·GJ.CTTiiT,..._~U.TirC~ .. yi;,cnACGCAtTGtTVU•~lGCCX-TGT«ATCT.UCJ.UC: i ATI; :i(Tl{;U.~UCC CCTJ,,;WGTTJ.CC""JJ;.UW---.... ~TA.C.U.C:.l TlG 1'14 I &TG U.CTTCTCATTG1'Tff--,(H.CTTOU-TTK"r'eUTOCi1.GT-TCC'ATACOCl(TGTlGTA--CAOCOC'l'C'IIGCJ.TCrur.u..1.c T1GUG.1.TCC ,a 1001 tl(CIXCC ibil(Wil I.., GG-TA.C.U.C 1'1,9 ... ... , TNlOU : ll'G CTTCTCl'TTGTT1T--G~rl.-Tm:'TC11TGCJ.GT•TOC-ATACGC1(TGT1'T.J.-~lGCOCTGT«AT'Cf.U.TJJ.J.CCTCA~TGC~liG1TCCTJ,;TGCf&CNGTTJ.CC.UCCTGUT'4a.LTATilGG-TACAJ.CACTS.G : t.49 : 1,1'(, TltTCATf01T1T--Ot.tTI'OTA .. 'Tl'TC'.TCTA'NC.I.G'f•TKATACG(MT6"'TTGTA•--C:A~OCT'O'N('.&TCl'AACAUCc;'N;'ATGro('1'11;A.Mll.KC'M'-CCCTN;1"QTI'A,CUCC:MioU'fOO-UTUMCO--T~A.l,(:ACJ..O t Ill~ : .I.TG CTT('TCATTGTMT--G~rA.-TTTCTc-UTCC.1.GT-TGC'ATACOC'l.('l'GTTGTA.--e.&.OCOCTC'NCJ.TCfU.T1UC(1CAf'G"roCT1GUCJ.TCCTTGTCCr.&(T«iTT1CC.liCCTGJ.J.T-(;G.A..I.T.lTilGG-TlC..U.CA.Cta.G : 149 P:tt : 1.'l"C TT"l.:TCJ.TfeT'TAT--C.M:TTC'U.-TA.ttTCTTTCC't«-'l'CTATIICCC'A('l"C~TA.--CA.CCCC~J.TCfilTJ.CJ.CC'TTA.TCT'K'MCA..U.l.C-CCJ.f'eTCCC.M:''NCTTAKJ.CCC'MaCTCe..UCATTJ.CC--TJ.1':UCJ.C"TiC : J. 0 ,m.r nc : I.TO .U.CTTCTC.lTTOTTTT .... QKTTO'!'l--TTI'CTClTTOc:TO(-TGT.lTACGT.l(Ul'~Tl ... ClOC(M;TGNTATCT.&J.Tl.(J.CCTTATGl'T(1'1G.l.U .. t:TI;.<;J.NT('«lo('l'GGTTAr<:OOC'C.&GA~UC.lTT.lGO-TATUCACT10 a 1"9 •tv1i : 1,1'(; CTTCTCA"mi1TffAOGXT'Cfl'liCl!'CiG?GflT'TGC'A.TGC.lTA.U.~UTOC1i:.TATJ.CA..W;ll1'CTTC1'Gl.U.1Tt.UCCJ.TT«U.CC'!GllCK'C<:.lMTCCU.TA.(.lTTJ.M'lT'TtGGJ.CTGGTTTGTilU.1'M'GG.l(CTC«.l : 157 •rv1 : LTG U.CTTCTCATT0'1'T1T1GG&CTCTTU.CAK.AGWT.t.TT«.LTCC.lTAAAG.UTGC..IGTlTU!UGUTGTTGTG.UU'Kl.t.TCJncC.UCnc:UGJ.CCClf'GTCCUT.LCATTAC'T.t.TTCOG.t.CT'GCTTTGTil,U.lTTGCJ.(,CTCGCJ. : L5'1 UC014 : lT4 CntTC.lTTGTTTTAGG.l(.TtnucA.n:..I.OTGT.lTTGC.J.n(.t.1'J..U0.UTGCUTJ.T.lCUGUTGTTGTGUJ.l'K.UCC.lTTCCU(nGUG1CCCl'l'GTC'CUTJ..(ATTJ.n.&TTCOCJ.CT04TT'l'iT4.lU.1tTGGUC:TC«.l 1 1.$'7 :IU672 : 11"..i ClTCTC.lTroTTTl'.lGGICTCTfilCATC.lGlATATTGC.a.TGC:.lTJ.A.lG.ll.TGCJ.GT.lTAC.l.a.o;uTGTTuTGil.lACCAJ.CCJ.TTCC.UCTlGJ.AGJ.roCUG~CU.TACA.Tr.l(TJ.TTCGGACTGGTTTTT.l.UliTTGGl.(CTC'G!A : 15"1 Tt>c-2 ; "'°TUTACTTJ.TlCCI.C-toCTTOQCTT?O-TOCTCTACC.a.•••.LiOGTTTT.lCCT1TTC'I.T•.a.GJ.TOOCACo\.CT.lTOGTl"CJ.J.M;ATOCACACCTAA1GTrA<TATCUCTCTC'UGATCC&GCT<»TOG'l-cc«TTATAOCT&QGTGTWCTAtC : 2"11 I.JOl : GGGT&ATJ..CTT.lT&K.lCTGCTTQGCTT'N-~TAG<;.l-•-U<;GTTTUCCTITTC:At"•.lG.lTGGC'ACA.Ci.LTCXiTI'C.\UCJ.TG<.J.CJ.OCTUWTI.lOAT'C-U.CTGT'(J..lliA,l«~T<ZTGGTGCGCTilTJ.GCTA.GGT<iTTGGTA{C : i72 •oe• : I.OGTJ.lT.lClTATAGC.lC'NCTT'OGiCTTN,,,T'OCTCTNN.l••-UOCT.lTrJ.C:CTlTTCU-.1(;J.TOQC'ACA('TlTGGTrCUAC.lTOC.lC.lC'CTU.NTI'.UT.lTCUC~TC'UClATCC.I.GTTQQTiiG7GCQCTT.lTJ.G(;T¥,,GTvfWGTACC ' a, GIO~ ; ~T.UT.t.CTI'.lTAGC:A~TntrTQC;TCTNJ!vA-·-A.AOGTTrfAl:-~At'•WAT'OOC'IIClt.'CTlTGvJTC.u..KATCXACACCTlilwTT.l(TATCUCTGTC.U.GATCClGC1WTW1GCG<;TTATAG<CTMiO'NTIGGTKC : JOl SZ3 ; GCNTilUClTJ.T1'C.AC1"TTINCTTT'G-TiiGTCT~l--.UGHTTTJ.OCTTTTC1T•~1~.IC.lCT.I.TGG1TCJ.UCAT«ACACCTU~TTJ.CTlT'C-UCTGTCUClATCC-1GCTGGTG.t;lGCGCTTUJ.GCTM.GTc;TTGGTJ&C : )01 40Jl,2 , t0CTUTAeTT.lTAGC'J.C1'Cc:~'l"T'TG-TOC"l'<T.M)Cl.-·-l..l.dG-Tl.rtl.CC~l.t-LC.lf'OCCl,Cl~l'IICOTTC,UACA'l"Ot.lCI.QCT.ll."IC"fTA(TI.TCU(TCttl.a.G.ITCC~TGIC1CCGIC'TTI.T.l"'r~TG'MCoTl.tC: , )01 mo11 ~ lGGTilT.lCTTA.CJ.G<'iCTGGTTOGCTl'TG-TGCTCTJ.GGJ.-·-ilGG11Tt.lCCTTTTe.lT-1G11'GGCJC.lC'tl.TGGTT'C.u.J.CiTG(.lCAO:TUTI.TT.l<T.lTCU.CT(;TCJ.J.GATCC1GCTO:t'roG1GCGCTT.lT.lOCT.JGGTGT'roGUCC : )01 ,.11 : QGGTilTA.CTTA.T.UCAG~T A.<:.M:TGT'C-TOCTT'l'.10(;1---a.u,nnATCTITTC.lT-.\C<il'<iGCM:ACT.LTOGlTCA.MXA WTJ.CAIXTU lGTCAC'C.t.TCUTTOKUCll'T«&QTAOOn;.G,lGClC-rrof.lQCGA.QTGTJGOTKT : )0 l 'ffitr )lC l GGCTilT.lC1TlT.l!C.lCTGCT.lGGCTG1'G-T'ClC'TTTJ.GC;i--llGGTTT'roTCffl'TC.lf-J.G11'00C'JrC.lCTll'OG1TCilGC.&.NUCJ.CCT1.lTATC-.lttATTUTTC:'K.l.lG.lTCC~'T«>TG(ti:'f'"l"GT.lGCC:JJaTGT'MGT.ltT t 101 •tv1i : lGTC1"...CT'C'OCCT-"T1C.llCTTTGTGCT'GGTGllT.lTGGJ.CJ.CAG.lGTTCC.UT.l(A.TT1TCilJ.1'(;1TTCGC.UTTAT.lCT.lTTT'CJ.MTCllGC.l.C-TTGil.lT.l.UTT(;K'A.U.1(CC.&.«.lC'f"NGJ.lGTCTC..i~MXTTGTTC.lT.l·· : )11 WlYJ I lOTCT.K:oc.«CT»-l'A1'AM.:TTNTO'tT'OOTOU.T.t.10Q.,t,(;l,TA0.t.GTTCC.t.ATAC.t.lTAfOA.AANTfl'OOC.AI.TTAT.\CTA~TI:ATOTOAM.CAC•Tf'Qu.ATJ.l.lTIOTCAAAM'.«A«AOTTWAJ.OttTCA.TCGTMOTN°'-ATA•• ; )I l 311:014 : lGTC1"'..('l'CGCG-YM1'1CileTnGTGCTGaTGU.TllGGiClC.a.GiGflCCUTACAff.&.rG.l.llTGTTiGC',,C.1.\.TT.lTlCTATTTC.l'TGTG.UCC.l.C-~i.UTlllTTGTC1.lilCCC1CC1GTTGG.U.GTC'Tti!"mTACG'M'(;f1CJ.T.1·- : 111 a..C">l : tCTCTGC~l!T.ltT.lCUC~TJ.'JCC,.lC~.l.C.lCT'ttC:J.J.T.l.CJ.TT.l'TCU.l'TC'M"fCOC-.li.TTATAC1'.lTTT(C,C'f'C1.l.eCJ.'T-'TTC.UJ.TJ.UTTCTC'.l.l!ACC,C~J.~J.J.CTC1'C.lT'TC"f'~~.ITA•· : )ll ORFSbofToT2 < Tori : rKATGAAGGT(;lCC-U.AaGCl'QCATTI4GAG.tCGT"'""'GTT'GTITTA.UT.l.l.t.CG.UC1llffiB.l : )40 D.JOl : rrC-A1'44A90~'-~UKr0¢1'«1.'nTAOl.'kCQT~Tl'OTT9TTTT.t..UTMM0.-.K.\,U A,\ : J .. (11 4084 ; f'T'C.l'NAJM:TC.lCCU.l.CTGC:TOUTTnGAG.M:J.T.M:TTOfflnTTUCT.l.U(O.U.CUJ. TU I tH C.!02 : tT'C.l.1t:.l,.lQCTC.lCC.J..lJ.CTGCTOCJ.TTT'AC.lG.l.CCTJ.TTTGT1':T1'TT.lJ..lT.lJ.lCCJ.J.C1U.TYAi- : 3,9 SU : fTC.llGA~TC.lC(l.U(TGCT'GC..lTTTMi-J.YKGTJJ;TTG~TTTT.UC:T.l.UCQUC.l.UTIU• 1 Jf9 41092 ; M'('ANUG<iTC.lCC.l.AACTGCTOC.lTTT.l.GlGJ.CI..TJCTTGTTG'MTfu.JT.u.A(G.U.C:U.1Cl'lJ.- : l69 mou ; ff<'.t.Q;Uf'..GTCACC.l.UCTGCTOCATTT.lGJ.GlCGTJ.Cff'GTWTT'CTUJTillCGllCU .. A.C1'.U- : 3'9 au I MT.&a;U.OGTC<:Ct.lGA.C'COC-TM;.lTTT.I.G.lG.a.C.lTJ.CAl'OTJ.<OATCTQ.TTCTTT.U.lCQU(TTU 1 ):69 'ffiLr l1C : fT'C.lC.l.J..lOGTTCC<J.OACTGC'Ti~ATT'l'.l.GAGJ.C.lTJ.C'.V:O'U.GATtl"G--TTCTTTA..UCG.U.CTTJ..l ; .369 Wtv1i : ---~ATCTroJ.CT--TTlf'(l(;l.GTJ.TC..l.CG.t.CG~TGTTGTTCT~AT--TTC.t.TCT.U.---·- : SU V.tvt 1 .-- ...... ~J.NTTOJ.eT---..Tr•~TU<".lCG.ltoT1CGTGTNTTC1'..0.lT-nc..lTCtil•--- : SH 31«:0H : ----W~TGTTG.lCT---tT.lTGGiGTAT'CJ..CGACCT'TCGTG~TlGJ.T--T'J'C.lT'tTU----- : 1H bf72 1 ••-•1CJ.'l'OTTOACT•--1Tl~AOTATCACGlCGTICC,TGTr.;TTCT~.lT•-TTC.AT'CT.U••--- 1 )U Fig 6. Alignment of ORF8 nucleotide sequences of SARS-CoV and bat SL-CoVs. The red box indicates the 29-nt deletion present in SARS-CoV of middle and late phase. RPPR Page 17

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Recombination analysis of the full-length genome sequences reveals frequent recombination events among different SL-CoV strains circulating in this bat population. For example, WIV16 appears to be a recombination product of WIVl and Rs4231. An important breakpoint is identified between the Nterminal domain (NTD) and RBD region in the S gene (Fig 7 A). Consequently, WIV16 is identical to Rs4231 and WIVl in NTD and RBD of the spike protein, respectively, and is highly homologous to SARSCoV in both NTD and RBD. This makes it the SL-CoV most closely related to the direct progenitor of SARS-CoV discovered to date. Moreover, evidence is found to support the hypothesis that the direct progenitor of SARS-CoV was generated from recombination of WIV16 with Rf4092 at the site near ORF8. This work, which identifies diverse SL-CoVs highly homologous to SARS-CoV in different regions of the genome, suggests that rhinolophid bats are an evolutionary epicenter of SL-CoV and offers more insights into the evolutionary origin of SARS-CoV. A. tm .. .. .. ., ,. Rs4231 "' 70 WIVl Q) (I) .. 1:; Rs4237 ""O .. 2 .. ~ .. E <ii a. .. 0 ,. ~ .. " "' ,. . . t 2.000 a,m &1'00 31/YJ 10.000 12.IOO 1410) IS.0,0 18.«IO 10J1Jt> 12.llOO 1'.JXIO lll.000 )e.(t)O XI.COO B. V) , .. " .. • " " Q) " ~ . " .._.. ,~-.Ar -.. w,~O.."--- ,...,,_~(1.,,,_..,,,J.1"1'rto,t,t~ -WIV16 - Rf4092 - Rs4081 0 )r,,» '\000 l,000 8000 IOJJl)f 12,000 14,(WXI 11,000 te.Q)O lOJ,ICIO »..co> 26,(0) ».i:,oo ~,000 10,G(IO .,..._. ,,.,.... - ..,., C-.0Vf" ~~ 1'.qe. 1..,#_ ,-_,.,). ~ 1• ..... "'-'-"I Fig 7 Bootscan analysis offull-length genome sequences of SL-Co Vs. (A) WIV16 is used as query sequence. (B) SARS-CoV SZ3 is used as the query sequence. (Kimura model, window size, lS00bp, step size, 300bp) RPPR Page 18

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B.2 (Year 2 NIAID CoV Report Final.pdf) 1R01Al110964 Year 2 Report Pl: Daszak, Peter Additional Year 2 items for Specific Aim 3: • The infectious clone of WIVl was successfully constructed using reverse genetic methods; • Two chimeric bat SARS-like coronavirus strains were constructed by replacing the S gene in the backbone of WIVl; • Permission to import mice with human ACE2 to China was obtained, so as to conduct the experimental infections proposed in our R0l specific aims. Specific Goals Not Met. • Comparative cophylogenetic analyses of bat host and CoV RdRp and Spike gene phylogenies, to assess patterns of evolutionary congruence and frequency of cross-species transmission (This will be conducted in year 3); • Animal infection experiments of SARS-like coronaviruses were not done, because of the unavailability of mice with human ACE2 in Year 2. We now have secured these mice and will begin this work in year 3. • Sampling of bat and other mammalian species in markets to screen for CoVs. We will begin this work in year 3. Section C: Accomplishments: Publications PUBLISHED Xing-Yi Ge, Ning Wang, Wei Zhang, Ben Hu, Bei Li, Yun-Zhi Zhang, Ji-Hua Zhou, Chu-Ming Luo, Xing-Lou Yang, Li-Jun Wu, Bo Wang, Yun Zhang, Zong-Xiao Li, and Zheng-Li Shi. Coexistence of multiple coronaviruses in several bat colonies in an abandoned mineshaft. Virologica Sinica 31, 31-40 {2016). Mei-Niang Wang, Wei Zhang, Yu-Tao Gao, Ben Hu, Xing-Yi Ge, Xing-Lou Yang, Yun-Zhi Zhang, Zheng-Li Shi. Longitudinal surveillance of SARS-like coronaviruses in bats by quantitative real-time PCR, Virologica Sinica 31(1): 78-80 (2016). Cristin C. W. Young and Kevin J. Olival. Optimizing Viral Discovery in Bats. PLoS ONE 11(2) (2016). Kevin J. Olival. To Cull, or Not To Cull, Bat is the Question. Ecohealth 13, 6-8 {2015). Xing-Lou Yang, Ben Hu, Bo Wang, Mei-Niang Wang, Qian Zhang, Wei Zhang, Li-Jun Wu, Xing-Yi Ge, YunZhi Zhang, Peter Daszak, Lin-Fa Wang, Zheng-Li Shi. Isolation and characterization of a novel bat coronavirus closely related to the direct progenitor of Severe Acute Respiratory Syndrome Coronavirus, Journal of Virology 90(6): 3253-6 (2015). Ben Hu, Xingyi Ge, Lin-Fa Wang, Zhengli Shi. Bat origin of human coronaviruses. Virology Journal 12 (1): 221 (2015) ACCEPTED, IN PRESS Lei-Ping Zeng, Yu-Tao Gao, Xying-Yi Ge, Qian Zhang, Cheng Peng, Xinglou Yang, Bin Tan, Jing Chen, Aleksei Chmura, Peter Daszak, and Zheng-Li Shi. Bat SARS-like coronavirus WIVl encodes an extra accessory protein ORFX involving in modulation of host immune response. Journal of Virology (in press, 2016) RPPR Page 19

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B.4 (Year 2 NIAID CoV Report Professional Development.pdt) 8.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? 1R01Al110964 Year 2 Report Pl: Daszak, Peter B.4 What opportunities for training and professional development has the project provided? We presented our project to graduate students, laboratory personnel, directors, and doctors from three Hospitals in Yunnan Province: Yunnan Provincial Institute of Endemic Diseases Control & Prevention (YNCDC); Dali Provincial Hospital; and The Third People's Hospital of Kunming. Select doctors at YNCDC (1) and Dali Provincial Hospital (3) were trained in the passive Hospital surveillance project protocols. We trained graduate students from Dali School of Public Health (1) and the Wuhan University School of Public Health (3) in qualitative behavioral risk data collection methodologies and data collection technologies, survey data collection and analysis. These were also enrolled in and passed the Human Subjects Research Course provided by the Collaborative Institutional Training Initiative (CITI Program) at the University of Miami (http://citiprogram.org). The CITI Program is a leading provider of research education content with web based training materials serving millions of learners at academic institutions, government agencies, and commercial organizations in the U.S. and around the world. RPPR Page 20

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RPPR FINAL C. PRODUCTS C.1 PUBLICATIONS Are there publications or manuscripts accepted for publication in a journal or other publication (e.g., book, one-time publication, monograph) during the reporting period resulting directly from this award? Yes Publications Reported for this Reporting Period ·i• ~:..,,,._...,_.,._;,,.,• ,------------------------------ Complete Yang XL, Hu B, Wang B, Wang MN, Zhang Q, Zhang W, Wu LJ, Ge XY, Zhang YZ, Daszak P, Wang LF, Shi ZL. Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus. J Virol. 2015 Dec 30;90(6):3253-6. PubMed PMID: 26719272; PubMed Central PMCID: PMC4810638. Complete Olival KJ. To Cull, or Not To Cull, Bat is the Question. Ecohealth. 2016 Mar;13(1):6-8. PubMed Pl\,11D: 26631385; PubMed Central PMCID: PMC4833651. Non-compliant Publications Previously Reported for this Project 0 .. •;a • "°\< ""''r"•'"<:·. ,,v•~~ .y.-,'"'t II,''•• •·•••;-:-,; ~I •~ v-_ - • ; .: --• Non-Compliant (b) (4) C.2 WEBSITE(S) OR OTHER INTERNET SITE(S) NOTHING TO REPORT C.3 TECHNOLOGIES OR TECHNIQUES NOTHING TO REPORT C.4 INVENTIONS, PATENT APPLICATIONS, AND/OR LICENSES Have inventions, patent applications and/or licenses resulted from the award during the reporting period? No C.5 OTHER PRODUCTS AND RESOURCE SHARING C.5.a Other products NOTHING TO REPORT C.5.b Resource sharing NOTHING TO REPORT RPPR Page 21

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RPPR FINAL D. PARTICIPANTS D.1 WHAT INDIVIDUALS HAVE WORKED ON THE PROJECT? .;,.-'>--,cv~.~•"·•-~!,,:.ot --;;:,:-~;4-w,,;f:'. )"""/•?,< ~:---::-::11-<·~., '"'"-,;'v:-;-.f-'. • 1rr~ ~i'Y.Jr;, 1:,;,_,, '.-:,..~.-: ,>,,:.-,:,-,.,, .. ~;,·11~· ~i~~,~ (.;>.,,l ~--,.,., 1~· ,!,..,_-,_,J 'I:'· .• ,,, ~~$• .. ~;\ r~";-·.~ • 1~1••->'.i< Plr.<- ,~::,;, I (b) (6)) y DASZAK, (b)('1 I (b)(6) BS,PHD PD/Pl NA (b) (4) (b) (6) PETER N HOSSEINI, (b)(~ (b)(6) BS,PHD Co- NA PARVIEZ Investigator RANA I (b)(6)J y Ross, (b) (6)1 PhD Co- NA Noam Investigator Martin N OLIVAL, (b)('1 (b) (6)] PHD Co- NA KEVIN J Investigator N KE, PHD Co- Center CHINA NA CHANGWE Investigator for N Disease Control and Preventio n of Guangdo ng Province N ZHANG. (b)(6)J PHD Co- East CHINA NA SHUYI Investigator China Normal Universit y N ZHANG, (b)(6)J PHD Co- Yunnan CHINA NA YUNZHI Investigator Provincia I Institute of Endemic Diseases Control & Preventio n N ZHU, (b) (6): PHD Co- East CHINA NA GUANGJIA Investigator China N Normal Universit y N GE, XINGYI PHD Co- Wuhan CHINA NA Investigator Institute of Virology N EPSTEIN, (b)(~ (b) (6)] MPH,DV Co- NA JONATHAN M,BA,PH Investigator H D N CHMURA, (b)(~ (b) (6)1 BS Non- NA ALEKSEI A Student Research Assistant N SHI, (b)(6)J PhD Co- Wuhan CHINA NA RPPR Page 22

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RPPR FINAL ZHENGLI Investigator Institute of Virology Glossary of acronyms: Foreign Org - Foreign Organization Affiliation S/K - Senior/Key SS - Supplement Support DOB - Date of Birth RE - Reentry Supplement Cal - Person Months (Calendar) DI - Diversity Supplement Aca - Person Months (Academic} OT- Other Sum - Person Months (Summer) NA - Not Applicable D.2 PERSONNEL UPDATES D.2.a Level of Effort Will there be, in the next budget period, either (1) a reduction of 25% or more in the level of effort from what was approved by the agency for the PD/Pl(s) or other senior/key personnel designated in the Notice of Award, or (2) a reduction in the level of effort below the minimum amount of effort required by the Notice of Award? No D.2.b New Senior/Key Personnel Are there, or will there be, new senior/key personnel? Yes File uploaded: Noam Ross CV 2016.pdf D.2.c Changes in Other Support Has there been a change in the active other support of senior/key personnel since the last reporting period? No D.2.d New Other Significant Contributors Are there, or will there be, new other significant contributors? No D.2.e Multi-Pl (MPI) Leadership Plan Will there be a change in the MPI Leadership Plan for the next budget period? NA RPPR Page 23

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D.2.b (Noam Ross CV 2016.pdf) (b)(6) Noam Ross http://www.noamross.net '#@noamross EDUCATION University of California Davis, CA Doctoral Candidate in Ecology Expected Completion Summer 2015 • Dissertation Committee: Alan Hastings (major professor, Ecology), David Rizzo (Plant Pathology), Jim Sanchirico (Natural Resource Economics) • Dissertation Research: "Managing Emerging Forest Disease Under Uncertainty" Brown University Providence, RI Bachalor of Scianca in Enuironmantal Scienca, J.,fagna Cum Lauda May 2006 • Honors Thesis: "Soil Organic Matter in Northern Mongolia: Permafrost and Land-Use interactions" ■ Phi Beta Kappa, Sigma Xi, Environmental Science Honors, Rosenberger Prize for Outstanding Service SCIENTIFIC PUBLICATIONS • Carl Boettiger*, Noam Ross*, Alan Hastings (2013) Early Warning Signals: The Charted And Uncharted Territories. Theoretical Ecology http://dx.doi.org/10.10o7/s12o8o-o13-0192-6 ■ Fuller, Kate, David Kling, Kaelin Kroetz, Noam Ross, and James N. Sanchirico (2013) Economics and Ecology of Open-Access Fisheries. In: Shogren, J.F., (ed.) Encyclopedia of Energy, NaturaJ Resource, and Environmental Economics, Vol. 2 Encyclopedia of Energy, Natural Resource, and Environmental Economics p.39-49. Amsterdam: Elsevier. http://dx.doi.org/10.1016/B978-o-12-375o67-9.oo114-5 In preparation (b) (4) •Co-equal author.ship POSTERS • Ross, Noam. "Optimal Control of Disease in Space: An Approach Using Individual-based Models," June 1-4, 2014. 12th Annual Conference of Ecology and Evolution of Infectious Disease, Fort Collins, Colorado. • Ross, Noam. "Designing Protective Treatments for Forest Disease Using a Spatial Point Process Model," November 20-21, 2014. California Forest Pest Council Annual Meeting, McClellan, CA. ■ Ross, Noam. "Optimal Control of Forest Disease Under Changing Community and Spatial Stmcture," November 4-18, 2013. Sustainable Management of Natural Resources Workshop, Mathematica] Biosciences Institute, Columbus, OH. PRESENTATIONS • Ross, Noam, "Fungal Disease Mortality: Modeling for Management of Sudden Oak Death." Dec 1, 2014 Invited talk at Eco Health Alliance, New York, NY. ■ Ross, Noam, "Modeling forest disease using a macroparasite framework ," Agust 13, 2014. 99th Annual Ecological Society of America Meeting, Sacramento, CA. • Ashander, Jamie, Kelly Gravuer, Megan Kelso, Mary E. Mendoza and Noam Ross "Managing RiverFloodplains Systems: A Historical and Ecological Perspective" September 14, 2002. Presentation at NSF REACH IGERT Floodplains Workshop RPPR Page 24

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D.2.b (Noam Ross CV 2016.pdf) AW ARDS + FE LL OW SH IP S (Total received $225,429) ■ Don Dahlsten Memorial Grant ($325) California Forest Pest Council, 2012 Designing Protective Treatments for Forest Disease Using Spatial Point Process Models ■ NSF IGERT Bridge Fellowship ($57,500) UC Davis, CA, 2012 Managing Emerging Forest Disease Under Uncertainty ■ NSF !GERT Traineeship in Rapid Environmental Change ($115,00) UC Davis, CA, 2010 Modifying River-Floodplain Systems: A Historical and Ecological Approach ■ UC Davis Graduate Group in Ecology Fellowship ($40,604) UC Davis, CA, 2010 ■ NSF Research Experience for Undergraduates Fellowship ($8,000) Acad. of Natural Sciences, PA, 2005 ■ Undergraduate Research Fellowship ($4,000) Brown University, RI, 2003 SERVICE + PROJECTS ■ Workshop Instructor, Software Capentry and Data Carpentry Foundations Jan 2015-Present ■ Student Rep, UC Davis Graduate Group in Ecology Executive Committee Sep 2013-Present ■ Reviewer: Theoretical Ecology (4 reviews) Feb 2013-Present ■ Web Developer and Technology Chair, Ecology Graduate Student Association June 2013-Present Creator+ Maintainer of graduate student blog, resources, and news site (egsa.ucdavis.edu) ■ Founder+ Organizer, Davis R Users' Group Sep 2012-Present Created users group that provides tutoring and seminars to graduate students in 10+ departments • Contributor, Rpackages knitr, knitcitations, rcrossref, rethinking ■ Organizer: NSF REACH I GERT Workshop on Multiple Goals in Floodplain Restoration ■ Organizer, UC Davis Conference on Ecology and the Business Sector ■ Organizer, UC Davis Graduate Group in Ecology Symposium ■ External Reviewer, World Resources Institute Corporate Ecosystem Services Review ■ External Reviewer, McKinsey-Clinton Global Initiative Forestry Project ■ Business Stewardship Volunteer, NY Coastal Marine Resources Center OTHER WORK EXPERIENCE 2012-Present Sep 2012 Apr2011 May 2010-2011 Jan 2008 Mar2008 Feb-Apr 2007 Green Order Analyst, Senior Analyst: Corporate Environmental Strategy+ Governance New York, NY Sep 2006-Oct 2009 ■ Conducted environmental performance analysis for products in energy, transportation, and water sectors ■ Created green product metrics system R&D stage-gating system for construction products manufacturer ■ Managed engagement with equipment rental company to identify growth opp01tunities in green building ■ Performed market and competitive analyses for a wide array of clients in retail, real estate financial and cleantech sectors; prepared and delivered client presentations; managed projects ■ Managed analysts performing environmental product certifications and market research ■ Developed firm seminar series and analyst training materials; conducted trainings on topics including auditing, statistical analysis, and environmental performance benchmarking ■ Audited certifications for environmental products and facility performance Wal-Mart Contract Researcher/Consultant: Energy Efficient Products Initiative Providence, RI May-Sep 2006 ■ Developed forecasting model for sales of energy-efficient lamps at Wal-Mart stores ■ Created guidelines for design oflamp recycling program RPPR Page 25

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D.2.b (Noam Ross CV 2016.pdf) Brown University Facilities Management Administrative, Research, + Teaching Assistant: Energy and Design Providence, RI Jan 2003-May 2006 ■ Developed energy-use and financial projections for university energy usage scenarios ■ Performed background research and feasibility analysis for university energy efficiency projects ■ Provided tutoring, logistical support and web design for two courses in sustainable design • Responsible for maintenance of energy efficient, low-impact building Hovsgol Lake Global Environmental Facility and Brown University Mongolia+ Providence RI National Science Foundation REU Fellow, Thesis Research June 2005-May 2006 Advisor: Clyde Goulden ■ Independent research on climate-land use interactions on permafrost soil carbon storage Plant surveys, soil pit excavation, soil physical and chemical analysis, soil microbial process incubations Marine Biological Laboratory Ecosystems Center Semester in Environmental Science Student Woods Hole, MA Aug-Dec 2004 Advisor: Charles Hopkinson ■ Examined effects of nitrogen pollution on structure of microplankton food webs ■ Microcosm experiments, fluorescence microscopy, dissolved nutrient analysis, planktonic growth incubations Brown Center for Environmental Studies Undergraduate Research Fellow Providence, RI ,Jun-Aug 2003 Advisor: Steven Hamburg ■ Conducted research in biogeochemistry at Hubbard Brook Experimental Forest and surrounding region; oversaw soil pit excavation by undergraduate and graduate field crew • Plant surveys, forest floor measurements, litter collection, soil pit excavation, soil physical and chemical analysis, GIS analysis in ESRI ArcMap PUBLICATIONS IN POPULAR PRESS ■ "Extinction Debt,"(Initial author) Wikipedia. Wikimedia Foundation, Inc., February 23, 2011 http:/ /en.wikipedia.org/wiki/Extinction_debt ■ "If Everyone Moves to the City, What Gets Left Behind?" Good.is, January 17, 2011. http://·www.good.is/post/if-everyone-moves-to-the-city-what-is-left-behind/ ■ "Why the Ethanol Debate Isn't Helping Anyone," GreenBiz.com, Jun 3, 2009. http://www.greenbiz.com/blog/ 2009 / 06 / 03/why-ethanol-de bate-isnt-hel ping-anyone ■ "Four Lean, Green Strategies for an Uncertain Economy," (with Andrew Shapiro) Harvard Business Review's Leading Green, Oct 29, 2008. http://blogs.hbr.org/2008/10/4-lean-green-strategies-for-an/ • "What a Silent Spring Means for Business Risk," GreenBiz.com, Mar 6, 2007. http://www.greenbiz.com/blog/ 2007 / o 3/ 05/ what-silent-spring-means-business-risk RPPR Page 26

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RPPR E. IMPACT E.1 WHAT IS THE IMPACT ON THE DEVELOPMENT OF HUMAN RESOURCES? Not Applicable FINAL E.2 WHAT IS THE IMPACT ON PHYSICAL, INSTITUTIONAL, OR INFORMATION RESOURCES THAT FORM INFRASTRUCTURE? NOTHING TO REPORT E.3 WHAT IS THE IMPACT ON TECHNOLOGY TRANSFER? Not Applicable E.4 WHAT DOLLAR AMOUNT OF THE AWARD'S BUDGET IS BEING SPENT IN FOREIGN COUNTRY(IES)? RPPR Page 27

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RPPR FINAL F.CHANGES F.1 CHANGES IN APPROACH AND REASONS FOR CHANGE Not Applicable F.2 ACTUAL OR ANTICIPATED CHALLENGES OR DELAYS AND ACTIONS OR PLANS TO RESOLVE THEM NOTHING TO REPORT F.3 SIGNIFICANT CHANGES TO HUMAN SUBJECTS, VERTEBRATE ANIMALS, BIOHAZARDS, AND/OR SELECT AGENTS F.3.a Human Subjects No Change F.3.b Vertebrate Animals No Change F.3.c Biohazards No Change F.3.d Select Agents No Change RPPR Page 28

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RPPR FINAL G. SPECIAL REPORTING REQUIREMENTS G.1 SPECIAL NOTICE OF AWARD TERMS AND FUNDING OPPORTUNITIES ANNOUNCEMENT REPORTING REQUIREMENTS NOTHING TO REPORT G.2 RESPONSIBLE CONDUCT OF RESEARCH Not Applicable G.3 MENTOR'S REPORT OR SPONSOR COMMENTS Not Applicable G.4 HUMAN SUBJECTS G.4.a Does the project involve human subjects? Yes Is the research exempt from Federal regulations? No Does this project involve a clinical trial? No G.4.b Inclusion Enrollment Data Report Attached: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 G.4.c ClinicalTrials.gov Does this project include one or more applicable clinical trials that must be registered in ClinicalTrials.gov under FDAAA? No G.5 HUMAN SUBJECTS EDUCATION REQUIREMENT Are there personnel on this project who are newly involved in the design or conduct of human subjects research? No G.6 HUMAN EMBRYONIC STEM CELLS (HESCS) Does this project involve human embryonic stem cells (only hESC lines listed as approved in the NIH Registry may be used in NIH funded research)? No G.7 VERTEBRATE ANIMALS Does this project involve vertebrate animals? Yes G.8 PROJECT/PERFORMANCE SITES RPPR Page 29

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RPPR FINAL [=======! ll~~ Primary: EcoHealth 077090066 NY-010 460 West 34th Street Alliance, Inc. 17th Floor New York NY 100012317 Wuhan Institute of 529027474 Xiao Hong Shan, No. 44 Virology Wuchang District Wuhan East China Normal 420945495 3663 Zhongshan Beilu University Shanghai ECOHEAL TH ALLIANCE 077090066 ECOHEAL TH ALLIANCE, INC. 460W 34TH ST NEW YORK NY 100012320 EcoHealth Alliance, Inc. 077090066 NY-010 460 West 34th Street 17th Floor New York NY 100012317 Wuhan Institute of 529027474 Xiao Hong Shan, No. 44 Virology Wuchang District Wuhan East China Normal 420945495 3663 Zhongshan Beilu University Shanghai G.9 FOREIGN COMPONENT Organization Name: Wuhan Institute of Virology Country: CHINA Description of Foreign Component: Principal Laboratory for all Research in China as per section GB (above) and detailed in our Specific Aims Organization Name: East China Normal University Country: CHINA Description of Foreign Component: Principal Coordinating Team for all project field work as per section GB (above) and detailed in our Specific Aims G.10 ESTIMATED UNOBLIGATED BALANCE G.1 0.a Is it anticipated that an estimated unobligated balance (including prior year carryover) will be greater than 25% of the current year's total approved budget? No G.11 PROGRAM INCOME Is program income anticipated during the next budget period? No G.12 F&A COSTS Is there a change in performance sites that will affect F&A costs? No RPPR Page 30

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Inclusion Data Record (IDR) #: 166195 Delayed Onset Study ?: No Enrollment Location: Foreign Inclusion Enrollment Report Using an Existing Dataset or Resource: No Clinical Trial: No NIH Defined Phase Ill Clinical Trial: No Study Title: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 Planned Enrollment Planned Enrollment Total: 2,460 NOTE: Planned enrollment data exists in the previous format; the PD/Pl did not enter the planned enrollment information in the modified format and was not required to do so. Only the total can be provided. Cumulative Enrollment Ethnic categories Racial Categories Unknown/Not Not Hispanic or Latino Hispanic or Latino Reported Ethnicity Total Female Male Unknown/ Female Male Unknown/ Female Male Unknown/ Not Reported Not Reported Not Reported American Indian/ Alaska Native 0 0 0 0 0 0 0 0 0 0 Asian 157 108 0 0 0 0 0 0 0 265 Native Hawaiian or 0 0 0 0 0 0 0 0 0 0 Other Pacific Is lander Black orAfrican American 0 0 0 0 0 0 0 0 0 0 Whne 0 0 0 0 0 0 0 0 0 0 More than One Race 0 0 0 0 0 0 0 0 0 0 Unknown or Not Reported 0 0 0 0 0 0 0 0 0 0 Total 157 108 0 0 0 0 0 0 0 265

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Notice of Award RESEARCH Federal Award Date: 05/26/2017 Department of Health and Human Services National Institutes of Health NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 5R01Al110964-04 FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Aleksei Chmura President 460 West 34th Street 17th Floor New York, NY 100012317 Award e-mailed to: (b)(6) ----------- Period Of Performance: Budget Period: 06/01/2017 - 05/31/2018 Project Period: 06/01/2014 - 05/31/2019 Dear Business Official: The National Institutes of Health hereby awards a grant in the amount of $597,112 (see "Award Calculation" in Section I and "Terms and Conditions" in Section Ill) to ECOHEALTH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the "Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01 Al 110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STIR) awards. Consult the NIH website http://qrants.nih.gov/grants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1 ID)

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Laura A. Pone Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2

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SECTION I -AWARD DATA- 5R01Al110964-04 Award Calculation (U.S. Dollars) Salaries and Wages Fringe Benefits Personnel Costs (Subtotal) Materials & Supplies Travel Other SubawardsfConsortium/Contractual Costs Federal Direct Costs Federal F&A Costs Approved Budget Total Amount of Federal Funds Obligated (Federal Share) TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS YR THIS AWARD CUMULATIVE TOTALS 4 $597,112 5 $581,646 $167,708 $54,168 $221,876 $7,000 $35,918 $9,800 $201,422 $476,016 $121,096 $597,112 $597,112 $597,112 $597,112 $597,112 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project Fiscal Information: CFDAName: CFDA Number: EIN: Document Number: PMS Account Type: Fiscal Year: IC CAN Al 8472350 Allergy and Infectious Diseases Research 93.855 1311726494A 1 RAl110964A P (Subaccount) 2017 2017 $597,112 2018 $581,646 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project NIH Administrative Data: PCC: M51 C / OC: 414E / Released: (b)(6) 05/25/2017 Award Processed: 05/26/2017 12:05: 11 AM SECTION 11- PAYMENTfHOTLINE INFORMATION- 5R01Al110964-04 For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS - 5R01Al110964-04 This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. c. 45 CFR Part 75. d. National Policy Requirements and all other requirements described in the NIH Grants Policy Statement, including addenda in effect as of the beginning date of the budget Page-3

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period. e. Federal Award Performance Goals: As required by the periodic report in the RPPR or in the final progress report when applicable. f. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. (See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain references cited above.) Research and Development (R&D): All awards issued by the National Institutes of Health (NIH) meet the definition of "Research and Development" at 45 CFR Part§ 75.2. As such, auditees should identify NIH awards as part of the R&D cluster on the Schedule of Expenditures of Federal Awards (SEFA). The auditor should test NIH awards for compliance as instructed in Part V, Clusters of Programs. NIH recognizes that some awards may have another classification for purposes of indirect costs. The auditor is not required to report the disconnect (i.e., the award is classified as R&D for Federal Audit Requirement purposes but non-research for indirect cost rate purposes), unless the auditee is charging indirect costs at a rate other than the rate(s) specified in the award document(s). An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the System for Award Management (SAM). Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://qrants.nih.gov/grants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01Al110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://qrants.nih.gov/grants/policy/awardconditions.htm for additional award applicability information. In accordance with P.L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OD-08-033 and the Public Access website: http:/ /pu blicaccess. nih. gov/. In accordance with the regulatory requirements provided at 45 CFR 75.113 and Appendix XII to 45 CFR Part 75, recipients that have currently active Federal grants, cooperative agreements, and procurement contracts with cumulative total value greater than $10,000,000 must report and maintain information in the System for Award Management (SAM) about civil, criminal, and administrative proceedings in connection with the award or performance of a Federal award that reached final disposition within the most recent five-year period. The recipient must also make semiannual disclosures regarding such proceedings. Proceedings information will be made publicly available in the designated integrity and performance system (currently the Federal Awardee Performance and Integrity Information System (FAPIIS)). Full reporting requirements and procedures are found in Appendix XII to 45 CFR Part 75. This term does not apply to NIH fellowships. Treatment of Program Income: Additional Costs SECTION IV- Al Special Terms and Conditions - 5R01Al110964-04 Page-4

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The Research Performance Progress Report (RPPR), Section G.9 (Foreign component), includes reporting requirements for all research performed outside of the United States. Research conducted at the following site(s) must be reported in your RPPR: San Pya Clinic, BURMA lnstitut Pasteur du Cambodge, CAMBODIA Primate Research Center at Bogor Agricultural University, INDONESIA Conservation Medicine, Ltd, MALAYSIA King Chulalongkorn Memorial Hospital, THAILAND Hanoi Agricultural University, VIETNAM National Animal Health Laboratory, LAOS ***••·····~••******1rlr***********************••·~••-1m••··1rlf•******** This Notice of Award (NoA) includes collaboration with Wuhan University School of Public Health, CHINA. This Notice of Award (NoA) includes funds for activity with Wuhan Institute of Virology, CHINA. This Notice of Award (NoA) includes funds for activity with (East China Normal University. This award may include collaborations with and/or between foreign organizations. Please be advised that short term travel visa expenses are an allowable expense on this grant, if justified as critical and necessary for the conduct of the project. This award is subject to the Clinical Terms of Award included in Monitoring of Clinical Trials and Studies - NIAID (see NIH Guide for Grants and Contracts, July 8, 2002, NOT Al-02-032). These terms and conditions are hereby incorporated by reference, and can be accessed via the following World Wide Web address: https://www.niaid.nih.gov/qrants-contracts/niaid-clinicalterms-award All submissions required by the NIAID Clinical Terms of Award must be forwarded electronically or by mail to the responsible NIAID Program Official identified on this Notice of Award. Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.selectagents.gov/Regulations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm). Research funded under this grant must adhere to the BMBL. including using the BM BL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Page-5

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Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. If work with a new or additional Agent(s)/Toxin(s) is proposed in the upcoming project period, provide: o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent(s), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted, with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. STAFF CONTACTS The Grants Management Specialist is responsible for the negotiation. award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Carine Normil Email: (b)(6) Phone: (b)(6) Fax: 301-493-0597 Program Official: Erik J. Stemmy Email: Cb) (6) Phone: (b)(6) ----- SPREADSHEET SUMMARY GRANT NUMBER: 5R01Al110964-04 INSTITUTION: ECOHEAL TH ALLIANCE, INC. Budget Year4 Year5 Salaries and Wages $167,708 $167,708 Fringe Benefits $54,168 $54,168 Personnel Costs (Subtotal) $221,876 $221,876 Materials & Suoolies $7,000 $3,500 Travel $35,918 $35,918 Other $9,800 $9,400 Subawards/Consortium/Contractual Costs $201,422 $191,576 TOTAL FEDERAL DC $476,016 $462,270 TOTAL FEDERAL F&A $121,096 $119,376 TOTAL COST $597,112 $581,646 Facilities and Administrative Costs Year4 Year5 F&A Cost Rate 1 44.1% 44.1% F&A Cost Base 1 $274,594 $270,694 F&A Costs 1 $121,096 $119,376 Page-6

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RPPR FINAL A. COVER PAGE Project Title: Understanding the Risk of Bat Coronavirus Emergence Grant Number: 5R01Al110964-04 Project/Grant Period: 06/01/2014- 05/31/2019 Reporting Period: 06/01/2016 - 05/31/2017 Requested Budget Period: 06/01/2017 - 05/31/2018 Report Term Frequency: Annual Date Submitted: 04/12/2017 Program Director/Principal Investigator Information: Recipient Organization: PETER DASZAK , BS PHD ECOHEAL TH ALLIANCE, INC. ECOHEAL TH ALLIANCE, INC. Phone number: I (b) (6)1 460 W 34TH ST Email:! (b) (6)] 17TH FLOOR NEW YORK, NY 100012320 DUNS: 077090066 EIN: 1311726494A1 RECIPIENT ID: Change of Contact PD/Pl: N/A Administrative Official: Signing Official: ALEKSEI CHMURA ALEKSEI CHMURA 460 W 34th St., 17th Floor 460 W 34th St., 17th Floor New York, NY 10001 New York, NY 10001 Phone number: (b) (6)' Phone number: (b)(6)~ Email:1 (bTI6}J Email:1 (b)(6) Human Subjects: Yes Vertebrate Animals: Yes HS Exempt: No Exemption Number: Phase Ill Clinical Trial: hESC:No Inventions/Patents: No RPPR Page 1

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RPPR FINAL B. ACCOMPLISHMENTS 8.1 WHAT ARE THE MAJOR GOALS OF THE PROJECT? Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-CoVs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel CoVs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2} how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses; screen people who have high occupational exposure to bats and other wildlife; and examine the genetics and receptor binding properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-CoVs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological opportunity for contact between species so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Gen bank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecological and phylogenetic traits to explain patterns of viral sharing between species. We will test for positive selection in market vs. wild-sampled viruses, and use data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. 8.1.a Have the major goals changed since the initial competing award or previous report? No B.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? File uploaded: 5R01Al110964-04.pdf B.3 COMPETITIVE REVISIONS/ADMINISTRATIVE SUPPLEMENTS For this reporting period, is there one or more Revision/Supplement associated with this award for which reporting is required? No B.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? File uploaded: 5R01Al110964-04 Professional Development.pdf RPPR Page2

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RPPR FINAL B.5 HOW HAVE THE RESULTS BEEN DISSEMINATED TO COMMUNITIES OF INTEREST? 1.Conference and University Lectures: Pl Daszak, and Co-investigators Shi, Epstein, Oliva!, and Zhang gave invited University and Conference lectures including Avoiding Catastrophe Meeting at Concordia Univ., Harvard Univ. Columbia Univ., National Academy of Sciences, World Humanitarian Summit in Turkey, NEIDL Symposium in Boston, Global Pandemic Policy Summit at Texas A&M Univ., One Health EcoHealth Congress in Australia, WHO briefing, Rockefeller Planetary Health meeting, 17th International Bats Conference, China National Global Virome Project Initiative Meeting, and others that included specific discussion of the current project and results. 2.Agency and other briefings: Pl Daszak and Co-investigator Shi introduced this project to potential collaborators within Rockefeller Foundation, WHO, FAO, International Collaboration Bureau of Chinese Academy of Sciences, Beijing Genomic Institute, National Natural Science Foundation of China, Institute of Pathogen Biology, Chinese Academy of Medical Science & Peking Union Medical College, and Chinese CDC. 3.Public outreach: Pl Daszak and Co-investigator Shi presented this work to members of NSF, NIH, U.S. CDC, the State of Forestry Administration of China, and the general public at the China National Virome Project Initiative Meeting hosted by Chinese CDC and Chinese Academy of Sciences (2017); Co-investigator Oliva! presented this work at the NYC Medtech Forum to the public (2016); Research Technician Dr. Guangjian Zhu presented this work at the China Conservation Expo to the conservation groups in China (2016). Co-Investigator Y-Z Zhang presented this project to the provincial infectious disease hospital Kunming No.3 People's Hospital in Yunnan province (2016). B.6 WHAT DO YOU PLAN TO DO DURING THE NEXT REPORTING PERIOD TO ACCOMPLISH THE GOALS? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. •To commence the analysis of data collected from the integrated biological behavioral surveillance questionnaires from Yunnan, Guangxi, and Guangdong provinces, linking to the viral and serological testing results of biological samples. •Following the successful pilot of wildlife trade network research in Lipu, Guilin, Guangxi province in Year 3, we will continue the Wild Animal Farms Survey in Guangxi, and expand to Yunnan and Guangdong in Year 4, with Institutional Review Board approvals from both Yunnan Institute of Endemic Diseases Control and Prevention and Hummingbird #2016-55, to: -Generate a network model of wildlife trade -Model trade flows in the wildlife farmer networks to identify locations of high potential for viral recombination -Update survey instrument for "second wave" network interviews •We will continue the passive hospital surveillance with anonymized, surveillance data collection from acutely ill hospital in-patients who 1) satisfy syndromic eligibility criteria; 2) have complete medical records; 3) non-normative laboratory confirmed diagnostic results; and suspected acute viral infection. Research has been successfully piloted in four hospitals in Yunnan province: 1) Dali College Affiliated Hospital; 2) Dali Prefecture Hospital; 3) Kunming No. 3 People's Hospital, and 4) Chuxiong Prefecture Hospital, 120 biological samples have been collected, with approval from the Institutional Review Boards of the School of Public Health of Wuhan University and Hummingbird IRB Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk •The genomic characterization of SL-CoVs in Year 3 was focused on Rhinolophus sinicus in Yunnan, our plan for Year 4 is to obtain complete S gene, RdRp gene or full-length genome sequences of more SL-CoVs from a broader range of bat species identified all over China and conduct a more comprehensive evolution study on SL-CoVs in bats. •To search for the receptor of SL-CoV with deletions in the homologous region of SARS-CoV RBD (i.e. Rp3, Rs672), and SL-CoVs which has been demonstrated to be unable to utilize bat ACE2 (i.e. Rs4231) whose receptors may be some molecules other than ACE2. •To conduct population genetics study of Rhinolophus sinicus ACE2s, which includes: the amplification of ACE2 genes from Rhinolophus sinicus samples of different origin, test of the usage efficiency of Rhinolophus sinicus ACE2s of different origins by SL-CoVs and kinetics study on the binding of SL-CoV RBD to different Rhinolophus sinicus ACE2s. •Phylogeographic study of bat-CoV to better understand the geographic distribution and evolution of bat-CoV genetic diversity in south China. •Phylogeographic study of bat host (Rhinolophus) species to assess the connectivity of bat populations and infer their historical movements and demographic history to improve our understanding of CoV transmission among bat populations in southern China. •Cophylogenetic analyses of bat host and CoV phylogenies to assess frequency of cross-species transmission. Comparison of Alphaand Beta-CoV cophylogenetic patterns building on Year 3 analyses using published sequences. Specific Aim 3: Testing predictions of CoV inter-species transmission. RPPR Page3

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RPPR FINAL •Using the reverse genetic method, we will construct chimeric viruses with the backbone of MERS-CoV and the S genes from diverse newly identified bat MERS-related coronaviruses, and examine the pathogenicity of bat MERS-related coronaviruses on cell and animal levels. ·The animal infection experiments are planned to be conducted in following years to study the pathogenicity of diverse SL-CoVs and MERS-related CoV that we identified in Chinese bats. •Surveillance of infection in human populations by bat-borne CoVs in Guangxi and Guangdong provinces in previously identified areas with human populations of high risk of exposure to bats. PCR and ELISA will be used, respectively, for detection of viral nucleic acids and antibodies against the viral nucleocapsid protein or spike protein. RPPR Page4

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B.2 (5R0IAII 10964-04.pdt) 8.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? 1R01Al110964 Year 3 Report Year 3 Report: Understanding the Risk of Bat Coronavirus Emergence Award Number: 5R01Al110964-04 B.2 What was accomplished under these goals? SUMMARY Pl: Daszak, Peter The results of the 3rd year of our R01 work are detailed below. They include: • Initial analysis of behavioral risk qualitative research in Yunnan and wildlife market observational data in Guangdong, that suggests a reduction in wildlife hunting, trade and consumption may be underway in southern China. • Results from a behavioral risk survey of over 1,000 people in two provinces of southern China that assesses exposure to wildlife and prior bouts of unusual illness, with concurrent taking of samples to test for evidence of exposure to SL-CoVs. • The finding of serological evidence of spillover of bat SARS-like CoVs in 6 people in Yunnan • Testing of over 1,000 bat samples to identify diverse alpha- and betacoronaviruses • Full genome characterization of 26 alphacoronaviruses. • Receptor binding domain sequences from 37 new bat SL-CoVs that shows S proteins re more diverse than previously thought. • Host-virus co-phylogeographic analysis of a diverse group of >1,300 bat CoVs showing that these viruses have a larger host range, weaker host specificity and higher frequency of cross-genera transmission than previously thought. • Use of our reverse genetics system to identify 3 more novel SL-CoVs with potential to directly infect people. Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces During Year 3 we began analyzing the qualitative research that was conducted in Year 2. In addition, we developed a digital application for a community-based integrated biological behavioral surveillance system and rolled this out in two provinces. The tool aims to identify specific animal exposure risk factors associated with biological evidence of exposure to SARSlike CoV (i.e. seropositive status). Qualitative Research Interviews conducted in Yunnan province during Year 2 were transcribed and translated into English. A total of 23 individuals (12 women; 11 men) were interviewed in rural regions where wildlife trade routes have been documented. Yunnan province was specifically selected for study because they have large wildlife populations, a diversity of wildlife species and numerous live animal markets. Individuals who were 18 years of age or older and who were able to provide informed consent were eligible to participate. The study was approved by the Institutional Review Boards of the School of Public Health at Wuhan University and Hummingbird IRB #2014-23. RPPR 1 Page5

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B.2 (5ROIAII 10964-04.pdt) 1R01Al110964 Year3 Report Pl: Daszak, Peter Participants were recruited primarily through local contacts that have been developed as part of wildlife conservation and health research that has been ongoing in these regions in China for the past decade. Contacts including wildlife conservationists and researchers, local government health outreach workers and wildlife farmers facilitated introductions and provided referrals. To achieve a sample with sufficient representation of categories of interest, participants were recruited using purposive sampling, which provides minimum quotas in terms of sex, age and wildlife exposure setting (e.g., live animal market, forest preserve). Educational attainment varied widely in the population; however, the majority of study participants reported limited schooling, primary education or less. This was further reflected in the occupational distribution of study participants (Fig. 1), while there was two respondents who reported more professional occupations, a doctor and an accountant, half (50%) were unskilled laborers or farmers, either agricultural or animal. There were one individuals who self-identified as animal farmers, farming wildlife, bamboo rat, civet, or nutria. Unemployed/ handyman 4" ~ anlmat furmer W>oror 15" 4" ShopOWner/ Office Woru,, 23" Figure 1. Occupation of Qualitative Research Participants (n=23) in Yunnan and Guangxi Provinces Thematic analysis provided the framework with which to code and analyze data from the ethnographic interviews and focus group. Five core themes were identified to form the basis for this: (1) human-animal contact, (2) unusual illness experience and response, (3) socioeconomics and daily living, (4) biosafety and (5) human environments and movement/travel. Individual interviews and field notes were studied to ensure familiarity with the data set in its entirety and to confirm narrative consistency within individual interviews prior to coding. Using these themes and a coding keyword guide allowed for a directed and consistent coverage of the domains that were the focus of the actual interviews. Qualitative data were reexamined to develop additional theoretical categories or typologies. This analysis aims to assess perceptions, knowledge and participation in the wildlife trade, as well as barriers to participation and observed changes over time. The data were coded for factors associated with wildlife consumption, socioeconomic drivers of wildlife trade, conservation and legal efforts, the prevalence and types of wildlife observed, and wildlife exposures that could transmit disease to humans (Table 1). RPPR 2 Page6

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Table 1. Topics covered in Ethnographic Interviews Theme Discussed in Ethnographic Interview No. of Respondents (%) (n=23) Work/Job Functions 22 96% Water& Food 22 96% Sanitation 22 96% Hygiene 22 96% Perceptions/Knowledge 22 96% Home Life 21 91% Education 20 87% Medical Care Treatment 20 87% Direct Contact with Animals 20 87% Travel 19 83% Observed Environment 19 83% Animal Responsibilities 19 83% Household Illness 19 83% Indirect Contact with Animals 19 83% Dally Routine 18 78% -= =,- - Family Economics 18 78% Illness from Animals 18 78% Animal Health 18 J 78% Animal Products/Rituals with Animal - 16 70% - Products Death 14 61% The data coding and analytic strategy was designed to avoid the need for expensive analytic software programs and to use standard word processing and spreadsheet programs readily available to in-country teams. These teams received training on qualitative data analysis, and they initiated the first phase of analyses. Analysis focused on wildlife trade and consumption in these two provinces, specifically on how respondents perceive and contact wildlife through the changing landscape around them. The aim was to identify motivations around animal consumption and practices. A number of participants reported that wildlife are purchased as a means to impress others as a symbol of wealth. Participants routinely reported that the cost of wildlife is double or triple that of regular livestock meat. Ironically, others reported that poorer individuals in these communities who continue to eat wildlife are sometimes scorned for their poverty, because this is a habit from an older time within China. Though there is a stigma to this habit, individuals did report opportunistically capturing and consuming wildlife when convenient. Participants also noted a decrease in wildlife over time: that in their childhood the forests would be full of the sounds of animals and birds, but this occurs no longer. This decrease was attributed to many factors, most commonly infrastructure development. Respondents discussed 3 RPPR Page 7

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter the government investing resources to build new roads and renovate local infrastructure with the intention of increasing tourism, and that this has had the impact of reducing forested habitat for wildlife. Hunting and selling of wildlife was not reported by any participant as a cause of observed wildlife depletion. However, participants did attribute a reduction in wildlife hunting and consumption to an increased enforcement of conservation laws. In particular, the story of one illfated hunter who killed a monkey-and was caught-was reported by a number of participants from the same village. Participants observed that the observed decrease in wildlife abundance and increased conservation law enforcement has made it more difficult to make a living from the wildlife trade. Participants reported choosing alternative forms of money making, indicating that only those people who belong to low socioeconomic classes continue to hunt secretly. The cost-benefit analysis that pits the threat of punitive consequences against the profits to be made through wildlife hunting are only feasible for those 'who have nothing to lose.' Table 2: S ecies Observed in Wet Markets in Guangdong Province from 2015 - 2016 Prionailurus bengalensis Nyctereutes procyonoides Sus scrofa Lepus sinensis Arctonyx collaris Hystrix brachyura Marmota sp. Rhizomes sinensis Erinaceus sp. Mustela putor/us Muridae Myocastor coypus Vu/pes sp. Mustela sibirica Paguma larvata Fe/is catus Canis lupus familiaris Cervinae Ovis aries Capra sp. Rattus norvegicus Leopard Cat Raccoon Dog Wild Boar Chinese Hare Hog Badger Porcupine Marmot Bamboo Rat Hedgehog Ferrets Rat (species unknown) Nutria Fox Siberian weasel Masked Palm Civet Domestic Cat Domestic Dog Sambar Deer Sheep Domestic Goat Common Rat Observations by research staff in live animal markets in Guangzhou found wildlife to be plentiful (Table 2), although no bats were seen for sale during the observation period. In contrast, wildlife 4 RPPR Page 8

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter was not found in live animal markets at the sites we visited in either Yunnan or Guangxi. This is a change from previous research visits to the same or similar communities, when bats, rodents and wild boar could be found. Locals in Yunnan and Guangxi attribute the change to conservation law enforcement. The success of conservation enforcement may have moved hunting and trapping underground and made the capture of local wildlife less economically feasible than other income generating activities. Integrated Biological Behavioral Surveillance in Yunnan and Guangdong Provinces To better assess the mechanisms of zoonotic viral spillover, and build on data acquired via ethnographic interview (above) we have designed a structured behavioral questionnaire to measure both exposure and outcome data. This behavioral risk survey assesses exposure to wildlife and bouts of unusual illness over a respondent's lifetime and in the past 12 months. In addition, participants were requested to provide serum to test for previous exposure to SARSlike CoV. The integrated surveillance was pilot-tested in October 2015 among residents living near bat caves or roosts where SL-CoVs have been previously detected in the bat population in Jinning County, Yunnan. After the questionnaire was pilot tested and optimized to fit the research aim, the survey was developed as a digital application (https :/ /www.dropbox.com/s/sv62neywuvl027 r/Questionna ire%20Com plete. docx?d I= 0% ). This allows standardization across all field teams and quality control. Four field team leads were trained on behavioral survey data collection, data collection technologies (the digital application) and analysis. The questionnaire was then administered in a follow-up survey in Yunnan province and then in Guangdong province. Surveillance in Guangxi is currently underway. Of 1089 participants who completed the behavioral questionnaire, 660 (61 % ) were women and 424 (39%) were men (5 missing for this variable). with a mean age of 50 (range: 10-99). Most reported being farmers (79%) (Fig. 2), a majority were long term residents (97%) and 41 % had a family income under 3000 RMB annually ($430). Almost three quarters (72%) of the respondents have had only primary level education or less. Occupation 14% 5% 2% 79% ■ Farmer ■ Construction ■ Migrant Farmer Other Figure 2. Occupation of Integrated Biological Behavioral Surveillance Participants in Yunnan and Guangdong Provinces RPPR 5 Page9

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Standardized syndromic case definitions informed questions concerning unusual illness experience (e.g. severe acute respiratory infections [SARI], influenza-like illness [ILi], febrile symptoms (Encephalitis]). Lifetime, 12 month, and unusual illnesses experienced in the family for the past 12 months were assessed for all participants. In the past year, SARI was reported by 55 (5.1%) respondents and 14 of those respondents also responded SARI symptoms in family members (Table 3). Table 3. Unusual Illness Experience In Respondents Lifetime, Past 12 months, Family members Influenza Like Illness (ILi) 305 (28.0%) 128 (11.8%) 142 (13.0%) Encephalitis 98 (9.0%) 52 (4.8%) 30 (2.8%) Hemorrhagic Fever 2 (0.2%) 2 (0.2%) 0 (0.0%) Fever with Diarrhea Nomiting 58 (5.3%) 25 (2.3%) I 21 (1.9%) Fever with Rash 10 (0.9%) 7 (0.6%) 7 (0.6%) Type of exposure and species exposed to are shown below (Table 4). Poultry was the most commonly contacted animal in almost all categories. Three quarters of respondents reported rodents or shrews entering their home in the past 12 months. Table 4: Animal Species Contact by Type of Contact "O "O - "O - Cl) Cl) Ill "O "O ... - "O ~ ..x CII "O Cl) CII - "O 0 CII CII C: Cl) CII Q. "C (V -c - .c CII - "O "C 1/) CII Cl) ... 0 (,) I.) s:::. c a. .!!! QI ::I ..x "O C: u "O .!!! iii ~ Cl ::I (V r/1 1/) 0 0 .:. C: :0 ::s ... .. "O C Cl) 0 ... ::c - Cl) C 'iij s:::. 0 11:1 - Cl) :::, I.) l!! a. 11:1 ~ C (.) .c CV "C 0 I.) Cl) Cl) :x: w C LL ::s Rodents/Shrews 0 33 5 834 38 2 1 1 28 26 Bats 0 5 0 180 8 lo 0 1 ] 5 5 Non-human 0 1 3 7 4 0 0 0 1 1 primates Birds 3 19 8 497 39 13 0 0 112 12 Carnivores 1 16 7 100 36 0 0 0 19 10 Ungulates 0 5 12 23 50 10 0 0 18 1 Poultry 5 514 843 134 719 5 8 6 425 7 Goats/Sheep 0 16 38 4 80 1 0 0 j 17 0 Swine 3 210 494 43 533 47 1 1 147 2 Cattle/Buffalo 0 12 77 10 102 5 1 0 ] 11 1 Dogs 342 40 303 252 62 0 0 22 16 2 Cats 163 10 137 275 18 0 0 11 11 0 RPPR 6 Page 10

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B.2 (SROIAII !0964-04.pdf) 1R01Al110964 Year 3 Report Pl: Daszak, Peter Animal exposures among those who reported unusual illness experiences in the past 12 months were evaluated, focusing on three high interest syndromes: SARI, ILi, and encephalitis. Of the 55 respondents who reported SARI symptoms, 49 reported: raising animals; animals in the home; preparing recently killed animals and buying live animals; 50% reported slaughter. Among the 16 respondents who reported ILi symptoms, 12 (75%) reported handling/preparing recently killed animals, 11 (69%) handling live animals or having animals in the home, 10 (63%) reported slaughtering/killing animals or buying live animals at wet market, 9 (56%) raised live animals, 7 (44%) reported a pet, and 1 (6%) reported animal feces near food or eating animal touched or damaged food, hunting, or eating raw/undercooked animal products. Among the four respondents who reported encephalitis symptoms, 3 (75%) reported hunting, handling or raising animals, 2 (50%) reported animals in the home, 1 (25%) reported having animals as pets, slaughtering/killing animals, or having bought live animals at a wet market. Table 5. Self-Reporting Symptoms of Syndromes and Sociodemographic and Animal Contact. SARI Positive Ill Positive Encephalitis Positive n=55 n=128 n=52 Sociodemographics n % n % n % Mother Primary education or less 54 98.2% 121 94.5% 50 96.2% Primary education or less 45 81.8% 94 73.4% 38 73.1% Female 32 58.2% 74 57.8% 29 55.8% - - Income <3000RMB 30 54.5% 45 35.2% 23 44.2% -- -- - Travel (past 12m) 30 54.5% 69 53.9% 34 65.4% Children < 5 yrs in Household 15 27.3% 38 29.7% 17 32.7% Household member with same 14 25.5% 46 35.9% 10 19.2% syndrome Respondent age <35 6 10.9% 24 18.8% 14 26.9% Animal Exposures n % n % n % Come in home 50 90.9% 117 91.4% 50 96.2% Raise animals 49 89.1% 113 88.3% 48 92.3% Prepare/cook recently killed 37 67.3% 95 74.2% 35 67.3% Handle live 36 65.5% 72 56.3% 38 73.1% Slaughtered 31 56.4% 57 44.5% 34 65.4% Animals as Pets 23 41.8% 55 43.0% 28 53.8% Buy Animals at Wet Market 16 29.1% 49 38.3% 4 7.7% Shared water source 9 16.4% 13 10.2% 12 23.1% Feces in/near food 8 14.5% 9 7.0% 8 15.4% Consume raw/undercooked 7 12.7% 10 7.8% 9 17.3% Scratch/bite 4 7.3% 2 1.6% 4 7.7% Consume food damaged by animals 3 5.5% 5 3.9% 2 3.8% Hunt or Trap 2 3.6% 4 3.1% 7 13.5% ---- Collect dead wildlife 1.8% 0.8% 1 1.9% Consume sick animals 0 0.0% 0.8% 0 0.0% 7 RPPR Page 11

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B.2 (SROIAII !0964-04.pdf) 1R01Al110964 Year 3 Report Pl: Daszak, Peter We examined the sociodemographic attributes and the types of contacts that were reported in those who reported SARI, ILi, or encephalitis-like symptoms in the past year (see Table 5). Over 65% of respondents these syndromes and also reported raising animals, animals coming in the home, or preparing meat or organs from a recently killed animal. A quarter of those who reported symptoms consistent with that of encephalitis were under the age of 35. Respondents were asked about the source of their unusual illnesses. None reported any kind of animal exposure as a potential source of infection and 11 % did not have any idea what may have caused their previous infection, despite the fact that a majority of respondents who reported SARI, ILi, or encephalitis symptoms also reported animal exposures (Table 5). Just over 30% of respondents reported purchasing live animals from a wet market in the past year. Over half (582; 53%) of respondents were worried about disease or disease outbreaks in animals at wet markets and 56% of people believe that animals spread disease. However, those who had purchased animals from markets in the last 12 months reported a great deal of behavior change being undertaken. In particular, respondents reported buying live animals less often 33%, only buying farmed wildlife 32% or buying meat at the supermarket 30% (Table 6). For those who participated in animal slaughter or were scratched or bitten in the past year, only 48 respondents (9.9%) reported visiting a doctor. Table 6: Behavior Change at Wet Market in the last 12 months Behavior Wash hands 119 - 33.4% Buy live animals less often 119 33.4% Buy only farmed wildlife 113 31.7% Sometimes shop for meat at supermarket 107 30.1% -- Wear gloves 7 1.9% -- -- Wear a mask 5 1.4% Serological Evidence of Bat SARS-Like CoV Infection in Humans Along with the behavioral survey questionnaire, respondents were also asked to provide a biological sample to assess SARS CoV spillover at the high-risk location where the questionnaire has been implemented. A sensitive and specific ELISA method was developed using the recombinant bat SL-CoV Rp3 NP protein to detect SL-CoV lgG antibodies. Six (2.8%) serum samples from 218 village residents who lived closely to the bat colonies in Yunnan where we isolated SL-CoV WIV1 and WIV16 were positive for SARS-like CoV antibodies (Fig. 3). The 6 ELISA positive samples were further confirmed as anti-SL-CoV NP lgG positive by western blot using recombinant Rp3-NP as antigen (Fig. 4). 8 RPPR Page 12

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report .i = 1.4 1.2 1.0 -; 0.8 > O 0.6 0 0.4 0.2 .. o.o..._--r--___ """T" ___ ----r __ Jinning Wuhan SARS patients human sera Pl: Daszak, Peter Figure 3. Serum samples from Jinning, Wuhan, and SARS patients were screened for reactivity of Rp3-NP. Bar in the diagram indicates optical density (OD) cutoff value (0.45) based on healthy blood donors in Wuhan. kDa "> #_ IJ,J.,,_,b' I:~ ~ ~ ~ ~</$ ~ Saniplcs (OD Value) !~ - JNl29 (0.71) ,~~~ Ii= - JN0.53 (0.70) !,:?-_- ;; -.,,., .. ,,, 43---==--= JNll7 (0.51) JS~ 1li-- 1 JN04 l (0.49) - SARS patient (1.19) ~ ~~ o",,_,<o"' ~ ~~ 4J? ~ Samples (OD Value) _ !N0l9 (0.48) _ JNOS9 (0.43) _ 1N036 (0.40) _ JNOJO (0.36) WH227 (0.41) Figure 4. Western blot analysis of reactivity of human sera to Rp3-NP. Linking Serological Findings with Respondent Questionnaire Data Of the 6 respondents in Jinning, Yunnan with serological evidence of SL-CoV infection, 4 had handled animals, 3 had raised or cooked meat from recently killed animals, 2 found animal feces near food stuffs, and 1 slaughtered or hunted an animal. Three of the individuals had contact with poultry in the past twelve months and 2 had contact with either birds, swine or buffalo. One individual reported having contact with a bat. Responses to the questionnaire show that in the last twelve months all of the respondents who have positive testing results, had animals in their dwelling and had contact with rodents or shrews. All 6 of the respondents had reported purchasing an animal from a wet market in the past twelve months. In addition, 215 oral swabs and 212 rectal swabs collected from human participants in Jinning and Yunnan province were tested for CoV RNA, and no positive results were found. 534 oral swabs, 526 rectal swabs from Xishuangbanna, Yunnan province; and 419 oral swabs, 412 rectal swabs from Ruyuan and Zengcheng, Guangdong province are being tested for CoV. RPPR 9 Page 13

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-Co V spillover risk Bat CoV PCR Detection and Sequencing from Live-Sampled Bat Populations We collected 893 rectal swab samples, 167 fecal samples and 33 blood samples from at least 17 bat genera in Yunnan, Guangdong, Guangxi, Hubei and Tibet provinces (Table 7) in Year 3. During this year, overall 1060 samples were tested for CoV RNA and 130 (12.3%) were positive (Table 8). Table 7. Bat samples collected for CoV surveillance in Year 3 Date of Sampling Sampling Locations Rectal Felclatl I Bl~od swab pe e specimen Ma 11 2016 Men la, Yunnan 32 9 Jingna, Yunnan 16 114 13 May 22" 2016 Lufen , Yunnan 53 June.July, 2016 Shixing county, Shaoguan, Guangdong 113 July 2016 Qin zhangshan, Shaoguan, Guan dong 101 16 26 July 2016 29 August Lengshuitang village, Guilin, Guangxi 135 August Nanxishan Park, Guilin, Gua 31 August 53 Liangkou twon, Conghhua, Guangdong 32 Jinning, Yunnan 34 Luten , Yunnan 25 Jingna, Yunnan 33 August, 2016 Men hai, Yunnan 125 ' • I • Yaoqu village, Mengla, Yunnan 30 -- • • I • Wuhan, Hubei 36 -- .. I • Motuo, Tibet Total 7 893 167 I 33 Genetically diverse alphacoronaviruses related to bat coronavirus 1 A/1 B, HKU7, HKU6 and HKU2 were identified in Miniopterus, Myotis and Rhinolophus bats, respectively. A novel alphacoronavirus related to human coronavirus NL63 was detected in Tylonycteris robustula in Yunnan. SARS-like coronaviruses were detected in 14 Chinese horseshoe bats (Rhinolophus sinicus) in Yunnan and Guangdong. Betacoronaviruses related to HKU5 were found in Pipistrellus abramus from Hubei, while two lineages of HKU4-related viruses were identified in two species of Tylonycteris bats in Yunnan (Fig. 5). 10 RPPR Page 14

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Table 8. CoV testing results for bat samples collected in Year 3 Aselliscus stoliczkanus 31 31 Rhinoluphus spp. 16/41 11 /136 6/60 5 33/242 Hipposideros spp. 17 1/126 6 8 1/157 Myotis spp. 7 6/34 7/69 1 13/111 Chaerephon spp. 8 8 Megaderma spp. 2 1 3 la io 1 1 Tylonycteris spp. 32/124 8 32/132 Pipistrel/us spp. 1 45 5/35 2 5/83 Eonycteris spelaea 1/29 1/29 Nyctalus velutinus 2 2 Coe/ops spp. 2 2 Miniopterus spp. 9/17 9/17 Taphozous 31 31 melanogopon Cynopterus sphinx 3 3 Murina spp. 1 1 Fecal pellets 35/167 35/167 Sub-total I 85/462 I 27 /370 I 13/166 I 5/36 I 0/26 I 130/1060 11 RPPR Page 15

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter 0.1 CAl60170/ Mlnlop/erus fullglnosus /Guangdor;;i/2016 1614341 Mlnfopterus schrelbersfl /Guangdong/2016 ol-coronoviru .. 1 B-stroin,WCF6-ORF1B•gcnc DQ666338 161460/ Hlpposlderos arm/ger /Guangdong/2016 CA 1600551 Mtnio,Xerus fuscus /Guangdong/2016 161454/ Mlnlopterus schrelbers/1 /Guangdong/2016 (61 8at-corona\/irus-1A-slrain-AFCD62 EU420136 Bat-coronavirus-HKU8-isolate•HKU8-1 RdRp DQ249228 161457/ Mm10p1erus sc reibers1i /Guangdong/2016" --j 1614471 Mlnlo,?terus fullglnosus /Gua_llQ9_Q!!QI~ Bat-coronavirus.+IKU7-lsolate-HKU7•1 RdRp DO249220 1619041 Myotis sll/gorens/s /Guangxi/2016 Myotis davidii corona1nrus isolate XYMd70 RdRp KF569991 ~-- ··1622171 Rhlnolophus slnicus /Yunnan/2016 (2) '-----1 _r 1 ~942/ Hipposlde10s Pomona /Gualljl!'i/2016 ( 4) L..{ Rousettus-bat-coronavirus-HKU10-isolale-175A JQ989271 Hippos,dero•-b@t-coron"'1rus-HKU10-isolot&- TT3AJQQ8Q266 1622801 Ty/onycterls robustu/a/Yunnan/2016 (2) '-------4 HCoV NL63 c=-:162191I Myotis pilosus /Yunnan/2016 L 161519/ Myotls ricketti /Guangdong/2016 161905/ Myotis siligoronsis /GuangxU2016 (61 1615141 Myotis p/losus /Guangdong/2016 (4) 1615151 Myot/s chlnensls /Guangdong/2016 e.,1.coronaviru"•HKU6-ioolatc-HKU6-1 RdRp DQ249224 r-- CA'l60"f957 Rh/nolophuspus,l/iis/CuangdongltOf6 m160888/ Rhino/ophus pus/llus/Guangxi/2016 (16) 160660/ Rhlnolophus sfnlcus /Yunnan/2016 AA160041/Yunnan/2016 (27) 162140/ Rhino10 nusamn1s1Guan Clong/2016 (4) 8at-coronavin1s-HKU2-strain-HKU2/HK/33/2006 EF203C67 ~A160258/Plp/srre/lus abramus/Hub~l/2016 (5) 6at-coronav1rus-HKU5-1 EF065S09 162331/Tylonycreris pachypus/Yunnan/2016 (24) Bat-coronavirus-HKU4-1 EF065S05 162270/Ty/onyercris robustu/a/Yunnan/2016 (4) -- Human-coronavirus-HKU111enotype-B AY88400t 162387/Rousettus sp./Yunnan/2016 (2) Bat-eo,onavirus-HKU0-2 EF065514 '-------1 e0768iEonycteris spelaea/Yunnan/2016 161465/Rhlnolophus s/n/cus/Guangdong/2016 (3) Bat-SARS-corooavi1us-Rf1 004 t 2042 Bat-SARS-corona1nrus-HKU3-1 00022305 '-----------H"162177/Rhinolophusstnlcus/Yunnanl2016 (11) Bat-SARS-coronavirus-Rp3 D0071615 SARS-coronavirus-BJ01 AY278488 SARS-coronavirus-S23 AY304486 SARS-coronavirus-GZ02 AY390556 RsSHC014 KC881005 Rs3367 KC881006 jYCaV a-~v Figure 5. Phylogenetic analysis of partial RdRp gene of CoV (440-nt partial sequence). Genomic Characterization of Novel Bat Alpha- Coronaviruses We generated full-length genome sequences of 26 novel alphacoronaviruses from multiple Hipposidoeros, Rhinolophus and Hypsugo bat species. These alphacoronaviruses grouped into 4 different lineages, including HKU10-like Co Vs and 3 novel species according to criteria generated by the International committee of Taxonomy of Viruses (ICTV) (Fig. 6). Strains belonging to the novel lineage from Rhinolophus share highly similar genome structures with each other but are distinct from all previously sequenced alphacoronaviruses. Putative 3b and 3c genes were identified at the upstream of the E gene, and a 7b gene at the downstream of the N gene was a homologue to Rhinolophus bat SARS-like CoV ?a gene. These results expand the understanding of genetic diversity of bat alphacoronaviruses. 12 RPPR Page 16

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter 100 92 S7 BtCoV 887 Hipposideros pomona Yunnan 2006 BtCoV 7345 Hipposideros pomona Yunnan 2014 BtCoV 3740 Hipposideros pomona Yunnan 2012 BtCoV QZ140937 Hi osiderQs mona Hainan 2014 Hipposideros HKU10 2 Rousettus HKU10 ........ __ 100-i BtCoV 141391 Hipposideros pomona Guangdong 2014 99 BtCoV 141402 Hipposideros pomona Guangdong 2014 100 BtCoV 3723 Hipposideros pomona Yunnan 2012 BtCoV 4996 Hipposideros pomona Yunnan 2013 4 BtCoV 7496 Aselliscus stoliczkanus Laos 2014 BtCoV 7549 Hipposideros pomona Laos 2014 BtCoV 7560 Hipposideros pomona Laos 2014 ,ij ~tCoV 7565 Hipposideros pomona Laos 2014 s14-1 BtCoV 7546 Hipposideros pomona Laos 2014 BtCoV 7557 Hipposiderospomona Laos 2014 BtCoV 2714 Hipposideros larvatus Yunnan 2011 -===""'i:w~B~t~C;,oV~27,_,2._,.7...,.H-'-'i,i;n:iosideros larvatus Yunnan 2011 ~ BtMf GD2012 BtCoV HD13591a Rhinolophus affinis Guangdong 2013 -~BtCoV 3376 Rhinolophus sinicus Yunan 2012 BtCoV 4125 Rhinolophus sinicus Yunan 2012 1 BtCoV 4259 Rhinolo hus sinicus Yunan 2013 t~vSZ2-013 88 BtCoV NL 140459 Hypsugo spp. Guangdong 2014 '----t----,,,,,-1 BtCoV NL 140460Alpha Hyps1.190 spp. Guangdong 2014 BtCoV NL140461 Hypsugo spp. Guangdong 2014 97 BtCoV NL140480 Hii:>J)Osideros arrnigerGuangdong 2014 ,-------....::::=---= RhBatCoV HKU2 ~,_,------- Mi BtCoV HKUS 0.02 Figure 6. Phylogenetic analysis based on full-length RdRp gene sequence of alpha-Co Vs Genetic Diversity of Receptor-Binding Domain (RBD) of SARS-Like Coronavirus in Chinese Bats RBD sequences from 37 newly identified SL-CoV from various horseshoe bat species and Hipposideros bat species in Yunnan, Guangdong, Guangxi, Hubei and Hunan provinces were amplified and sequenced in Year 3. Phylogenetic analysis revealed that SL-CoV circulating in bat populations in China are highly diverse in the RBD region (Fi.g 7). Some strains possessed an RBD sequence distinct from all currently known bat SL-CoVs and formed a new cluster in the phylogenetic tree. However, except for a few strains from Yunnan, most of these SL-CoVs contained nucleotide deletions and were relatively distant to SARS-CoV in the RBD region. These findings suggest that the S gene of SL-CoVs in Chinese bats is even more genetically diverse than expected. The genomic characterization of SL-CoVs in Year 3 was focused on Rhinolophus sinicus in Yunnan, our plan for Year 4 is to obtain complete S gene, RdRp gene or full-length genome sequences of more SL-CoVs from a broader range of bat species identified all over China and conduct a more comprehensive study of the evolution of SL-CoVs in bats. 13 RPPR Page 17

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Figure 7. Phylogenetic analysis of the RBD region of the S gene of bat SL-Co Vs detected in China (newly identified sequences were marked in red). Bat Coronavirus Host-virus Phylogeography in China To analyze the extent to which different bat species and genera are host to similar bat-CoVs, we reconstructed viral phylogenetic relationships and mapped host-species associations onto these phylogenies. Our dataset includes all CoV RdRp sequences isolated from bat specimens collected by our team from 2008-2015 (Alpha-CoVs: n = 491 - Beta-CoVs: n = 326), including those collected under prior NIAID funding (1 R01 Al079231), and funding from Chinese Federal Agencies. All Chinese bat CoV RdRp sequences available in GenBank were also added to our dataset (Alpha-CoVs: n = 226 - Beta-CoVs: n = 206). Phylogenetic trees were reconstructed for Alpha- and Beta-CoVs separately using Bayesian inference and Maximum Likelihood (ML) approaches. RAxML was used to perform ML analysis and Bayesian analyses were performed with MrBayes 3.2.6. 14 RPPR Page 18

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B.2 (5ROIAII 10964-04.pdt) 1R01Al110964 Year3 Report Pl: Daszak, Peter Beta-CoV sequences clustered into four main genetic lineages: B (SARS-CoV and SARS-like CoVs), C (MERS-CoV), D and a potential new lineage related to lineage B (Fig. 8). An important phylogenetic structure is observed within lineages C and D. Alpha-CoV sequences clustered into numerous closely related and less-differentiated lineages (Fig. 9). We observed significant CoV lineage sharing among bat genera in our phylogenetic trees. Importantly SARS-like CoVs (SL-CoVs in lineage B) have been detected in Hipposideridae bats in addition to Rhinolophidae bats which were thought to be the putative natural host taxa of SLCoV (Fig. 8). We found additional bat genera that also hosted CoVs in this clade (Fig. 8), expanding potential host targets for novel SL-CoV discovery. CoVs closely related to Bat coronavirus HKU9 (lineage D), which were thought to be specific to pteropodid bats, have also been detected in hipposiderid and vespertilionid bats (Fig. 8). Important lineage sharing across several bat families has also been observed among most Alpha-CoV lineages (Fig. 9). We used host DNA barcoding to confirm these findings - host mitochondrial sequences were generated to confirm the host species identity for most samples. These results indicate a larger host range, weaker host specificity and higher frequency of cross-genera transmission for most bat CoV lineages than previously thought. These findings will have important implication in our understanding of bat CoV emergence and spillover risk in China. In Year 4 we will expand these analyses to include more explicit co-evolutionary analyses to identify the frequency and timing of host switching events for each major clade. 15 RPPR Page 19

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Beta-CoV 532 sequences (206 Genbank + 326 WIV) Lineage D Pteropodidae Pl: Daszak, Peter Rousettus + Eonycteris _.(HKU9H~~u;ettus -!Rousettus HKU9 < Rousettus + Eonycteris + Megaerops Cynopterus + Rousettus + H1pposideros + Pipistreltus + M otis HKU3 Rhinotophus + Pipistreltus + Myolis + Hipposideros + Tytonyc1erts .. Mlniopterus + Assetiscus + Chaerophon Lineage B (SARS) ~, H1ppos1deros (] Hipposideros + Vesperti110 HKU4 Myotis + Tylonycteris -,. Scotophitus + Cyncpterus -rt io+ Pipistlellus 7 Vespertilio < Hypsugo + Pipistrelus + Myotis HKU5 Pipistreltus Lineage C (MERS) Vespertilionidae Figure 8. Maximum Likelihood tree of partial RdRp gene sequences of Beta-Co Vs. Bat host genera are indicated along each lineage. Bat genera listed in red correspond to minor and potential new bat hosts and may represent cross-genera/family transmission events. RPPR 16 Page 20

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Alpha-CoV 717 sequences (226 GB + 491 WIV) - - ,j HKU2 Rhinolophus + Pipistrellus + Hipposideros + Myotis "'· ::. Mini~terus + Murin a Pl: Daszak, Peter Miniopterus + Rhinolophvs + Taphozous + Myofis + Hipposideros + Tylonycteris + Pipistrellus + Submyotodon Miniopterus bat CoV 1A + 1B _ I ,:, HKU7 Miniopterus + Myotis [ i H~~~olophus / Miniopterus + Rhinolophus + Myotis \ + Hipposideros + Taphozous -N~talus --\ Hypsugo + H,pposideros · Rhinolophus + Miniopten.Js + Myotis l --ln. r <J Hipposideros + Aselliscus + Rhinolophus I , ·1 + Rouseltus , HKU10 , Rhinolophus ~ ·1 Myotis + Rhinolophus + Miniopterus -r,iyotis <IMyotis ~ ]Myotis :1 Scotophilus bat CoV 512 L.; I Scotophilus + Kerivoula + Pipistrellus ;i + Miniopterus + Myotis + Rhinolophus Myotis + Pipistrellus I HKU6 Myotis + Mlniopterus + Taphozous + Hipposideros + Vespertilio + Nyctalus + Aselliscus Figure 9. Maximum Likelihood tree of partial RdRp gene sequences of Alpha-Co Vs. Bat host genera are indicated along each lineage. Bat genera listed in red correspond to minor and potential new bat hosts and may represent cross-genera/family transmission events. RPPR 17 Page 21

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report Pl: Daszak, Peter Global analysis of bat viral sharing to identify key host species We curated and analyzed a global dataset of bat host-virus associations to better understand the frequency, and connectivity of viral sharing among bats. We also used this to examine the importance of cave-roosting bats species in harboring and sharing viruses with non caveroosting species, and to identify specific hosts that are central in the network (Fig. 10). Cave roosting bat species are host to most CoVs found in bats (orange). We identified global patterns of viral coinfection based on the number of connections between each virus in the network (Fig. 10). We will expand this approach to our China-Gov specific field data in Year 4 . • • □· 8 Figure 10. An analysis of global bat virus sharing using data from the published literature combined with field data. Network analysis includes 152 bat host species and 80 fCTV recognized viral species, with 273 host-viral associations. Unique viruses are represented in circles with known Co Vs shown in orange, and each square represents a unique bat species. Green squares = facultative cave-roosting bat species; Blue squares = obligate cave-roosting species; Yellow squares = non cave-roosting species. Viruses are linked in the network based on host species that have been observed harboring the same virus - as detected using PCR or viral isolation:.. 18 RPPR Page 22

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B.2 (5ROIAII 10964-04.pdt) 1R01Al110964 Year3 Report Pl: Daszak, Peter Specific Aim 3: Testing predictions of CoV inter-species transmission In Year 3 we established an effective and economic reverse genetics system for bat SL-CoV which can be applied to efficiently rescue SL-CoVs that are difficult to culture. This can be used to explore the functions of newly identified SL-CoV genes, as well as to assess pathogenesis of novel bat SL-CoVs. Using this system, we demonstrated that the unique ORFx in WIV1 and WIV16 is a functional gene involving modulation of the host immune response but not essential for in vitro viral replication (Zeng et al, 2016, J Virol). Identification of Three Novel SL-Co Vs with Potential for Direct Transmission to Humans In Y2, we conducted full-length genome sequencing of 11 novel SL-CoVs detected in a single bat habitat in Yunnan province, which included strains highly similar to human/civet SARS-CoV in the most variable genes (N-terminal domain and RBD in the S gene, ORF8 and ORF3) (under revision). Based on recombination analysis, we hypothesized that the direct progenitor of the pandemic SARS-CoV may originated from this location after sequential recombination events at multiple genomic positions. Among the 11 newly identified SL-CoVs, three different strains namely Rs4874, Rs7327 and Rs4231 contained no deletions in the RBD region but their RBD sequences varied from each other. Rs4874 has an S gene almost identical to that of WIV16. Rs7327's S protein varies from that of WIV1 and WIV16 at three aa residues in the receptor-binding motif, including one contact residue (aa 484) with human ACE2. Rs4231 shares similar NTD sequence with WIV1 and WIV16, but has a distinct RBD sequence. In Year 3, we successfully isolated Rs4874 from the single fecal sample. Using the reverse genetic system we previously developed, we constructed two chimeric viruses with the WIV1 backbone replaced with the S gene of Rs7327 and Rs4231, respectively. Vero E6 cells were respectively infected with Rs4874, WIV1-Rs4231S and WIV1- Rs7327S, and efficient virus replication was detected by immunofluorescence assay in all infections. To assess the usage of human ACE2 by the three novel SL-CoVs, we conducted virus infectivity studies using Hela cells with or without the expression of human ACE2. All viruses replicated efficiently in the human ACE2-expressing cells. The results were further confirmed by quantification of viral RNA using real-time RT-PCR (Fig.11). These finding suggests that diverse variants of SL-CoV S protein without deletions in their RBD are able to use human ACE2 as receptor for cell entry. Diverse SL-CoVs capable of direct transmission to humans are circulating in bats in southwestern China, which represents a potential risk of emergence given the opportunity to spillover to other animals and/or human populations. 19 RPPR Page 23

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B.2 (5R01AII 10964-04.pdf) 1 RO 1 Al 110964 Year 3 Report B A virus hACE2 Rs4874 WIV1• Rs4231S WIV1· Rs7327S ..J l.E+0S + + Hela-hACE2 E 0 1.E+04 +-----~,.._,__ __ _ II) 9 1.E+03 (.} ~ l.E+02 +----=.,,,.;...._ ____ _ 1.E+Ol -+----------- Oh 12h 24h 48h Hour post infection Pl: Daszak, Peter -Rs4874 _.,. WIV1-Rs4231 ~WIV1-Rs7327 Figure 11. Analysis of receptor usage by immunofluorescence assay (A) and real-time PCR (8). Additional Year 3 items for Specific Aim 3: • The full-length infectious cDNA clone of MERS-CoV has been successfully constructed. RPPR The full-length S gene of 12 different novel bat MERS-related coronaviruses have been amplified and cloned into the T-vectors. In Y4, we aim to use the reverse genetic method, and construct chimeric viruses with the backbone of MERS-CoV and the S genes from 20 Page 24

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B.2 (5ROIAII 10964-04.pdt) 1R01Al110964 Year3 Report Pl: Daszak, Peter diverse newly identified bat MERS-related coronaviruses, to examine the pathogenicity of bat MERS-related coronaviruses on cell and animal levels. • Establishment of animal infection models for bat SL-CoV and MERS-related CoV: Mice with human ACE2 have been imported to China and have been bred for one generation in Wuhan Institute of Virology. Transgenic mice that express human DPP4 have also been constructed and are being bred. The animal infection experiments are planned to be conducted in following years to study the pathogenicity of diverse SL-CoVs and MERS-related CoV that we identified in Chinese bats. Specific Goal Not Meet • Observations and animal sampling at wildlife markets were not done in Year 3 because the stricter law enforcement and subsequent cautiousness of traders make it difficult to access to wild animal in markets. Instead, we piloted the wild animal farm survey and will be focusing on it in Year 4, with evidence from pre-investigations that shows most wild animal farms serve as transit points during the wildlife trade. • The passive hospital surveillance has been piloted in Year 3 and will continue in Year 4 to collect and test samples for SL-CoV and other viral families • Cophylogenetic analyses of bat host and CoV phylogenies to assess patterns of evolutionary congruence and frequency of cross-species transmission to be continued in Year4 • Animal infection experiments of SL-CoVs and MERS-related CoV were not done in Year 3, as this is planned as part of work in Year 4. Significant Oral Presentations 1. Daszak P. Plenary talk, One Health-EcoHealth Congress, Melbourne, Dec. 2016 2. Daszak P. 2nd annual Global Pandemic Policy Summit, Scowcroft Ctr, Texas A&M Univ. 3. Daszak P. Global Health Security Agenda side event, UN World Humanitarian Summit: FAO/WHO/USAID/Global He@lth 2030 Innovation Task Force; Istanbul, Turkey. 4. Daszak P. Symposium at Ecole du Val-de-Grace, Paris 5. Daszak P. Plenary, Institute of Zoology symposium on Bushmeat and disease risks, London. 6. Daszak P. Duke University Provost's Forum on Conservation and Health 7. Oliva! KJ. The 17th International Bat Research Conference "Assessing the Risk of Disease Emergence from Bat Hunting: Overview and Implications for Risk Mitigation". Durban, South Africa, 2016 8. Daszak P. American Public Health Association Annual Meeting 2016 "Preliminary Results from An Innovative One Health Behavioral Surveillance System". Denver, 2016 RPPR 21 Page 25

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B.4 (5ROIAII 10964-04 Professional Development.pdt) lROlAil 10964 Year 3 Report PI: Daszak, Peter 8.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? We presented this work to the chief physicians, nurses, and directors from county-level clinics in Guangdong and Yunnan provinces during the implementation of Integrated Biological Behavioral Surveillance in Chuxiong and Guangzhou. All the research staff were trained and retrained for the biosafety and PPE use for human biological sampling. 11 graduate students from School of Public Health of Wuhan University and Wuhan Institute of Virology of CAS were trained for laboratory and field biosafety and PPE use, behavioral data collection methodologies and technologies, and data analysis. Research Technician Dr. Guangjian Zhu was invited by the Institute of Pathogen Biology, Chinese Academy of Medical Science & Peking Union Medical College to provide training to 10 field team members regarding biosafety and PPE use, bats and rodents sampling. RPPR Page 26

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RPPR FINAL C. PRODUCTS C.1 PUBLICATIONS Are there publications or manuscripts accepted for publication in a journal or other publication (e.g., book, one-time publication, monograph) during the reporting period resulting directly from this award? Yes Publications Reported for this Reporting Period ·i• ~:..,,,._...,_.,._;,,.,• ,------------------------------ Non-Compliant (b)(4) Complete Zeng LP, Gao YT, Ge XY, Zhang Q, Peng C, Yang XL, Tan B, Chen J, Chmura AA. Daszak P, Shi ZL. Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response. Journal of virology. 2016 July 15;90(14):6573-82. PubMed PMID: 27170748; PubMed Central PMCID: PMC4936131. Complete Olival KJ, Willoughby AR. Prioritizing the 'Dormant' Flaviviruses. EcoHealth. 2017 March;14(1 ):1-2. PubMed PMID: 28194584; PubMed Central PMCID: PMC5386397. C.2 WEBSITE($) OR OTHER INTERNET SITE(S) Nothing to report C.3 TECHNOLOGIES OR TECHNIQUES NOTHING TO REPORT C.4 INVENTIONS, PATENT APPLICATIONS, AND/OR LICENSES Have inventions, patent applications and/or licenses resulted from the award during the reporting period? No C.5 OTHER PRODUCTS AND RESOURCE SHARING NOTHING TO REPORT RPPR Page 27

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RPPR FINAL D. PARTICIPANTS D.1 WHAT INDIVIDUALS HAVE WORKED ON THE PROJECT? ~l'~lrl41~,.--n • <.->.: '' ' • • 1 ,, ' ~.. ,. " f l ' } ,., ! ~-~F7~7 , ~ 1.,.,. • ~" ' ' l ~ 1,.« ~ I '' • ' .. ~ '1:,_.,t,,M,,C,,.'<,,,.,_<-:'.,K.-~ '-"''"" ·-:-,t,~,:,,.,,. • r~~-,,,.,.,,,,..; ;I.,',.,.~,. r.,-.,J~<,; \:'.::f ·"'" ;t,,.~~--"'·'';,~,1 $;,:-c,:. (b) (6)] y N (b)(6>: N N N N N N N N N Glossary of acronyms: S/K - Senior/Key DOB - Date of Birth DASZAK, PETER KE, CHANGWEN Ross, Noam Martin SHI, ZHENGLI OLIVAL, KEVIN J ZHANG, YUNZHI ZHU, GUANGJIAN GE, XINGYI EPSTEIN, JONATHAN H CHMURA, ALEKSEI A ZHANG, SHUYI Cal - Person Months (Calendar) Aca - Person Months (Academic) Sum • Person Months (Summer) D.2 PERSONNEL UPDATES D.2.a Level of Effort RPPR BS,PHD PHO PhD PhD PHO PHO PHO PHO MPH,DVM ,BA.PHO BS PHO PD/Pl CoInvestigator CoInvestigator CoInvestigator CoInvestigator CoInvestigator CoInvestigator CoInvestigator CoInvestigator Non-Student Research Assistant CoInvestigator (b) (4) (b) (6) Center for CHINA Disease Control and Prevention of Guangdon g Province Wuhan CHINA Institute of Virology Yunnan CHINA Provincial Institute of Endemic Diseases Control & Prevention East China CHINA Normal University Wuhan CHINA Institute of Virology East China CHINA Normal University Foreign Org - Foreign Organization Affiliation SS - Supplement Support RE - Reentry Supplement DI - Diversity Supplement OT- Other NA - Not Applicable Page 28 NA NA NA NA NA NA NA NA NA NA NA

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RPPR FINAL Will there be, in the next budget period, either (1) a reduction of 25% or more in the level of effort from what was approved by the agency for the PD/Pl(s) or other senior/key personnel designated in the Notice of Award, or (2) a reduction in the level of effort below the minimum amount of effort required by the Notice of Award? No D.2.b New Senior/Key Personnel Are there, or will there be, new senior/key personnel? No D.2.c Changes in Other Support Has there been a change in the active other support of senior/key personnel since the last reporting period? No D.2.d New Other Significant Contributors Are there, or will there be, new other significant contributors? No D.2.e Multi-Pl (MPI) Leadership Plan Will there be a change in the MPI Leadership Plan for the next budget period? NA RPPR Page 29

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RPPR E. IMPACT E.1 WHAT IS THE IMPACT ON THE DEVELOPMENT OF HUMAN RESOURCES? Not Applicable FINAL E.2 WHAT IS THE IMPACT ON PHYSICAL, INSTITUTIONAL, OR INFORMATION RESOURCES THAT FORM INFRASTRUCTURE? NOTHING TO REPORT E.3 WHAT IS THE IMPACT ON TECHNOLOGY TRANSFER? Not Applicable E.4 WHAT DOLLAR AMOUNT OF THE AWARD'S BUDGET IS BEING SPENT IN FOREIGN COUNTRY(IES)? RPPR Page 30

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RPPR FINAL F.CHANGES F.1 CHANGES IN APPROACH AND REASONS FOR CHANGE Not Applicable F.2 ACTUAL OR ANTICIPATED CHALLENGES OR DELAYS AND ACTIONS OR PLANS TO RESOLVE THEM NOTHING TO REPORT F.3 SIGNIFICANT CHANGES TO HUMAN SUBJECTS, VERTEBRATE ANIMALS, BIOHAZARDS, AND/OR SELECT AGENTS F.3.a Human Subjects No Change F.3.b Vertebrate Animals No Change F.3.c Biohazards No Change F.3.d Select Agents No Change RPPR Page 31

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RPPR FINAL G. SPECIAL REPORTING REQUIREMENTS G.1 SPECIAL NOTICE OF AWARD TERMS AND FUNDING OPPORTUNITIES ANNOUNCEMENT REPORTING REQUIREMENTS NOTHING TO REPORT G.2 RESPONSIBLE CONDUCT OF RESEARCH Not Applicable G.3 MENTOR'S REPORT OR SPONSOR COMMENTS Not Applicable G.4 HUMAN SUBJECTS G.4.a Does the project involve human subjects? Yes Is the research exempt from Federal regulations? No Does this project involve a clinical trial? No G.4.b Inclusion Enrollment Data Report Attached: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 G.4.c ClinicalTrials.gov Does this project include one or more applicable clinical trials that must be registered in ClinicalTrials.gov under FDAAA? No G.5 HUMAN SUBJECTS EDUCATION REQUIREMENT Are there personnel on this project who are newly involved in the design or conduct of human subjects research? No G.6 HUMAN EMBRYONIC STEM CELLS (HESCS) Does this project involve human embryonic stem cells (only hESC lines listed as approved in the NIH Registry may be used in NIH funded research)? No G.7 VERTEBRATE ANIMALS Does this project involve vertebrate animals? Yes G.8 PROJECT/PERFORMANCE SITES RPPR Page 32

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RPPR FINAL [=======! ll~~ Primary: EcoHealth 077090066 NY-010 460 West 34th Street Alliance, Inc. 17th Floor New York NY 100012317 Wuhan Institute of 529027474 Xiao Hong Shan, No. 44 Virology Wuchang District Wuhan Wuhan University School 549376772 00-000 115 Donghu Road of Public Health Wuhan nullnull G.9 FOREIGN COMPONENT Organization Name: Wuhan Institute of Virology Country: CHINA Description of Foreign Component: Principal Laboratory for all Research in China as per section GB (above) and detailed in our Specific Aims Organization Name: Wuhan School of Public Health Country: CHINA Description of Foreign Component: Principal Coordinating Team for all project field work as per section GB (above) and detailed in our Specific Aims G.10 ESTIMATED UNOBLIGATED BALANCE G.10.a Is it anticipated that an estimated unobligated balance (including prior year carryover) will be greater than 25% of the current year's total approved budget? No G.11 PROGRAM INCOME Is program income anticipated during the next budget period? No G.12 F&A COSTS Is there a change in perfonnance sites that will affect F&A costs? No RPPR Page 33

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Inclusion Data Record (IDR) #: 166195 Delayed Onset Study ?: No Enrollment Location: Foreign Inclusion Enrollment Report Using an Existing Dataset or Resource: No Clinical Trial: No NIH Defined Phase Ill Clinical Trial: No Study Title: Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 Planned Enrollment Planned Enrollment Total: 2,460 NOTE: Planned enrollment data exists in the previous format; the PD/Pl did not enter the planned enrollment information in the modified format and was not required to do so. Only the total can be provided. Cumulative Enrollment Ethnic categories Racial Categories Unknown/Not Not Hispanic or Latino Hispanic or Latino Reported Ethnicity Total Female Male Unknown/ Female Male Unknown/ Female Male Unknown/ Not Reported Not Reported Not Reported American Indian/ Alaska Native 0 0 0 0 0 0 0 0 0 0 Asian 708 459 0 0 0 0 0 0 0 1167 Native Hawaiian or 0 0 0 0 0 0 0 0 0 0 Other Pacific Is lander Black orAfrican American 0 0 0 0 0 0 0 0 0 0 Whne 0 0 0 0 0 0 0 0 0 0 More than One Race 0 0 0 0 0 0 0 0 0 0 Unknown or Not Reported 0 0 0 0 0 0 0 0 0 0 Total 708 459 0 0 0 0 0 0 0 1167

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Notice of Award RESEARCH Federal Award Date: 06/18/2018 Department of Health and Human Services National Institutes of Health NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 5R01Al110964-05 FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Aleksei Chmura President 460 West 34th Street 17th Floor New York, NY 100012317 Award e-mailed to: (b)(6) ----------- Period Of Performance: Budget Period: 06/01/2018 - 05/31/2019 Project Period: 06/01/2014 - 05/31/2019 Dear Business Official: The National Institutes of Health hereby awards a grant in the amount of $581,646 (see "Award Calculation" in Section I and "Terms and Conditions" in Section Ill) to ECOHEALTH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the "Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01 Al 110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STIR) awards. Consult the NIH website http://grants.nih.gov/grants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1 ID)

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Tseday G Girma Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2

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SECTION I -AWARD DATA- 5R01Al110964-05 Award Calculation (U.S. Dollars) Salaries and Wages Fringe Benefits Personnel Costs (Subtotal) Materials & Supplies Travel Other SubawardsfConsortium/Contractual Costs Federal Direct Costs Federal F&A Costs Approved Budget Total Amount of Federal Funds Obligated (Federal Share) TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS YR I THIS AWARD I CUMULATIVE TOTALS 5 I $581,646 I Fiscal Information: CFDAName: CFDA Number: Allergy and Infectious Diseases Research 93.855 EIN: Document Number: PMS Account Type: Fiscal Year: 1311726494A1 RAl110964A P (Subaccount) 2018 I CAN 8472350 NIH Administrative Data: 12018 $581,646 PCC: M51 C / OC: 414E / Released: (b)(6) 06/15/2018 Award Processed: 06/18/2018 12:02:35 AM SECTION 11- PAYMENTfHOTLINE INFORMATION - 5R01Al110964-05 $167,708 $54,168 $221,876 $3,500 $35,918 $9,400 $191,576 $462,270 $119,376 $581,646 $581,646 $581,646 $581,646 $581,646 For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS - 5R01Al110964-05 This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. c. 45 CFR Part 75. d. National Policy Requirements and all other requirements described in the NIH Grants Policy Statement, including addenda in effect as of the beginning date of the budget period. e. Federal Award Performance Goals: As required by the periodic report in the RPPR or in the final progress report when applicable. f. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. Page-3

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(See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain references cited above.) Research and Development (R&D): All awards issued by the National Institutes of Health (NIH) meet the definition of "Research and Development" at 45 CFR Part§ 75.2. As such, auditees should identify NIH awards as part of the R&D cluster on the Schedule of Expenditures of Federal Awards (SEFA). The auditor should test NIH awards for compliance as instructed in Part V, Clusters of Programs. NIH recognizes that some awards may have another classification for purposes of indirect costs. The auditor is not required to report the disconnect (i.e., the award is classified as R&D for Federal Audit Requirement purposes but non-research for indirect cost rate purposes), unless the auditee is charging indirect costs at a rate other than the rate(s) specified in the award document(s). An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the System for Award Management (SAM). Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://qrants.nih.gov/qrants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01 Al 110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://grants.nih.gov/qrants/policy/awardconditions.htm for additional award applicability information. In accordance with P.L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OD-08-033 and the Public Access website: http://publicaccess.nih.gov/. This award represents the final year of the competitive segment for this grant. See the NIH Grants Policy Statement Section 8.6 Closeout for complete closeout requirements at: http://qrants.nih.gov/qrants/policy/policy.htm#qps. A final expenditure Federal Financial Report (FFR) (SF 425) must be submitted through the eRA Commons (Commons) within 120 days of the period of performance end date; see the NIH Grants Policy Statement Section 8.6.1 Financial Reports, http://grants.nih.gov/grants/policy/policy.htm#gps, for additional information on this submission requirement. The final FFR must indicate the exact balance of unobligated funds and may not reflect any unliquidated obligations. There must be no discrepancies between the final FFR expenditure data and the Payment Management System's (PMS) quarterly cash transaction data. A final quarterly federal cash transaction report is not required for awards in PMS B subaccounts (i.e., awards to foreign entities and to Federal agencies). NIH will close the awards using the last recorded cash drawdown level in PMS for awards that do not require a final FFR on expenditures or quarterly federal cash transaction reporting. It is important to note that for financial closeout, if a grantee fails to submit a required final expenditure FFR, NIH will close the grant using the last recorded cash drawdown level. If the grantee submits a final expenditure FFR but does not reconcile any discrepancies between expenditures reported on the final expenditure FFR and the last cash report to PMS, NIH will close the award at the lower amount. This could be considered a debt or result in disallowed costs. Page-4

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A Final Invention Statement and Certification form (HHS 568), (not applicable to training, construction, conference or cancer education grants) must be submitted within 120 days of the expiration date. The HHS 568 form may be downloaded at: http://qrants.nih.gov/grants/forms.htm. This paragraph does not apply to Training grants, Fellowships, and certain other programs-Le., activity codes C06, D42, D43, D71, DP7, G07, GOB, G11, K12, K16, K30, P09, P40, P41, P51, R13,R25,R28, R30,R90,RL5,RL9,S10,S14,S15,U13,U14,U41,U42,U45,UC6,UC7,UR2, X01, X02. Unless an application for competitive renewal is submitted, a Final Research Performance Progress Report (Final RPPR) must also be submitted within 120 days of the period of performance end date. If a competitive renewal application is submitted prior to that date, then an Interim RPPR must be submitted by that date as well. Instructions for preparing an Interim or Final RPPR are at: https://grants.nih.gov/grants/rppr/rppr instruction guide.pdf. Any other specific requirements set forth in the terms and conditions of the award must also be addressed in the Interim or Final RPPR. Note that data reported within Section I of the Interim and Final RPPR forms will be made public and should be written for a lay person audience. NIH strongly encourages electronic submission of the final invention statement through the Closeout feature in the Commons, but will accept an email or hard copy submission as indicated below. Email: The final invention statement may be e-mailed as PDF attachments to: NIHCloseoutCenter@mail.nih.gov. Hard copy: Paper submissions of the final invention statement may be faxed to the NIH Division of Central Grants Processing, Grants Closeout Center, at 301-480-2304, or mailed to: National Institutes of Health Office of Extramural Research Division of Central Grants Processing Grants Closeout Center 6705 Rockledge Drive Suite 5016, MSC 7986 Bethesda, MD 20892-7986 (for regular or U.S. Postal Service Express mail) Bethesda, MD 20817 (for other courier/express deliveries only) NOTE: If this is the final year of a competitive segment due to the transfer of the grant to another institution, then a Final RPPR is not required. However, a final expenditure FFR is required and should be submitted electronically as noted above. If not already submitted, the Final Invention Statement is required and should be sent directly to the assigned Grants Management Specialist. In accordance with the regulatory requirements provided at 45 CFR 75.113 and Appendix XII to 45 CFR Part 75, recipients that have currently active Federal grants, cooperative agreements, and procurement contracts with cumulative total value greater than $10,000,000 must report and maintain information in the System for Award Management (SAM) about civil, criminal, and administrative proceedings in connection with the award or performance of a Federal award that reached final disposition within the most recent five-year period. The recipient must also make semiannual disclosures regarding such proceedings. Proceedings information will be made publicly available in the designated integrity and performance system (currently the Federal Awardee Performance and Integrity Information System (FAPIIS)). Full reporting requirements and procedures are found in Appendix XII to 45 CFR Part 75. This term does not apply to NIH fellowships. Treatment of Program Income: Additional Costs SECTION IV - Al Special Terms and Conditions - 5R01Al110964-05 Clinical Trial Indicator: No This award does not support any NIH-defined Clinical Trials. See the NIH Grants Policy Statement Section 1.2 for NIH definition of Clinical Trial. Page-5

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If any experiments proposed in this award result in a virus with enhanced growth by more than 1 log compared to wild type strains, you must notify your NIAID Program Officer and Grants Management Specialist immediately. Further research involving the resulting virus(es) may require review by the Department of Health and Human Services in accordance with the Framework for Guiding Funding Decisions about Proposed Research Involving Enhanced Potential Pandemic Pathogens (https ://www .phe.gov/s3/dualuse/Documents/P3CO .pdf). The Research Performance Progress Report (RPPR), Section G.9 (Foreign component), includes reporting requirements for all research performed outside of the United States. Research conducted at the following site(s) must be reported in your RPPR: San Pya Clinic, BURMA lnstitut Pasteur du Cambodge, CAMBODIA Primate Research Center at Bogor Agricultural University, INDONESIA Conservation Medicine, Ltd, MALAYSIA King Chulalongkorn Memorial Hospital, THAILAND Hanoi Agricultural University, VIETNAM National Animal Health Laboratory, LAOS **************** This Notice of Award (NoA) includes collaboration with Wuhan University School of Public Health, CHINA. **************** This Notice of Award (NoA) includes funds for activity with Wuhan Institute of Virology, CHINA. **************** This Notice of Award (NoA) includes funds for activity with East China Normal University. **************** This award may include collaborations with and/or between foreign organizations. Please be advised that short term travel visa expenses are an allowable expense on this grant, if justified as critical and necessary for the conduct of the project. This award is subject to the Clinical Terms of Award included in Monitoring of Clinical Trials and Studies - NIAID (see NIH Guide for Grants and Contracts, July 8, 2002, NOT Al-02-032). These terms and conditions are hereby incorporated by reference, and can be accessed via the following World Wide Web address: https:/lwww.niaid.nih.gov/grants-contracts/niaid-clinicalterms-award All submissions required by the NIAID Clinical Terms of Award must be forwarded electronically or by mail to the responsible NIAID Program Official identified on this Notice of Award. Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.selectagents.gov/Requlations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm ). Research funded under this grant must adhere to the BMBL, including using the BM BL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. Page-6

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If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. If work with a new or additional Agent(s)/Toxin(s) is proposed in the upcoming project period, provide: o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent(s), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted. with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. STAFF CONTACTS The Grants Management Specialist is responsible for the negotiation, award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Adam Graham Email: (b)(6) Phone: (b)(6) Fax: 301-493-0597 Program Official: Erik J. Stemmy Email: (b)(6) Phone: (b)(6) ----- SPREADSHEET SUMMARY GRANT NUMBER: 5R01Al110964-05 INSTITUTION: ECOHEAL TH ALLIANCE. INC. Budaet Years Salaries and Waaes $167 708 Frinae Benefits $54,168 Personnel Costs /Subtotal) $221 876 Materials & Sunnlies $3 500 Travel $35,918 Other $9 400 Subawards/Consortium/Contractual Costs $191 576 TOTAL FEDERAL DC $462 270 TOTAL FEDERAL F&A $119 376 TOTAL COST $581 646 Facilities and Administrative Costs Years F&A Cost Rate 1 44.1% F&A Cost Base 1 $270,694 Page-7

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F&A Costs 1 1 $119,376 Page-8

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RPPR FINAL A. COVER PAGE Project Title: Understanding the Risk of Bat Coronavirus Emergence Grant Number: 5R01Al110964-05 Project/Grant Period: 06/01/2014- 05/31/2019 Reporting Period: 06/01/2017 - 05/31/2018 Requested Budget Period: 06/01/2018 - 05/31/2019 Report Term Frequency: Annual Date Submitted: 09/16/2020 Program Director/Principal Investigator Information: Recipient Organization: PETER DASZAK , PHD BS ECOHEAL TH ALLIANCE, INC. ECOHEAL TH ALLIANCE, INC. Phone number: I (b) (6)1 460 W 34TH ST Email:! (b) (6)] 17TH FLOOR NEW YORK, NY 100012320 DUNS: 077090066 EIN: 1311726494A1 RECIPIENT ID: NIAID Coronavirus Change of Contact PD/Pl: N/A Administrative Official: Signing Official: ALEKSEI CHMURA ALEKSEI CHMURA 460 W 34th St., 17th Floor 460 W 34th St., 17th Floor New York, NY 10001 New York, NY 10001 Phone number: (b)(6)~ Phone number: (b)(6)~ Email:1 (b)(6) Email:1 (b)(6) Human Subjects: Yes Vertebrate Animals: Yes HS Exempt: No Exemption Number: Phase Ill Clinical Trial: hESC:No Inventions/Patents: No RPPR Page 1

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RPPR FINAL B. ACCOMPLISHMENTS 8.1 WHAT ARE THE MAJOR GOALS OF THE PROJECT? Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of severe acute respiratory syndrome coronavirus (SARS-CoV) in 2002, and the recent emergence Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then hundreds of novel bat-CoVs have been discovered (including >260 by our group). These, and other wildlife species, are hunted, traded, butchered and consumed across Asia, creating a largescale human-wildlife interface, and high risk of future emergence of novel CoVs. To understand the risk of zoonotic CoV emergence, we propose to examine 1) the transmission dynamics of bat-CoVs across the human-wildlife interface, and 2} how this process is affected by CoV evolutionary potential, and how it might force CoV evolution. We will assess the nature and frequency of contact among animals and people in two critical human-animal interfaces: live animal markets in China and people who are highly exposed to bats in rural China. In the markets we hypothesize that viral emergence may be accelerated by heightened mixing of host species leading to viral evolution, and high potential for contact with humans. In this study, we propose three specific aims and will screen free ranging and captive bats in China for known and novel coronaviruses; screen people who have high occupational exposure to bats and other wildlife; and examine the genetics and receptor binding properties of novel bat-CoVs we have already identified and those we will discover. We will then use ecological and evolutionary analyses and predictive mathematical models to examine the risk of future bat-CoV spillover to humans. This work will follow 3 specific aims: Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. We will examine if: 1) wildlife markets in China provide enhanced capacity for bat-CoVs to infect other hosts, either via evolutionary adaptation or recombination; 2) the import of animals from throughout Southeast Asia introduces a higher genetic diversity of mammalian CoVs in market systems compared to within intact ecosystems of China and Southeast Asia; We will interview people about the nature and frequency of contact with bats and other wildlife; collect blood samples from people highly exposed to wildlife; and collect a full range of clinical samples from bats and other mammals in the wild and in wetmarkets; and screen these for CoVs using serological and molecular assays. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk. We propose two competing hypotheses: 1) CoV host-range in bats and other mammals is limited by the phylogenetic relatedness of bats and evolutionary conservation of CoV receptors; 2) CoV host-range is limited by geographic and ecological opportunity for contact between species so that the wildlife trade disrupts the 'natural' co-phylogeny, facilitates spillover and promotes viral evolution. We will develop CoV phylogenies from sequence data collected previously by our group, and in the proposed study, as well as from Gen bank. We will examine co-evolutionary congruence of bat-CoVs and their hosts using both functional (receptor) and neutral genes. We will predict host-range in unsampled species using a generalizable model of host and viral ecological and phylogenetic traits to explain patterns of viral sharing between species. We will test for positive selection in market vs. wild-sampled viruses, and use data to parameterize mathematical models that predict CoV evolutionary and transmission dynamics. We will then examine scenarios of how CoVs with different transmissibility would likely emerge in wildlife markets. Specific Aim 3: Testing predictions of CoV inter-species transmission. We will test our models of host range (i.e. emergence potential) experimentally using reverse genetics, pseudovirus and receptor binding assays, and virus infection experiments in cell culture and humanized mice. With bat-CoVs that we've isolated or sequenced, and using live virus or pseudovirus infection in cells of different origin or expressing different receptor molecules, we will assess potential for each isolated virus and those with receptor binding site sequence, to spill over. We will do this by sequencing the spike (or other receptor binding/fusion) protein genes from all our bat-CoVs, creating mutants to identify how significantly each would need to evolve to use ACE2, CD26/DPP4 (MERS-CoV receptor) or other potential CoV receptors. We will then use receptor-mutant pseudovirus binding assays, in vitro studies in bat, primate, human and other species' cell lines, and with humanized mice where particularly interesting viruses are identified phylogenetically, or isolated. These tests will provide public health-relevant data, and also iteratively improve our predictive model to better target bat species and CoVs during our field studies to obtain bat-CoV strains of the greatest interest for understanding the mechanisms of cross-species transmission. 8.1.a Have the major goals changed since the initial competing award or previous report? No B.2 WHAT WAS ACCOMPLISHED UNDER THESE GOALS? File uploaded: Year 4 NIAID CoV Report_Final for eRA Commons.pdf B.3 COMPETITIVE REVISIONS/ADMINISTRATIVE SUPPLEMENTS For this reporting period, is there one or more Revision/Supplement associated with this award for which reporting is required? No B.4 WHAT OPPORTUNITIES FOR TRAINING AND PROFESSIONAL DEVELOPMENT HAS THE PROJECT PROVIDED? File uploaded: Year 4 NIAID CoV Training and Prof Devlp.pdf RPPR Page2

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RPPR FINAL B.5 HOW HAVE THE RESULTS BEEN DISSEMINATED TO COMMUNITIES OF INTEREST? 1.Conference and University Lectures: Pl Daszak, and Co-investigators Shi, Epstein, Oliva!, and Zhang gave invited University and Conference lectures including Harvard Univ. Columbia Univ., Tufts Univ., Mt. Sinai, the 2nd International Symposium on Emerging Viral Disease in China, the 2nd International Symposium on the Infectious Diseases of Bats in Colorado, Cell Symposia: Emerging and Reemerging Viruses 2017 in Virginia, The International Union of Microbiological Societies 2017 National Academy of Sciences in Singapore, 2018 Borneo Quality of Life Conference in Malaysia, 2017 Chemical and Biological Defense Science and Technology (CBD S&T) in California, Prince Mahidol Award Conference in Bangkok, Collaboration for Environmental Evidence Meeting in Paris, US-China NSF Ecology and Evolution of Infectious Disease (EEID) Meeting, and others that included specific discussion of the current project and results. 2.Agency and other briefings: Pl Daszak and Co-investigator Shi introduced this project and discussed new opportunities about predicting and preventing zoonoses within National Institute of Allergy and Infectious Disease Office, Defense Advanced Research Projects Agency, National Natural Science Foundation of China, Chinese Center for Disease Control and Prevention, US NASEM Forum on Microbial Threats, Chinese Academy of Sciences, and the Health Working Group at the US Embassy in Beijing. 3.Public outreach: Pl Daszak and Co-investigator Shi, Epstein, Olival, have presented this work to the general public in a series of meetings over Year 4 including at Cosmos Club briefings that EcoHealth Alliances hosts in Washington DC, over 10 meetings on the China National Virome Project and the Global Virome Project in China, Europe, Australia, Southeast Asia and Latin America. Coinvestigator Oliva I presented this work at a public event on Disease Transmission and Technologies in New York, co-investigator Ross presented this work at EcoHealth Webinar on wildlife trade network research. Zhu broadly introduced this work to the conservation and ecological research community in China through field training workshops. B.6 WHAT DO YOU PLAN TO DO DURING THE NEXT REPORTING PERIOD TO ACCOMPLISH THE GOALS? Specific Aim 1: Assessment of CoV spillover potential at high risk human-wildlife interfaces. •To commence an in-depth analysis of data collected from the integrated biological behavioral surveillance from Yunnan, Guangxi, and Guangdong provinces, incorporating questionnaires and serological testing results. •To initiate lab analysis of human samples collected from the passive hospital surveillance from four hospitals in Yunnan province: 1) Dali College Affiliated Hospital; 2) Dali Prefecture Hospital; 3) Kunming No. 3 People's Hospital, and 4) Chuxiong Prefecture Hospital. The goal will be to identify examples of CoV spillover events in China that may lead to illness. Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk •To repeat and continue in vivo experiments of SARSr-CoVs with spike variants on hACE-expressing transgenic mice (survival rate, histopathological analysis, etc) to evaluate the risk of cross-species infection of different SARSr-CoVs to humans; •Continue searching for the receptor of SARSr-CoVs with deletions in the homologous region of SARS-CoV RBD (i.e. Rp3, Rs672), and SARSr-CoVs that are unable to utilize bat ACE2 (e.g. Rs4231 ). •Continue the phylogeographic study of bat-CoV with newly collected samples to better understand the geographic distribution and evolution of bat-CoV genetic diversity in south China and SE Asia. Specific Aim 3: Testing predictions of CoV inter-species transmission. •Using the full-length infectious cDNA clone of MERS-CoV, chimeric viruses with the spikes of newly identified MERSr-CoVs will be constructed. The pathogenesis of these MERSr-CoVs will be tested on the human DPP4-expressing mouse model that has already been developed and validated in Y4. •To conduct a population genetics study of Rhinolophus sinicus ACE2s, including the amplification of ACE2 genes from R. sinicus samples of different origin, test of the usage efficiency of R. sinicus ACE2s of different origins by SL-CoVs and kinetics study on the binding of SL-CoV RBD to different R. sinicus ACE2s. •In collaboration with South China Agrricultural University, gather data on the spatial structure and barn-level mortality records to parameterize our mathematical model of virus spread that incorporates a meta-population structure in individual and use this to fit the model on a training set of farms and validate it on a hold-out set. •Using the intra-farm transmission model, we will (a) determine the characteristics of a farm that determine the likelihood and size of an outbreak given a spillover event, and (b) determine whether SADS and PEDV outbreaks on farms can be distinguished by differing dynamics, as measured by transmission parameters in our intra-farm transmission model. 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B.2 (Year 4 NIAID CoV Report_Final for eRA Commons.pdf) 1 R01Al110964 Year 4 Report Pl: Daszak, Peter Year 4 Report: Understanding the Risk of Bat Coronavirus Emergence Award Number: R01Al110964-03 Reporting Period: 06/01/2017 - 05/31/2018 ************* B.2 What was accomplished under these goals? Summary The results of the 4th year of our R01 work are detailed below. They include: • Completed behavioral risk survey questionnaires and biological sample data collection for 1,585 people in Yunnan, Guangxi, and Guangdong provinces. • Preliminary analysis of behavioral survey responses exploring key risk factors relating to potential viral zoonotic disease spillover in China, indicating notable differences among the respondents in Guangdong, Guangxi, and Yunnan. • Completed serologic testing of collected human samples for MERS-CoV, SARSr-CoV, HKU9 CoV and HKU10 CoV, showing the serologic evidence of spillover of bat SARSrelated CoVs (7 people in Yunnan province) and HKU9 CoV (2 people in Guangxi province). • Testing of samples from 671 individual bats to identify diverse alpha- and betacoronaviruses. • Genetic diversity and genomic characterization of beta-coronaviruses in fruit bats and characterization of the full-length genome sequence of a novel HKU9-related CoV. • Analysis of host-virus phylogeography for all bat CoV RdRp sequences collected by our group in China from 2008-2015 (Alpha-CoVs: n = 491; Beta-CoVs: n = 326) to identify the geographic areas that are likely sources of origin/diversity for this important group of viruses. • Identification of two novel MERS-related Co Vs that use DPP4 receptor. • In vivo infection of SARSr-CoVs with variants of S protein in human ACE2 (hACE2) expressing mice. • Identification of a novel bat-origin CoV (swine acute diarrhea syndrome coronavirus, SADSCoV) causing a multi-farm outbreak of fatal acute diarrhea in piglets in Guangdong (published in Nature in April 2018). • Development of an intra-farm transmission model to understand SADS-CoV spread and help predict and prevent future outbreaks. Specific Aim 1: Assessment of CoV spillover potential at high-risk human-wildlife interfaces During Year 4 we completed behavioral risk surveys and biological sample collection from people at selected sites in three provinces in southern China (Guangdong, Guangxi, and Yunnan) and began analyzing the results. 1 RPPR Page4

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Behavioral Survey We administered 1,585 surveys in Guangdong, Guangxi, and Yunnan provinces. Questions explored respondent health-seeking behavior, experiences with unusual illnesses, contact with wildlife and livestock, and general background information. Blood samples were collected from respondents and tested for SAS-related Co Vs (SARSr-CoVs) and HKU10-CoV using serological assays. Survey data was analyzed by province to examine patterns among respondent characteristics and behavioral risk factors across provinces. Respondent General Background Information Of the 1,585 respondents who completed the survey, 420 were from Guangdong, 412 were from Guangxi, and 753 were from Yunnan. More females than males completed the survey in all provinces. The mean age of the overall survey sample was 52 years (Figs. 1, 2) . ... ,..,,,. (f'l•QO} _J_ 260 6 - Figure 1: Gender of respondents ,..,.., 1"11•nl1 0 25 Xl 7!- ,oo, i, so 15 11k:O ~ 60-,; 1'KI Years Figure 2: Age distribution of respondents. Across all provinces, most respondents had lived in their respective locales for more than 5 years (96.3%) (Fig. 3) and earned less than 10,000 renminbi (RMB) annually (84.6%) (Fig. 4). In 2016, the updated poverty standard in China was 3,000 RMB as defined by Poverty Alleviation Office of State Council. More families in Guangxi (61.8%) lived at or below the poverty level as compared to those in Guangdong (36.9%) and Yunnan (43.3%). ' - Figure 3: Duration of residency. RPPR I-"- ·- 122 .... ,,_-.,, .... --... ., .... ,_.,o, ·-•-IV -·-1· ,. .... ,.~ I· Figure 4: Family annual per capita income (RMB). 2 Page5

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter In Guangdong, Guangxi, and Yunnan, 73.9%, 57.0% and 69.6% of respondents, respectively, had a primary school-level education or less (Fig. 5). Across all provinces the most common livelihood was crop production. In Yunnan, 699 out of 753 (92.8%) individuals from the province identified crop production as a livelihood activity. In comparison, 237 out of 420 (56.4%) individuals from Guangdong, and 260 out of 412 (63.1%) individuals from Guangxi (Fig. 6) named crop production as a livelihood in the last year. Respondents, however, where not restricted to defining a single livelihood, many indicated engaging in multiple types of livelihoods. c.n,,g.,.,. Iii• 41~ _, n•405J -I· 1(1! ,.,.,.,.J_.t;llOOI- 211 129 --·---1·· 163 C91tg-fJ\Jf~rs..t i;tor.i&IOOJI 7 21 3 25 sc 7S , .. 25 sc ,, ,oo 0 ,i, Pffcffl ('lr.J Figure 5: Highest level of education completed Types of activities done to earn livelihood since this time lost year• Etlrndhn orin1n11illl1 gu oi\ t,motfCtopprodUdlon • - 237 Wlldllft ,utaurant bn•ntn - I ·1 WUClltxo•e animal 11.lat~lii:et business - Rancher.farmer animal p1odudion b11slnass... I 1 Me.fl Pr0<:es,:1no, s1au'1\lerrioust, ao-al'lolr-j 1 zoc11anau.1ryen1m11 ht11lh car•- P,oteded are.awot...,_,. 13 Htmto-ntr.appttJfi~h~rf'OtJ-ift1Q3111tr6flnon t1moerror&11 ptOCIUCl collooorMlgrallttaborer~ I 21 !Junie doctor heJ.ler, comrnumt:, IIHltl'I WOOef I 1 ConstructJon - I 1 00,o,• - 162 250 500 150 eu,mgo (• a C90) 4 260 3 3 22 3 3 2 2 80 3 19 86 250 500 150 Count ,_ (n.,.150t 292 so 75 ,oo ~unn~n (n • i25) ------- 43 45 zso 500 699 750 Figure 6: Types of activities conducted to earn a livelihood since this time last year (above) 3 RPPR Page6

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter In Guangdong, Guangxi, and Yunnan, 41.7%, 50.7% and 59.6% of respondents, respectively, indicated that they traveled outside of their village town or city in the past year. Among those who traveled, the average number of trips was 5 in Guangdong and Guangxi, and 6 in Yunnan. The average distance traveled by respondents in Guangdong and Yunnan were 113 Km and 118 Km, respectively, compared to 66 Km by respondents in Guangxi. Health-Seeking Behavior and Experiences with Unusual Illnesses When asked where they usually get treatment for illness or infection, the top 3 responses across all provinces in aggregate were hospitals, clinics, and pharmacies/dispensaries in descending order (Fig. 7). However, within Yunnan, most respondents went to hospitals, followed by pharmacies, then clinics. Mob1e Clime- 15 Commumty health worlcor-16 Tr.ad,tion:;J healer -14 Pho1moo1ldio•on•o,y-172 Do not receive medical treatment -122 Olhor- 13 Guangaong (n = 527) 356 200 400 16 21 '7 ' ' 600 0 Guangxj (n = 522) 213 252 200 400 Count 600 0 Yunnan (n = 8<4) 238 200 400 Figure 7: Location where care was usually received for illness or infection. 600 All survey respondents were asked whether they had experienced an unusual illness in their lifetime and in the past year, defined by a series of the most common symptoms associated with encephalitis, hemorrhagic fever (HF), severe acute respiratory infection (SARI), and influenzalike illness (ILi). Additional symptoms that were asked about included: fever with diarrhea or vomiting; fever with rash; and, persistent rash or sores on skin. Respondents were not restricted to selecting one illness and could provide multiple responses. The proportion of respondents who had an unusual illness with any of the above-mentioned symptoms in their lifetime varied slightly by province. Between the three provinces, Yunnan had the fewest number of respondents who reported experiencing the symptoms provided {38.8%), compared to Guangdong and Guangxi (51.9% and 51.3%, respectively). Yunnan was also the only province where less than half of the respondents reported experiencing the symptoms provided (Fig. 8). 4 RPPR Page 7

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Figure 8: Respondent's experience of unusual illnesses . .. ,. .._ :!1•415) 451 .:. Across all three provinces, among those who had experienced any symptoms of unusual illness in their lifetimes, those associated with ILi were the most commonly reported. In Guangdong province, this was followed by symptoms associated with SARI, then by other symptoms not mentioned in the survey. In Guangxi province, the second most reported symptoms were ones associated with encephalitis, followed by other symptoms not mentioned in the survey. Similarly, in Yunnan, symptoms associated with encephalitis were the second most commonly reported, but this was followed by symptoms associated with SARI (Fig. 9). Figure 9: Symptoms reported by people who had experienced unusual illness in their lifetime. Fevorwrth headach• and s9Wre fatigue or weakness (encephalitis)- 115 Fever with bleedng or bruising not ,elated to tnJur)"(HF)- Fever "'ith cough and shome"3 of breath or difficully breathing (SARJ)-163 Guangd«lg (n = 300) Fe,,., w.111 musele acnes, ooug~. or sore 11roa1 (lLI)· ■ 172 Feverwilh diarrhea orvomnlng- 115 reverw,urn,sh-13 "'•"'••tent r•<h or oor<o on olan-18 Had symptoms but none ofthes•-124 49 24 40 13 4 41 Guangi<I Ill ='.306) 135 49 Yunnan (n~435) 166 • ' ' • + • ' ' • + • ' ' • o 100 2oe 300 .ioo soo o 100 200 300 •oo soo o 100 200 300 •oo soo Count In each province, just under one-third of respondents who experienced the symptoms associated with an unusual illness in their lifetime indicated experiencing any of the symptoms in the past year - 22_2% in Guangdong, 32.8% in Guangxi and 23.0% in Yunnan (Fig. 10). RPPR 5 Page 8

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Figure 10: Whether respondents had experienced symptoms associated with an unusual illness, in the past year. to· 322 to· 132 92 ,O· 270 172 577 Of the respondents who reported having symptoms of unusual illness in the past year, across all three provinces, symptoms associated ILi were the most commonly reported. In Guangdong province, this was followed by symptoms associated with SARI then by other symptoms not provided in the survey. In Guangxi, symptoms associated with ILi were followed by symptoms associated with encephalitis, then by fever with diarrhea or vomiting. In Yunnan, symptoms associated with ILi were followed by symptoms associated with encephalitis, then by both SARI and other symptoms not provided in survey (Fig. 11). Figure 11: Symptoms experienced by those reporting unusual illness in the past year. Guangdong Guan11XI Yunn311 (n: 113) (n = 197) ln=239) F~9< Wllh h•~-• and ,e..,,e lai.vo,e or w"" .ness (encephol~• • I 3 30 50 f-wl#l blttC,"9 or lll\U$1ng nol <llill•d lo 11\)Ur;{HF)· l2 Fevtr w,L, COlllJh ar10 snonness 01 Ofulll or dll\lcult1 Ofealh,ng (SARI)· 117 16 40 Fevai will! mus dot ames. cough or sore ttvoat IIU) • 9 84 74 l'ovorwilh dlonh•• orvomollng· 17 29 118 Fe.or w11n rasn • 12 10 P~rsisienl rash or sores on sl<ln •~ 3 Had s,mptoms DUI none or tilts•· 111 25 40 ' 0 25 !50 75 100 1250 25 50 75 100 1250 25 50 75 100 125 Count When respondents were asked what caused the symptoms associated with unusual illness experienced in the past year, 64.4% in Guangxi (85 of 132 respondents), and 50.0% in both Guangdong and Yunnan (46 of 92 respondents and 86 of 172, respectively), said they did not know the cause (Fig. 12). Only one respondent in Guangxi said their symptoms were due to 6 RPPR Page9

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter contact with animals (wild animals, specifically). Two respondents in Guangdong and one respondent in Guangxi said their symptoms were due to contact with animals (non-wild animals, specifically), whereas none of the respondents in Yunnan attributed their cause to contact with animals. Contad 1Mlh slCk pooplo - Conlactwitn wtld animals - Contact ,.;th olher animals· 12 Bao 1000 or water· 16 eao splnts/Wlta>cran • Wound or injury· I 2 ldonHnow· Other· Guang<long (n=90) 0 50 100 ' 150 13 2 26 ' 0 50 CuanllXi (n= 128) 85 100 Count Figure 12: Reported cause of sickness in the past year. 13 I 1 150 0 Yunnan (n = 155) 86 48 50 100 150 Respondents reporting an unusual illness in the past year were asked if any of the people they lived with in the past year had symptoms similar to theirs, to assess possibilities of transmission among household members. Most respondents did not, across all three provinces: 82.4% in Guangdong, 83.6% in Guangxi and 79.9% in Yunnan (Fig. 13). Figure 13: Whether household members had similar symptoms of unusual illness, in the past year 80· 60· io· 73 o· 341 . No Yes GU-ll1QXI (o•.f03) 337 No 150 YES ' Yunnan (n • 7-48) 598 No 7 RPPR Page 10

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Of the household members who experienced symptoms of unusual illness in the past year, the most commonly reported symptoms were those associated with ILi (Fig. 14). Femw,th htadadle and some laHgue or wnkness (encephahti>) • ~ FO\/Ot with blHdi-ng Or bl'lllzlng not rolatodlo 11'1/W"J(HF} • Fev&rwllh couah ano snc<1ness or ueatn o, <ltllcu111bre,lh1n0 (SARI)· 116 Guar,goong (n = 84) Fwer wilh muscle aches. cough, orsore throat (IU)· .57 f ...,er 'Mlh dlanhea or -.mmno · 12 Fel9r with rashPersislenl rash or sores on sijn • 12 Hao symptOms lklt ~ne Of In He· 13 0 50 ' ' ' 100 150 0 11 26 8 19 ' ' 5-0 100 150 0 Count Figure 14: Symptoms of household members who were ill, in past year. Yurman (n = 193) ;o 100 15-0 Respondents were also asked if any members of their household who experienced symptoms of unusual illness died as a result of their illness in the past year. Across all the three provinces, almost none had died from these illnesses (Fig. 15). 100• 75• g t! ., ~ 0.. 50· 2~· D· Yes ' Ou1nodong (ns 73) 72 137 2 3 No Yea No Yes No Figure 15: Whether household members died from illness, in the past year. RPPR 8 Page 11

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B.2 (Year4 NIAID CoV Report_Final for eRA Commons.pelt) 1 R01 Al 110964 Year 4 Report Pl: Daszak, Peter Contact with Animals All respondents were asked about various types of animal contacts in their lifetime and in the past year. More than two-thirds of the respondents across all provinces, as well as in each of the provinces, reported raising an animal within their lifetime (71.2% in Guangdong, 77.7% in Guangxi, and 97.7% in Yunnan). More than half of the respondents in each province reported having animals come inside their dwellings (83.1 % in Guangdong, 60.2% in Guangxi, and 92.5% in Yunnan). More than half of respondents in each province reported handling live animals (51.5 % in Guangdong, 56.9% in Guangxi, and 62.9% in Yunnan) (Table 1). Respondents from Yunnan had more types of contact with animals in their lifetime than those from Guangdong and Guangxi. With the exception of cooking or handling meat, organs, or blood from a recently killed animal and being scratched or bitten by an animal, the proportion of respondents from Yunnan who engaged in all types of animal activities was higher than the other provinces. Guangdong Guangxi Yunnan Type of animal contact (past year) (n) (%) (n) (%) (n) (%) Lived with an animal as a pet 43 100 % 72 98.6% 335 100 % Handled live animals 212 100 % 226 98.3% 332 99.7% Raised a live animal 296 100 % 312 99.4% 518 99.8 % Shared water source with animals for washing 47 100 % 19 95.0% 97 100% Seen animal feces in or near food before you have eaten it 18 100 % 15 93.8% 43 100% Eaten food after an animal has touched or damaged it 6 100 % 6 100 % 29 100 % Animals come inside the dwelling where you live 345 100 % 239 98.0% 493 100% Cooked or handled meat, organs, or blood from a recently 333 100 % 144 97.3% 412 100% killed animal Eaten raw or undercooked meat or organs or blood 2 100 % 25 89.3% 65 98.5% Eaten an animal that was not well/sick -- -- 1 100% 6 100 % Found a dead animal and collected it to eat, share, or sell -- -- 3 100% 10 100 % Been scratched or bitten by an animal 1 100 % 31 100% 28 96.6 % Slaughtered an animal 145 100 % 69 98.6% 303 100% Hunted or trapped an animal 9 100 % 4 100% 22 95.7% Table 1: Types of animal contact, within a respondent's lifetime. Respondents who reported having animal contact in their lifetime were also asked to indicate if they had the same type of animal contact in the past year (Table 2). In the past year, across all three provinces and in each province, almost all respondents engaged in all contact types with the exception of eating an animal that was not well/sick, and finding a dead animal and collecting it to eat, share, or sell (0% for both in Guangdong). 9 RPPR Page 12

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Guangdong Guangxi Yunnan Type of animal contact (lifetime) (n) (%) (n) (%) (n) (%) Lived with an animal as a pet 43 10.4% 73 18.1 % 335 62.9% Handled live animals 212 51.5 % 230 56.9 % 334 62.8% Raised a live animal 296 71.2 % 314 77.7% 521 97.7% Shared water source with animals for washing 47 11.5 % 21 5.2% 97 18.2% Seen animal feces in or near food before you have eaten it 18 4.4% 16 3.9% 43 8.1 % Eaten food after an animal has touched or damaged it 6 1.5% 6 1.5% 29.0 5.4% Animals come inside the dwelling where you live 345 83.1 % 244 60.2% 493 92.5% Cooked or handled meat, organs, or blood from a recently 333 80.4% 148 36.7% 413 77.5% killed animal Eaten raw or undercooked meat or organs or blood 2 0.5% 28 6.9% 68 12.8% Eaten an animal that was not well/sick -- -- 1 0.3% 6 1.1 % Found a dead animal and collected it to eat, share, or sell -- -- 3 0.7% 10 1.9% Table 2: Types of animal contact, in past year. Respondents who had animal contact in the past year were asked to identify the animals involved in the interaction. (Figs. 16-26. below: the first two figures are enlarged to show row labels, which are identical for all). Cats and dogs were the most common pets reported across all provinces and in each province (Fig. 1Gb). 43 72 335 ' ' ' tlo ns leS No Figure 16a (top) & b (below): (a) Whether respondents had lived with an animal as a pet, in the past year, and (b) among those who had, types of animal kept as pets. 10 RPPR Page 13

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Rodents/Shrews - Bats - NHPBirosCarnivoresungulatesPoultry- 1 Goats/Sheep - Guangdong (n=50) 5 Guangxl (n = BJ) Yunnan (n =436) Swine - 2 3 Gattie/Buffalo - Oons-131 Gats-118 I I I f I I 0 100 200 300 400 0 60 15 288 I I I f ! I I I I 100 200 300 400 0 100 200 300 400 Count Poultry was the most common type of animal handled across all provinces as well as in each province, with 96.2%, 90.3%, and 92.8% of respondents handling animals in Guangdong, Guangxi and Yunnan, respectively (Fig. 17b). ~':r.~ ....... Cuano®no a...,,,,, Vuman l/'l•nl (f\ool 240> n •'2li)J (n - soa, ,., 212 229 332 Roo•nlalShltwS - lo 2 22 e•1 - 5 "HP .. " S1tds.· 12 2 114 C:.imlvOfes• fe 3 17 i unou1a1H •i ~- ~ Pwlir,·-204 204 308 .. C0.111'1Sl'IH0-12 2 Is :'I• S'Wln•• r 10 105 Cl'lll..,&.tl'alo 2 le Oogs• le 10 24 • 1 O· - ta.1;• 1 3 . . ... • I • • • • . ' .. "' ,;, '" ,.. '" 0 1()0 211"0 300 400 500 0 100 200 .JOO 400 5-30 0 100 200 100 400 500 Couil Figure 17a & b: ( a) Whether respondents had handled live animals, in the past year, and (b) among those who had, types of live animals handled. 11 RPPR Page 14

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Poultry was also the most commonly raised animal in each of the three provinces; 95.3%, 87.5%, 95.4% in Guangdong, Guangxi, and Yunnan, respectively (Fig. 18b). ,~· ...... , ..... ~~ :':'31, , ...... ,.:u ,,. ~11 ~ ntai "" 216 312 618 '4od!n~l!''I\' ~ s .. •-· 13 II ...... b 1 C11'1\lnl"• :e t IJl'>9Willlu 1 j. f'OI.IIIJ) -Z82 27J ~tit ~ 12, II -· ~ 28 ~m,u.o ~ ~ .,.. 157 10 2 """ l:12 120 '' I 0 200 •OO 600. ,.;. 100 ooco ,.. •oo Counl ◄ ~'° !00 Figures 18a & b: (a) Whether respondents had raised Jive animals in the past year, and (b) among those who had, types of animals raised. In all three of the provinces, the most common type of animals found in respondent dwellings were rodents or shrews. In Guangdong and Yunnan, birds were the second most common animal type found in dwellings. In Guangxi province, birds along with poultry were the second most common animal type. Respondents in Guangdong and Yunnan reported that all 12 animal taxa had come inside their dwellings in the past year. Taxa seen in the dwellings of respondents from Guangdong and Yunnan and not Guangxi were non-human primates, ungulates, goats or sheep, swine, and cattle or buffalo (Fig. 20b). --,..,., G.iJ-0""19 ,. • 630) 'tu'Vi.n (1'•1~ R~t.11$111t•1· -31& Bm· 138 451 6 - " -·11 Ill,..,112 C.lfflttlOf"e•· 168 IJnO •••· '2 ....., I® Goillll/Sf'IHP" 11 jt\'lt,·• ~ C,lllot<J• q• 11 Dogs 125 Colt 128 ◄ 18 10 6 2 o :>oo ,oo •oo o 200 '°' ~on Collnt 130 318 3 1,0 91 138 180 22$ .&O ,00 Figure 19a & b: (a) Whether respondents had animals come inside dwelling, in the past year, and (b) among those who had, types of animals in dwelling. RPPR Mit 12 Page 15

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Almost all of the respondents who said they have cooked or handled meat, organs, or blood in their lifetime reported doing so in the past year. Common animal types that were cooked handled included poultry and swine in all three provinces (Fig. 20) . • - 412 ...... •b -.....t.~ 111 "' t ... .. 1,0 ~ .. In u,,-..iu 11 c.o ....... ll1 ....... r.u (,1 --• 111 . , .. -' ·" ~-~- .._ \11•1Nl '" !a I" 1,2 2 )al ,,. -"" I" 1 .. 1,2 "'' '" "" . Coon Figure 20a & b: (a) Whether respondents had cooked or handled meat, organs or blood from a recently killed animal, in the past year, and (b) among those who had, types of animals whose meat, organs or blood was cooked or handled. ;. More respondents in Yunnan reported eating raw or undercooked meat compared to respondents in Guangdong and Guangxi (Fig. 21). In Yunnan, 96% of respondents who ate raw or undercooked meat in their lifetime did so in the past year. The types of animal products that were eaten raw or undercooked by respondents in Yunnan were mostly from swine. In Guangxi, the most commonly reported type of animal meat that had been eaten raw or undercooked was that of carnivores. ,. " - ·-.... I -.; q, ... ,...__ - ... - - --I• --I• ---""" ca1, -II I• J2 1, ;, .- ., ' • ., Coon Figure 21 a & b: (a) Whether respondents had eaten raw or undercooked meat or organs or blood, in the past year, and (b) among those who had, types of animals whose meat, organs or blood were eaten raw or undercooked. 13 RPPR Page 16

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Across all provinces, a total of 13 respondents in Guangxi and Yunnan indicated that they collected an animal that was found dead to eat, share or sell. In Guandong, no respondents reported finding a dead animal and collecting it to eat, share, or sell. The most common type of animal collected across all provinces in aggregate was poultry. In Yunnan, poultry was the most common type of animal found dead and collected to eat, share or sell (80.0%), whereas dogs were the most common type in Guangxi (66.7%) (Fig. 22) . ...... , .. , ·- #•II] - ,.,,,., .. ,. ..., "-"11) t:~ ... -· I• •-· ,,14'- " .,. t -- 1 I,. -,....,. ,, ~, .... ~-- .. -- I• I• CM&.'-· °'" 2 .... II ' ~ ·~ .. Con Figure 22 a & b: (a) Whether respondents had found a dead animal and collected it to eat, share, or sell, in the past year, and (b) among those who had, types of animals that were found dead and collected to eat, share, or sell. In each province, almost all of the respondents who indicated being scratched or bitten by an animal in their lifetime said it occurred in the past year (100% in Guangdong, 98.6% in Guangxi, and 100% in Yunnan). In both Guangxi and Yunnan, dogs were the common type of animal that respondents said they were scratched or bitten by (64.5% in Guangxi and 50.0% in Yunnan). Cats were the second most common in Guangxi and Yunnan (9.6% in Guangxi, and 28.5% in Yunnan). Across all three provinces, only one respondent from Yunnan said that they were scratched or bitten by a bat (Fig. 23). - ...... ,, . !ll'i•Jll '""' -- -'"' 21 lloWICI, ........ .. -·· ! ,.. __ I .. I• . ,.. .5 -- I• (;;1tc.9idl.,1 .... I - c .. 20 -H •· ;. c..... Figure 23 a & b: ( a) Whether respondents had been scratched or bitten by an animal, in the past year, and (b) among those who had, types of animals that scratched or bit respondents. RPPR 14 Page 17

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Poultry was the most common type of animal slaughtered during the past year across all provinces as well as in each province (95.8% in Guangdong, 79.7% in Guangxi, and 94.1% in Yunnan). In addition to poultry, respondents in Yunnan also commonly only slaughtered swine (43.9%), compared to 1.4% in Guangdong and 7.3% in Guangxi (Fig. 24). .. _ - -~ """"'"'''"' Guano• VuM.lln ..... ...,., ... - t'I• 171U 11'1•71) u,~ 76) ... •"l JI AoOfflb.r.::l!rt-..a I 10 16 -- 5 .. fUros· - 13 18 Camwrea I U 6 6 , ung~l~fl• 11 2 IM ,,,, • .., .• ,39 155 285 .. c,au~ .. o· l2 6 h't!"11'1" 12 5 133 " .,.,...a,.., •. , 2 3 18 Ooo, 13 5 --J..- Cm ' ' ' .. 0 100 200 30Q 400 0 100 20 100 AOO 0 100 200 ">l •00 CO\Jffl Figure 24 a & b: (a) Whether respondents had slaughtered an animal, in the past year, and (b) among those who had, types of animals slaughtered. Carnivores were the most common taxa of animals hunted or trapped in the past year, in Guangdong and Guangxi. In Yunnan, rodents or shrews and birds were reported as the most common. Bats, non-human primates and dogs were animal types hunted by respondents in Yunnan but not by respondents in Guangdong and Guangxi (Fig. 25). -- - ,_ - GU;al\lll ... . .. .,,, ~I\# 1!' 11'1 = 41 -tn>lt) ,.,. ~,t.tlStttM· 4 10 - s " .... I• ~ 1, --11 ~3 10 Cfll'MO'H•-6 l Un:,!J .. 1 .. ·l1 J ,0 ,.....,.1, Go:11"9\tto· -6 I< ii ,.. .... ,..,, I· C:1<11-."3.ibi.• 11 .... 2 ... C.I> '• .. ,. " , .. ,. ,, ... • .. , . Courc Figure 25 a & b: (a) Whether respondents had hunted or trapped an animal, in the past year, and (b) among those who had, types of animals hunted or trapped. , . 15 RPPR Page 18

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter In examining bat-specific contact, across all provinces and within each province, the most common interaction with bats was finding them inside their houses. Respondents in Yunnan also hunted/trapped and handled bats, and were scratched/bitten by bats, whereas these did not occur in Guangdong or Guangxi (Fig. 26). Guangdong Cuangxi Yunnan (n = 39) (n = 15} tn = 173) Hunted/lrapped - 5 s1auol\tere<1 - 5 Saatche-dlb1tlen - 11 Cool<edlhan<lted • 11 la In house -138 14 149 Handlecl- 5 0 50 100 150 200 0 50 100 150 200 0 50 100 150 200 Count Figure 26: Types of bat contact. After respondents were asked about their contact with wildlife and livestock, they were asked about their knowledge of whether animals can spread diseases and whether they were worried about diseases and disease outbreaks at wet markets. The proportion of respondents who thought that animals can spread disease was highest in Guangdong province (72.3%). In Guangxi and Yunnan, the proportion of those who thought animals could spread disease compared to those who thought that they did not were roughly equivalent - 47.5% versus 50.7% in Guangxi and 49.2% versus 49.3% in Yunnan (Fig. 27). »I •• 110 /0 ,., N• ,., RPPR 371 ... , .. Figure 27: Whether respondents thought that animals can spread disease. 16 Page 19

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Similarly, when respondents were asked about whether they were worried about diseases or disease outbreaks in animals at wet markets, Guangdong had the highest proportion of respondents who said they were worried (67.3%). In both Guangxi and Yunnan, the proportion of respondents that was not worried (57.5% and 51.5%, respectively) was higher than the proportion that was worried (Fig. 28) 280 YU CU&noOOIO (n~41f) 382 No YU Figure 28: Whether respondents were worried about diseases or disease outbreaks in animals at wet markets. Serological Evidence of Bat SARS-related CoV Infection in Humans Respondents were asked to provide a biological sample to assess whether SARS-CoV spillover had occurred at the high-risk location where the survey has been implemented. A total of 1,530 serum samples were collected from 2016 to 2017 from individual residents in villages close to bat caves where coronaviruses were previously detected. We developed an ELISA serology test using the purified NP protein of MERS-CoY, SARSr-CoV, HKU9 CoV and HKU1 o CoV as coating antigen respectively and using Anti-Human lgG Monoclonal antibody as secondary antibody. All sera were screened for antibodies against these 4 bat-origin coronaviruses. Anti-SARSr-CoV NP lgG was detected in 10 samples, and 6 samples were positive for lgG against HKU1 0 NP. The 16 ELISA positive samples were further tested by confirmatory western blot, 7 samples from Yunnan province were confirmed positive for anti-SARSr-CoV, two samples (one from Guangdong province and one Guangxi province) were confirmed positive for anti-HKU10 (Table 3). 17 RPPR Page 20

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B.2 (Year4 NIAID CoV Report_Final for eRA Commons.pelt) 1 R01 Al 110964 Year 4 Report Pl: Daszak, Peter NP Antibody Positive No. Locations Sample No. MERS HKU9 CoV CoV SARSr-CoV HKU10 CoV Jinning 209 *6 Yunnan Mengla 168 2 (*1) (2016) Jing hong 212 2 Lufeng 144 Guangdong Zengcheng 234 1 2 (2016) Ruyuan 179 Guangxi Mashan 160 1 (2017) Guilin 224 *2 Total 1,530 0 0 *7 *2 Table 3 Results of ELISA testing of human sera for antibodies to 4 different bat Co V species (*confirmed with western blot). Links Between ELISA Results and Behavior Only one out of the seven SARS-related CoV seropositive respondents said that they had an unusual illness in their lifetime with reported symptoms similar to encephalitis or neural involvement. Two of the respondents said they had experienced symptoms in the past year with only one respondent specifying that they experienced epigastric pain and dizziness. The seven seropositive SARSr-CoV respondents reported various types of animal contacts in the past year. Three had lived with an animal as a pet, four handled a live animal, four raised a live animal, five saw animals inside their dwellings, five had cooked or handled meat, organs, or blood from a recently killed animals, one ate an animal that they knew was not well or sick, one was scratched or bitten by an animal, and four had slaughtered an animal. The only bat contact reported was by one respondent who saw a bat in their dwelling. Both of the respondents who tested positive for HKU10-CoV antibodies said they had experienced an unusual illness in their lifetime, with symptoms associated with encephalitis and SARI. Neither respondent had experienced any symptoms of unusual illness in the past year. Both had reported handling and raising animals, with one indicating they saw animals come inside their dwelling, and one indicating cooking or handling meat, organs, or blood from a recently killed animal. No bat contact was reported by either of the respondents. Overall, five of the total nine SARS-related CoV and HKU10-CoV seropositive respondents reported being worried about disease or disease outbreaks at wet markets. Seven of the nine reported purchasing live animals from a wet market. Specific Aim 1: Summary of Key Findings Our analysis of the key risk factors relating to potential viral zoonotic disease spillover in China indicated some notable differences among the respondents in Guangdong, Guangxi, and Yunnan. With respect to demographic factors, Guangxi fared the lowest on key socio-economic 18 RPPR Page 21

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B.2 (Year 4 NIAID CoV Report_Final for eRA Commons.pdf) 1 R01 Al 110964 Year 4 Report Pl: Daszak, Peter status indicators when compared to Guangdong and Yunnan provinces as reflected by the higher proportion of respondents in Guangxi living under the poverty level. When assessing the type of animal contact and the associated animal taxa over the course of a respondent's lifetime, the results show that respondents in Yunnan engaged in greater contact with animals then those from Guangdong and Guangxi. For example, for 12 of the 14 animal contact types, a higher proportion of Yunnan respondents engaged in these respective activities than in Guangdong and Guangxi. Respondents in Yunnan also reported hunting bats, dogs, and non-human primates which were not reported to being hunted in Guangdong and Guangxi. Swine contact was higher in Yunnan for handling, raising, and slaughtering activities. When examining the various types of animal contact associated with bats only, our results also show that Yunnan respondents reported more varied types of contact with bats. Respondents in Yunnan indicated handling, being scratched by, slaughtering, and hunting bats, but these interactions did not occur in Guangdong or Guangxi. Additional analyses that examine predictors of animal contact in each province will be the focus of human behavioral analyses in Year 5 of the study. Even though our sample population lives in areas that have dense and diverse bat populations, our results show an overall low proportion of respondents reporting hunting and trapping bats in all three provinces. The low proportion of hunting practice could be attributed to the success of conservation enforcement efforts undertaken by the government. These efforts may have effectively reduced the illegal practice of hunting wildlife or, as a consequence, moved the activity underground which made respondents less forthcoming about revealing their engagement in such practices. Further investigation into the potential causes is also warranted. Our analyses also reveal differences in perceptions associated with zoonotic disease spillover between Guangdong, and Guangxi and Yunnan. For example, the proportion of respondents who thought that animals can spread disease was highest in Guangdong province at 72.3%, as compared to Guangxi (48.3%) and Yunnan (49.9%). Moreover, about two-thirds of respondents in Guangdong were worried about diseases and disease outbreaks in wet markets. These differences in perception observed in Guangdong compared to Guangxi and Yunnan could potentially be attributable to a heightened awareness of zoonotic disease emergence due to the 2001 SARS outbreak. Finally, our serological testing results provide the first evidence ever of a bat SARSr-CoV spilling over into people in the wild. All of the SARSr-CoV positive individuals were from Yunnan province, which is the site of a cave in which we have identified a large diversity of SARSr-CoVs within the virome of which every genetic element of SARS-CoV can be identified. These findings warrant further investigations into the type of exposures that may have contributed to bat SARSrelated Co Vs to infect humans in this particular region. They also highlight this region as a hotspot for SARSr-CoV future spillover risk. 19 RPPR Page 22

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Specific Aim 2: Receptor evolution, host range and predictive modeling of bat-CoV emergence risk Bat CoV PCR Detection and Sequencing from Live-Sampled Bat Populations We collected rectal swab and oral swab samples from 671 individual bats from 20 species in Guangdong and Guangxi provinces in southern China in Year 4 (Table 4). 671 rectal swab samples were tested for CoV RNA and 154 (23.0%} were positive (Table 5). Date of Sampling Sampling Locations Rectal swabs Oral swabs May 10th 2017 Hezhou, Guangxi 6 6 May11-12 th 2017 Chongzuo, Guangxi 67 67 May 13th 2017 Nanning, Guangxi 66 66 May 17th, 2017 Beihai, Guangxi 23 23 May 19th 2017 Chongzuo, Guangxi 36 36 May 21st 2017 Yangshan, Qingyuan, Guangdong 46 46 May 22nd , June 7th 2017 Huidong, Huizhou, Guangdong 103 103 June 91 h 2017 Nanning, Guangxi 71 71 June 91 h 2017 Ningming, Chongzuo, Guangxi 63 63 September 10th 2017 Huidong, Huizhou, Guangdong 100 100 September 11th 2017 Yingde, Guangdong 90 90 Total 671 671 Table 4. Bat samples collected for CoV surveillance in Year 4 Species Guangdong Guangxi Total Rhino/ophus sinicus 9/27 6 9/33 Rhinolphus rex 4 4 Rhinolophus pusilus 1 2 3 Rhinolophus pearsoni 5 5 Hipposideros armiger 24 8 32 Hipposideros laNatus 9 9 18 Hipposideros pomona 20 20 Hipposideros pratti 26 26 Aselliscus stoliczkanus 1 1 Miniopterus fuliginosus 1 1 Miniopterus pusillus 29/39 29/39 Myotis chinensis 2/27 2/27 Myotis daubentonii 2 2 Myotis ricketti 86/178 86/178 Pipistrel/us abramus 2 2 Pipistrellus pipistrel/us 2 2 Scotophi/us kuh/i 24/137 24/137 Tylonycteris pachypus 4/115 4/115 Tylonycteris robustula 3 3 Cynopterus sphinx 23 23 Total 126'339 28/332 154/671 Table 5. Number of bat specimens tested and positive (bold) in Year 4 RPPR 20 Page 23

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter A high prevalence of HKU6-related coronaviruses (48.3%), Scotophilus coronavirus 512 (17.5%), and coronavirus 1 B (71.8%) was detected in Myotis ricketii, Schotophilus khulii and Miniopterus pusillus, respectively. SARS-related coronaviruses and HKU2-related coronaviruses were discovered in 4 and 5 Rhino/ophus sinicus samples respectively from Guangdong. HKU4 coronaviruses were identified in 4 Tyfonycteris pachypus from Guangxi (Fig. 29). 1516 MyotischinensiS1GD 2017(2) Bat-coronavirus-HKU6,,isolate-HKU6-1 RdRp DQ249224 613 Myotls ricketlVGOZ017(85) '-----661 Scotophllus kulii/GX 2017_CoV 612(M} 1539-2 Mytois chinensisJGD 2017 ,_ ___ Rousettus-bat-coronavirus-HKU1 0-isolate-175A JQ989271 Hipposideros-bat-coronavirus-HKU10-isolate-TT3A JQ989266 ------Hcov NL63 555 Myotis rickotii/GO 2017 Myotis davidii coronavirus isolate XYMd70 RdRp KF569991 1439-2 Minlopteruspus/1/us/GD 2017@) Bat-coronavirus-HKUB solate-HKUS-1 RdRp DQ249228 Bat-coronavirus-HKU7 -isolale-HKU7 -1 RdRp DQ249226 Bat-coronavirus-1A-strain.AfCD62 EU420138 1464 Minioprerus pui//us/GD 2017(28) Bat-coronavirus-1B-strain-WCF6-ORF18-gene DQ666338 1593 Rhinolophussinicus/GD 2017(5J Bal,coronavirus-HKU2-strain-HKU2/HK/33/2006 EF203067 735 Tylonycteris pacllypus/GX 2017~) Bat-coronavirus-HKU4-1 EF065505 '----Bat-coronavirus-HKUS-1 EF065509 '------- Human-coronav,rus-HKU 1-genotype-B AY884001 '-----Bat-coronavirus-HKU9-2 EF065514 t-----1 01 1572 Rhinolophus sinicus/GD 2017(4J Bat-SARS-coronavirus-HKU3-1 DQ022305 Bat-SARS-coronavirus-Rf1 DO412042 Bat-SARS-coronavirus-Rp3 DQ071615 RsSHC014 KC881005 Rs3367 KC881006 SARS-coronavirus-SZ3 AY304486 SARS-coronavirus-GZ02 AY390556 SARS-coronavirus-BJ01 AY278488 Figure 29: Phylogenetic analysis of partial RdRp gene of CoV (440-nt partial sequence) Genetic Diversity and Genomic Characterization of Betacoronaviruses in Fruit Bats In Year 4, we analyzed the genetic diversity of betacoronaviruses we have detected since 2009 in different species of fruit bats in Yunnan province, including Eonycteris spefaea, Rousettus leschenaultia and an unclassified Rousettus species. These viruses are classified into two betacoronavirus species, HKU9-CoV and GCCDC1-CoV. All HKU9-related viruses (n=46) were found in Rousettus spp. bats while GCCDC1-related viruses (n=13) from E. spelaea. Phylogenetic analysis of the full-length N gene suggests that HKU9-related Co Vs are highly diverse and divided into 5 lineages with previously reported strains, and the GCCDC1 -related Co Vs were more similar between each other (Fig. 30). The full-length genome sequence of a novel HKU9-related CoV termed 2202 was determined. It shares 83% nt identity with other HKU9 strains, with the most divergent regions located in the S 21 RPPR Page 24

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter protein, but shares only 68% aa identity with those of other HKU9 strains. Virus quantification revealed that intestine was the primary infected organ for HKU9-related Co Vs while kidney and lungs could also be target tissues, suggesting potential for spillover through oral-fecal, respiratory, or uro-genitary routes. B 10 BatCoV5767_Ro.le_Chuxiong_2013 BatCoV 5739_Ro.le_ Chuldong_2013 BatCoV HKUS-5-2 HM211099 BatCoV 5766_Ro.le_Chwciong_2013 Lineage1 1 Ba1CoV 5746 Ro,le_Chuxiong 2013 ~--BatCoV HKU9-10-2_HM211101 BatCoV HKIJ9·2 EF065514 BatCoV 2180_Ro.sp_Jlnghong_2009 I BatCoV HKU9-4 EFffi5516 Lineage2 '----BatCoV 2171_R>.sp_Jinghoog_2009 '-------BatCoY 2202_Ro.sp_Jlnghong_2009 I Uneage3 57 BatCoV HKl.9-5-1 HM211098 I 10 BatCoV HKU9.J EF065515 Lineaged 6 BatCoV HKU9-10-1 HM211100 BatCoV 6300 R>.e OiulQOl1g_2J14 I BatCoY 140197 Ro.I,; Mm;la __ 2014 Lineages BatCoV HKUS. IEF065513 BatCoV 160765 Eo Sp Mengla 2016 BatCoV 18l769_Eo.sp_Mengla_2016 6;,tCoV9444 EHp ~&.,ngla 2015 BatCoV 160760_Eo.sp_Mengla_2016 BatCoV GCCDC1-346 KU762337 Bl!!CoV J463_Eo.sp_M,ngla_201'5 BaCoVGCCDCl-356 NC030886 ----------Human-CoVtt<U1 NC006577 ~--------Murine-CoV NC001846 74'----------Bovine-CoVU00735 -------------SARS-CoV NC004718 ~--------BetaCoV Erinaoeos NC022643 99,...._ __ --l --------MERS-CoV NC019843 0.05 --------BatCoV HKUS NC009020 '---------BatCoVHKU4 EF065505 Figure 30. Phylogenetic analysis of full-length N gene of HKU9 and GCCDC1 Co Vs Bat Coronavirus Host-Virus Phylogeography in China We used discrete ancestral character state reconstruction to estimate viral history and reconstructed the inferred bat host genus for each node within the phylogenetic tree (Figs. 31, 32). The color of tree branches indicates the inferred ancestral host bat genus for the reconstructed phylogeny. Rhinofophus is the inferred ancestral host of lineages B and C (SARSlike CoVs and MERS-like CoVs, respectively). This genus played an important role in the diversification of Beta-Co Vs. A larger host diversity is observed for Alpha-CoVs. Our dataset for this analysis includes all CoV RdRp sequences isolated from bat specimens collected by our team from 2008-2015 (Alpha-CoVs: n = 491 - Beta-CoVs: n = 326), including those collected under prior NIAID funding (1 R01 AI079231), funding from Chinese Federal Agencies, and a large majority from our current NIAID project. All Chinese bat CoV RdRp sequences available in GenBank were also added to our dataset (Alpha-CoVs: n = 226 - Beta-CoVs: n = 206). 22 RPPR Page 25

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Phylogenetic trees were reconstructed for Alpha- and Beta-Co Vs separately using Bayesian inference (BEAST 1.8). 0•11tU1 • Rhinolophus • • • • • • • • • P,p-:-. n Rousettus ■ Rhltlel-, .. 1111 • P'H\tffln ■ Sut,11-, 11'1 • t . " ■ u"'" ■ v-., Rhinolophus Rhinolophus Beta-Co Vs Tylonycteris Pipistrellus Vespertilio Rousettus L Cynopterus Hipposideridos 20 Lineage C (MERS-like) Lineage D New lineage Figure 31. Ancestral host reconstruction for Beta-Co Vs, at a host genus level. Oot.11 . .... • • • • • Rhinolophus ■ -Myo~ • • • • ■ ...... ~ Miniopterus -c.....=--.-----. ...... ~; ... ::::j~~~~~ ■ Stet p~II n. ■ .,..., . , Sc ophilus Mlnlopterus . ,.,. . -, .. " . '■ •ao•nJ.t• Scotophilus c::=- Rhinolophus Myotis Myotis Alpha-Co Vs Scotophilus 10 Lineage 1 HKU2 Lineage 4_A HKUlO Lineage 2 lineage 3 HKU7 + HKU8 + lA + 1B Lineage 4_8 HKU10 Lineage 5 HKU6 + 512 Figure 32. Ancestral host reconstruction for Alpha-Co Vs, at a host genus level. RPPR 23 Page 26

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter To better understand the geographic origins and extent of specific CoV clades, we also used discrete ancestral character state reconstruction in BEAST to reconstruct the ancestral location of each branch of the tree. We used SPREAD to visualize the tree in its geographic context and infer CoV spatial spread in China (Fig. 33). These analyses allow us to identify the geographic areas that are likely sources of origin/diversity for this important group of viruses. The common ancestor of most Beta-Co Vs lineages is located in Hong Kong and Guangdong. The common ancestor of most Alpha-CoV lineages was located in Yunnan province, and our results suggest they spread to other provinces from Yunnan. Beta-Co Vs Alpha-Co Vs Figure 33. Ancestral location reconstruction for Beta- and Alpha-Co Vs. The bigger the circle is, the more ancestral the corresponding node is. Specific Aim 3: Testing Predictions of CoV Inter-Species Transmission Identification of two novel MERS-related CoVs that use DPP4 receptor Two novel MERSr-CoVs, BtCoV/li/GD/2013-845 and BtCoV/li/GD/2014-422, were identified from great evening bats (/a io) in Guangdong province. Phylogenetic analysis of polyprotein 1 and the E, M, and N proteins suggests that the two novel strains are more closely related to MERS-CoV than to other lineage C Beta-CoVs. Their RdRp sequences are closely related to those of MERS-CoY and other MERSr-CoVs, with 94.4-97.0% aa identities. In contrast, they are divergent from MERS-CoV and other MERSr-CoVs in the spike protein, with only 58.9- 64. 7% aa identities. However, in the receptor-binding domain (RBD) of the spike protein, the two novel MERSr-CoVs are identical to MERS-CoV at six out of the 13 residues that directly interact with human DPP4 receptor, making them more similar to MERS-CoV than any other known lineage C BetaCoVs (Fig. 34a). Protein-protein interaction assays demonstrated that the spike proteins of the novel MERSr-CoVs bind to both human and bat DPP4 (Fig. 34b). Moreover, bat cells exogenously expressing human DPP4 support the entry of the retrovirus pseudotyped with BtCoV/li/GD/2014-422 spike, while the pseudovirus fails to enter cells that do not express DPP4. The results demonstrate that the spike protein of the newly identified MERSr-CoV recognizes the human DPP4 receptor. 24 RPPR Page 27

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter A MER$ 422 845 HKU4 SC2013 Neo POF-2180 HKU5 MERS 422 845 HKU4 SC201J Neo PDF-2180 HKU5 B fl° C :, 0 ~ (/) ~ !Z' (/) C ~ (J u, ~ "' ~ 467 FNYKQSFSNP TCLILATVPH NLTT---ITK PLKYSY!NKC SRLLSDD-RT 515 YNYKQSFANP TCRIFATAPA NLT----ITK PSSYSF!SKC SRLTGDNSBI 516 FNYKQSFANP TCRIFATAPA NLT----ISK PSSYSYISKC SRLTGDNQHI 517 YNYKQSFANP TCRVMASVLA NVT----ITK PHAYGY!SKC SRLTGANQOV 517 FNYKQDFSNP TCRILATVPA Nl.SASGLLPK PSNYVWLSEC YQNSFTG--- 488 FNYNQDYSNP SCRIHSKVNS SIG----ISY AGAYSYITNC NYGATNK--- 512 FNYNQOYSNP SCRIHSKVNS SVG----ISY SGLYSY!TNC NYGGFNK--- 513 FNYKQDFSNP TCRVLATVPQ NLTT---ITK PSNYAYLTEC YKTSAYG--- 518 513 EVPQLVNANQ YSPCVSIVPS TVWEDGDYYR KQLSPLEGGG WLVASGSTVA 562 ETPIVINPGE YSICKNFAPN GFSQDGDYFT RQLSQLEGGG ILVGVGSVTP 566 ETPITINPGE YSICRGFAPN GLSEDGQVFT RQLSDYEGGG TLVGVGNTVP 567 ETPLYINPGE YSICROFSPG GFSEDGQVFK RTLTQFEGGG LLIGVGTRVP 567 KNFQYVKAGQ YTPCLGLI\AN GFEKSYQTHR OPV------S KLAVTGWTP 532 DDVVKPGGRA SOQCITGALN S-PTTGQLWA YNF-----GG VPYRVSRLTY 556 DDVVKPGGRA SQPCVTGALN S-PTNGQVWS FNF-----GG VPYRTSRLTY 557 KNYLYNAPGA YTPCLSLI\SR GFSTKYQSHS D--------G ELTTTGYIYP 561 • 160k 140k 120k 100k 80k 60K 40k 20k 0 Human DPP4 Bat DPP4 ■ MERS-CoV ■ 422-CoV HKU5-CoV HumanACE2 Figure 34. BtCoV/li/GD/2014-422 RBD analysis (a) and DPP4-binding assay (b) In Vivo Infection of Human ACE2 (hACE2) Expressing Mice with SARSr-CoV S Protein variants Using the reverse genetic methods we previously developed, infectious clones with the WIV1 backbone and the spike protein of SHC014, WIV16 and Rs4231, respectively, were constructed and recombinant viruses were successfully rescued. In Year 4, we performed preliminary in vivo infection of SARSr-CoVs on transgenic mice that express hACE2. Mice were infected with 105 pfu of full-length recombinant virus of WIV1 (rWIV1) and the three chimeric viruses with different spikes. Pathogenesis of the 4 SARSr-CoVs was then determined in a 2-week course. Mice challenged with rW IV1-SHC014S have experienced about 20% body weight loss by the 6th day post infection, while rWIV1 and rWIV-4231 S produced less body weight loss. In the mice infected with rWIV1-WIV16S, no body weight loss was observed (Fig. 35a). 2 and 4 days post infection, the viral load in lung tissues of mice challenged with rWIV1-SHC014S, rWIV1-WIV16S and rWIV1-Rs4231 S reached more than 106 genome copies/g and were significantly higher than that in rWIV1-infected mice (Fig. 35b). These results demonstrate varying pathogenicity of SARSr-CoVs with different spike proteins in humanized mice. 25 RPPR Page 28

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter .c 10 C) ·.; 1.1 C) E3 WIV-1 'iii 3: ..,. Control .9! 8 fD SHC014 >, 1.0 ,._ WIV-1 CL t 0 liiiil WIV-16 0 ~ SHC014 u Ill ~ 6 IIID 4231 't: 0.9 _.,_ WIV-16 .. 0 cii ..,_ 4231 C 4 a, 0 0.8 0 "' ., c, 2 C) .. 0.7 0 c ..J a, 0 0 0.6 '), "' "' ~ ! 0 2 3 4 5 6 i§ DPI l:>q be,i; Figure 35. In vivo infection of SARSr-CoVs in hACE2-expressing mice. {a, left) Body weight change after infection; {b, right) Viral load in lung tissues Additional Year 4 Results for Specific Aim 3: Identification of a HKU2-related Coronavirus of Bat Origin that Caused Fatal Acute Diarrhea in Piglets From October 2016, a series of fatal swine diarrhea disease outbreaks occurred in Guangdong province. By May 2017, it had resulted in death of 24,693 piglets across four farms. We identified a novel coronavirus as the etiological agent of the disease by metagenomic analysis, viral isolation and experimental infection, and named this "Swine Acute Diarrhea Syndrome coronavirus (SADS-CoV). During Year 4, we submitted and published a paper on this finding to Nature (Zhou et al., 2018). The full-length genome of SADS-CoY shares 95% sequence identity to bat CoY HKU2. However, the S gene sequence identity is only 86%, suggesting that the previously reported HKU2-CoV is not the direct progenitor of SADS-CoV, but that they may have originated from a common ancestor. Using a SADS-CoV specific qPCR assay based on its RdRp gene, SADS-related coronaviruses (SADSr-CoVs) were detected in rectal swabs of Rhinolophus bats collected from 2013 to 2016 in Guangdong. Full-length genome sequencing of 4 bat SADSr-CoVs revealed 96% to 98% overall genome sequence identity between SADSr-CoVs and SADS-CoV. Most importantly, the S protein of SADS-CoV shared more than 98% sequence identity with those of the two SADSrCoVs (162149 and 141388), compared to 86% with HKU2-CoV (Fig. 36a). The phylogeny of S1 protein sequence showed strong co-evolutionary relationships with bat alphacoronavirus and their hosts, with swine SADS-CoV more closely related to SADSr-CoVs from Rhino/ophus affinis than strains from Rhinolophus sinicus in which HKU2-CoV was found (Fig. 3Gb). Analysis of the 33 SADS-CoV full genome sequences we were able to characterize from pigs suggests that viruses from the four farms may have been transmitted from their reservoir hosts independently. These findings highlight the importance of identifying coronavirus diversity and distribution in bats to mitigate future outbreaks that threaten livestock and public health. 26 RPPR Page 29

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter A SADS•CoV: 27,173bp NS3a liS7a C ORF la j 1 0Rf1b rs IIIEE:::.I, --1-62-14-~-V:-27-.17-7 bp-.-98-.48-%_iclen_tify--~ .. -.. _ ... -._ .. --.. _-~-_:-_-~ ...~;_··:__-_·--·_~~----~:-_.·-=,:M::E:_M~-1_ 1 N__,~: 14138$-CoV: 27, 174 bp, 98.0S% identify 8462.CoV: 27,200 bp, 96.36% identify 8495.COV: 27,198 bp, 96.28% identify HKU2.GO.COV: 27,165 bp, QS.09% Klonafy NS7b NS7e NS7a NS7b NS7, I!\:] NS7• lb B - Rhinok,plw-s affltis - ~inolophus sinlrus - Rhinolophus r&X - Pig o. 0.99 Figure 36. Genome organization and comparison (a) and Phylogenetic analysis of S1 protein (b) of SADS-CoV and bat SADSr-CoVs Intra-Farm Transmission Model to Understand to Predict Future Transmission and Outbreak To better understand amplification dynamics and assess the potential for future transmission resulting in large outbreaks, we developed an intra-farm, age-structured, stochastic transmission model for SADS-CoV (Fig. 37). We developed multiple versions of this model to represent different hypotheses of disease transmission mechanisms and fit them to time-series data of reported deaths on multiple SADS-infected farms. SADSModel gPs Os wOs It.Or no0,+w0 1 ro, o. .,o. Piglets < 4 Days Old Piglets > 4 Days Old I\ = /J (P1+n,+S,) N' ,s /1.Ss rs, SH Sows Figure 37: Schematic of intra-farm transmission mode. ;s, 27 RPPR Page 30

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B.2 (Year4 NIAID CoV Report_Final foreRA Commons.pdt) 1 R01 Al110964 Year 4 Report Pl: Daszak, Peter Our first model structure, which assumed equal mixing of animals across farms (Fig. 38) showed that age structure alone was insufficient to generate the temporal pattern of reported deaths on SADS-infected farms. Our second model structure (Fig. 39) represented individual barns on a farm as a series of pig-virus meta-populations. This structure was sufficient to recreate the dynamics of the series of rapid "mini-epidemics" that progressed in SADS-infected farms. 300 100 0 20 40 Tme{Days) Figure 38: Best-fit simulations (red) from an equal-mixing transmission model and actual reported death time series {black) on a SADS-infected farm. f/) .c 300 :,ii 200 0 ~ ~ a: 100 20 40 time s1mula11on -data Figure 39: Best-fit simulations (grey) from an metapopulation transmission model and actual reported death time series (black) on a SADS-infected farm. 28 RPPR Page 31

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B.2 (Year 4 NIAID CoV Report_Final for eRA Commons.pdf) 1 R01 Al 110964 Year 4 Report Pl: Daszak, Peter Specific Goals Not Meet • The wild animal farm survey was piloted in early Y4, with data collected from seven wild animal farms, it was postponed due to the emergence of SADS-CoV where our group had focused on instead in Y4, but will be resumed in Y5 to continue collecting and analyzing data. • The passive hospital surveillance has been piloted will continue in Year 4 to collect and test for CoVs. 29 RPPR Page 32

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B.4 (Year4 NIAID CoV Training and ProfDevlp.pdf) 1R01Al110964 Year4 Report Pl: Daszak, Peter B. 4 What opportunities for training and professional development has the project provided? 1. Conference and University lectures: We provided human subject research trainings to chief physicians and nurses at local clinics, staff from Yunnan Institute of Endemic Diseases Control and Prevention, students from Dali College and Wuhan University for both qualitative and quantitative research. 2. Agency and other briefing: Dr. Guangjian Zhu was invited by the Guangdong Institute of Applied Nature Resources, Guangdong Academy of Sciences to provide training to 8 field team members regarding biosafety and PPE use, bats and rodents sampling. Dr. Zhengli Shi participated in the US National Science Foundation-funded EcoHealthNet (grant to EcoHealth Alliance - Epstein Pl) that provides research exchange opportunities to undergraduate and graduate-level students. 3. Public outreach: Pl Daszak, and Co-investigators Shi, Epstein, and Olival presented the results of this project to the public via interviews with national central and local television, social media, newspaper and journals in China and the US. 1 RPPR Page 33

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RPPR FINAL C. PRODUCTS C.1 PUBLICATIONS Are there publications or manuscripts accepted for publication in a journal or other publication (e.g., book, one-time publication, monograph) during the reporting period resulting directly from this award? Yes Publications Reported for this Reporting Period ~·· ~-~ ......,..,_-(.,:,1'_•,V , ______________________________ Complete Luo CM, Wang N, Yang XL, Liu HZ, Zhang W, Li B, Hu B, Peng C, Geng QB, Zhu GJ, Li F, Shi ZL. Discovery of Novel Bat Coronaviruses in South China That Use the Same Receptor as Middle East Respiratory Syndrome Coronavirus. Journal of virology. 2018 July 1 ;92(13). PubMed PMID: 29669833; PubMed Central PMCID: PMC6002729; DOI: 10.1128/JVl.00116-18. Complete Field HE. Evidence of Australian bat lyssavirus infection in diverse Australian bat taxa. Zoonoses and public health. 2018 September;65(6):742-748. PubMed PMID: 29785730; PubMed Central PMCID: PMC6249124; DOI: 10.1111 /zph.12480. Complete Eskew EA, Oliva! KJ. De-urbanization and Zoonotic Disease Risk. EcoHealth. 2018 December;15(4):707-712. PubMed PMID: 30120670; PubMed Central PMCID: PMC6265062; DOI: 10 .1007 /s 10393-018-1359-9. Complete Cui J, Li F, Shi ZL. Origin and evolution of pathogenic coronaviruses. Nature reviews. Microbiology. 2019 March;17(3):181-192. PubMed PMID: 30531947; PubMed Central PMCID: PMC7097006; DOI: 10.1038/s41579-018-0118-9. Complete Li HY, Zhu GJ, Zhang YZ, Zhang LB, Hagan EA, Martinez S, Chmura AA, Francisco L, Tai H, Miller M, Daszak P. A qualitative study of zoonotic risk factors among rural communities in southern China. International health. 2020 February 12;12(2):77-85. PubMed PMID: 32040190; PubMed Central PMCID: PMC7017878; DOI: 10.1093/inthealth/ihaa001. C.2 WEBSITE(S) OR OTHER INTERNET SITE(S) Nothing to report C.3 TECHNOLOGIES OR TECHNIQUES NOTHING TO REPORT C.4 INVENTIONS, PATENT APPLICATIONS, AND/OR LICENSES Have inventions, patent applications and/or licenses resulted from the award during the reporting period? No If yes, has this information been previously provided to the PHS or to the official responsible for patent matters at the grantee organization? No C.5 OTHER PRODUCTS AND RESOURCE SHARING Nothing to report RPPR Page 34

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RPPR FINAL D. PARTICIPANTS D.1 WHAT INDIVIDUALS HAVE WORKED ON THE PROJECT? ~l'~lrl41~,.--n ~-~F7~7 • <.->.: '' ' • • 1 ,, ' ~.. ,. " f l ' } ,., ! , ~ 1.,.,. • ~" ' ' l ~ 1,.« ~ I '' • ' .. ~ '1:,_.,t,,M,,C,,.'<,,,.,_<-:'.,K.-~ '-"''"" ·-:-,t,~,:,,.,,. • r~~-,,,.,.,,,,..; ;I.,',.,.~,. r.,-.,J~<,; \:'.::f ·"'" ;t,,.~~--"'·'';,~,1 $;,:-c,:. (b) (4) (b) (6) (b)(6)l y (b) (6)] N (b)(6)] N lblc6ll y (b) (6)1 y N N N y (b)(6)l N (b) (6)1 N Glossary of acronyms: S/K - Senior/Key DOB - Date of Birth DASZAK, PETER Chmura, Aleksei Ross, Noam Martin Olival, Kevin J. Zhang, Shu-yi ZHU, GUANGJIAN GE, XINGYI KE, CHANGWEN ZHANG, YUNZHI EPSTEIN, JONATHAN H SHI, ZHENGLI Cal - Person Months (Calendar) Aca - Person Months (Academic) Sum - Person Months (Summer) D.2 PERSONNEL UPDATES D.2.a Level of Effort RPPR BS,PHD PD/Pl BS,PHD Non-Student Research Assistant PhD CoInvestigator PHO CoInvestigator PHO CoInvestigator PHO CoInvestigator PHO CoInvestigator PHO CoInvestigator PHO CoInvestigator MPH,DVM Co- ,BA,PHD Investigator PhD CoInvestigator East China CHINA Normal University East China CHINA Normal University Wuhan CHINA Institute of Virology Center for CHINA Disease Control and Prevention of Guangdon g Province Yunnan CHINA Provincial Institute of Endemic Diseases Control & Prevention Wuhan CHINA Institute of Virology Foreign Org - Foreign Organization Affiliation SS - Supplement Support RE - Reentry Supplement DI - Diversity Supplement OT - Other NA - Not Applicable Page35 NA NA NA NA NA NA NA NA NA NA NA

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RPPR FINAL Will there be, in the next budget period, either (1) a reduction of 25% or more in the level of effort from what was approved by the agency for the PD/Pl(s) or other senior/key personnel designated in the Notice of Award, or (2) a reduction in the level of effort below the minimum amount of effort required by the Notice of Award? No 0.2.b New Senior/Key Personnel Are there, or will there be, new senior/key personnel? No 0.2.c Changes in Other Support Has there been a change in the active other support of senior/key personnel since the last reporting period? No D.2.d New Other Significant Contributors Are there, or will there be, new other significant contributors? No 0.2.e Multi-Pl (MPI) Leadership Plan Will there be a change in the MPI Leadership Plan for the next budget period? NA RPPR Page 36

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RPPR E. IMPACT E.1 WHAT IS THE IMPACT ON THE DEVELOPMENT OF HUMAN RESOURCES? Not Applicable FINAL E.2 WHAT IS THE IMPACT ON PHYSICAL, INSTITUTIONAL, OR INFORMATION RESOURCES THAT FORM INFRASTRUCTURE? NOTHING TO REPORT E.3 WHAT IS THE IMPACT ON TECHNOLOGY TRANSFER? Not Applicable E.4 WHAT DOLLAR AMOUNT OF THE AWARD'S BUDGET IS BEING SPENT IN FOREIGN COUNTRY(IES)? RPPR Page 37

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RPPR FINAL F.CHANGES F.1 CHANGES IN APPROACH AND REASONS FOR CHANGE Not Applicable F.2 ACTUAL OR ANTICIPATED CHALLENGES OR DELAYS AND ACTIONS OR PLANS TO RESOLVE THEM NOTHING TO REPORT F.3 SIGNIFICANT CHANGES TO HUMAN SUBJECTS, VERTEBRATE ANIMALS, BIOHAZARDS, AND/OR SELECT AGENTS F.3.a Human Subjects No Change F.3.b Vertebrate Animals No Change F.3.c Biohazards No Change F.3.d Select Agents No Change RPPR Page 38

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RPPR FINAL G. SPECIAL REPORTING REQUIREMENTS G.1 SPECIAL NOTICE OF AWARD TERMS AND FUNDING OPPORTUNITIES ANNOUNCEMENT REPORTING REQUIREMENTS NOTHING TO REPORT G.2 RESPONSIBLE CONDUCT OF RESEARCH Not Applicable G.3 MENTOR'S REPORT OR SPONSOR COMMENTS Not Applicable G.4 HUMAN SUBJECTS 58010 Understanding the Risk of Bat Coronavirus EmergencePROTOCOL-001 G.5 HUMAN SUBJECTS EDUCATION REQUIREMENT NO NO Are there personnel on this project who are newly involved in the design or conduct of human subjects research? No G.6 HUMAN EMBRYONIC STEM CELLS (HESCS) NO Does this project involve human embryonic stem cells (only hESC lines listed as approved in the NIH Registry may be used in NIH funded research)? No G.7 VERTEBRATE ANIMALS Does this project involve vertebrate animals? Yes G.8 PROJECT/PERFORMANCE SITES (!,--.z-.f.:, ·t ~,. .~, .. , . .,, .,._"f-,., 1~t>fi·-: ,,,_.,,,--,--•,>l.,.. s r· .<.t~-.~- j'["'\·t-"",,,~w. }n~ ~ < ~ ·t-" ~- , / ; 1\ : .. .,. • d, -: r~ ~ ·. r._ ,..\ , . ' ... 5 11 ~ ....... 1.~ .,_ ·, ~ t ~ Z .. -----·---------· --------------------- Primary: 077090066 NY-010 460 West 34th Street EcoHealth 17th Floor Alliance, Inc. New York NY 100012317 Wuhan Institute of 529027474 Xiao Hong Shan, No. 44 Virology Wuchang District Wuhan NONE East China Normal 420945495 3663 Zhongshan Beilu University Shanghai NONE ECOHEALTH 077090066 ECOHEAL TH ALLIANCE, INC. ALLIANCE 460 W 34TH ST RPPR Page 39

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RPPR FINAL I I I I NEW YORK NY 100012320 I G.9 FOREIGN COMPONENT Organization Name: Wuhan Institute of Virology Country: CHINA Description of Foreign Component: Principal Laboratory for all Research in China as per section GB (above) and detailed in our Specific Aims Organization Name: Wuhan School of Public Health Country: CHINA Description of Foreign Component: Principal Coordinating Team for all project field work as per section GB (above) and detailed in our Specific Aims G.10 ESTIMATED UNOBLIGATED BALANCE G.1 0.a Is it anticipated that an estimated unobligated balance (including prior year carryover) will be greater than 25% of the current year's total approved budget? No G.11 PROGRAM INCOME Is program income anticipated during the next budget period? No G.12 F&A COSTS Is there a change in performance sites that will affect F&A costs? No RPPR Page40

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Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001) Section 1 - Basic Information (Study 58010) 1 . 1 . Study Title • Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001 1.2. Is this study exempt from Federal Regulations • 1.3. Exemption Number 1.4. Clinical Trial Questionnaire • 1.4.a. Does the study involve human participants? O Yes 0 1 02 1.4.b. Are the participants prospectively assigned to an intervention? e No 03 1.4.c. Is the study designed to evaluate the effect of the intervention on the participants? 1.4.d. Is the effect that will be evaluated a health-related biomedical or behavioral outcome? 1.5. Provide the ClinicalTrials.gov Identifier (e.g. NCT87654321) for this trial, if applicable RPPR 04 05 • Yes 0 Yes O Yes O Yes 06 07 0 No e No e No e No 0MB Number: 0925-0001 and 0925--0002 Expiration Date: 03/31/2020 08 Page41

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DASZAK,PETER Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001) Section 2 - Study Population Characteristics (Study 58010) 2.1. Conditions or Focus of Study 2.2. Eligibility Criteria 2.3. Age Limits 2.4. Inclusion of Women, Minorities, and Children 2.5. Recruitment and Retention Plan 2.6. Recruitment Status 2.7. Study Timeline RPPR Min Age: Not yet recruiting Max Age: Page42

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DASZAK, PETER Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-DOI) Inclusion Enrollment Reports IER ID# Enrollment Location Type Enrollment Location IER 58010 Foreign RPPR Page 43

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DASZAK, PETER Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-DOI) Inclusion Enrollment Report 58010 Using an Existing Dataset or Resource* : O Yes • No Enrollment Location Type* : O Domestic • Foreign Enrollment Country(ies): CHN: CHINA Enrollment Location(s): Comments: Planned Ethnic Categories Racial Categories Total Not Hispanic or Latino Hispanic or Latino Female Male Female Male American Indian/ 0 0 0 0 0 Alaska Native Asian 1230 1230 0 0 2460 Native Hawaiian or 0 0 0 0 0 Other Pacific Islander Black or African 0 0 0 0 0 American White 0 0 0 0 0 More than One Race 0 0 0 0 0 Total 1230 1230 0 0 2460 Cumulative (Actual) Ethnic Categories Unknown/Not Racial Categories Not Hispanic or Latino Hispanic or Latino Reported Ethnicity Total Unknown/ Unknowni Unknown/ Female Male Not Female Male Not Female Male Not Reported Reported Reported American Indian/ 0 0 0 0 0 0 0 0 0 0 Alaska Native Asian 980 616 0 0 0 0 0 0 0 1596 Native Hawaiian or 0 0 0 0 0 0 0 0 0 0 Other Pacific Islander Black or African 0 0 0 0 0 0 0 0 0 0 American White 0 0 0 0 0 0 0 0 0 0 More than One Race 0 0 0 0 0 0 0 0 0 0 Unknown or 0 0 0 0 0 0 0 0 0 0 Not Reported Total 980 616 0 0 0 0 0 0 0 1596 RPPR Page 44

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DASZAK,PETER Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-001) Section 3 - Protection and Monitoring Plans (Study 58010) 3.1. Protection of Human Subjects 3.2. Is this a multi-site study that will use the same protocol to O Yes conduct non-exempt human subjects research at more than one domestic site? If yes, describe the single IRB plan 3.3. Data and Safety Monitoring Plan 3.4. Will a Data and Safety Monitoring Board be appointed for this study? 3.5. Overall structure of the study team RPPR o Yes 0 No 0 N/A O No Page 45

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DASZAK, PETER Human Subject Report (Understanding the Risk of Bat Coronavirus Emergence-PROTOCOL-DOI) Section 4 - Protocol Synopsis {Study 58010) 4.1. Brief Summary 4.2. Study Design 4.2.a. Narrative Study Description 4.2.b. Primary Purpose 4.2.c. Interventions j Type j Name ] Description 4.2.d. Study Phase Is this an NIH-defined Phase Ill Clinical Trial? O Yes 4.2.e. Intervention Model 4.2.f. Masking o Participant Yes o Care Provider 4.2.g. Allocation 4.3. Outcome Measures Type Name 4.4. Statistical Design and Power 4.5. Subject Participation Duration Time Frame 4.6. Will the study use an FDA-regulated intervention? 4.6.a. If yes, describe the availability of lnvestigational Product (IP) and lnvestigational New Drug (IND)/ lnvestigational Device Exemption (IDE) status 4.7. Dissemination Plan RPPR O Yes e No O No o Investigator o Outcomes Assessor Brief Description 0 No Page 46

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Notice of Award RESEARCH Federal Award Date: 07/13/2020 Department of Health and Human Services National Institutes of Health NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Grant Number: 2R01Al110964-06 REVISED FAIN: R01Al110964 Principal lnvestigator(s): PETER DASZAK,PHD Project Title: Understanding the Risk of Bat Coronavirus Emergence Dr. Daszak, Peter PD/Pl 460 West 34th Street Suite 1701 New York, NY 100012320 Award e-mailed to: (b)(6) ----------- Period Of Performance: Budget Period: 07/24/2019 - 06/30/2021 Project Period: 06/01/2014 -06/30/2025 Dear Business Official: The National Institutes of Health hereby revises this award to reflect an increase in the amount of $369,819 (see "Award Calculation" in Section I and "Terms and Conditions" in Section Ill) to ECO HEAL TH ALLIANCE, INC. in support of the above referenced project. This award is pursuant to the authority of 42 USC 241 42 CFR 52 and is subject to the requirements of this statute and regulation and of other referenced, incorporated or attached terms and conditions. Acceptance of this award including the ''Terms and Conditions" is acknowledged by the grantee when funds are drawn down or otherwise obtained from the grant payment system. Each publication, press release, or other document about research supported by an NIH award must include an acknowledgment of NIH award support and a disclaimer such as "Research reported in this publication was supported by the National Institute Of Allergy And Infectious Diseases of the National Institutes of Health under Award Number R01Al110964. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health." Prior to issuing a press release concerning the outcome of this research, please notify the NIH awarding IC in advance to allow for coordination. Award recipients must promote objectivity in research by establishing standards that provide a reasonable expectation that the design, conduct and reporting of research funded under NIH awards will be free from bias resulting from an Investigator's Financial Conflict of Interest (FCOI), in accordance with the 2011 revised regulation at 42 CFR Part 50 Subpart F. The Institution shall submit all FCOI reports to the NIH through the eRA Commons FCOI Module. The regulation does not apply to Phase I Small Business Innovative Research (SBIR) and Small Business Technology Transfer (STIR) awards. Consult the NIH website http://grants.nih.gov/grants/policy/coi/ for a link to the regulation and additional important information. If you have any questions about this award, please contact the individual(s) referenced in Section IV. Sincerely yours, Page-1 NIH NGAR I Version 5€-12.126120182 2200 PMI Generated on 7/1512020 120048AM ID)

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Emily Linde Grants Management Officer NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES Additional information follows Page-2 NIH NGA RI Version 56 -12/2612018 2 22 00 PMI Genera1ed on. 7115/2020 1200.48 AM

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SECTION I -AWARD DATA- 2R01Al110964-06 REVISED Award Calculation (U.S. Dollars) Salaries and Wages Fringe Benefits Personnel Costs (Subtotal) Consultant Services Materials & Supplies Travel SubawardsfConsortiumfContractual Costs Federal Direct Costs Federal F&A Costs Approved Budget Total Amount of Federal Funds Obligated (Federal Share) TOTAL FEDERAL AWARD AMOUNT AMOUNT OF THIS ACTION (FEDERAL SHARE) SUMMARY TOTALS FOR ALL YEARS $170,325 $53,654 $223,979 $49,809 $20,170 $15,045 $229,923 $538,926 $123,054 $661,980 $661,980 $661,980 $369,819 YR THIS AWARD CUMULATIVE TOTALS 6 $661,980 $661,980 7 $637,980 $637,980 8 $637,980 $637,980 9 $637,980 $637,980 10 $637,980 $637,980 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project Fiscal Information: CFDA Name: Allergy and Infectious Diseases Research CFDA Number: 93.855 EIN: 1311726494A1 Document Number: RAl110964B PMS Account Type: P (Subaccount) Fiscal Year: 2019 IC CAN 2019 2021 Al 8472364 $661,980 $637.980 2022 $637,980 2023 2024 $637,980 $637,980 Recommended future year total cost support, subject to the availability of funds and satisfactory progress of the project NIH Administrative Data: PCC: M51C B / OC: 41022 / Released: CbH6J 07/13/2020 Award Processed: 07/15/2020 12:00:48 AM SECTION 11- PAYMENTIHOTLINE INFORMATION - 2R01Al110964-06 REVISED For payment and HHS Office of Inspector General Hotline information, see the NIH Home Page at http://qrants.nih.gov/qrants/policy/awardconditions.htm SECTION Ill -TERMS AND CONDITIONS - 2R01Al110964-06 REVISED This award is based on the application submitted to, and as approved by, NIH on the above-titled project and is subject to the terms and conditions incorporated either directly or by reference in the following: a. The grant program legislation and program regulation cited in this Notice of Award. b. Conditions on activities and expenditure of funds in other statutory requirements, such as those included in appropriations acts. Page-3 NIH NGAR I Vers,on 5€-12.126120182 2200 PMI Generated on 7/1512020 120048AM

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c. 45 CFR Part 75. d. National Policy Requirements and all other requirements described in the NIH Grants Policy Statement, including addenda in effect as of the beginning date of the budget period. e. Federal Award Performance Goals: As required by the periodic report in the RPPR or in the final progress report when applicable. f. This award notice, INCLUDING THE TERMS AND CONDITIONS CITED BELOW. (See NIH Home Page at http://grants.nih.gov/grants/policy/awardconditions.htm for certain references cited above.) Research and Development (R&D): All awards issued by the National Institutes of Health (NIH) meet the definition of "Research and Development" at 45 CFR Part§ 75.2. As such, auditees should identify NIH awards as part of the R&D cluster on the Schedule of Expenditures of Federal Awards (SEFA). The auditor should test NIH awards for compliance as instructed in Part V, Clusters of Programs. NIH recognizes that some awards may have another classification for purposes of indirect costs. The auditor is not required to report the disconnect (i.e., the award is classified as R&D for Federal Audit Requirement purposes but non-research for indirect cost rate purposes), unless the auditee is charging indirect costs at a rate other than the rate(s) specified in the award document(s). An unobligated balance may be carried over into the next budget period without Grants Management Officer prior approval. This grant is subject to Streamlined Noncompeting Award Procedures (SNAP). This award is subject to the requirements of 2 CFR Part 25 for institutions to receive a Dun & Bradstreet Universal Numbering System (DUNS) number and maintain an active registration in the System for Award Management (SAM). Should a consortium/subaward be issued under this award, a DUNS requirement must be included. See http://grants.nih.gov/grants/policy/awardconditions.htm for the full NIH award term implementing this requirement and other additional information. This award has been assigned the Federal Award Identification Number (FAIN) R01Al110964. Recipients must document the assigned FAIN on each consortium/subaward issued under this award. Based on the project period start date of this project, this award is likely subject to the Transparency Act subaward and executive compensation reporting requirement of 2 CFR Part 170. There are conditions that may exclude this award; see http://grants.nih.gov/grants/policy/awardconditions.htm for additional award applicability information. In accordance with P.L. 110-161, compliance with the NIH Public Access Policy is now mandatory. For more information, see NOT-OD-08-033 and the Public Access website: http://publicaccess.nih.gov/. In accordance with the regulatory requirements provided at 45 CFR 75.113 and Appendix XII to 45 CFR Part 75, recipients that have currently active Federal grants, cooperative agreements, and procurement contracts with cumulative total value greater than $10,000,000 must report and maintain information in the System for Award Management (SAM) about civil, criminal, and administrative proceedings in connection with the award or performance of a Federal award that reached final disposition within the most recent five-year period. The recipient must also make semiannual disclosures regarding such proceedings. Proceedings information will be made publicly available in the designated integrity and performance system (currently the Federal Awardee Performance and Integrity Information System (FAPIIS}). Full reporting requirements and procedures are found in Appendix XII to 45 CFR Part 75. This term does not apply to NIH fellowships. Page-4 NIH NGA RI Version 56 -12/2612018 2 22 00 PMI Genera1ed on. 7115/2020 1200.48 AM

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Treatment of Program Income: Additional Costs SECTION IV - Al Special Terms and Conditions - 2R01Al110964-06 REVISED Clinical Trial Indicator: No This award does not support any NIH-defined Clinical Trials. See the NIH Grants Policy Statement Section 1.2 for NIH definition of Clinical Trial. REVISED AWARD: Pursuant to the letter to EcoHealth Alliance, Inc. dated July 8, 2020, this award has been reinstated; however, all activities are suspended until such time as these concerns in the letter have been addressed to NIH's satisfaction. Supersedes previous Notice of Award dated 04/27/2020. All other terms and conditions still apply to this award. REVISED AWARD: This award is revised to adjust the budget in accordance with the letter from Aleksei Chmura/ECOHealth Alliance. Supersedes previous Notice of Award dated 07/24/2019. This Notice of Award (NoA) includes funds for activity with The University of North Carolina at Chapel Hill in the amount of $77,750 ($50,000 direct costs+ $27,750F&A costs). This Notice of Award (NoA) includes funds for activity with Wuhan Institute of Virology in the amount of $76,301 ($70,649 direct costs + $5,652 F&A costs). This Notice of Award (NoA) includes funds for activity with Institute of Pathogen Biology in the amount of $75,600 ($70,000 direct costs + $5,600 F&A costs). The Research Performance Progress Report (RPPR), Section G.9 (Foreign component), includes reporting requirements for all research performed outside of the United States. Research conducted at the following site(s) must be reported in your RPPR: Wuhan Institute of Virology, CHINA Institute of Pathogen Biology, CHINA East China Normal University, CHINA Duke-NUS Medical School, SINGAPORE This award reflects current Federal policies regarding Facilities & Administrative (F&A) Costs for foreign grantees including foreign sub-awardees, and domestic awards with foreign subawardees. Please see: Chapter 16 Grants to Foreign Organizations, International Organizations, and Domestic Grants with Foreign Components, Section 16.6 "Allowable and Unallowable Cost" of the NIH Grants Policy. This award may include collaborations with and/or between foreign organizations. Please be advised that short term travel visa expenses are an allowable expense on this grant, if justified as critical and necessary for the conduct of the project. The budget period anniversary start date for future year(s) will be July 1. ******************** Page-5 NIH NGA RI Vers,on 56. 12/2612018 2 22 00 PMI Generated on 7/1512020 noo 48 AM

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Dissemination of study data will be in accord with the Recipient's accepted genomic data sharing plan as stated in the page(s) 203 of the application. Failure to adhere to the sharing plan as mutually agreed upon by the Recipient and the NIAID may result in Enforcement Actions as described in the NIH Grants Policy Statement. ******************** This award is subject to the Clinical Terms of Award referenced in the NIH Guide for Grants and Contracts, July 8, 2002, NOT Al-02-032. These terms and conditions are hereby incorporated by reference, and can be accessed via the following World Wide Web address: https://www.niaid.nih.gov/grants-contracts/niaid-clinical-terms-award All submissions required by the NIAID Clinical Terms of Award must be forwarded electronically or by mail to the responsible NIAID Program Official identified on this Notice of Award. ******************** Awardees who conduct research involving Select Agents (see 42 CFR 73 for the Select Agent list; and 7 CFR 331 and 9 CFR 121 for the relevant animal and plant pathogens at http://www.selectagents.gov/Regulations.html) must complete registration with CDC (or APHIS, depending on the agent) before using NIH funds. No funds can be used for research involving Select Agents if the final registration certificate is denied. Prior to conducting a restricted experiment with a Select Agent or Toxin, awardees must notify the NIAID and must request and receive approval from CDC or APHIS. ******************** Select Agents: Awardee of a project that at any time involves a restricted experiment with a select agent, is responsible for notifying and receiving prior approval from the NIAID. Please be advised that changes in the use of a Select Agent will be considered a change in scope and require NIH awarding office prior approval. The approval is necessary for new select agent experiments as well as changes in on-going experiments that would require change in the biosafety plan and/or biosafety containment level. An approval to conduct a restricted experiment granted to an individual cannot be assumed an approval to other individuals who conduct the same restricted experiment as defined in the Select Agents Regulation 42 CFR Part 73, Section 13.b (http://www.selectagents.gov/Regulations.html). Highly Pathogenic Agent: NIAID defines a Highly Pathogenic Agent as an infectious Agent or Toxin that may warrant a biocontainment safety level of BSL3 or higher according to the current edition of the CDC/NIH Biosafety in Microbiological and Biomedical Laboratories (BMBL) (http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm). Research funded under this grant must adhere to the BMBL, including using the BMBL-recommended biocontainment level at a minimum. If your Institutional Biosafety Committee (or equivalent body) or designated institutional biosafety official recommend a higher biocontainment level, the highest recommended containment level must be used. When submitting future Progress Reports indicate at the beginning of the report: If no research with a Highly Pathogenic Agent or Select Agent has been performed or is planned to be performed under this grant. If your IBC or equivalent body or official has determined, for example, by conducting a risk assessment, that the work being planned or performed under this grant may be conducted at a biocontainment safety level that is lower than BSL3. If the work involves Select Agents and/or Highly Pathogenic Agents, also address the following points: Any changes in the use of the Agent(s) or Toxin(s) including its restricted experiments that have resulted in a change in the required biocontainment level, and any resultant change in location, if applicable, as determined by your IBC or equivalent body or official. If work with a new or additional Agent(s)ffoxin(s) is proposed in the upcoming project period, provide: Page-6 NIH NGA RI Version 56 -12/2612018 2 22 00 PMI Genera1ed on. 7115/2020 1200.48 AM

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o A list of the new and/or additional Agent(s) that will be studied; o A description of the work that will be done with the Agent( s ), and whether or not the work is a restricted experiment; o The title and location for each biocontainment resource/facility, including the name of the organization that operates the facility, and the biocontainment level at which the work will be conducted, with documentation of approval by your IBC or equivalent body or official. It is important to note if the work is being done in a new location. STAFF CONT ACTS The Grants Management Specialist is responsible for the negotiation, award and administration of this project and for interpretation of Grants Administration policies and provisions. The Program Official is responsible for the scientific, programmatic and technical aspects of this project. These individuals work together in overall project administration. Prior approval requests (signed by an Authorized Organizational Representative) should be submitted in writing to the Grants Management Specialist. Requests may be made via e-mail. Grants Management Specialist: Shaun W Gratton Email: (b)(6) Phone: (b)(6) Program Official: Erik J. Stemmy Email: (b)(6) Phone: (b)(6) ----- SPREADSHEET SUMMARY GRANT NUMBER: 2R01Al110964-06 REVISED INSTITUTION: ECOHEAL TH ALLIANCE, INC. Budqet Year6 Year 7 Year8 Year 9 Year10 Salaries and Waqes $170,325 $170,123 $170,123 $170,123 $170,123 Frinqe Benefits $53,654 $53,590 $53,590 $53,590 $53,590 Personnel Costs (Subtotal) $223,979 $223,713 $223,713 $223,713 $223,713 Consultant Services $49,809 $49,750 $49,750 $49,750 $49,750 Materials & Suoolies $20,170 $14,850 $14,850 $14,850 $14,850 Travel $15,045 $15,027 $15,027 $15,027 $15,027 Subawards/Consortium/Contractual $229,923 $229,651 $229,651 $229,651 $229,651 Costs Publication Costs $6,000 $6,000 $6,000 $6,000 TOTAL FEDERAL DC $538,926 $538,991 $538,991 $538,991 $538,991 TOTAL FEDERAL F&A $123,054 $98,989 $98,989 $98,989 $98,989 TOTAL COST $661,980 $637,980 $637,980 $637,980 $637,980 Facilities and Administrative Costs Year6 Year7 Year8 Year9 Year 10 F&A Cost Rate 1 32% 32% 32% 32% 32% F&A Cost Base 1 $384,547 $309,340 $309,340 $309,340 $309,340 F&A Costs 1 $123,054 $98,989 $98,989 $98,989 $98,989 Page-7 NIH NGAR I Version 5€-12.126120182 2200 PMI Generated on 7/1512020 120048AM

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Pl: DASZAK, PETER Title: Understanding the Risk of Bat Coronavirus Emergence Received: 11/05/2018 FOA: PA 18-484 Council: 05/2019 Clinical Trial:Not Allowed Competition ID: FORMS-E FOA Title: NIH Research Project Grant (Parent R01 Clinical Trial Not Allowed) 2 R01 Al110964-06 Dual: Accession Number: 4237214 IPF: 4415701 Organization: ECOHEAL TH ALLIANCE, INC. Former Number: Department: IRG/SRG: CRFS AIDS:N Expedited: N Subtotal Direct Costs Animals: Y New Investigator: N (excludes consortium F&A) Humans: Y Early Stage Investigator: N Year 6: 515,358 Clinical Trial: N Year?: 515,358 Current HS Code: (b) ( 4) Year 8: 515,358 HESC: N Year 9: 515,358 Year 10: 515,358 Senior/Key Personnel: Organization: Role Category: PETER DASZAK ECOHEAL TH ALLIANCE, INC. PD/Pl Zheng Li Shi Wuhan Institute of Virology Co-Investigator Kevin Olival EcoHealth Alliance Co-Investigator Ralph Barie University of North Carolina Co-Investigator Noam Ross EcoHealth Alliance Co-Investigator Alice Latinne EcoHealth Alliance Other (Specify)-Research Scientist HongYing Li EcoHealth Alliance Other (Specify)-Research Scientist Leilani Francisco EcoHealth Alliance Co-Investigator Amy Sims University of North Carolina at Chapel Co-Investigator Hill Emily Hagan EcoHealth Alliance Other (Specify)-Research Scientist Guangjian Zhu East China Normal University Co-Investigator Linfa Wang Duke-NUS Medical School Co-Investigator Lili Ren Institute of Pathogen Biology Co-Investigator Li Guo Institute of Pathogen Biology Co-Investigator Peng Zhou Wuhan Institute of Virology Co-Investigator Ben Hu Wuhan Institute of Virology Co-Investigator Aleksei Chmura EcoHealth Alliance Other (Specify)-Research Scientist

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APPLICATION FOR FEDERAL ASSISTANCE SF 424 (R&R) 1. TYPE OF SUBMISSION* 3. DATE RECEIVED BY STATE 4.a. Federal Identifier Al110964 0MB Number: 4040-0001 EJ<pira1ion Date: 1013112019 I State Application Identifier 0 Pre-application • Application 0 Changed/Corrected b. Agency Routing Number Application 2. DATE SUBMITTED I Application Identifier c. Previous Grants.gov Tracking Number 5. APPLICANT INFORMATION Organizational DUNS*: 0770900660000 Legal Name·: ECOHEAL TH ALLIANCE, INC. Department: Division: Street1 •: ECOHEAL TH ALLIANCE, INC. Street2: 460W 34TH ST City•: NEW YORK County: State*: NY: New York Province: Country*: USA: UNITED STATES ZIP / Postal Code*: 100012320 Person to be contacted on matters involving this application Prefix: Dr. First Name·: Peter Middle Name: Last Name·: Daszak Suffix: Position/Tille: PD/Pl Street1 •: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES ZIP/ Postal Code•: 100012320 Phone Number·:I (b)(~ Fax Number: 2123804465 Email:! (b) (6)] 6. EMPLOYER IDENTIFICATION NUMBER (EIN) or (TIN)* 311726494 7. TYPE OF APPLICANT* M: Nonprofit with 501 C3 IRS Status (Other than Institution of Higher Education) Other (Specify): Small Business Organization Type 0 Women Owned 0 Socially and Economically Disadvantaged 8. TYPE OF APPLICATION' ONew 0 Resubmission • Renewal 0 Continuation 0 Revision Is this application being submitted to other agencies?* 9. NAME OF FEDERAL AGENCY' National Institutes of Health 11. DESCRIPTIVE TITLE OF APPLICANT'S PROJECT' Understanding the Risk of Bat Coronavirus Emergence 12. PROPOSED PROJECT Start Date· Ending Date· 06/01/2019 05/31/2024 Tracking Number: GRANT12743073 If Revision, mark appropriate box(es). 0 A. Increase Award 0 8. Decrease Award 0 C. Increase Duration 0 D. Decrease Duration O E. Other (specify) : OYes eNo What other Agencies? 10. CATALOG OF FEDERAL DOMESTIC ASSISTANCE NUMBER TITLE: 13. CONGRESSIONAL DISTRICTS OF APPLICANT NY-010 Page 1 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER SF 424 (R&R) APPLICATION FOR FEDERAL ASSISTANCE Page 2 14. PROJECT DIRECTOR/PRINCIPAL INVESTIGATOR CONTACT INFORMATION Prefix: Dr. First Name•: PETER Middle Name: Last Name*: DASZAK Suffix: Position/Title: President Organization Name*: ECOHEAL TH ALLIANCE, INC. Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State·: NY: New York Province: Country•: USA: UNITED STATES ZIP/ Postal Code*: 100012317 Phone Number*:( (b) (6)1 Fax Number: +12123804465 Email*:( (b)(~ 15. ESTIMATED PROJECT FUNDING a. Total Federal Funds Requested* b. Total Non-Federal Funds* c. Total Federal & Non-Federal Funds* d. Estimated Program Income* $3,586,760.00 $0.00 $3,586,760.00 $0.00 16.IS APPLICATION SUBJECT TO REVIEW BY STATE EXECUTIVE ORDER 12372 PROCESS?* a. YES O THIS PREAPPLICATION/APPLICATION WAS MADE AVAILABLE TO THE STATE EXECUTIVE ORDER 12372 PROCESS FOR REVIEW ON: DATE: b.NO e PROGRAM IS NOT COVERED BY E.O. 12372; OR 0 PROGRAM HAS NOT BEEN SELECTED BY STATE FOR REVIEW 17. By signing this application, I certify (1) to the statements contained in the list of certifications• and (2) that the statements herein are true, complete and accurate to the best of my knowledge. I also provide the required assurances * and agree to comply with any resulting terms if I accept an award. I am aware that any false, fictitious, or fraudulent statements or claims may subject me to criminal, civil, or administrative penalties. (U.S. Code, Title 18, Section 1001) e I agree• • The est of ~rtifications and assurances, or an fntemet site where you may obtain tNs list. is contained in the announcement or agency specific instructions. 18. SFLLL or OTHER EXPLANATORY DOCUMENTATION 19. AUTHORIZED REPRESENTATIVE Prefix: Dr. First Name•: Aleksei Middle Name: Position/Title*: Authorized Organizational Representative Organization Name*: EcoHealth Alliance, Inc. Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State•: Province: Country*: ZIP / Postal Code*: NY: New York USA: UNITED STATES 100012320 File Name: Phone Number·:1 ___ Cb_)_(6J_,I Fax Number: 2123804465 Signature of Authorized Representative' Aleksei Chmura 20. PRE-APPLICATION File Name: Last Name*: Chmura Suffix: Emai1•:1 _________ Cb_>_C6J_,I Date Signed* 11/05/2018 21. COVER LETTER ATTACHMENT File Name:NIAID_COV_2018_Cover_Letter_Final.pdf Tracking Number: GRANT12743073 Page 2 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 424 R&R and PHS-398 Specific Table Of Contents SF 424 R&R Cover Page .....................................•...................................................................... 1 Table of Contents •........................................................•.................•....................................... 3 Performance Sites ...................................................................•................................................. 4 Research & Related Other Project lnformation ...................................................................... 6 Project Summary/ Abstract(Description) .............................................................................. 7 Project Narrative .................•.........................•.......................................................................• 8 Facilities & Other Resources ................................................................................................ 9 Equipment. ............................................................................................................................ 12 Research & Related Senior/Key Person ................................................................................ 14 Research & Related Budget Year - 1 ...................................................................................... 81 Research & Related Budget Year - 2 ...................................................................................... 84 Research & Related Budget Year - 3 ...................................................................................... 87 Research & Related Budget Year - 4 ...................................................................................... 90 Research & Related Budget Year - 5 ...................................................................................... 93 Budget Justification ................................................................................................................ 96 Research & Related Cumulative Budget. ............................................................................ 100 Research & Related Budget - Consortium Budget (Subaward 1) ..................................... 101 Research & Related Budget - Consortium Budget (Subaward 2) ..................................... 119 Research & Related Budget - Consortium Budget (Subaward 3) ..................................... 137 Total Direct Costs Less Consortium F&A ........................................................................... 155 PHS398 Cover Page Supplement. ........................................................................................ 156 PHS 398 Research Plan ......................................................................................................... 158 Specific Aims ...................................................................................................................... 159 Research Strategy ...........................................•.................................................................. 160 Progress Report Publication List ..................................................................................... 172 PHS Human Subjects and Clinical Trials lnformation .......................................•............ 173 Study 1: Understanding the Risk of Bat Coronavirus Emergence: Community and clinic-based surveillance to capture SARSr-CoV spillover, routes of exposu ......... 175 Inclusion Enrollment Reports .................................................................................... 180 Vertebrate Animals .............•.....•...................•...................•.....•......•............•............•......... 187 Select Agent Research ...................................................................................................... 190 Bibliography & References Cited ..................................................................................... 193 Consortium/Contractual Arrangements ........................................................................... 197 Letters of Support .............................................................................................................. 198 Resource Sharing Plan(s) ................................................................................................. 202 Authentication of Key Biological and/or Chemical Resources ..................................... 203 Table of Contents Page 3

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Contact PD/Pl: DASZAK, PETER 0MB Number: 4040-0010 Expiration Oate: 10/31/2019 Project/Performance Site Location(s) Project/Performance Site Primary Location 0 I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: Duns Number: Street1 •: Street2: City*: County: State*: Province: Country*: Zip/ Postal Code*: ECOHEAL TH ALLIANCE, INC. 0770900660000 ECOHEAL TH ALLIANCE, INC. 460 W 34TH ST NEW YORK NY: New York USA: UNITED STATES 100012320 Project/Performance Site Congressional District*: NY-010 Project/Performance Site Location 1 0 I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: DUNS Number: Street1 *: Street2: City*: County: State*: Province: Country*: Zip/ Postal Code*: University of North Carolina at Chapel Hill 6081952770000 McGavran-Greenberg Hall Campus Box 7435 Chapel Hill NC: North Carolina USA: UNITED STATES 275997435 Project/Performance Site Congressional District*: NC-004 Page 4 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Project/Performance Site Location 2 Organization Name: DUNS Number: Street1 *: Street2: City*: County: State*: Province: Country•: Zip/ Postal Code*: Wuhan Institute of Virology 5290274740000 Xiao Hong SHan, No. 44 Wuchang District Wuhan CHN: CHINA 430071 Project/Performance Site Congressional District*: Project/Performance Site Location 3 0 I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. 00-000 0 I am submitting an application as an individual, and not on behalf of a company, state, local or tribal government, academia, or other type of organization. Organization Name: DUNS Number: Institute of Pathogen Biology 5281563570000 Street1 *: Street2: City*: County: State*: Province: Country•: Zip/ Postal Code*: Dong Dan San Tiao, No. 9 Dongcheng District Beijing CHN: CHINA 100730 Project/Performance Site Congressional District*: Additional Location(s) File Name: Tracking Number: GRANT12743073 00-000 Page 5 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED Other Project Information 1. Are Human Subjects Involved?* • Yes O No 1.a. If YES to Human Subjects Is the Project Exempt from Federal regulations? 0 Yes e No If YES, check appropriate exemption number: - 1 - 2 - 3 _4 - 5 - 6 If NO, is the IRB review Pending? • Yes O No IRB Approval Date: 03-15-2019 Human Subject Assurance Number None 2. Are Vertebrate Animals Used?* • Yes o No 2.a. If YES to Vertebrate Animals Is the IACUC review Pending? • Yes 0 No IACUC Approval Date: 03-15-2019 Animal Welfare Assurance Number None 3. Is proprietary/privileged information included in the application?* o Yes e No 4.a. Does this project have an actual or potential impact• positive or negative • on the environment?* 4.b. If yes, please explain: - 7 0MB Number. 4040-0001 Expiration Dale: 10/3112019 - 8 0 Yes e No 4.c. If this project has an actual or potential impact on the environment, has an exemption been authorized or an O Yes 0 No environmental assessment (EA) or environmental impact statement (EIS) been performed? 4.d. If yes, please explain: 5. Is the research performance site designated, or eligible to be designated, as a historic place?* o Yes e No 5.a. If yes, please explain: 6. Does this project involve activities outside the United States or partnership with international • Yes 0 No collaborators?* 6.a. If yes, identify countries: China 6.b. Optional Explanation: Filename 7. Project Summary/Abstract* NIAID_COV _2019_PROJECT _SUMMARY _final.pd! 8. Project Narrative* N IAI D _ COV _2019_NARRATIVE_Fina I. pdf 9. Bibliography & References Cited NIAID_COV_2019_REFERENCES.pdf 10.Facilities & Other Resources NIAID_COV _2019_FACILITIES_v01_PD.pdf 11.Equipment NIAID_COV _2019_EQUIPMENT _v01.pdf Page 6 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Project Summary: Understanding the Risk of Bat Coronavirus Emergence Novel zoonotic, bat-origin CoVs are a significant threat to global health and food security, as the cause of SARS in China in 2002, the ongoing outbreak of MERS, and of a newly emerged Swine Acute Diarrhea Syndrome in China. In a previous R01 we found that bats in southern China harbor an extraordinary diversity of SARSr-CoVs, some of which can use human ACE2 to enter cells, infect humanized mouse models causing SARS-like illness, and evade available therapies or vaccines. We found that people living close to bat habitats are the primary risk groups for spillover, that at one site diverse SARSr-CoVs exist that contain every genetic element of the SARS-CoV genome, and identified serological evidence of human exposure among people living nearby. These findings have led to 18 published peer-reviewed papers, including two papers in Nature. and a review in Ce//. Yet salient questions remain on the origin, diversity, capacity to cause illness, and risk of spillover of these viruses. In this R01 renewal we will address these issues through 3 specific aims: Aim 1. Characterize the diversity and distribution of high spillover-risk SARSr-CoVs in bats in southern China. We will use phylogeographic and viral discovery curve analyses to target additional bat sample collection and molecular CoV screening to fill in gaps in our previous sampling and fully characterize natural SARSr-CoV diversity in southern China. We will sequence receptor binding domains (spike proteins) to identify viruses with the highest potential for spillover which we will include in our experimental investigations (Aim 3). Aim 2. Community, and clinic-based syndromic, surveillance to capture SARSr-CoV spillover, routes of exposure and potential public health consequences. We will conduct biological-behavioral surveillance in high-risk populations, with known bat contact, in community and clinical settings to 1) identify risk factors for serological and PCR evidence of bat SARSr-CoVs; & 2) assess possible health effects of SARSr-CoVs infection in people. We will analyze bat-CoV serology against human-wildlife contact and exposure data to quantify risk factors and health impacts of SARSr-CoV spillover. Aim 3. In vitro and in vivo characterization of SARSr-CoV spillover risk, coupled with spatial and phylogenetic analyses to identify the regions and viruses of public health concern. We will use S protein sequence data, infectious clone technology, in vitro and in vivo infection experiments and analysis of receptor binding to test the hypothesis that % divergence thresholds in S protein sequences predict spillover potential. We will combine these data with bat host distribution, viral diversity and phylogeny, human survey of risk behaviors and illness, and serology to identify SARSr-CoV spillover risk hotspots across southern China. Together these data and analyses will be critical for the future development of public health interventions and enhanced surveillance to prevent the re-emergence of SARS or the emergence of a novel SARSr-CoV. Page 7 Project Summary/Abstract

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Renewal: Understanding the Risk of Bat Coronavirus Emergence Project Narrative Most emerging human viruses come from wildlife, and these represent a significant threat to public health and biosecurity in the US and globally, as was demonstrated by the SARS coronavirus pandemic of 2002-03. This project seeks to understand what factors allow coronaviruses, including close relatives to SARS, to evolve and jump into the human population by studying viral diversity in their animal reservoirs (bats), surveying people that live in high-risk communities in China for evidence of bat-coronavirus infection, and conducting laboratory experiments to analyze and predict which newly-discovered viruses pose the greatest threat to human health. Page 8 Project Narrative

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Facilities, Equipment, and Other Resources EcoHealth Alliance, New York, USA (Ors. Daszak, Olival, Francisco, Ross) EcoHealth Alliance is a New York-based 501 (c) 3 non-profit institution that conducts scientific research on emerging zoonoses and global health capacity building. EcoHealth Alliance New York headquarters has Cb> <4> square feet of office space including a meeting room and basic laboratory - freezer storage and light microscopy. The scientific staff (34 core scientists, 100+ field staff) is supported by a core ad min staff of 18 who are available for work on this project and funded through private donor and federal support. EcoHealth Alliance does not support diagnostic facilities at its core headquarters and works in partnership with a network of leading diagnostic labs both in the USA and around the world. EcoHealth Alliance is equipped with fiber optic Internet access and video conferencing facilities to facilitate easy communication between collaborators. EcoHealth Alliance employees have around-the-clock access to servers, VPNs, encryption software, IT support, and all necessary software including Git and Github (Hosted software revision/audit service), Sublime and Vim text editors, Vagrant and Oracle Virtualbox virtual machines, Google Apps (Hosted email and collaboration web based software), Ansible (Server provisioning software framework), Python, NodeJS, and R programming languages, Meteor (Javascript framework), Bash shell scripts, Jenkins (Continuous Integration server), Microsoft Office and Adobe CS6 running on both Apple Mac OS X, Ubuntu linux, and Windows Operating Systems. EcoHealth Alliance has a dedicated quad-core Linux server and another dedicated dual quad-core Mac Pro Server - each with 4TB hard drives. Either server individually or in combination may be used for intensive computational modeling and/or database processing by all the grantees. Access to the cloud and supercomputing services (Amazon) is provided by core funding to EcoHealth Alliance. EcoHealth Alliance is the headquarters of a global network of over 70 partners that provides exceptional leverage for the core scientists. This network includes staff from: academic institutions at leading national universities; intergovernmental agencies (WHO, OIE, FAO, DIVERSITAS, IUCN); infectious disease surveillance laboratories including BSL-3 and -4 laboratories; national government agency offices and labs; locally-based wildlife conservation organizations in Asia, Africa and Latin America. EcoHealth Alliance is the headquarters of: The Consortium for Conservation Medicine (CCM); the journal EcoHealth; an NSF Research Coordination Network (EcoHealthNET); the IUCN Wildlife Health Specialist Group; and the OIE Wildlife Health Network. EcoHealth Alliance is a voting member of the IUCN and a partner in Columbia University's Earth Institute Center for Environmental Sustainability (EICES) and all senior scientific staff members are Adjunct Faculty at Columbia University's Department of Ecology, Evolution, and Environmental Biology or at the Mailman School of Public Health. Institute of Pathogen Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China (Drs. Ren, Guo) The Institute of Pathogen Biology (IPB) is a key (flagship) institute within the Chinese Academy of Medical Sciences & Peking Union Medical College. IPB's mission is to conduct high quality research in basic and applied biology of critically important human pathogens. The ultimate goal is to conduct research and develop technology for better diagnosis, treatment, and prevention of infectious diseases. IPB specializes in multidisciplinary approaches to pathogen research and technological development focused on improving China's capability to diagnose, treat and prevent infectious diseases. Human Resources. The department involved in this project consists of 30 staff members: 10 with a clinical medicine background, 12 biological research scientists, 3 bioinformaticists, 3 animal technicians, and 2 biochemists working on protein expression and purification. Lab Facilities. The IPB includes the Ministry of Health Key Laboratory of Systems Biology of Pathogens, Christophe Merieux Laboratory, the AIDS Research Center, laboratories focused on Bacteriology, Virology, Mycology, Parasitology, and the Epidemiological Information Analysis Department. The institute has established platforms in metagenomics, transcriptome, morphology, molecular biology, and immunology. All of these are funded centrally and available to conduct the research proposed in the current R01. Page 9 Facilities & Other Resources

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BSL2 Facility. The institute has three laboratories of (bH 4 >equipped as BSL2 space for virology, immunology and clinical sample preparation. Equipment includes an lllumina Hiseq 2500, Miseq and BGI 500, gel electrophoresis, power supplies, thermal cyclers, a programmable heat block, heat blocks, water baths, CO2 incubators (2), several -70°C freezers, one -140°C freezer, refrigerators, DNA documentation system, DNA sequencing and computer assisted sequence analysis programs, several microfuges, Nikon and Zeiss microscopes with photographic and fluorescent capabilities, several class 2 environmental hoods, refrigerated water baths, real time thermocyclers, and spectrophotometers. The laboratory has an ELISA plate reader, an illuminometer, ELISA plate washer, spectrophotometers, and other equipment that is routinely used in characterizing antibody-protein interactions. (b) ( 4) BSL 3 Facility. The institute shares an additional ___ of BSL3 facilities equipped with sterile hoods (BSCIIA), CO2 incubators, -70C freezer, an inverted Nikon fluorescent microscope, and equipment for virus isolation and culture, and molecular genetics research. University of North Carolina at Chapel Hill, USA (Barie and Sims) The Department of Epidemiology is an internationally recognized leader in epidemiologic research and training. The department offers research training in most specialized areas including cancer, cardiovascular diseases, environmental and occupational health, health services/clinical epidemiology, reproductive health and infectious diseases. The department's current faculty consists of 51 regular full-time faculty and 151 adjunct faculty members. The department has 218 graduate students enrolled, including 20 in the MPH program, 5 in the MSPH program, 20 in the MSCR program and 173 in the Ph.D. program. The Department of Epidemiology is headquartered in the four-story McGavran-Greenberg Building. The epidemiology administrative and office space occupies (b) (4) square feet and provides additional classroom space. Most of the department's research staff occupies a research annex consisting of approximately (b)(4) square feet of contiguous rental space in a commercial office building. Dr. Barie has three laboratories of (bH 4> square feet each equipped as BL2 space for molecular biology, virology, immunology and recombinant DNA techniques, as laid out in the current R01 proposal. Equipment is available for gel electrophoresis, PCR, and BSL2 sample storage and handling facilities. It includes a DNA documentation system, DNA sequencing and computer assisted sequence analysis programs, several microfuges, a microscopy suite, 10+ IBM and Apple Pentium 11/111 computers with accompanying software, three thermocyclers, a fume hood, Nuclisens reader, hybridization oven, real time thermocyclers, three fluorescent inverted scopes with computer software (Olympus IX51 ), and a spectrophotometer. A Roche Light Cycler 48011 is available for real time measurements. The laboratory has an ELISA plate reader, an illuminometer, 200 cages for animal maintenance and breeding in Seal-Safe housing, Bio Rad low pressure chromatography system, ELISA plate washer, spectrophotometers, and other equipment that is routinely used in characterizing antibody-protein interactions. The Barie laboratory contains an additional (b)(4J square feet of newly renovated BSL3 facilities with enhanced features including shower in/shower out facility; dual anteroom access; Hepa filtered exhaust; redundant exhaust fans; card key access; an alarm system to Public Health/Campus Police; laboratory controlled combination lock; and Techniplast Sealsafe™ Hepa filtered animal housing for 300+ rodents. PAPR and tyvek suits are worn at all times in the BSL3 facility. The BL3 facilities are in an adjacent and attached building (b)( 4J or in (bH 4>, the latter space is directly adjacent to Dr. Baric's BSL2 laboratory resources. Each facility is equipped with sterile hoods (BSCIIA), four CO2 incubators, gel electrophoresis equipment, thermal cyclers and power supplies, and related equipment necessary for virus cultivation and molecular genetic research. The facilities each house a -70°C freezer, an inverted Nikon fluorescent microscope with a digital camera, an ELISA plate reader and illuminometer. Both facilities contain rodent-sized Seal-Safe systems (~192 cages) for maintaining animals in a Hepa-filtered Air in/out environment, exhausted into the BSL3 Hepa-filtered exhaust system. An 8 chamber Buxco plethysmography system that allows for repetitive, noninvasive measures of the number of breaths, tidal volume, airway responsiveness, enhanced pause, and respiratory gases from live control and infected mice in a contained system is housed in the main BSL3 laboratory in (b) <4> 2 Page 10 Facilities & Other Resources

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter The Department of Epidemiology provides cold-room, autoclave, centralized dishwashing and a darkroom with an automated developer. The campus has central facilities for DNA oligonucleotide synthesis, histopathology, DNA sequencing, EM, light and confocal microscopy, automated PCR genotyping and Taqman facilities, and Fluorescent activated cell sorter facilities (FAC). As a member of the Department of Microbiology and Immunology and UNC Cancer center, Dr. Barie and his team have access to these facilities at a discounted cost. The University provides a variety of core services including: sequencing and deep sequencing cores, genomics cores, oligonucleotide synthesis cores, hybridoma cores, transgenic cores, structural biology cores, etc. typical of any world class research institution. Campus wide core facilities are available for oligonucleotide synthesis, Sanger and 454 sequencing, RNAseq, pathology and histology services, and Flow Cytometry. Approximately, 40,000 cages are available for CC RIX production in the CbH4) on UNC Campus. Wuhan Institute of Virology, Chinese Academy of Sciences, Hubei, China (Shi 1 Zhou, and Hu) The Wuhan Institute of Virology (WIV), Chinese Academy of Sciences (GAS) is the only institute specializing in virology, viral pathology and virus technology among 19 other biological and biomedical research institutes in CAS. WIV is China's premier institute for virologic research. It consists of three research departments and one center: the Departments of Molecular Virology, of Bio-control, of Analytical Biochemistry and Biotechnology, and the Virus Resource and Bioinformation Center. It contains the Key Laboratory of Molecular Virology of CAS, the Joint-laboratory of Invertebrate Virology, an HIV Pre-screening Lab and the Hubei Engineering and Technology Research Center for Viral Diseases. The institute is further divided into 14 research groups, one of which (the Emerging Virus Laboratory) is headed by Dr. Zhengli Shi. The supporting system of the institute consists of an analytical equipment center, an experimental animal center, the editorial office of Virologica Sinica and a computer network center. The virus resource and bio-information center of China contains the largest virus bank in Asia, curating around 800 viral strains. The Wuhan Institute of Virology is a World Health Organization collaborating center. It also has partnerships, research collaborations and contracts with universities and research institutes in more than 30 counties and regions including a long-time (>15 years) partnership with EcoHealth Alliance. There are 14 professors, 36 associated professors, and 47 assistant professors conducting research on virology and five of these have been awarded honors in the "Hundred Talents Project". In 2013, the first BSL-4 lab in China was opened at this Institute in a bespoke facility which was designed with the assistance of the US CDC and L'lnstitut Pasteur of France. The WIV Emerging Virus Laboratory, headed by Dr. Shi, was set up to carry out exactly the sort of experimental activities on emerging viruses listed in the current R01 proposal. This lab possesses all necessary facilities for molecular biology and virology including a bank of -ao·c freezers, PCR machines, gel electrophoresis and imaging systems, biosafety cabinets, super-clean benches, and cell culture rooms. A Core Facility Center was established at WIV to provide technological services to faculty, students, and visiting researchers. Core Facility Center equipment includes: a transmission electron microscope, ultracentrifugation machines, small animal in vivo imaging systems, confocal laser scanning microscopes, flow cytometry, a realtime qPCR system, and a high-throughput sequencing and analyzing system. In addition, WIV owns a complete biosafety research platform, which consists of the first national BSL-4 laboratory in China, and a cluster of BSL-3 and BSL-2 labs. 3 Page 11 Facilities & Other Resources

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Equipment EcoHealth Alliance (Daszak, Francisco, Olival, Ross) EcoHealth Alliance is equipped with fiber optic Internet access and video conferencing facilities to facilitate easy communication between collaborators. EcoHealth Alliance employees have around the clock access to servers, VPNs, encryption software, IT support, and all necessary software including Git and Github (Hosted software revision/audit service), Sublime and Vim text editors, Vagrant and Oracle Virtualbox virtual machines, Google Apps (Hosted email and collaboration web based software), Ansible (Server provisioning software framework), Python, NodeJS, and R programming languages, Meteor (Javascript framework), Bash shell scripts, Jenkins (Continuous Integration seNer), Microsoft Office and Adobe CS6 running on both Apple Mac OS X, Ubuntu linux, and Windows Operating Systems. Additionally, EcoHealth Alliance has a dedicated quadcore Linux server and another dedicated dual quad-core Mac Pro Server - each with 4TB hard drives. Either server individually or in combination may be used for intensive computational modeling and/or database processing by all the grantees. Access to the cloud and supercomputing services (Amazon) is provided by core funding to EcoHealth Alliance. Institute of Pathogen Biology (Ren, Guo) The Institute of Pathogen Biology laboratories have equipment required for general microbiological, molecular, and biochemical work including microcentrifuges, agarose and polyacrylamide electrophoresis equipment, spectrophotometer, rocking and shaking platforms, bead-beater cell disruptor, and incubators (shaking and static). Major equipment relevant to this proposal which are available include: BSL2 Facility. The institute has three laboratories of CbH4 )equipped as BSL2 space for the virology, immunology and clinical samples pretreatment. Equipment includes lllumina Hiseq 2500, Miseq and BGI 500, gel electrophoresis equipment, power supplies, thermal cyclers, a programmable heat block, heat blocks, water baths, CO2 incubators (2), several -70°C freezers, one -140°C freezer, refrigerators, DNA documentation system, DNA sequencing and computer assisted sequence analysis programs, several microfuges, Nikon and Zeiss microscopes with photographic and fluorescent capabilities, several class 2 environmental hoods, refrigerated water baths, real time thermocyclers, and spectrophotometer. The laboratory has an ELISA plate reader, an illuminometer, ELISA plate washer, spectrophotometers, and other equipment that is routinely used in characterizing antibody-protein interactions. BSL 3 Facility. The institute shares an additional CbH4 )of BSL3 facilities equipped with sterile hoods (BSCIIA), CO2 incubators, -7o·c freezer, an inverted Nikon fluorescent microscope with an assortment of filters, magnifications and digital camera, and related equipment necessary for virus cultivation and molecular genetic research. Wuhan Institute of Virology (Shi, Zhou, Hu) Institute of Virology's Emerging Virus Laboratory has equipment required for general microbiological, molecular, and biochemical work including microcentrifuges, agarose and polyacrylamide electrophoresis equipment, spectrophotometer, rocking and shaking platforms, bead-beater cell disruptor, and incubators (shaking and static). Major equipment relevant to this proposal which are available include: -so·c freezers, PCR machines, gel electrophoresis and imaging system, biosafety cabinets, super-clean benches, and cell culture rooms. A Core Facility Center was established at Wuhan Institute of Virology to provide technological services to faculty, students, and visiting researchers. The equipment installed in the Core Facility Center include: transmission electron microscope, ultracentrifugation machines, small animal in vivo imaging systems, confocal laser scanning microscopes, flow cytometry, a real-time qPCR system, and a high-throughput sequencing and analyzing system. In addition, the Wuhan Institute of Virology owns a complete biosafety research platform, which consists of the first national BSL-4 laboratory in China, and a cluster of BSL-3 and BSL-2 labs. These labs contain gel electrophoresis equipment, power supplies, thermal cyders, programmable heat blocks, heat blocks, water Page 12 Equipment

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter baths, CO2 incubators, -70°C freezers, -140°C freezers, refrigerators, DNA documentation system, DNA sequencing and computer assisted sequence analysis programs, microfuges, Nikon and Zeiss microscopes with photographic and fluorescent capabilities, several class 2 environmental hoods, refrigerated water baths, real time thermocyclers, and spectrophotometers. The laboratory also has an ELISA plate reader, an illuminometer, ELISA plate washer, spectrophotometers, and other equipment that is routinely used in characterizing antibody-protein interactions. University of North Carolina at Chapel Hill Barie Laboratory (Barie, Sims) The three laboratories of the Barie Lab in the Department of Epidemiology have equipment required for general microbiological, molecular, and biochemical work including microcentrifuges, agarose and polyacrylamide electrophoresis equipment, spectrophotometer, rocking and shaking platforms, bead-beater cell disruptor, and incubators (shaking and static). Major equipment relevant to this proposal which are available include: gel electrophoresis equipment, power supplies, thermal cyclers, a programmable heat block, heat blocks, water baths, CO2 incubators {2), several -70°C freezers, one -140°C freezer, refrigerators, DNA documentation system, DNA sequencing and computer assisted sequence analysis programs, several microfuges, two Nikon microscopes with photographic and fluoresr.ent r..apabilities. several class 2 environmental hoods. refrigerated water baths, 10+ IBM and Apple Pentium 11/111 computers with accompanying software, three thermocyclers, a fume hood, Nuclisens reader, hybridization oven, real time thermocyclers, three fluorescent inverted scopes with computer software (Olympus IX51 ), and a spectrophotometer. A Roche Light Cycler 48011 is available for real time measurements. The laboratory has an ELISA plate reader, an illuminometer, 200 cages for animal maintenance and breeding in Seal-Safe housing, Bio Rad low pressure chromatography system, ELISA plate washer, spectrophotometers, and other equipment that is routinely used in characterizing antibody-protein interactions. BSL3 Facility features include: shower in/shower out facility; dual anteroom access; Hepa filtered exhaust; redundant exhaust fans; card key access; an alarm system to Public Health/Campus Police; laboratory controlled combination lock; and Techniplast Sealsafe™ Hepa filtered animal housing for 300+ rodents. PAPR and tyvek suits are worn at all times in the BSL3 facility. The BL3 facilities are in an adjacent and attached building (b)(4 or in (b)(4l, the latter space is directly adjacent to Dr. Baric's BSL2 laboratory resources. Each facility is equipped with sterile hoods (BSCI IA), four CO2 incubators, gel electrophoresis equipment, thermal cyclers and power supplies, and related equipment necessary for virus cultivation and molecular genetic research. The facilities each house a -70°C freezer, an inverted Nikon fluorescent microscope with an assortment of filters, magnifications and digital camera, an ELISA plate reader and illuminometer. Both facilities contain rodent-sized Seal-Safe systems (~192 cages) for maintaining animals in a Hepa-filtered Air in/out environment, exhausted into the BSL3 Hepafiltered exhaust system. An 8 chamber Buxco plethysmography system that allows for repetitive, noninvasive measures of the number of breaths, tidal volume, airway responsiveness, enhanced pause, and respiratory gases from live control and infected mice in a contained system is housed in the main BSL3 laboratory in (b) (4) 2 Page 13 Equipment

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED Senior/Key Person Profile (Expanded) PROFILE . Project Director/Principal Investigator Prefix: Dr. First Name*: PETER Middle Name Last Name•: DASZAK Position/Title*: President Organization Name*: ECOHEALTH ALLIANCE, INC. Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State•: NY: New York Province: Country•: USA: UNITED STATES Zip / Postal Code*: 100012317 Phone Number*:( (b) (6)] Fax Number: +12123804465 E-Mail*:( (b) (6)1 Credential, e.g., agency login:! (b)(6)1 Project Role*: PD/Pl Other Project Role Category: Degree Type: PHO Degree Year: 1993 Attach Biographical Sketch*: File Name: DASZAK_Peter _Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 14 0MB Number: 4040-0001 Expiration Dato: 10/31/2019 Suffix: Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018·11·05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Zheng Li Middle Name Last Name•: Shi Suffix: Position/Title•: Senior Scientist Organization Name*: Wuhan Institute of Virology Department: Division: Street1 *: Xiao Hong Shan, no. 44 Street2: City*: Wuhan County: State*: Province: Country*: CHN: CHINA Zip/ Postal Code•: 430071 Phone Number*:I (b) (6)] Fax Number: E-Mail*:r (b) (6)] Credential. e.g .. agency login:! (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2000 Attach Biographical Sketch*: File Name: SH I_Zhengli_Biosketch_Final .pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Kevin Middle Name J. Last Name*: Oliva! Suffix: Position/Title*: Senior Research Scientist Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip / Postal Code': 100012317 Phone Number*:I (b) (6)] Fax Number: E-Mail*:I (b) (6)] Credential, e.g., agency login:ICb> <6?] Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2008 Attach Biographical Sketch*: File Name: OLIVAL_Kevin_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 15 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Ralph Middle Name S Last Name•: Barie Suffix: Position/Title•: Professor Organization Name*: University of North Carolina Department: Division: Street1 *: UNIVERSITY OF NORTH CAROLINA Street2: DEPT EPIDEMIOLOGY City*: CHAPEL HILL County: State*: NC: North Carolina Province: Country*: USA: UNITED STATES Zip/ Postal Code•: 275997435 Phone Number*J (b)(6jJ Fax Number: +19199662089 E-Mail*:I (b) (6)] Credential. e.g .. agency login: I (b) (6)] Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 1977 Attach Biographical Sketch*: File Name: SARIC _Ralph_Biosketch_Final .pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Noam Middle Name Last Name*: Ross Suffix: Position/Title*: Disease Ecologist Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip / Postal Code': 100012317 Phone Number*:! (b) (6)] FaxNumbe~+12123804465 E-Mail*:I (b) (6)] Credential, e.g., agency login: noamross Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2015 Attach Biographical Sketch*: File Name: ROSS_Noam_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 16 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018·11·05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Alice Middle Name Last Name•: Latinne Suffix: Position/Title•: Research Scientist Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip/ Postal Code•: 100012317 Phone Number*J (b)(6)l Fax Number: +12123804465 E-Mail*:( (b)(6) Credential. e.g .. agency login:( (b)(~ Project Role*: Other (Specify) Other Project Role Category: Research Scientist Degree Type: PHO Degree Year: 2012 Attach Biographical Sketch*: File Name: LATI NNE_Alice_Biosketch_Final.pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Ms. First Name*: HongYing Middle Name Last Name*: Li Suffix: Position/Title*: Research Scientist & China Programs Coord. Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip / Postal Code': 100012317 Phone Number*J (b) (6)] FaxNumbe~+12123804465 E-Mail*:I (b) (6)] Credential, e.g., agency login: I (b) (6)] Project Role*: Other (Specify) Other Project Role Category: Research Scientist Degree Type: MPH Degree Year: 2015 Attach Biographical Sketch*: File Name: LI_Hongying_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 17 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Leilani Middle Name Last Name•: Francisco Suffix: Position/Title•: Senior Behavioral Risk Surveillance Coord. Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip/ Postal Code•: 100012317 Phone Number*:I (b)(6jJ Fax Number: +12123804465 E-Mail*:I (b) (6)] Credential. e.g .. agency login: r (b) (6)] Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2010 Attach Biographical Sketch*: File Name: FRANCESCO _Leilani_Biosketch_Final .pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Amy Middle Name C Last Name*: Sims Suffix: Position/Title*: Associate Professor Organization Name*: University of North Carolina at Chapel Hill Department: Division: Street1 *: University of North Carolina Street2: 3304 MHRC, School of Public Health City*: Chapel Hill County: State*: NC: North Carolina Province: Country*: USA: UNITED STATES Zip / Postal Code': 275997290 Phone Number*:[ (b)(6)j Fax Number: (919) 966-0584 E-Mail*:I (b)(6)] Credential, e.g., agency login:I (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2001 Attach Biographical Sketch*: File Name: SIMS_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 18 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Ms. First Name*: Emily Middle Name E Last Name•: Hagan Suffix: Position/Title•: Behavioral Research Scientist Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip/ Postal Code•: 100012317 Phone Number*:! (b)(~ Fax Number: +12123804465 E-Mail*:( (b) (6)] Credential. e.g .. agency login: r (b)(6)j Project Role*: Other (Specify) Other Project Role Category: Research Scientist Degree Type: MPH Degree Year: 2013 Attach Biographical Sketch*: File Name: HAGAN_Emily _Biosketch_Final. pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Guangjian Middle Name Last Name*: Zhu Suffix: Position/Title*: Research Scientist & China Field Coordinator Organization Name*: East China Normal University Department: Division: Street1 *: School of Life Science, B327 Street2: Science building, 3663 Zhongshan Beilu City*: Shanghai County: State*: Province: Country*: CHN: CHINA Zip / Postal Code': 200062 Phone Number*:r (b)(6)j Fax Number: E-Mail*:! (b)(6)] Credential, e.g., agency login: r (b) (6)] Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2012 Attach Biographical Sketch*: File Name: ZHU_ GuangJian_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 19 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Linfa Middle Name Last Name•: Wang Suffix: Position/Title•: Professor & Director Organization Name*: Duke-NUS Medical School Department: Division: Street1 *: 8 College Road Street2: City*: Singapore County: State*: Province: Country*: SGP: SINGAPORE Zip/ Postal Code•: 169857 Phone Number*:I (b) (6)] Fax Number: E-Mail*:I (b)(6)j Credential. e.g .. agency login: r (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 1986 Attach Biographical Sketch*: File Name: WANG_Linfa_Final.pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Lili Middle Name Last Name*: Ren Suffix: Position/Title*: Research Scientist Organization Name*: Institute of Pathogen Biology Department: Division: Street1 *: No. 9 Dong Dan San Tiao Street2: Dongcheng District City*: Beijing County: State*: Province: Country*: CHN: CHINA Zip / Postal Code': 100730 Phone Number*:[ (b)(6)j Fax Number: E-Mail*:I (b) (6)] Credential, e.g., agency login: [ (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2005 Attach Biographical Sketch*: File Name: REN_Lili_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 20 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Li Middle Name Last Name•: Guo Suffix: Position/Title•: Professor Organization Name*: Institute of Pathogen Biology Department: Division: Street1 *: No. 9 Dong Dan San Tiao Street2: Dongcheng District City*: Beijing County: State*: Province: Country*: CHN: CHINA Zip/ Postal Code•: 100730 Phone Number*:I (b) (6)] Fax Number: E-Mail*:r (b) (6)] Credential. e.g .. agency login: r (b)(~ Project Role*: Co-Investigator Other Project Role Category: Degree Type: MD Degree Year: 2006 Attach Biographical Sketch*: File Name: GUO_Li_Biosketch_Final.pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Peng Middle Name Last Name*: Zhou Suffix: Position/Title*: Principal Investigator Organization Name*: Wuhan Institute of Virology Department: Division: Street1 *: Xiao Hong Shan, No. 44 Street2: City*: Wuhan County: State*: Province: Country*: CHN: CHINA Zip / Postal Code': 430071 Phone Number*:[ (b)(~ Fax Number: E-Mail*:I (b) (6)] Credential, e.g., agency login: [ (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2011 Attach Biographical Sketch*: File Name: ZHOU_Peng_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 21 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11·05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER PROFILE - Senior/Key Person Prefix: Dr. First Name*: Ben Middle Name Last Name•: Hu Suffix: Position/Title•: Research Scientist Organization Name*: Wuhan Institute of Virology Department: Division: Street1 *: Xiao Hong Shan, No. 44 Street2: City*: Wuhan County: State*: Province: Country*: CHN: CHINA Zip/ Postal Code•: 430071 Phone Number*:I (b)(~ Fax Number: E-Mail*:I (b)(~ Credential. e.g .. agency login: I (b)(6)j Project Role*: Co-Investigator Other Project Role Category: Degree Type: PHO Degree Year: 2015 Attach Biographical Sketch*: File Name: HU_Ben_Biosketch_final.pdf Attach Current & Pending Support: File Name: PROFILE - Senior/Key Person Prefix: Dr. First Name*: Aleksei Middle Name Last Name*: Chmura Suffix: Position/Title*: Research Scientist Organization Name*: EcoHealth Alliance Department: Division: Street1 *: 460 West 34th Street Street2: Suite 1701 City*: New York County: State*: NY: New York Province: Country*: USA: UNITED STATES Zip / Postal Code': 100012317 Phone Number*:I (b)(6j] FaxNumbe~+12123804465 E-Mail*:I (b) (6)J Credential, e.g., agency login: I (b) (6)) Project Role*: Other (Specify) Other Project Role Category: Research Scientist Degree Type: PHO Degree Year: 2018 Attach Biographical Sketch*: File Name: CHMURA_Aleksei_Biosketch_Final.pdf Attach Current & Pending Support: File Name: Page 22 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:3122.000-05:00

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator (Last, Rrst, Middle): Daszak, P. NAME: Peter Daszak BIOGRAPHICAL SKETCH DO NOT EXCEED FIVE PAGES. eRA COMMONS USER NAME (credential, e.g., agency login): (b)(6J POSITION TITLE: President & Chief Scien1ist EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) DEGREE Completion INSTITUTION AND LOCATION (if Date FIELD OF STUDY applicable) MMNYYY Bangor University, UK B.S (hons) 07/1986 Zoology University of East London, UK Ph.D. 03/1993 Infectious Diseases A. Personal Statement I have the broad expertise in emerging viral zoonoses, and scientific management experience to support this proposed work that involves an international interdisciplinary team working on field collection of wildlife and human samples, human behavioral risk surveys, modeling and analytics, and viral characterization in vitro and in vivo. I am President and Chief Scientist of EcoHealth Alliance, a US-based 501 (c) 3 institution that conducts research on emerging zoonoses and global health capacity building. My 20+ years of NIH-funded research focuses on understanding the links among disease emergence in wildlife, livestock and people, particularly viral zoonoses. This includes identifying the bat origin of SARS-CoV and SADS-CoV, analyzing the ecology of West Nile, Nipah and Hendra virus emergence, publishing the first unbiased analysis of global emerging disease hotspots, and developing the scientific rationale for the Global Virome Project (GVP). Over the past 18 years I have been the Pl on 4 multidisciplinary R01s that use modeling, epidemiology, laboratory and field science to test hypotheses on the emergence of wildlife-origin viral zoonoses, including SARS-CoV, Nipah and Hendra virus, Avian influenza and novel viruses from bats. I have successfully managed teams of virologists, field biologists, mathematical modelers, veterinarians, epidemiologists, laboratorians and anthropologists. Much of the groundwork for the current proposal has developed from my previous collaborative research with each member of the team assembled in the current R01 renewal proposal. 1. Li W, Shi Z, Yu M, Ren W, Smith C, Epstein JH, Wang H, Crameri G, Hu Z, Zhang H, Zhang J, McEachern J, Field H, Daszak P, Eaton BT, Zhang S & Wang L-F (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science 310: 676-679. 2. Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, and Daszak P* (2008). Global trends in emerging infectious diseases. Nature 451 :990-993 3. Olival KJ*, Hosseini PR, Zambrana-Torrelio C, Ross N, Bogich TL, 0aszak P* (2017). Host and viral traits predict zoonotic spillover from mammals. Nature 546, 646-650. 4. Carroll D, Daszak P*, Wolfe ND, Gao GF, Morel C, Morzaria S, Pablos-Mendez A, Tomori 0, Mazet JAK (2018). The global virome project. Science 359: 872-87 4. PHS 398/2590 (Rev. 06/09) Pagel Biographical Sketch Page 23 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator (Last, First, Middle): Daszak, P. B. Positions and Honors Positions and Employment 1993 -98 Senior Faculty Research Scientist, Kingston University UK 1998 Guest Researcher, Centers for Disease Control and Prevention (CDC) 1999 -01 Faculty Research Scientist, University of Georgia 2001 - Sr. Adjunct Faculty, Columbia University 2001 - 09 Executive Director, Consortium for Conservation Medicine, EcoHealth Alliance, New York 2009 - President & Chief Scientist, EcoHealth Alliance New York Other Experience and Professional Membership 2003 - 7 NIH: ad hoc member, ZRG1 IDM-G 90 (2003-5) ZRG1 IRAP-Q (2005-7) 2004 - Editorial Board, Conserv. Biol. 2005 NIAID: Steering Committee, workshop on virus-host shifts & emergence of new pathogens 2010 - Editor-in-Chief, EcoHealth; Member of IOM Forum on Microbial Threats; External Advisory Board, OHS and Kansas State Univ. Ctr. of Excellence for Emerg. & Zoonotic Animal Diseases (CEEZAD) 2011 Steering Committeee, NIAID Workshop on Arboviruses 2014 - Member NRG Advisory Committee to advise the US Global Change Research Program (USGCRP) 2015 - Member of Supervisory Board, One Health Platform; Editorial Board One Health 2016 - Member, WHO Expert group on Public Health Emergency Disease Prioritization 2016- Member, Core Steering Committee & Co-Chair, Science & Technol WG, Global Virome Project 2017 External Review Committee, CSIRO Health & Biosecurity Business Unit 2017 - Chair, Forum on Microbial Threats, National Academies of Science, Engineering & Medicine Honors 1999 2000 2002 2003 2007 2008 2012 2013 2013 2015 2018 - Meritorious service award, CDC CSIRO silver medal for collaborative research Honored by the naming of a new species of centipede, Cryptops daszaki (J Nat Hist 36: 76-106) 6th Annual Lecturer, Medicine & Humanities, Texas A&M Finalist, Director's Pioneer Award Presidential Lecturer, University of Montana Elected member of the Cosmos Club, Washington DC Honored by the naming of a new parasite species, lsospora daszaki (Parasit. Res. 111 :1463-1466) Hsu-Li Distinguished Lectureship in International Epidemiology, Univ. Iowa Robert Leader Endowed Lecture in Food Safety, Michigan State Univ. Member, National Institute of Medicine (NAM), USA. C. Contribution to Science 1. Research on the bat origins of emerging viruses. A range high impact emerging viruses appear to have bat reservoirs (e.g. SARS-CoV, EBOV, NiV, HeV, MERS-CoV, SADS-CoV). As Pl on four prior R01 s, my work has helped demonstrate the bat-origin for some of these (SARS-CoV, SADS-CoV), analyze the drivers of emergence and risk factors for spillover. Collaborating with virologists in China, we have isolated and characterized SARS-like Co Vs from bats that use the same human host cell receptor (ACE-2) as SARSCoV. This work provides critical reagents and resources that have helped advance understanding of virushost binding and may contribute to vaccine development. My other work identified factors underlying the emergence of NiV from Pteropus bats in Malaysia and Bangladesh; that MERS-CoV likely originated in bats; that SADS-CoV originates in bats; and that bats harbor a significantly higher proportion of zoonoses than all other mammalian groups after correcting for reporting biases. PHS 398/2590 (Rev. 06/09) Pageg Biographical Sketch Page 24 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator (Last, First, Middle): Daszak, P. a. Pulliam JRC, Epstein JH, Dushoff J, Rahman SA, Bunning M, HERG, Jamaluddin AA, Hyatt AD, Field HE, Dobson AP & Daszak P* and the Henipavirus Ecology Research Group (HERG). {2012). Agricultural intensification, priming for persistence, and the emergence of Nipah virus: a lethal batborne zoonosis. J Roy Soc Interface 9:89-101 b. Ge X-Y, Li J-L, Yang X-L, Chmura AA, Zhu G, Epstein JH, Mazet JK, Hu B, Zhang W, Peng C, Zhang Y-J, Luo C-M, Tan B, Wang N, Zhu Y, Crameri G, Zhang S-Y, Wang L-F, Daszak P*, Shi ZL* (2013). Isolation and characterization of a bat SARS-like Coronavirus that uses the ACE2 receptor. Nature 503: 535-538. c. Memish ZA, Mishra N, Olival KJ, Fagbo SF, Kapoor V, Epstein JH, Al Hakeem R, Durosinloun A, Al Asmari M, Islam A, Kapoor A, Briese T, Daszak P, Al Rabeeah A, Lipkin WI. (2013). Middle East respiratory syndrome coronavirus in bats, Saudi Arabia. EID 19(11 ): 1819-1823. d. Zhou P, Fan H, Lan T, Yang X-L, Shi W-F, Zhang W, Zhu Y, Zhang Y-W, Xie Q-M, Mani S, Zheng X-S, Li B, Li J-M, Guo H, Pei G-O, An X-P, Chen J-W, Zhou L, Mai K-J, Wu Z-X, Li D, Anderson DE, Zhang L-B, Li S-Y, Mi Z-O, He T-T, Cong F, Fuo P-J, Huang R, Luo Y, Liu X-L, Chen J, Huang Y, Sun Q, Zhang X-L-L, Wang Y-Y, Xing S-Z, Chen Y-S, Sun Y, Li J, Daszak P*, Wang L-F*, Shi ZL*, Tong Y-G*, Ma J-Y* (2018). Fatal Swine Acute Diarrhea Syndrome caused by an HKU2-related Coronavirus of Bat Origin. Nature 556: 255-258. 2. Analyzing the process of disease emergence. Emerging infectious diseases are a significant threat to global health. However, their emergence is sporadic, complex, and seemingly unpredictable. In the early 2000s I started to use analytical approaches to see if there are patterns in disease emergence, and if these are predictable. By collating a database of all known prior EID events, identifying their point origins, and correcting for reporting biases, I published the first ever predictive 'hotspots' maps of where disease emergence is most likely. Under various grants that I have led, or been a co-investigator on, I have published spatial analyses of the drivers of disease spread, and strategies to predict pandemic emergence. a. Kilpatrick AM, Chmura AA, Gibbons DW, Fleischer RC, Marra PP & Daszak P {2006). Predicting the global spread of H5N1 avian influenza. PNAS 103: 19368-19373. b. Morse SS, Mazet JAK, Woolhouse M, Parrish CR, Carroll D, Karesh WB, Zambrana-Torrelio C, Lipkin WI, Daszak P* (2012). Prediction and prevention of the next pandemic zoonosis. Lancet 380:1956-1965. c. Daszak P*, Zambrana-Torellio C, Bogich TL, Fernandez M, Epstein JH, Murray KA, Hamilton H (2013). Interdisciplinary approaches to understanding disease emergence: The past, present and future drivers of Nipah virus emergence. PNAS 110: 3681-3688 d. Allen T, Murray KA, Zambrana-Torrelio C, Morse SS, Rondinini C, Di Marco M, Breit N, Olival KJ, Daszak P* (2017). Global hotspots and correlates of emerging zoonotic diseases. Nature Comm 8: 1124 3. Studies of wildlife disease ecology to understand emerging zoonoses. The majority of EIDs are zoonotic, with the majority of these originating in wildlife. In the 1990s, new collaborations among ecologists and medical researchers began to show that understanding disease dynamics in wildlife can allow better forecasting of disease risk in people. I reviewed this field in a paper in Science in 2000 and in a more recent paper in Nature on the links among biodiversity and health. During the last two decades, I have led collaborative research programs on how the ecology of specific wildlife-origin zoonoses can help explain patterns of risk to people. This includes my work in 4 R01 s and as EHA institutional lead for USAID-EPT-PREDICT, and Chief of Party for USAID-IDEEAL. This work has led to strategies to estimate the diversity of yet-to-be discovered viruses, and a program to identify them (the Global Virome Project). PHS 398/2590 (Rev. 06/09) Page~ Biographical Sketch Page 25 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator (Last, First, Middle): Daszak, P. a. Daszak P*, Cunningham AA, Hyatt AD (2000). Emerging infectious diseases of wildlife - threats to biodiversity and human health. Science 287: 443-449 b. Keesing F, Belden LK, Daszak P, Dobson A, Harvell CD, Holt RD, Hudson P, Jolles A, Jones KE, Mitchell CE, Myers SS, Bogich T & Ostfeld RS. (2010). Impacts of biodiversity on the emergence and transmission of infectious diseases. Nature 468:647-652. c. Anthony SJ, Epstein JH, Murray KA, Navarrete-Macias I, Zambrana-Torrelio CM, Solovyov A, Ojeda-Flores R, Arrigo NC, Islam A, Ali Khan S, Hosseini P, Bogich TL, Olival KJ, Sanchez-Leon MD, Karesh W, Goldstein T, Luby SP, Morse SS, Mazet JAK, Daszak P, Lipkin WI. (2013). A strategy to estimate unknown viral diversity in mammals. MBio 4(5): e00598-13. D. Additional Information: Research Support and/or Scholastic Performance Ongoing Research Support USAID Emerging Pandemic Threats PREDICT-2 Mazet (Pl) 10/01 /14 - 09/30/19 The goal of this work is to conduct surveillance for novel pathogens in wildlife, livestock and people; characterize human risk behavior; analyze EID risk; and design interventions in >20 countries Role: Pl on Subcontract 1R01 Al110964 Daszak (Pl) 06/01/14-05/31/19 Understanding the Risk of Bat Coronavirus Emergence The goal of this work is to conduct ecological and virological studies on bats in China that harbor SARS-like coronaviruses, and conduct behavioral risk surveys and testing in people, with a goal of identifying risk factors for further spillover of SARS-like CoVs, and help identify the likely drivers of the SARS-CoV outbreak in 2003. Role: Pl USAID 1414374 (ROMA, Thailand) Daszak (CoP) 10/01/13- 03/30/19 Infectious Disease Emergence and Economics of Altered Landscapes (IDEEAL) The goal of this cooperative agreement is to analyze how land use change affects disease risk in SE Asia, and how economic costs of disease can be used to develop novel intervention policies. Role: Chief of Party Completed Research Support NSF DEB 1414374 Perrings (Pl) 10/15/14 - 04/14/18 US-UK Collab: Risks of Animal and Plant Infectious Diseases through Trade (RAPID Trade) The goal of this NSF-NIH-USDA EEID award, joint with a UK BBSRC grant is to analyze and model how policy changes to trade affect emerging disease risk globally Role: Co-Investigator HDTRA 1 Allen (Pl) 04/15/15 - 04/14/17 Global Rapid Identification of undiagnosed EID Events The goal of this project was to design software that can be used in the DoD biosurveillance ecosystem (BSVE) to rapidly diagnose novel EID events. Role: Co-Investigator 1 R01 GM100471 (NIGMS) Perrings (Pl) 09/15/11-06/30/15 MASpread: Modeling Anthropogenic Effects in the Spread of Infectious Disease The goal of this project was to develop novel approaches to modeling and analyzing disease spread and the social decisions involved in control Role: Co-Investigator NSF Daszak (Pl) 07/01/10-06/30/15 PHS 398/2590 (Rev. 06/09) Page 1, Biographical Sketch Page 26 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator (Last, First, Middle): Daszak, P. EcoHealthNet - a Research Coordination Network Funding for student exchange and workshops to fuse veterinary science, ecology and human medical sciences Role: Pl USAID Emerging Pandemic Threats PREDICT-1 Mazet (Pl) 10/01/09 - 09/30/14 The goal of this work was to conduct surveillance for novel pathogens in wildlife, livestock and people in developing countries Role: Pl on Subcontract 2 R01TW005869 Daszak (Pl) 09/01/08 - 08/31/13 The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh This project involved mathematical modeling and fieldwork on the dynamics of Nipah virus in Bangladesh Role: Pl NSF DEB-1257513 Daszak (Pl) 08/15/12-07/31/13 US-China Ecology and Evolution of Infectious Diseases Collaborative Workshop; Kunming, China The goal of this work was to organize a workshop among NIH, NSF, leading US and Chinese scientists to discuss potential for a jointly funded NIH-NSF-China funding mechanism Role: Pl 1 R01 AI079231 (NIAID) Daszak (Pl) 09/18/08 - 08/31 /13 Risk of viral emergence from bats. The goal was to model hotspots for bat viral diversity, identify & characterize new bat viruses & understand their pathology Role: Pl NSF BCS 0826779 Daszak (Pl) 10/01/08- 03/31/12 AOC - HSD - Collaborative Research: Human-related factors affecting emerging infectious diseases The goal of this work was to analyze how socio-economic and environmental drivers predict risk of EIDs Role: Pl on lead proposal R01 TW005869 - supplemental Daszak (Pl) 09/01/08 - 08/31 /11 Supplemental funding: Predicting the risk of global H5N1 spread This project involved mathematical modeling and fieldwork in Bangladesh and China to understand risk of H5N1 spread. Role: Pl NSF EF-062239 Kilpatrick (Pl) 09/01 /06 - 08/30/11 Predicting spatial variation in West Nile virus transmission The goal was to study interaction among WNV vector, reservoir host populations across an urban-to-rural gradient. Role: Co-Pl R01 TW05869 (Fogarty Intl. Ctr.) Daszak (Pl) 08/01/02 - 05/31/07 Anthropogenic change & emerging zoonotic paramyxoviruses The goal was to identify the cause of emergence of Nipah and Hendra viruses in Malaysia and Australia. Role: Pl NSF HSD 0525216 Daszak (Pl) 10/15/05 - 10/14/06 Collaborative Research: Socio-Economic and Environmental Drivers of Emerging Diseases The goal of this work was to analyze patterns of disease emergence globally leading to development of a global hotspots map of disease emergence. Role: Pl PHS 398/2590 (Rev. 06/09) Page~ Biographical Sketch Page 27 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Zhengli Shi eRA COMMONS USER NAME (credential, e.g., agency login) (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Department of Biology, Wuhan University, China Wuhan Inst. Virol., Chinese Acad. Sci., China University Montpellier II, Montpellier, France A. Personal Statement POSITION TITLE Co-Investigator DEGREE BS MS Ph.D. MM/YY FIELD OF STUDY 1987 Genetics 1990 Virology 2000 Virology I have been working on the discovery and characterization of novel viruses from bats and other wildlife since 2004. This included the discovery that Chinese horseshoe bats are the natural reservoir of SARSr-CoVs and the likely origin of SARS-CoV. My group then isolated SARSr-CoVs from bats sharing high homology with human SARS-CoV and demonstrated their interspecies transmission risk, largely confirming bats as the source of SARs. My lab has carried out systematic studies on the epidemiology, genetic evolution, interspecies infection mechanism and pathogenesis of a series of bat-borne emerging viruses including SARSr-CoV, MERS-CoV, EBOV and others. This work has involved collaboration on all other scientists on this R01 renewal proposal, in particular Drs. Daszak and Linfa Wang, who I have collaborated with since 2003, publishing 2 papers in Nature and one in Science together on our bat-virus work, as well as dozens of others. Recently, this collaborative team discovered that an outbreak of fatal Swine Acute Diarrhea Syndrome in southern China that killed more than 24,000 piglets was caused by spillover of bat HKU2-related coronaviruses. In this proposed work, my group will be responsible for CoV testing in bat samples, serological testing in human samples, and virus characterization work such as cell entry analysis and receptor identification. B. Positions and Honors. Positions and Employment 1990-93 1993- 95 2000- Research assistant, Wuhan Institute of Virology, Chinese Academy of Sciences, China Research scientist, Wuhan Institute of Virology, Chinese Academy of Sciences, China Senior Scientist, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China Other Experience and Professional Memberships 2011 - 2013- 2014- 2014- 2015- 2016-18 2017 - 19 2017-2019 Honors Biosketches Director, Center for Emerging Infectious Diseases, Wuhan Inst. Virology, Chinese Acad. Sci. Director, BSL-3 laboratory at Wuhan Institute of Virology, Chinese Acad. Sci. Director, Committee of Biosafety, Wuhan Institute of Virology, Chinese Acad. Sci. Director, CAS Key Laboratory of Special Pathogens and Biosaf ety Vice Director, BSL-4 laboratory, Wuhan Institute of Virology, Chinese Acad. Sci. Associate Editor of Virology Journal Editorial Board of Virology Editor in Chief, Virologica Sinica Page 28

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 2003 Natural Science Award (the Second Prize) of Hubei Province, China. 2004 Outstanding supervisor of graduate student of Hubei Province, China. 2006 Outstanding scientist of the Chinese Academy of Sciences. 2006 Outstanding Research Article on Natural Science (the First Prize), Hubei Province, China 2014 Young and Middle-aged Scholar with Distinguished Contribution in Hubei Province, China 2014 Outstanding Research Article on Natural Science (the Grand Prize), Hubei Province, China 2016 Palm Knight Medal for Education, Government of the Republic of France 2017 Natural Science Award (the First Prize) of Hubei Province, China. C. Selected peer-reviewed publications most relevant to the current application *=Co-corresponding or first author Li W*, Shi Z*, Yu M, Ren W, Smith C, Epstein HJ, Wang H, Crameri G, Hu Z, Zhang H, Zhang J, Mceachern J, Field H, Daszak P, Eaton TB, Zhang S, Wang LF (2005). Bats are natural reservoirs of SARS-like coronaviruses. Science, 310: 676-679. Ren W, Qu X, Li W, Han Z, Yu M, Zhang S, Wang LF, Deng H, Shi Z (2008) Difference in receptor usage between SARS coronavirus and SARS-like coronavirus of bat origin. Journal of Virology 82(4): 1899-1907. Yuan J, Hon CC, Li Y, Wang D, Xu G, Zhang H, Zhou P, Poon LM, Lam TT, Leung FC. Shi Z (2010). Intra-species Diversity of SARS-Like Coronaviruses {CoVs) in Rhinolophus sinicus and Its Implications on the Origin of SARS-CoVs in human. Journal of General Virology, 91(4):1058-1062. Ge XY, Li JL, Yang X-L, Chmura AA, Zhu G, Epstein JH, MazetJK, Hu B, Zhang W, Peng C, Zhang YJ, Luo CM, Tan B, Wang N, Zhu Y, Crameri G, Zhang SY, Wang LF, Daszak P*, Shi Z* (2013). Isolation and characterization of a bat SARS-like Coronavirus that uses the ACE2 receptor. Nature 503: 535-538. Menachery VD, Yount BL, Debbink K, Agnihothram S, Gralinski LE, Plante JA, Graham RL, Scobey T, Ge XY, Donaldson EF, Randell SH, Lanzavecchia A, Marasco WA, Shi Z*, Barie RS* {2015). A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence. Nature Medicine, 21 :1508-1513. Yang XL, Hu B, Wang B, Wang MN, Zhang Q, Zhang W, Wu LJ, Ge XY, Zhang YZ, Daszak P, Wang LF*, Shi Z* (2016). Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus. Journal of Virology, 90: 3253-3256. Zeng L, Ge X, Peng C, Yang X, Tan B, Gao Y, Chen J, Chmura AA, Daszak P*, Shi Z* (2016) Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response. Journal of Virology, 90( 14): 6573-6582. Hu B, Zeng LP, Yang XL, Ge XY, Zhang W, Li B, Xie JZ, Shen XR, Zhang VZ, Wang N, Luo OS, Zheng XS, Wang MN, Daszak P, Wang LF, Cui J*, Shi Z* (2017). Discovery of A Rich Gene Pool of Bat SAAS-related Coronaviruses Provides New Insights into the Origin of SARS Coronavirus. PLOS Pathogens, 13(11 ): e1006698. Zhou P, Fan H, Lan T, Yang XL, Shi WF, Zhang W, Zhu Y, Zhang YW, Xie QM, Mani S, Zheng XS, Li B, Li JM, Guo H, Pei GO, An XP, Chen JW, Zhou L, Mai KJ, Wu ZX, Li D, Anderson D, Zhang LB, Li SY, Mi ZQ, He TT, Cong F, Guo PJ, Huang R, Luo Y, Liu XL, Chen J, Huang Y, Sun Q, Zhang XLL, Wang YY, Xing SZ, Chen YS, Sun Y, Li J, Daszak P, Wang LF, Shi Z, Tong YG, Ma JY (2018) Fatal swine acute diarrhea syndrome caused by an HKU-2 related coronavirus of bat origin. Nature, 556: 255-258. Luo CM, Wang N, Yang XL, Liu HZ, Zhang W, Li B, Hu B, Peng C, Geng QB, Zhu G, Li F*, Shi Z* (2018). Discovery of Novel Bat Coronaviruses in South China That Use the Same Receptor as Middle East Respiratory Syndrome Coronavirus. Journal of Virology, 92 (13): e00116-18. Additional recent publications of importance to the field (in chronological order) 2 Page 29 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Ge X, Li Y, Yang X, Zhang H, Zhou P, Zhang Y, Shi Z (2012). Metagenomic analysis of viruses from bat fecal samples reveals many novel viruses in insectivorous bats in china. Journal of Virology, 86, 4620-4630. Yuan J, Zhang Y, Li J, Zhang Y, Wang LF*, Shi Z* (2012). Serological evidence of ebolavirus infection in bats, China. Virology Journal, 9: 236. Yang XL, Zhang YZ, Jiang RD, Guo H, Zhang W, Li B, Wang N, Wang L, Waruhiu C, Zhou JH, Li SY, Daszak P, Wang LF*, Shi Z* (2017). Genetically Diverse Filoviruses in Rousettus and Eonycteris spp. Bats, China, 2009 and 2015. Emerging Infectious Diseases, 23(3) :482-486. Zeng LP, Ge XV, Peng C, Tai WB, Jiang SB, Du LY*, Shi Z* (2017). Cross-neutralization of SAAS coronavirus-specific antibodies against bat SAAS-like coronaviruses. Science China Life Sciences, 60(12):1399-1402. Wang N, Li SY, Yang XL, Huang, HM, Zhang YJ, Guo H, Luo CM, Miller M, Zhu G, Chmura AA, Hagan E, Zhou JH, Zhang YZ, Wang LF, Daszak p•, Shi z• (2018). Serological Evidence of Bat SARS-Related Coronavirus Infection in Humans, China. Virologica Sinica, 33(1):104-107. D. Research Support Ongoing Research Support Geographical distribution and genetic variation of pathogens in Africa Role: Pl (b) (4) 31770175 National Natural Science Foundation of China 01/01/2018-12/31/2021 Evolution mechanism of the adation of bat SAAS-related coronaviruses to host receptor molecules and the risk of interspecies infection Role: Pl Genetic evolution and transmission mechanism of important bat-borne viruses Role: Pl R01 Al110964 Daszak (Pl) Understanding Risk of Bat Coronaviruses (b) (4) 06/01/14-05/31/19 The goal of this study is to analyze the risk of coronavirus spillover from bats to humans in Southern China Role: Co-Investigator Emerging Pandemic Threat Program, USAID Mazet (Pl) 10/01/14-09/30/19 PREDICT2 The goal of this project is to create and implement a global virus surveillance system in animals and humans and analyze spillover risk. Role: China Country Coordinator Completed Research Support Metagenomic analysis of bat intestinal viruses Role: Pl Mechanism of interspecies transmission of zoonotic viruses 3 Biosketches (b) (4) (b) (4) Page 30

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Role: Co-Pl (b) (4) Genetic diversity, identification and pathogenesis of bat viruses (b) (4) 4 Page 31 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Kevin J. Olival eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Colorado State University, Fort Collins, CO Columbia University, New York, NY Columbia University, New York, NY American Museum of Natural History, New York NIH Fogarty US Global Health Fellow, New York A. Personal Statement POSITION TITLE Co-Investigator DEGREE BS MA PhD Post Doc Post Doc MM/YY FIELD OF STUDY 05/1997 Biology 10/2003 Conservation Biology 05/2008 Ecology & Evolution 08/2009 Molecular Parasitology 08/2011 EIDs The goal of this proposal is to understand the current and future threat of bat-borne coronavirus spillover in Southern China, by identifying which viruses, host species, and human behaviors are associated with the highest risk of CoV exposure. Specifically, we will use a combination of targeted bat sampling, human behavioral risk analyses, mathematical modeling, and phylogenetic and molecular methods to test several hypotheses related to zoonotic spillover risk of ~-CoVs, with specific attention paid towards SARSr-CoVs. My research experience over the last 16 years on bat-borne disease evolution, ecology, dynamics, population genetics, and viral discovery is strongly complementary to these aims. Our current proposal builds upon the findings of an ongoing NIAID R01 grant (ending 5/31/19), for which I was a co-investigator. Prior to this I coordinated research efforts under a NIAID award (2011-2016), investigating the risk of viral emergence from bats. This included sample collection and testing of thousands of bats from 8 countries globally. As an NIH Fogarty Global Health Post-Doc Fellow, I gained invaluable experience working internationally with a project focused on the ecology and evolution of Nipah virus in Bangladesh. My work over the last decade includes leading field investigations and bat viral surveillance in a wide range of countries, including: Bangladesh, Cambodia, India, Indonesia, Malaysia, Thailand, Philippines, Saudi Arabia, Georgia, Jordan, and Turkey. Discoveries include the first viral isolation of Nipah virus from the large flying fox in Malaysia; evidence of MERS-CoV in bats in Saudi Arabia; and the first serological evidence of Ebola Zaire virus in bats in Asia. I currently serve as the Modeling & Analytics coordinator under the USAID PREDICT-2 project, working with a team of analyst to develop new approaches to predict and prevent zoonoses. As part of this effort, I developed a new approach that combines phylogenetic, ecological, and life-history traits to predict viral diversity, host range, and spillover potential, leading to a recent first author paper in Nature. 1. (b) (4) 2. Memish ZA, Mishra N, Olival KJ, Fagbo SF, Kapoor V, Epstein JH, AIHakeem R, Al Asmari M, Islam A, Kapoor A, Briese T, Daszak P, Al Rabeeah AA, Lipkin WI. (2013). Middle East Respiratory Syndrome Coronavirus in Bats, Saudi Arabia. Emerging Infectious Diseases. 19(11): 1819-1823. 3. Olival KJ*, Hosseini P, Zambra-Torrellio C, Ross N, Bogich T, Daszak P*. (2017). Host and viral traits predict zoonotic spillover from mammals. Nature 546(7660): 646-650. *corresponding author B. Positions and Honors Page 32 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Positions and Employment 1999 - 02 Research Associate, Kewalo Marine Laboratory, University of Hawaii 2003 - 07 US Environmental Protection Agency STAR Fellow 2006 - 13 Instructor, Columbia University Secondary School Summer Program 201 0 - 15 Senior Research Scientist, EcoHealth Alliance 2015 - 17 Associate Vice President for Research, EcoHealth Alliance 2009 - Visiting Scientist, American Museum of Natural History 2009 - Adjunct Faculty, Earth Institute Center for Environmental Sustainability, Columbia University 2017 - Vice President for Research, EcoHealth Alliance Other Experience and Professional Memberships 1998 - 00 Member, AAAS 2000 - 02 Mentor, NSF Undergraduate Mentoring in Environmental Biology (UMEB), University of Hawaii 2003 - 05 Member, American Society of Mammalogists 2005 - 06 Member, New York Academy of Sciences 2011 - Scientific Steering Committee Member, Southeast Asian Bat Conservation Research Unit 2011 - Scientific Advisory Board Member, Lubee Bat Conservancy, FL 2011 - Scientific Advisor, Bat Conservation International 2011 - Review Editor, EcoHealth 2015 - US White-Nose Syndrome Stakeholder Committee and Communications Committee Member 2015 - Island and Seas, Board Member 2017 - DoD DTRA: Steering Committee Member, Bat One Health Research Network Honors 1993-97 2003 2005-07 2004-07 2008 2013 2013-14 2016 2017-18 Colorado State University Distinguished Scholar Award NSF Graduate Student Fellowship, Honorable Mention Bat Conservation International Student Award and Scholarship US EPA STAR Fellowship Award PhD with Distinction, Columbia University Plenary talk on bat virus modeling at 11th Annual ASM Biodefense and EID Research Meeting Institute of Medicine, Forum on Microbial Threats. Invited speaker, briefings on MERS-CoV and Emerging Viral Diseases Plenary Speaker, NYC Medtech conference - Global Virome Project Three papers awarded the lnCites Highly Cited Paper™ designation (top 1 % in field) for Immunology and Microbiology C. Contribution to Science 1. Viral Discovery and Characterization in Bats A large body of my research has focused on understanding the distribution and diversity of viruses in wildlife populations to better understand the ecological risk of viral emergence. This includes the first use of species accumulation curves to estimate viral diversity using data from longitudinal surveillance of fruit bats in Bangladesh, and a large meta-analysis of viral prevalence in bats to optimize discovery strategies. Two field studies highlighted below include a broad geographic survey of bat coronaviruses in Thailand, and the first isolation and full genome characterization of Nipah virus from the large flying fox in Malaysia. a. Rahman SA, Hassan SS, Oliva! KJ, Mohamed M, Chang L-Y, Hassan L, Saad NM, Shohaimi SA, Mam at ZC, Nairn MS, Epstein JH, Suri AS, Field HE, Daszak P and HERG. (2010). Characterization of Nipah virus from Naturally Infected Pteropus vampyrus Bats, Malaysia. Emerging Infectious Disease 16(12): 1990-1993. 2 Page 33 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter b. Anthony SJ, Epstein JH, Murray KA, Navarrete-Macias I, Zambrana-Torrelio CM, Solovyov A, Ojeda-Flores R, Arrigo NC, Islam A, Khan SA, Hosseini P, Bogich TL, Olival KJ, Sanchez-Leon MD, Karesh WB, Goldstein T, Luby SP, Morse SS, Mazet JAK, Daszak P, Lipkin WI. (2013). A Strategy To Estimate Unknown Viral Diversity in Mammals. Mbio. 4(5): e00598-13. c. Wacharapluesadee S, Duengkae P, Rodparn A, Kaewpom T, Maneeorn P, Kanchanasaka B, Yinsakmongkon s, Sittidetboripat N, Chareesaen C, Khlangsap N, Pidthong A, Leadprathom K, Ghai S, Epstein JH, Daszak P, Olival KJ, Blair PJ, Callahan MV, Hemachudha T. (2015). Diversity of Coronavirus in Bats from Eastern Thailand. Virology Journal 12:57. d. Young CC and Olival KJ*. (2016). Optimizing Viral Discovery in Bats. PLO$ ONE 11(2): e0149237. 2. Serological Surveillance Bats are believed to harbor a unique and large diversity of viruses, including a number of pathogens that pose a risk to human health (e.g. Ebola, Nipah, SARS-CoV). I have been involved with field and laboratory investigations of several bat-borne pathogens that pose the greatest risk to humans over the years, including Filoviruses, Henipaviruses, and SARS and MERS-related Coronaviruses. Collection and analysis of serological data was critical to each of these studies. Using serological and PCR data we discovered that bats are reservoirs of Ebola Reston virus in the Philippines. Extensive, proactive surveillance of wild bat and primate populations in Thailand for Ebola viruses importantly showed that several suspected species are likely not important reservoirs. The work in Thailand was predicated by my own investigations in Bangladesh where we discovered the first evidence for Ebola Zaire virus infection in a wildlife species outside of Africa- changing our paradigm as to where these viruses can be found globally. Lastly, I have been involved with extensive work to identify the natural reservoir host of Reston virus in Philippines that included both molecular and serological findings. a. Olival KJ*, Islam A, Yu M, Anthony SJ, Epstein JH, Khan SA, Khan SU, Crameri G, Wang LF, Lipkin WI, Luby SP, and Daszak P. (2013). Ebolavirus Antibodies in Fruit Bats, Bangladesh. Emerging Infectious Diseases 19(2): 270-273. b. Wacharapluesadee S, Olival KJ, Kanchanasaka B, Duengkae P, Kaewchot S, Srongmongkol P, leamsaard G, Maneeorn P, Sittidetboripat N, Kaewpom T, Petcharat S, Yingsakmongkon S, Rollin PE, Towner JS , Hemachudha ,. (2015). Surveillance for Ebola Virus in Wildlife, Thailand. Emerging Infectious Diseases 21 (12): 2271-2273. c. Jayme S, Yu M, Jong Cd, Olival KJ, Tagtag A, Hughes T, Foard A, Marsh G, Crameri G, Epstein JH, Santos I, Catbagan D, Lim M, Benigno C, Wang L, Daszak P, Field H, Newman S. (2015). Molecular evidence of Ebola Reston virus infection in Philippine bats. Virology Journal. 12(1): 107. d. (b) (4) 3. Modeling Disease Emergence and Spillover Risk I have used my applied ecology background working with analyses of wildlife and their pathogens to develop new models to improve our global understanding of zoonotic spillover and disease circulation. In addition to my previously mentioned Nature paper, this includes studies that examined the environmental drivers of bat virus spillover to humans, cross-species transmission among bat species, spatial analysis of emerging zoonotic disease hotspots, and host-specific determinants of fungal infection in bats. These modeling approaches explicitly use data from PCR- and serology-based field studies, combined with an understanding of wildlife biology and ecology, to assess the environmental and demographic drivers of disease transmission -- bridging the gap between field investigations and modeling transmission risk. 3 Page 34 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter a. Brierley L, Vonhof MJ, Olival KJ, Daszak P, Jones KE. (2016). Quantifying global drivers of zoonotic bat viruses: a process-based perspective. American Naturalist 187: E53-64 b. Willoughby AR, Phelps K, PREDICT Consortium, Olival KJ*. (2017). "A Comparative Analysis of Viral Richness and Viral Sharing in Cave-Roosting Bats". Diversity 9 (35). c. Allen T, Murray KA, Zambrana-Torrelio C, Morse SS, Rondinini C, Di Marco M, Breit N, Olival KJ, Daszak P. (2017). Global hotspots and correlates of emerging zoonotic diseases. Nature Communications. 8( 1124): 1-1 o d. Verant ML, Bohuski EA, Richgels KLD, Olival KJ, Epstein JH, and Blehert DS. (2018). Determinants of Psudogymnoascus destructans within bat hibernacula: implications for surveillance and management of white-nose syndrome. Journal of Applied Ecology 55: 820-829. D. Additional Information: Research Support and/or Scholastic Performance Ongoing Research Support HDTRA11710064 Olival (Pl) 10/02/17-10/01/22 Understanding the Risk of Bat-Borne Zoonotic Disease Emergence in Western Asia The goal of this project is to characterize pathogen diversity, strengthen zoonotic disease surveillance capacity, and test key hypotheses about the risk of bat-borne zoonotic disease emergence in Western Asia. Role: Pl R01 Al110964 Daszak (Pl) 06/01 /14-05/31 /19 Understanding Risk of Bat Coronaviruses The goal of this study is to analyze the risk of coronavirus spillover from bats to humans in Southern China Role: co-Pl Emerging Pandemic Threat Program, USAID Mazet (Pl) 10/01 /14-09/30/19 PREDICT 2 The goal of this project is to create and implement a global virus surveillance system in animals and humans and analyze spillover risk. Role: Modeling and Analytics Coordinator; Country lead for Indonesia, South Sudan, and Thailand. Completed Research Support Emerging Pandemic Threat Program, USAID Mazet (Pl) 1 0/01/09-09/30/ 14 PREDICT The goal of this project was to conduct zoonotic virus surveillance in wildlife in 20 countries, and modeling hotspots and drivers for disease emergence. Role: Key Personnel: Modeling Team; Country lead for Thailand and Indonesia Service Award, US Fish and Wildlife Epstein (Pl) 09/01 /12-09/30/14 Characterization of Climatic Parameters within Bat Hibernacula, their Influence on Environmental Loads of Geomyces destructans, and Implications for the Migration of White-Nose Syndrome in Bats. The goal of this project was to identify environmental and other factors that influence the progression and severity of White Nose Syndrome in bats. Role: co-Pl R01 AI079231 Daszak (Pl) 09/18/08-08/31 /13 4 Page 35 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Risk of viral emergence from bats Modeled hotspots for viral diversity and emergence in bats, discovery of new viruses, and in vitro test of infectiousness for novel pathogens. Role: Key Personnel: led project implementation, study design, and phylogenetic modeling Endangered Species grant, USGS Russell, Vonhof, and Olival (Pl) 06/18/12-06/17/13 Genetic Approaches to Defining Taxonomic and conservation Units for the Hawaiian Hoary Bat The goal of this project was to determine the phylogenetic position and conservation genetic units for endangered hoary bats. Role: co-Pl 3R01 TW005869-06S1 Daszak (Pl) 09/01/09 - 8/31/11 NIH Fogarty Ecology of Infectious Diseases ARRA award The goal of this project was to conduct Nipah virus surveillance in wild bat populations and use genetic methods to understand viral circulation in Bangladesh. Role: Fogarty US Global Health Fellow 5 Page 36 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Ralph Steven Barie eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION North Carolina State University, Raleigh, NC North Carolina State University, Raleigh, NC University of Southern CA, School of Med, (Los Angeles, CA) POSITION TITLE Co-Investigator DEGREE BS Ph.D. Post-Doc MM/YY FIELD OF STUDY 1977 Zoology 1982 Microbiology 1986 Microbiology A. Personal Statement: The Barie laboratory uses genetic, biochemical, molecular and immunologic approaches to study the molecular mechanisms regulating viral evolution, virus immunity, virus-host interactions and vaccine mediated protective immunity using coronaviruses (CoV), noroviruses and flaviviruses (Dengue) as models. SARS-CoV and MERS-CoV are used as models to address fundamental questions in genetics, structure-function analyses, entry and cross species transmission, fidelity regulation, host susceptibility allele mapping, pathogenesis as well as therapeutic design and testing. Synthetic genomics and reverse genetics are used to create a panel of CoV molecular cDNA clones for SARS-CoV, SARS-like bat coronaviruses (SL-CoV), MERS-CoV, several human coronavirus, Dengue 1-4 and Zika virus. The Barie laboratory has developed key animal models of human disease, including SARS-CoV and SL-CoV pathogenesis in young and aged mice, and CRISPR gene edited mice encoding permissive mutations in the murine dipeptidyl peptidase receptor, making the animals permissive for MERS-CoV infection and disease. The Barie laboratory has longstanding expertise in CoV evolution and emergence, replication, virus-receptor interactions, genetics, animal model development and pathogenesis. Not only has the Barie laboratory made fundamental breakthroughs in all aspects of CoV genetics, biology and immunology, but it has designed, developed and tested small molecule inhibitors and vaccines against emerging Co Vs. Our group has collaborated with Ors. Daszak, Shi and Wang on SARSr-CoVs for the past 3 years, and this R01 is a natural development of this collaboration. Qualifications by Publication:: >314 total publications, >120 since 2013, H-index: 84. http://www.ncbi.nlm.nih.gov/sites/myncbi/ralph.baric.1 /bibliography/40583903/public/?sort=date&direction=ascending. Key Manuscripts 1. Sheahan TP, Sims AC, Graham RL, Menachery VD, Gralinski LE, Case JB, Leist SR, Pyre K, Feng JY, Trantcheva I, Bannister R, Park Y, Babusis D, Clarke MO, Mackman RL, Spahn JE, Palmiotti CA, Siegel D, Ray AS, Cihlar T, Jordan R, Denison MR, Barie RS (2017). Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses. Science Translational Medicine, 9(396). eaal3653. PMC5567817. 2. Scobey T, Yount BL, Sims AC, Donaldson EF, Agnihothram SS, Menachery VD, Graham RL, Swanstrom J, Bove PF, Kim JD, Grego S, Randell SH, Barie RS (2013). Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus. Proceedings of the National Academy of the Sciences, 110(40):16157-62. PMC3791741. 3. Menachery, VD, Yount, BL, Debbink, K, Agnihothram, S., Gralinski, LE, Plante, JA, Graham, RL, Scobey T, Ge SY, Donaldson EF, Randell SH, Lanzavecchia A, Marasco WA, Shi Z, Barie RS (2015). A SARSlike cluster of circulating bat coronaviruses shows potential for human emergence. Nature Medicine, Nov 9. doi: 10.1038/nm.3985. [Epub ahead of print]. PMID:26552008. Page 37 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 4. Cockrell AS, Yount· BL, Scobey T, Jensen K, Douglas M, Beall A, Tang XC, Marasco WA, Heise MT, Barie RS (2016). A Mouse Model for MERS Coronavirus Induced Severe Respiratory Distress Syndrome. Nature Microbiology, 2:16226. PMC5578707. B_ Positions and Honors_ Employment Experience: 1986-92 Assistant Professor, Department of Parasitology and Laboratory Practice and Department of Epidemiology, University of North Carolina (UNC), Chapel Hill, NC 1992-2001 Associate Professor, Departments of Epidemiology and Microbiology & Immunology, UNG Chapel Hill 2001- Professor, Departments of Epidemiology and Microbiology and Immunology, UNG Chapel Hill Selected Awards/Honors: 2018 US Natl. Acad. Sci. "China-US Workshop on Challenges of Emerging Infections, Laboratory Safety 2015 2015 2015 2014 2005-15 2008-15 2008 2007-08 2005-09 2003 1989-94 1984-86 and Global Health Security, Jan 2018, Galveston, Tx. US Natl. Acad. Sci./UK Royal Society Workshop: Sackler Scientific Forum on the Trends in Synthetic Biology and Gain of Function and Regulatory Implications, U.K. US Natl. Acad. Sci. "China-U.S. Workshop on the Challenges of Emerging Infections. Laboratory Safety, and Global Health Security" September 28-30 in Beijing, China MERS-CoV Stakeholders Workshop, invited panelist, NIH National Academy of Sciences: Working Group on Risks and Benefits of Gain of Function Research Review Board, J. Virology Senior Editor, Plos Pathogens US Natl. Acad. Sci. Working Group: Gene Sequence Methods for Classification of Select Agents Associate Editor, Plos Pathogens Permanent Member, NIH VirB Study Section FinalisVRunner-up, World Technology Award Established Investigator: American Heart Association Harvey Weaver Scholar, National Multiple Sclerosis Society C. Contributions to Virology: The Barie laboratory has made significant contributions to our understanding of all aspects of CoV biology, including: i) CoV genetics and reverse genetics for SARS-CoV, MHV, MERSCoV, HCoV NL63, PEDV, TGEV, bat SARS-like CoV (SL-CoV), BtCoV HKU-5 and others, ii) demonstration of proof-reading activities in the CoV genome, iii) identification and characterization of bat SL-CoV with prepandemic potential, iii) coronavirus transcription mechanisms, iv) mechanisms of interferon antagonism and interferon stimulated gene expression control, v) virus host susceptibility allele mapping, vi) epitope mapping of human monoclonal antibodies, vii) identification of broad spectrum human monoclonal antibodies against SARS-CoV and MERS-CoV, viii) mouse models of human disease (MERS-CoV and SARS-CoV), ix) aging and emerging coronavirus vaccine efficacy, and x) live and attenuated vaccine design in young and aged animal models of human disease. The Barie laboratory has also made major contributions to norovirus immunology and flavvirus reverse genetics and the human immune responses after infection. Some representative major contributions outside and within the Co V field include: 1. (b)(4) 2. Gralinski LE, Ferris MT, Aylor DL, Whitmore AC, Green R, Frieman MB, Deming D, Menachery VD, Miller DR, Buus RJ, Bell TA, Churchill GA, Threadgill DW, Katze MG, McMillan L, Valdar w, Heise MT, PardoManuel de Villena F, Barie RS (2015) Genome Wide Identification of SARS-CoV Susceptibility Loci Using the Collaborative Cross. PLOS Genetics, 11(10): e1005504. PMID:26452100. 3. Lindesmith L, Moe C, Marionneau S, Ruvoen N, Jiang X, Lindblad L, Stewart P, LePendu J, Barie R (2003). Human susceptibility and resistance to Norwalk virus infection. Nature Medicine, 9(5):548-53. PMID:12692541. 2 Page 38 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 4. Lindesmith LC, Donaldson EF, Lobue AD, Cannon JL, Zheng DP, Vinje J, Barie RS (2008). Mechanisms of Gll.4 norovirus persistence in human populations. PLO$ Medicine, 5(2):e31. PMC2235898. C.1. Coronavirus Pathogenesis and Virus Immunity. Our group has studied the role of virus-immune interactions in coronavirus and other emerging virus pathogenesis mechanisms. 1. Rasmussen AL, Okumura A, Ferris MT, Green R, Feldmann F, Kelly SM, Scott DP, Safronetz D, Haddock E, Lacasse R, Thomas MJ, Sova P, Carter VS, Weiss JM, Miller DR, Shaw GD, Korth MJ, Heise MT, Barie RS, de Villena FP, Feldmann H, Katze MG (2014). Host genetic diversity enables Ebola hemorrhagic fever pathogenesis and resistance. Science, 2014 346(6212) :987-91. PMC4241145. 2. Gralinski LE, Sheahan TP, Morrison TE, Menachery VD, Jensen K, Leist SR, Whitmore A, Heise MT, Barie RS (2018). Complement Activation Contributes to Severe Acute Respiratory Syndrome Coronavirus Pathogenesis. mBio, 9(5). e01753-18. PMC6178621. 3. Menachery VD, Eisfeld AJ, Schafer A, Josset L, Sims AC, Prall S, Fan S, Li C, Neumann G, Tilton SC, Chang J, Gralinski LE, Long C, Green R, Williams CM, Weiss J, Matzke MM, Webb-Robertson BJ, Schepmoes AA, Shukla AK, Metz TO, Smith RD, Waters KM, Katze MG, Kawaoka Y, Barie RS (2014). Pathogenic influenza viruses and coronaviruses utilize similar and contrasting approaches to control interferon-stimulated gene responses. mBio, 5(3): e01174-14. PMC4030454. 4. Graham RL, Becker MM, Eckerle LO, Bolles M, Denison MR, Barie RS (2012). A live, impaired-fidelity coronavirus vaccine protects in an aged, immunocompromised mouse model of lethal disease. Nature Medicine, 18(12):1820-6. PMCID: PMC3518599. C.2. Coronavirus Innate Immunity/Animal Models. The Barie laboratory group has studied CoV host range expansion using experimental evolution and SARS-CoV, MERS-CoV, civet SL-CoV, bat SL-CoV, and bat CoV HKU5 as models. This includes synthetic reconstruction of civet and bat CoV from in silica sequence, the first reported recovery of recombinant bat viruses, and characterization of host range phenotypes in vitro and in vivo. Applications of experimental evolution have focused on molecular mechanisms associated with virusreceptor interactions in viral persistence, virus innate immune interactions, and increased virulence in mice. 1. Agnihothram S, Yount BL, Donaldson EF, Huynh J, Menachery VD, Gralinski LE, Graham RL, Becker MM, Tamar S, Scobey TD, Osswald HL, Whitmore A, Gopal R, Ghosh AK, Mesecar A, Zambon M, Heise M, Denison MR, Barie RS (2014). A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKU5 variant. mBio, 5(2): e00047-14. PMC3977350. 2. Sheahan T, Rockx B, Donaldson E, Corti D, Barie R (2008). Pathways of cross-species transmission of synthetically reconstructed zoonotic severe acute respiratory syndrome coronavirus. Journal of Virology, 82(17):8721-32. PMC2519660 3. Becker MM, Graham RL, Donaldson EF, Rockx B, Sims AC, Sheahan T, Pickles RJ, Corti D, Johnston RE, Barie R*, Denison MR* (2008). Synthetic recombinant bat SAAS-like coronavirus is infectious in cultured cells and in mice. Proceedings of the National Academy of the Sciences, 105(50):19944-9. PMC2588415. (* = co-first authors) 4. Menachery VD, Schafer A, Burnum-Johnson KE, Mitchell HD, Eisfeld AJ, Walters KB, Nicora CD, Purvine SO, Casey GP, Monroe ME, Weitz KK, Stratton KG, Webb-Robertson BM, Gralinski LE, Metz TO, Smith RD, Waters KM, Sims AC, Kawaoka Y, Barie RS (2018). MERS-CoV and H5N1 influenza virus antagonize antigen presentation by altering the epigenetic landscape. Proceedings of the National Academy of the Sciences, 115(5): E1012-E1021. PMID: 29339515. C.3. Virus Genetic Platforms. The Barie laboratory has pioneered reverse genetic analyses of CoVs and DENVs. Several CoV infectious cDNA clones are available in the lab, including SARS-CoV, MERS-CoV, conventional human and model Co Vs, and several bat Co Vs with pandemic potential. The availability of these genetic platforms allows for detailed studies into the role of viral genes in pathogenesis, innate immune antiviral immunity, vaccine performance and design, virus-receptor interactions, entry and virus evolution. 1. Yount B, Curtis, K, Fritz L, Hensley L, Jahrling P, Prentice E, Denison M, Geisbert T, Barie RS (2003). Reverse Genetics with a full length infectious cDNA for the SARS Coronavirus. Proceedings of the National Academy of the Sciences, 100(22): 12995-13000. PMCID: PMC240733. 3 Page 39 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 2. Rockx B, Sheahan T, Donaldson E, Harkema J, Sims A, Heise M, Pickles R, Cameron M, Kelvin D, Barie R (2007). Synthetic reconstruction of zoonotic and early human severe acute respiratory syndrome coronavirus isolates that produce fatal disease in aged mice. Journal of Virology 81(14):7410-23. PMC1933338. 3. Widman DG, Young E, Yount BL, Plante KS, Gallichotte EN, Carbaugh DL, Peck KM, Plante J, Swanstrom J, Heise MT, Lazear HM, Barie RS (2017). A Reverse Genetics Platform that Spans the Zika Virus Family Tree. mBio, 8(2): e02014-16. PMC5340872 4. Donaldson EF, Yount B, Sims AC, Burkett S, Pickles RJ, Barie RS (2008). Systematic assembly of a fulllength infectious clone of human coronavirus NL63. Journal of Virology, 82(23):11948-57. PMC2583659. C4. Virus Vaccine Design and Antiviral lmmunotherapy. Viruses are major causes of morbidity and mortality globally. The Barie laboratory has used structure-guided immunogen design and epitope exchange to build multivalent immunogens to increase vaccine breadth and diagnostic potential. 1. Deming DJ, Sheahan T, Heise M, Yount B, Davis N, Sims A, Suthar M, Whitmore JH, Pickles R, West A, Donaldson E, Curtis K, Johnston, RE, Barie RS (2006). Vaccine efficacy in senescent mice challenged with recombinant SARS-CoV bearing epidemic and zoonotic spike variants. PLOS Medicine, 3(12): e525 PMCID: PMC1716185. 2. Tang XC, Agnihothram SS, Jiao Y, Stanhope J, Graham RL, Peterson EC, Avnir Y, Tallarico AS, Sheehan J, Zhu Q, Barie RS, Marasco WA (2014). Identification of human neutralizing antibodies against MERSCoV and their role in virus adaptive evolution. Proceedings of the National Academy of the Sciences, 111 (19):E2018-26. PMC4024880 3. Lindesmith LC, Ferris MT, Mullan CW, Ferreira J, Debbink K, Swanstrom J, Richardson C, Goodwin RR, Baehner F, Mendelman PM, Bargatze RF, Barie RS (2015). Broad blockade antibody responses in human volunteers after immunization with a multivalent norovirus VLP candidate vaccine: immunological analyses from a phase I clinical trial. PLOS Medicine, 12(3) :e1001807 PMC4371888. 4. Bolles M, Deming D, Long K, Agnihothram S, Whitmore A, Ferris M, Funkhouser W, Gralinski L, Totura A, Heise M, Barie RS (2011 ). A double-inactivated severe acute respiratory syndrome coronavirus vaccine provides incomplete protection in mice and induces increased eosinophilic proinflammatory pulmonary response upon challenge. Journal of Virology, 85(23) :12201-15. PMC3209347 D.Research Support. U19 Al 100625 Barie/Heise (MPI) 09/01/17-08/31/22 Systems lmmunogenetics of Biodefense Pathogens in the Collaborative Cross The Collaborative Cross is a mouse resource for study of complex genetic interactions in diverse populations, to identify novel polymorphic genes regulating immune responses to SARS, influenza and WNV, analyze genetic underpinning of immune phenotypes in mice and humans, and generate panels of genetically defined mice to probe polymorphic gene control of immune responses against a pathogens or other immune stimuli. R01 A1108197 Denison/Barie (MPI) 05/01/18-04/30/23 Determinants of Coronavirus Fidelity in Replication and Pathogenesis Experiments in this aim will test the hypothesis that nsp14 functions in maintaining high replication fidelity and viral RNA synthesis are coupled and that targeted engineered mutations across nsp14 alter: a) RNA fidelity outcomes; b) sensitivity to nucleoside mutagens and polymerase inhibitors; c) sensitivity to innate immunity. HHSN2722010000191-HHSN27200003 Barie (Pl) 09/30/17-03/31/24 MERS-CoV Mouse Model for Vaccine & Therapeutic Testing (Task Order A57) Use generation of transgenic mice and modifications to the MERS-CoV genome to identify a mouse model for MERS-CoV that recapitulates human disease phenotypes for evaluating vaccine platforms and therapeutics. U19 Al 109680 Whitley (Pl) 03/01/14-02/28/19 Antiviral Drug Discovery and Development Center The specific aims of the proposal will identify small molecule inhibitors of CoV fidelity and RNA capping, define their mechanism of action, and determine their efficacy against SARS-CoV and across CoV families using in vivo mouse models of acute and persistent CoV disease. Role: Co-Investigator 4 Page 40 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter U19 Al 109761 Lipkin (Pl) 03/01/14-02/28/19 Diagnostic and Prognostic Biomarkers for Severe Viral Disease The goal is to develop new platform technologies that use functional genomics as diagnostic and prognostic indicators of severe end stage lung disease, systemic and enteric diseases following virus infection, including coronaviruses, flaviviruses and noroviruses. Role: Project Leader R01 Al110700 Barie (Pl) 04/20/15-03/31/20 Mechanisms of MERS-CoV Entry, Cross-species Transmission and Pathogenesis The overall goal is to build a comprehensive understanding of the molecular mechanisms guiding group 2c CoV receptor recognition, entry and pathogenesis. (b)(4J Barie (Pl) (b)(4) --.,...,..---,-,,,..,..- Breadth of Blockade Antibody Responses Following Norovirus Vaccination. (b) <4> and UNC will collaborate to evaluate the breadth of the antibody blockade response following norovirus vaccination in various human volunteer populations. P01 Al106695 Harris (Pl) 07/1/2015-6/30/20 Protective immunity following dengue virus natural infections and vaccination Project 2: Aravinda deSilva and Ralph S. Barie (Co-Pl). The goal is to identify natural correlates of protective immunity following natural infection and or vaccination. Role: Co-Investigator R01-Al125198 de Silva (Pl) 05/01/16 - 04/30/21 Preclinical assays to predict dengue vaccine efficacy We use samples from DENV tetravalent Sanofi Pasteur vaccine clinical trials to identify mechanisms and correlates of protective immunity or breakthrough infections in vaccinees. Role: Co-investigator. R01 1Al132178 Barie/Sheahan( MPI) 08/15/17-8/14/22 Broad-spectrum antiviral GS-5734 to treat MERS-CoV and related emerging CoV:. The goal of this proposal is collaborate with Gilead Inc. and obtain GS-5734 preclinical data for IND development and translational studies, all designed to move the therapeutic into human trials. (bH 4> Breuer (Pl) (b)(4) .,.,..,,,----,--.,...,....--,-----,---,,----....,...,.----- Why do Norovirus pandemics occur and how can we control them? The program uses hospital and community cohorts of NoV infected individuals to ask fundamental questions into the molecular and evolutionary epidemiology of human NoV infections, focusing on the Gll.4 strains, leading to new models of virus emergence and disease prevention. Role: Co-Investigator: R01 Al 089728 Li (Pl) 07 /01 /16-06/30/21 University of Minnesota/NIAID Receptor recognition and cell entry of coronaviruses The program studies receptor usage and cell entry mechanisms of emerging coronaviruses, focused on PEDV, MHV and SARS-like Coronaviruses. Role: Co-Investigator R21 Al135682 UT Austin/NIAID Georgiou (Pl) Molecular Analysis of Serum Antibody Constituents in Zika Virus Infection. 04/01 /18-03/30/20 The goal of this application is to identify antibodies that make up the serologic repertoire after Zikv infection of naive and DENV preimmune individuals. Role: Co-investigator. R21 Al137887 Moorman/Heise (MPI) 02/05/18-01/31/20 NIH/NIAID $150,000 Molecular Characterization of Functional RNA Structures in the ZikV genome The goal of this project is to study the RNA Structure of Zika virus. Proposed studies will identify new viral virulence determinants that can be targeted to generate safer and more effective Zika virus vaccines and therapeutics. Role: Co-Investigator. 5 Page 41 Biosketches

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Contact PD/Pl: DASZAK, PETER NAME Noam Ross eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Brown University (US) University of California-Davis, (US) EDUCATION/TRAINING INSTITUTION AND LOCATION Brown University, Providence, RI University of California-Davis, Davis, CA A. Personal Statement Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH POSITION TITLE Co-Investigator DEGREE MM/VY FIELD OF STUDY BS 05/2006 Environmental Sci. PhD 09/2015 Ecology DEGREE MM/YYYY FIELD OF STUDY BS 05/2006 Environmental Science Ph.D 09/2015 Ecology The goal of our proposal is to identify and quantify the drivers of bat-borne coronavirus spillover in Southern China, by identifying which host traits, viral characteristics, and human behaviors are associated with the highest risk of CoV exposure. This will require statistical and mathematical modeling approaches that can integrate the separate ecological, evolutionary, and behavioral processes into a robust framework. My background in quantitative disease ecology makes me a natural fit to work on the statistical and mathematical aspects of this project. My research has consisted of developing both statistical and theoretical models for emerging diseases in both plants, mammals, and humans. I have developed dynamic models of diseases such as MERS and Ebola virus in wildlife populations in order to support targeting field surveillance, and applied predictive empirical and mechanistic modeling techniques to the study of Nipah virus emergence and circulation in bats. My statistical work has included analysis of survey-based evidence of new disease emergence in Uganda, global predictive models of anthrax emergence, and large-scale macroecological patterns in host-virus associations which captured previously unmodeled heterogeneity in disease burden. Importantly, this work included the creation of methods and open-source tools for simulating, fitting, and performing optimization using such models, ensuring that I will be able to support the creation of robust and reproducible statistical models this project. a. Oliva! KJ, Hosseini PR, Zambrana-Torrelio C, Ross N, Bogich TL, Daszak P (2017). Host and viral traits predict zoonotic spillover from mammals. Nature 546: 646-650 b. Salerno J, Ross N, Ghai R, Mahero M, Travis DA, Gillespie TR, Hartter J (2017) Human-wildlife interactions predict febrile illness in park landscapes of western Uganda. EcoHealth 14(4):675-690. c. Carlson CJ, Kracalik I, Ross N, Alexander K, Hugh-Jones ME, Fegan M, Elkin B, Epp T, Shury T, Bagirova M, Getz WM, Blackbum JK (2018} The global distribution of Bacillus anthracis and associated anthrax risk to humans, livestock, and wildlife. Nature Microbiology In Review. B. Positions and Honors Positions and Employment 2006 Contract Market Researcher: Energy Efficient Products Initiative, Wal-Mart, Providence, RI 2006 - 07 Analyst, Environmental Markets and Performance, GreenOrder, New York, NY Page 42 Biosketches

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Contact PD/Pl: DASZAK, PETER 2007 - 09 2010 - 15 2015 - 17 2017 - Senior Analyst, Environmental Markets and Performance, GreenOrder, New York, NY Graduate Researcher, University of California-Davis Disease Ecologist, EcoHealth Alliance, New York, NY Senior Research Scientist, EcoHealth Alliance, New York, NY Other Experience and Professional Memberships 2012 - 13 Member, NSF IGERT.org advisory board 2012 - 15 Founder and Organizer, Davis R Users' Group 2013 - Member, Ecological Society of America 2014 - Contributor and reviewer, ROpenSci 2014 - Meeting Session Organizer, Ecological Society of America 2015 - Instructor, Software Carpentry Foundation 2015 - Instructor, Data Carpentry Foundation 2015 - Associate Editor, ROpenSci 2016 - Member, R Epidemics Consortium Reviewer: Ecology Letters, Theoretical Ecology, EcoHealth, Conservation Letters, Biological Reviews, Journal of Open Source Software Awards and Fellowships 201 0 NSF IGERT Trainees hip in Rapid Environmental Change 201 0 UC Davis Graduate Ecology Fellowship 2012 Don Dahlsten Memorial Grant, California Forest Pest Council 2012 NSF IGERT Bridge Fellowship C. Contribution to Science 1. Modeling Dynamics of Heterogeneity: I have worked on both theoretical and applied approaches of dealing with heterogeneity when modeling ecological-epidemiological dynamics. This work focused on fungal disease epidemics using a framework traditionally used for parasites of stable populations in order to capture the role of individual variation in infection level. While the mathematical basis of these models for populations at or approximately at equilibrium is well established, their dynamic properties are less well known due to analytical intractability, and this they are little-used in emerging diseases and epidemics. My work showed how and where these models diverged from other, traditional models in their dynamical properties, and identified statistical patterns that could be used to identify where these models are appropriate. I developed numerical tools for their simulation, modeling and control, which have been used in applied disease management studies. a. Schreiber S, Ross N (2016) Individual-based Integral Projection Models: The role of size-structure on extinction risk and establishment success. Methods in Ecology and Evolution. http://dx.doi.org/10.1111 /2041-21 0X.12537 b. Cobb RC, Ross N, Hayden JK, Eyre CA, Dodd RS, Frankel SJ, Garbelloto M, Rizzo DM (2018) Promise and pitfalls of endemic resistance when cultural resources are threatened by exotic tree pathogens. Phytopathology. https://doi.org/10.1094/PHYTO-04-18-0142-R c. Cobb RC, Hartsough P, Ross N, Klein J, LaFever DH, Frankel SJ, Rizzo DM (2017) Resiliency or restoration: management of sudden oak death before and after outbreak. Forest Phytophthoras. https://doi.orq/10.5399/osu/fp.7.1.4021 d. Ross N (2015). Disease with Multiple Infections: Population Structure, Dynamics, and Control. University of California, Davis. Dissertation. 2. Modeling decision-making in complex systems: A long-standing theme of my work has been linking ecological dynamics to social systems and decision-making under uncertainty. This has included determining whether statistical signals of ecological changes are sufficient to justify management changes in fisheries, and has recently extended to optimizing investment in disease surveillance and intervention. 2 Page 43 Biosketches

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Contact PD/Pl: DASZAK, PETER a) Machalaba C, Smith KM, Awada L, Berry K, Berthe F, Bouley TA, Bruce M, Abrahantes JC, Turabi EL, Feferholtz Y, Flynn L, Fournie G, Andre A, Grace D, Jonas 0, Kimani T, Gall FL, Jose J, Peyre MM, Pinto J, Ross N, Ruegg SR, Salerno RH, Seifman R, Zambrana-Torrelio C, Karesh WB. (2017) One Health Economics to confront disease threats. Transactions of the Royal Society of Tropical Medicine and Hygiene https://doi.org/10.1093/trstmh/trx039 b) Boettiger C*, Ross N*, Hastings A (2013) Early Warning Signals: The Charted And Uncharted Territories. Theoretical Ecology http:/ /dx .doi .org/10 .1007 /s 12080-013-0192-6 (*Co-equal authors) c) Fuller K, Kling D, Kroetz K, Ross N, Sanchirico JN (2013) Economics and Ecology of Open-Access Fisheries. In: Shogren JF (ed.) Encyclopedia of Energy, Natural Resource, and Environmental Economics, Vol. 2 p.39-49. Amsterdam: Elsevier. http://dx.doi.org/10.1016/B978-0-12-375067- 9.00114-5 3. Statistical software and reproducibility: As associate editor of the ROpenSci project, and a member of the Software Carpentry foundation, I develop, evaluate, and set standards and develop training materials for open-source statistical software overseeing the publication of over 30 scientific software packages in the past two years. I have also worked in the development and dissemination of tools for the use of nonlinear modeling methods. a. Ross N (2016) fasterize: high performance raster conversion for modern spatial data. https:/ /g ithub .com/ecohealth al liance/f asterize b. c. d. Ross N (2018) Nonlinear Modeling in R with GAMs: An Interactive Course. DataCamp https://www.datacamp.com/courses/nonlinear-modelinq-in-r-with-qams D. Research Support Ongoing USAID EPT PREDICT-2 Mazet (Pl) 10/01 /14 - 09/30/19 Conducting surveillance for novel pathogens in wildlife, livestock and people; characterizing human risk behavior; modeling risk of novel disease emergence; identifying mitigation strategies Amount: $35 Million subcontract from a $100 Million award Role: Disease Ecologist 1 R01Al110964 Daszak (Pl) 06/01/14 - 05/31/19 NIAID: Understanding the Risk of Bat Coronavirus Emergence Bat ecological, human risk behavioral and virological studies to understand the risk of bat coronavirus emergence Role: Key Personnel H DTRA 1-14-1-0029 Karesh ( Pl) Understanding Rift Valley Fever in Republic of South Africa Role: Key Personnel Completed 5/17/16 - 5/16/18 W911 NF-13-1-0305 Hastings (Pl) 9/1/13-8/31/16 Army Research Office Mathematical Sciences Core Program Dynamics at Intermediate Time Scales and Management of Ecological Populations 3 Page 44 Biosketches (b) (4)

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Contact PD/Pl: DASZAK, PETER Role: Supported Graduate Student EF-0622770 Rizzo (Pl) 8/23/06-8/31/11 NSF Ecology of Infectious Disease Program Collaborative Research: Sudden Oak Death: Feedback Between a Generalist Pathogen, Hosts, and Heterogeneous Environments at Multiple Spatial and Temporal Scales Role: Supported Graduate Student 4 Page 45 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter NAME Alice Latinne eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION University of Namur, Namur, Belgium University of Liege, Liege, Belgium University of Liege, Liege, Belgium A. Personal Statement BIOGRAPHICAL SKETCH POSITION TITLE Research Scientist DEGREE MM/YY BSC 06/2004 MSC 06/2006 PHD 12/2012 FIELD OF STUDY Biology Animal Biology Molecular Biology My research focuses on understanding the dynamics of pathogens within and among wildlife populations, livestock, and humans. I have conducted fieldwork in Asia for the past 6 years, focused on the evolutionary dynamics of host-pathogen (rodent-virus; bat-virus) interactions, the phylogenetics of co-evolution, and analysis of phylogeographic scale. My main interest is to analyze he risk of zoonotic pathogen emergence at high-risk human-wildlife interfaces. My published work analyzes patterns and likelihood of pathogen sharing among species, and to determine how the host phylogenetic and phylogeographic structure affects pathogen distribution and cross-species transmission. Prior to my current position at EcoHealth Alliance, I was a Marie Curie COFUND fellow conducting postdoctoral research at the lnstitut des Sciences de !'Evolution in Montpellier (ISEM, France) and at the Kasetsart University in Thailand. B. Positions and Honors Positions and Employment 2013-2013 Research Assistant, University of Liege, Liege, Belgium 2014- Research Associate, University of Liege, Liege, Belgium 2015- Research Scientist, EcoHealth Aliiance, New York Honors 2007 2008 2013 2013 Belgian Government graduate scholarship, Belgian Fund for Research in Industry and Agriculture, Belgium Belgian Government graduate scholarship, Belgian Fund for Scientific Research, Belgium Award "VOCATIO" (Vocation grant) from the Belgian Foundation of Vocation (VOCATIO) Marie Curie COFUND fellowship from European Union C. Contribution to Science: Selected peer-reviewed publications most relevant to the current application 1. Latinne A, Beze F, Delhaes L, Pottier M, Gantois N, Nguyen J, Blasdell K, Dei-Cas E, Morand S, Chabe M (2017). Genetic diversity and evolution of Pneumocystis fungi infecting wild Southeast Asian murid rodents. Parasitology, 145(7): 885-900. PMID: 29117878 Page 46 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 2. Olival KJ, Latinne A, Islam A, Eng strand R, Hersch R, Amato G, Epstein JH, Daszak P (2016). Using bat population genetics to understand Nipah virus dynamics and cross-species transmission in south and southeast Asia. International Bat Research Conference, Durban. 3. Morand S, Bordes F, Chen H, Claude J, Gosson J, Galan M, Czirjak GA, Greenwood AD, Latinne A, Michaux J, Ribas A (2015) Global parasite and Rattus rodent invasions: the consequences for rodentborne diseases. Integrative Zoology, 10(5), 409-423. PMID: 26037785 4. Latinne A, Meynard CN, Herbreteau V, Waengsothorn S, Morand S, Michaux J (2015). Influence of past and future climate changes on the distribution of three Southeast Asian murine rodents. Journal of Biogeography, 42(9), 1714-1726. doi.org/10.1111/jbi.12528 5. Blasdell K, Bordes F, Chaval Y, Claude J, Gosson J, Latinne A, Michaux J, Morand S, Pages M, Tran A (2015). Progress on research on rodents and rodent-borne zoonoses in South-east Asia. Wildlife Research, 42(2), 98-107. doi.org/10.1071/WR14201 Additional recent publications 1. Mouton A, Mortelliti A, Grill A, Sara M, Krystufek B, Juskaitis R, Latinne A, Amori G, Randi E, Buchner S, Schulz B, Ehlers S, Lang J, Adamik P, Verbeylen G, Dorenbosch M, Trout R, Elmeros M, Aloise G, Mazzoti S, Matur F, Poitevin F, Michaux JR (2017). Evolutionary history and species delimitations: a case study of the hazel dormouse, Muscardinus avellanarius. Conservation Genetics, 18(1): 181-196. doi.org/10.1007/s 10592-016-0892-8 2. Smitz N, Cornelis D, Chardonnet P, Caron A, de Garine-Wichatitsky M, Jori F, Mouton A, Latinne A, Pigneur L, Melletti M, Kanapeckas KL, Marescaux J, Lopes-Pereira C, Michaux J (2014). Genetic structure of fragmented southern populations of African Cape buffalo ( Syncerus caffer caffer). BMC Evolutionary Biology, 14: 203. doi.org/10.1186/s12862-014-0203-2 3. Latinne A, Galan M, Waengsothorn S, Rojanadilok P, Eiamampai K, Sribuarod K, Michaux J (2014). Diet analysis of Leopoldamys neilli, a cave-dwelling rodent in Southeast Asia, using Next-Generation Sequencing from feces. Journal of Cave and Karst Studies, 76(2): 139-145. doi.org/10.4311/2013LSC0100 4. Latinne A, Chaval Y, Waengsothorn S, Rojanadilok P, Eiamampai K, Sribuarod K, Herbreteau V, Morand S, Michaux J (2013). Is Leopoldamys neilli (Rodentia, Muridae) a synonym of Leopoldamys herberti? A reply to Balakirev et al. (2013). Zootaxa, 3731 (4): 589-598. doi.org/10.11646/zootaxa.3731.4.10 5. Latinne A, Waengsothorn S, Rojanadilok P, Eiamampai K, Sribuarod K, Michaux J (2013). Diversity and endemism of Murinae rodents in Thai limestone karsts. Systematics and Biodiversity, 11 (3): 323-344. doi.org/10.1080/14772000.2013.818587 6. Pauwels OSG, Sumontha M, Latinne A, Grismer LL (2013). Cyrtodacty/us sanook (Squamata: Gekkonidae), a new cave-dwelling gecko from Chumphon Province, southern Thailand. Zootaxa, 3635(3): 275-285. PMID: 26097949 7. Latinne A, Waengsothorn S, Rojanadilok P, Eiamampai K, Sribuarod K, Michaux J (2012). Combined Mitochondrial and Nuclear Markers Revealed a Deep Vicariant History for Leopoldamys neilli, a CaveDwelling Rodent of Thailand. PLOS One, 7(10), e47670. PMID: 23118888 2 Page 47 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Hongying Li eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION School of Life Sciences, Sun Yat-Sen University, China School of Public Health, Emory University School of Life Sciences, Kingston University, UK A. Personal Statement POSITION TITLE Research Scientist DEGREE MM/YY BS 06/2012 MPH 05/2015 Ph.D Candidate 2018- FIELD OF STUDY Biosciences Health Policy Infectious Diseases I have an interdisciplinary background in ecology, public health, and human behavior, coupled with extensive on-the-ground experience working with communities, governmental and academic partners in China. For the past 3 years I have worked as China Programs Coordinator at EcoHealth Alliance, acting as the key point-of-contact among EcoHealth staff and our partners in China. I have coordinated fieldwork to conduct bat sampling, and human behavioral risk assessments across 5 provinces in southern China. I have also liaised directly with all key partners on this proposal. Additionally, I coordinate EcoHealth Alliance's wildlife trade research in China and SE Asia focusing on analyzing incentives to trade and consume wildlife. I work closely with Chinese Health and Forestry governmental departments, research institutes, and local organizations to foster collaboration and communication as part of my PhD research on "Policy and Human Behavioral Strategies to Mitigate Zoonotic Disease Emergence in Southern China". B. Positions and Honors. Positions and Employment 2011 - 12 2013-14 2015- 2017- Research Assistant of HIV Prevention Program, Yunnan Maternity and Children's Hospital, China Program Assistant of School HIV/AIDS & School Education, UNESCO Beijing, China China Programs Coordinator & Research Scientist, EcoHealth Alliance, USA Coordinator of the Initiative of National Virome Project in China Other Experience and Professional Memberships 2018- Member, IUCN SSC Pangolin Specialist Group 2018- Member, Society for Applied Microbiology 2017- Member, China Health Policy and Management Society 2016- Member, International Association for Ecology & Health 2016- Columnist, China Environment 2016- Asian Representative, Conservation Leadership Programme Honors 201 0 National Scholarship, Ministry of Education, the People's Republic of China. 2012 Outstanding Graduate Award, Sun Vat-sen University, China 2016 Invited speaker, China Conservation Network workshop. "Impacts of wildlife trade on public health" 2017 Invited Speaker, International Association for Ecology & Health. "Understanding the wildlife trade in China" Page 48 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter C. Selected peer-reviewed publications most relevant to the current application Liang X, Zhang L, Wan Y, Yu X, Guo Y, Chen X, Li H (2012). Changes in the diurnal rhythms during a 45-day head-down bed rest. PLOS One, 7(10), e47984. Wu Z, Lu L, Du J, Yang L, Ren X, Liu B, Li H, Zhu Y (2018). Comparative analysis of rodent and small mammal viromes to better understand the wildlife origin of emerging infectious diseases. Microbiome, 6(1 }, 178. Additional recent publications of importance to the field (in chronological order) Li H, Zhu G, Zhang Y, Daszak P (2018). Qualitative Approach to Developing a One Health Intervention Strategy for Zoonosis Risk Mitigation in Southern China. Poster Presentation at One Health Congress 2018. Li H, Chmura AA, Ma C, Gabriel G, Daszak P (2018). Attitudes Towards Wildlife Trade and Disease Risk in China. Poster presentation at One Health Congress 2018. Li H, Zhu G, Zhang Y, Daszak P (2018). Viral Pathogen Discovery in China: Understanding the Risks of Bat Coronaviruses. Poster presentation at USAID EPT-2 PREDICT Meeting. D. Research Support Ongoing Research Support R01 Al110964 Daszak (Pl) 06/01/14-05/31/19 Understanding Risk of Bat Coronaviruses The goal of this study is to analyze the risk of coronavirus spillover from bats to humans in Southern China Role: Project Coordinator & Human Research Lead Emerging Pandemic Threat Program, USAID Mazet (Pl) 10/01 /14-09/30/19 PREDICT2 The goal of this project is to create and implement a global virus surveillance system in animals and humans and analyze spillover risk. Role: Country Coordinator for China Completed Research Support (b) (4) Zhang (Pl) 01/01/16-12/31/17 The goal of this study is to understand the current population and distribution of the critically endangered Chinese pangolin (Manis pentadactyle) in mainland China Role: Community Research Lead 2 Page 49 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Leilani V. Francisco eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION University of Maryland, College Park, Maryland POSITION TITLE Co-Investigator DEGREE MM/YY BA 05/1995 FIELD OF STUDY Anthropology (Sociocultural) University of South Florida, Tampa, Florida MA 05/2002 Applied Anthropology (Medical) Johns Hopkins University, Baltimore, Maryland Ph.D 05/2010 Public Health (lnt'I Health) Project Manaoement Institute, Pennsylvania PMP Current Project Manaoement A. Personal Statement I have a Bachelors and Masters degree in anthropology and applied anthropology, and over 20 years of professional experience managing human behavioral research projects in public health, the majority of which has been in developing countries. I have extensive experience in the private sector, managing research projects and evaluating public health interventions for infectious diseases. I have worked extensively on: quantitative, qualitative, and mixed-methods study design, data collection, and analysis; management of behavioral intervention projects, public health assessments, and behavioral research study design. My work has focused on HIV/AIDS and other zoonotic infectious diseases, and sociocultural behavior change interventions. Previously, I managed a portfolio of global health contracts valued at over $20 million in service to the U.S. Agency for International Development (USAID), (b)(4) (b)(4), and the Centers for Disease ---------------------------- Cont r o I and Prevention (CDC). While my private sector career meant that I was not able to publish much of my work, I generated over 80 high-profile technical reports for federal and international health agencies. At EcoHealth Alliance I lead a behavioral risk team for USAID/EPT PREDICT (project ending 2019) characterizing behavioral risk in 28 countries with high-risk human-animal disease transmission interfaces. I have been B. Positions and Honors Positions and Employment 2017-Present 2017-Present 2017-Present 2013-2017 2010-2012 2010 2010 2007-2008 2007-2008 2005-2007 2004-2005 2003 2002-2004 Biosketches Senior Scientist, EcoHealth Alliance, NY USAID PREDICT-2 Global Director for Behavioral Risk Surveillance, NY USAID PREDICT-2 Partner Lead for Ecological and Biological Human Surveillance, NY Lead Associate/ Senior Lead Scientist, Booz Allen Hamilton, Washington, DC Associate I Lead Scientist, Boaz Allen Hamilton, Washington, DC Research Consultant, Johns Hopkins Bloomberg School of Public Health, Center for Communication Programs, Baltimore, MD Research Consultant, Academy for Educational Development, Washington, DC Research Fellow in Social Epidemiology, London School of Hygiene and Tropical Medicine, London, UK and Kampala, Uganda SASA! Study Baseline Project Leader, London School of Hygiene and Tropical Medicine, London, UK and Kampala, Uganda Senior Research Analyst, American Institutes for Research, Washington, DC Research Analyst, American Institute for Research, Washington, DC Research Consultant, International Center for Research on Women, Washington, DC Health Research Scientist, Battelle Memorial Institute, Arlington, VA Page 50 (b)(4)

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Contact PD/Pl: DASZAK, PETER 1999-2001 1998-1999 1998 1997-1998 1996-1997 1995-1996 Health Researcher, Battelle Memorial Institute, Arlington, VA Graduate RA, Center for Urban Transportation Research, Tampa, FL Graduate RA, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL Graduate RA, University of South Florida, Department of Anthropology, Tampa, FL Project Manager, Cultural Systems Analysis Group, Univ Maryland, College Park, Maryland RA, Cultural Systems Analysis Group, University of Maryland, College Park, Maryland Other Experience and Professional Memberships Member, American Public Health Association (APHA) Member, American Evaluation Association (AEA) Member, Global Health Council (GHC) Member, American Anthropological Association (AAA) Member, Society for Applied Anthropology (Sf AA) Honors 2007-2010 2003 Johns Hopkins University Tuition Scholarship Distinguished Service Award, Latin American Youth Center 1999 1999 Center for Urban Transportation Research Graduate Assistantship Latin American and Caribbean Studies Passport Scholarship 1998 1997-1998 Latin American and Caribbean Studies Research Grant Department of Anthropology Graduate Assistantship C. Contribution to Science 1. Ethical and Robust Human Subjects Research: My advanced training and experience in designing, carrying out, and evaluating mixed-methods research projects with vulnerable human populations, has allowed me to contribute to the body of literature and recommended practices around balancing robust study design with the ethical treatment of human subjects. With public health research and evaluation experience spanning Africa, Asia, Central America, the Caribbean, and North America, my contributions within this subject area have added to the discourse of building, implementing, and measuring scientific exploration in the name of human health improvements without compromising human privacy, dignity, and respect. a. Francisco LV, Abramsky T, Kiss L Michau L, Musuya T, Kerrigan D, Kaye D, Watts C (2013). Violence against Women and HIV Risk Behaviours in Kampala, Uganda: Baseline Findings from the SASA! Study. Violence Against Women, 19(7): 814-832. b. Wagman J, Francisco LV, Glass N, Sharps PW, Campbell JC (2008). Ethical challenges of research on and care for victims of intimate partner violence. Journal of Clinical Ethics, 19(4):371- 80. c. Campbell JC, Baty ML, Ghandour RM, Stockman JK, Francisco LV, Wagman J (2008). The intersection of intimate partner violence against women and HIV/AIDS: a review. International Journal of Injury Control and Safety Promotion, 15(4), 221-31. d. Campbell, JC, Baty ML, Ghandour RM, Stockman JK, Francisco LV, Wagman J (2008). The Intersection of Violence against Women and HIV/AIDS. In Scott KA (Rapporteur) Violence Prevention in Low- and Middle- Income Countries: Finding a Place on the Global Agenda, pp.149-166. Washington, DC: Institute of Medicine, National Academies Press. 2. Scientific approaches to behavioral intervention: Through the example of my work as a scientist with subject matter expertise in behavior change, I have built a strong case that scientific evidence can and should make its way into the hands of decision-makers and the community. This evidence-action gap is one that is often recognized, but regularly left unaddressed. My work in Kampala Uganda using a cluster randomized controlled trial to understand the impact of an intervention in preventing violence against 2 Page 51 Biosketches

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Contact PD/Pl: DASZAK, PETER women and reducing their HIV risk was recognized by Harvard University as a program that closes gender gaps in economic opportunity, politics, health, and education. It was also added to the Women and Public Policy Program's Gender Action Portal, a hub of scientific evidence providing insights on the impact of policies, strategies and practices aimed at closing gender gaps, and taking promising interventions to scale. Additionally, I led the development of a behavioral intervention resource in the form of a moderated picture book, "Living Safely with Bats," based upon feedback from communities living in countries and in areas of regular bat-human contact in their homes. This resource became a key component in ministerial and community outreach by the USAID PREDICT consortium following the announcement of the discovery of the Bombali ebolavirus in 2018, and reflects my continued efforts to translate research to practice. a. Abramsky T, Devries K, Kiss L, Nakuti J, Kyegombe N, Starmann E, Cundill B, Francisco LV, Kaye D, Musuya T, Michau L, Watts C (2014). Findings from the SASA! Study: a cluster randomised controlled trial to assess the impact of a community mobilisation intervention to prevent violence against women and reduce HIV risk in Kampala, Uganda. BMC Medicine, 12:122. b. Francisco LV, Sullivan A, Goley J, Martinez S, Saylors K, Euren J, Epstein JH, Bird B, Goldstein T, Walking D, Johnson C, Hagan E, Olival KJ, Karesh WB, Daszak P, Mazet JK (2018). Living Safely with Bats: a risk-reduction resource to help communities in developing countries change behavior to minimize zoonotic spillover from bats. USAID Washington, DC. c. Campbell JC, Baty ML, Ghandour RM, Stockman JK, Francisco LV, Wagman J (2008). The Intersection of Violence against Women and HIV/AIDS. In Scott KA (Rapporteur). Violence Prevention in Low- and Middle- Income Countries: Finding a Place on the Global Agenda, pp.149-166. Washington, DC: Institute of Medicine, The National Academies Press. 3. Applied behavioral research: Through my advanced training and experience in quantitative, qualitative, and mixed-methods research methodology I have focused on promoting the application of behavioral research to on-the-ground problems. My authorship of over 80 technical reports and publications reinforces my track record of commitment to making robustly-generated methodologies available and accessible to those who affect policy and programming. a. Francisco LV, et al. (2015). DTRA CBEP Country Assessment Manual: Guidance for Implementation of CBEP Assessments of Country Capabilities in Biosurveillance, Biosafety, and Biosecurity. Booz Allen Hamilton, Lorton, VA. b. Francisco LV, et al. (2011 ). Resilience and Prevention Study: Program Evaluation Framework for the Never Leave a Marine Behind (NLMB) Program. For the Defense Centers of Excellence for Psychological Health and Traumatic Brain Injury, US Department of Defense. Booz Allen Hamilton, Rockville, MD. c. Francisco LV (2010). Operational Plan for Ethnographic and Network Assessment Research Project. For Centers for Disease Control and Prevention HIV prevention project in Cote d'Ivoire and Zambia. Academy for Educational Development, Washington, DC. Complete List of Published Work in MyBibliography: D. Additional Information: Research Support and/or Scholastic Performance Ongoing Research Support R01 Al110964 Daszak (Pl) 06/01/14-05/31/19 NIAID: Understanding the Risk of Bat Coronavirus Emergence Bat ecological, human risk behavioral and virological studies to understand the risk of bat coronavirus emergence Role: Research Scientist USAID EPT PREDICT-2 Mazet (Pl) 10/01 /14 - 09/30/19 3 Page 52 Biosketches

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Contact PD/Pl: DASZAK, PETER Conducting surveillance for novel pathogens in wildlife, livestock and people; characterizing human risk behavior; modeling risk of novel disease emergence; identifying mitigation strategies The goal of this project is to assist focal countries in monitoring viruses with pandemic potential, as well as the behaviors, practices, and conditions that are associated with viral evolution, spillover, amplification, and spread. Role: Research Scientist Completed Research Support CDC CGH DGHT Zambia ART Bell (Project Director) 03/28/16 - 01/15/17 Centers for Disease Control and Prevention (CDC), Center for Global Health (CGH), Division of Global HIV/AIDS and Tuberculosis (DGHT), ART Readiness in HIV-infected Pregnant Women: From Formative Qualitative Research to Individual Randomized Trial - Zambia Trial monitoring visits to evaluate accuracy of screening instrument and effectiveness of enhanced adherence package through early data on virologic response, mother to child transmission (MTCT) rates of HIV, and renal function. Role: Project Manager PFSCM Projects McLaughlin (Officer in Charge) 06/01/2014- 01/15/17 Partnership for Supply Chain Management (PFSCM) Projects: USAID Supply Chain Management System (SCMS); Global Fund Pooled Procurement Mechanism (PPM); 3MDG Regional Supply Chain Strengthening (RSCS) Led and oversaw all company-wide team members and activities associated with these three projects, as part of a 16-member consortium, known as the Partnership for Supply Chain Management (PFSCM). All projects focused on increasing regular and consistent HIV/AIDS treatment through health systems strengthening, performance management, country strategic planning, and technical assistance provision. Role: Program Manager Page 53 Biosketches

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Contact PD/Pl: DASZAK, PETER NAME Amy Catherine Sims eRA COMMONS USER NAME (b) (4) EDUCATION/TRAINING INSTITUTION AND LOCATION University of Alabama at Birmingham Vanderbilt University, Nashville, TN Duke University, Durham, NC BIOGRAPHICAL SKETCH POSITION TITLE Co-Investigator DEGREE BS PhD Postdoctoral University of North Carolina at Chapel Hill (US) Postdoctoral A. Personal Statement Program Director/Principal Investigator: Daszak, Peter MM/VY FIELD OF STUDY 05/1995 Molecular Biology 05/2001 Microbiology & lmmuno 08/2002 RNA/Protein Interaction 10/2005 Virology The identification of highly pathogenic human coronaviruses (SARS-CoV and MERS-CoV) underscored the importance of understanding how viruses emerge from zoonotic reservoirs and how these emergent viruses replicate and cause pathogenesis in the new host. My research has focused on several key aspects of these questions by working to understand the cellular tropism of SARS-CoV and MERS-CoV in primary human lung cells, how host genetic pathways and gene networks affect virus replication and pathogenesis and how manipulating the coronavirus genome changes the host innate immune response to virus infection.Dr. Sims created the humanized transgenic mice that facilitate bat coronavirus replication in coronavirus small animal models and has significant expertise using the coronavirus reverse genetics platform established at UNG.She pioneered the use of primary human lung cell cultures for understanding coronavirus cellular permissivity, in vitro replication kinetics, and therapeutic treatment options within the Barie laboratory. Relevant publications: My most relevant work to date focuses on using primary human lung cells as culture models for human and human-like bat coronavirus strains. 1. Menachery VD, Yount BL, Sims AC, Agnihothram S, Gralinski LE, Plante JA, Graham RL, Scobey T, Royal s, Pickles RJ, Randell SH, Lanzavecchia A, Marasco WA, Shi z, Barie RS (2016). SAAS-like WIV1-CoV poised for human emergence. Proceedings of the National Academy of the Sciences 15:113(11 ): 3048- 53. PMC4801244 2. Becker MM, Graham RL, Donaldson EF, Rockx B, Sims AC, Timothy Sheahan, Raymond Pickles, Davide Corti, Robert E. Johnston, Ralph S. Barie, Mark R. Denison (2008). Platforms for the Synthetic Reconstitution of Noncultivable Zoonotic Viruses. Proceedings of the National Academy of the Sciences PMC2588415 3. Sims AC, Barie RS, Yount B, Burkett SE, Jeffers L, Pickles RJ (2005). SARS-CoV infection of human ciliated airway epithelium: the role of the ciliated cell in viral spread in the conducting airways of the lung. Journal of Virology 79(24):15511-15524, 2005. PMC1316022 4. Scobey T, Yount BL, Sims AC, Donaldson EF, Agnihothram SS, Menachery VD, Graham RL, Swanstrom J, Bove PF, Kim JD, Grego S, Randell SH, Barie RS. Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus.Proceedings of the National Academy of the Sciences USA. 2013 Oct 1;110(40):16157- 62. PMID: 24043791. PMC3791741 5. Sims AC, Sheahan TP, Graham RL, Menachery VD, Gralinski LE, Case JB, Leist SR, Pyre K, Feng JY, Trantcheva I, Bannister R, Park Y, Babusis D, Clarke MO, Mackman RL, Siegel D, Ray AS, Cihlar T, Jordan R, Denison MR, Barie RS (2017). Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses. Science Translational Medicine 28;9(396). PMC5567817 Page 54 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter B. Positions and Honors 1993 American Society of Microbiology Undergraduate Research Award, University of Alabama 1994 Albert Einstein College of Medicine Summer Student Award 1996 - 01 Graduate Student, Laboratory of Mark Denison, Vanderbilt University, Nashville, TN 1999 Dissertation Enhancement Award, Vanderbilt University 2001 - 02 Postdoctoral Fellow, Laboratory of Jack Keene, Duke University, Durham, NC 2002 - 05 Postdoctoral Fellow, Laboratory of Ralph Barie, UNG at Chapel Hill 2002 - 04 Infectious Disease Pathogenesis Training Grant Fellow (NIH/NIAID 5T32Al07151-27) 2005 - 17 Research Assistant Professor, Department of Epidemiology, UNG, Chapel Hill, NC 2017 - Research Associate Professor, Department of Epidemiology, UNG, Chapel Hill, NC C. Contributions to Science 1. In vitro models for viral infection. Finding suitable in vitro models for studying newly identified or emerged human respiratory viruses can be a challenge. Primary cells isolated from the human conducting airway can be cultured at an air liquid interface and following maturation recapitulate the morphology of the airway epithelium. These cultures provide a unique in vitro model and for one human coronavirus, HKU1, provide the only in vitro model for studying this virus. a) Sims AC, Pyre K, Dijkman R, Jebbink M, Long C, Deming D, Donaldson E, Vabret A, Barie RS, van der Hoek L, Pickles R (2010). Culturing the unculturable: human coronavirus HKU1 infects, replicates, and produces progeny virions in human ciliated airway epithelial cell cultures. Journal of Virology, 84(21 ): 11255-63. PMC2953148 b) Sims AC, Barie RS, Yount B, Burkett SE, Jeffers L, Pickles RJ (2005). SARS-CoV infection of human ciliated airway epithelium: the role of the ciliated cell in viral spread in the conducting airways of the lung. Journal of Virology, 79(24): 15511-15524. PMC1316022 2. Gene pathways to regulate viral replication. In collaboration with researchers at the University of Wisconsin Madison and Pacific Northwest National Laboratories, I have been working to identify specific host gene networks and pathways that regulate lethal human respiratory virus replication and pathogenesis. Specifically, I was interested in determining genes that regulate SARS-CoV and MERS-CoV replication in human cell lines, models of the human conducting airway and mouse models. a) Sims AC, Tilton SC, Menachery VD, Gralinski LE, Schafer A, Matzke MM, Webb-Robertson BM, Chang J, Luna ML, Long CE, Shukla AK, Bankhead AR, Burkett SE, Zornetzer G, Tseng CK, Metz TO, Pickles R, Mcweeney S, Smith RD, Katze MG, Waters KM, and Barie RS (2013). Release of SARS-CoV Nuclear Import Block Enhances Host Transcription in Human Lung Cells. Journal of Virology, 87(7): 3885-902. PMC3624188 b) Mitchell HD, Eisfeld AJ, Sims AC, Waters KM. A Network Integration Approach to Identify Highly Conserved Regulatory Targets Related to Pathogenicity for Influenza and SARS-CoV Respiratory Viruses. PLoS ONE 8(7): e69374. PMC3723910 c) Menachery VD, Eisfeld AJ, Josset L, Sims AC, Schaefer A, Proll S, Fan S, Li C, Neumann G, Tilton SC, Chang J, Gralinski LE, Long C, Green R, Matzke MM, Webb-Robertson BJ, Shukula AK, Burkett S, Metz TO, Pickles R, Smith RD, Waters KM, Katze M, Kawaoka Y, Barie RS (2014) .Pathogenic influenza and coronaviruses utilize similar and contrasting approaches to control global ISG responses. mBio, 5(3). PMC4030454 d) Aevermann BO, Pickett BE, Kumar S, Sims AC, Sova P, Tam VC, Tchitchek N, Thomas PG, Tilton SC, Totura A, Wang J, Webb-Robertson B, Wen J, Weiss J, Yang J, Yount B, Zhang Q, Mcweeney S, Smith RD, Waters KM, Kawaoka Y, Barie RS, Aderem A, Katze MM, Scheuermann R (2014). A Comprehensive Collection of Systems Biology Data Characterizing the Host Response to Viral Infection. Nature's Scientific Data, 1(10). 1038/sdata.2014.33. PMC4410982 Complete List of Published Work in NCBI MyBiblioqraphy: 2 Page 55 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter http://www.ncbi. nlm .nih.gov/myncbi/collections/biblioqraphy/49189460/ D. Additional Information: Research Support and/or Scholastic Performance Ongoing Research Support U19-Al106772-01 (Pl: Kawaoka) 06/01/13-05/31/19 Univ. of Wisconsin/NIH MERS-CoV Supplement for OMICs Proposal The proposed studies will provide a more detailed look at the intracellular environment by taking "snapshots" of the lipids, metabolytes, and proteins present during viral infection time courses.These assays will allow us to determine the innate immune response occurring immediately following virus infection and to determine how the virus and cell interact over a 72 hour window. Role: Project Pl U19 Al 109680 CETR (Pl: Whitley) 03/01/14-02/28/19 UAB/NIH/NIAID Antiviral Drug Discovery and Development Center The specific aims of the proposal will identify small molecule inhibitors of CoV fidelity and RNA capping, define their mechanism of action, and determine their efficacy against SARS-CoV and across CoV families using in vivo mouse models of acute and persistent CoV disease. Role: Investigator U19 Al109761 CETR (Pl: Lipkin) 03/01/14-02/28/19 Columbia/NIH/NIAID Diagnostic and Prognostic Biomarkers for Viral Severe Lung Disease The overall goal of this program is to develop new platform technologies that use functional genomics as diagnostic and prognostic indicators of severe end stage lung disease following virus infection of the lung. Role: Investigator R01 Al110700 (Pl: Barie) 04/01/15-03/31/20 NIH Mechanisms of MERS-CoV Entry, Cross-species Transmission and Pathogenesis The overall goal is to build a comprehensive understanding of the molecular mechanisms guiding group 2c CoV receptor recognition, entry and pathogenesis. Role: Investigator 1 R01 Al132178-01 (MPl:Sheahan/Baric) 08/06/17-07/31/22 NIH Broad-spectrum antiviral GS-5734 to treat MERS-CoV and related emerging CoV In partnership with Gilead Sciences, we aim to accelerate the preclinical development of GS-5734 and promote IND licensure. We define the pharmacokinetics, pharmacodynamics, resistance profile, efficacy breadth and mechanism of action of GS-5734 against MERS-CoV and related emerging CoV. Role: Investigator Completed Research Support Contract 576652 (Pl:Katze) 09/26/08-09/25/13 University of Washington/NIAID Systems Biology of Lethal and Attenuated SARS-CoV Infection The overall hypothesis is that highly pathogenic respiratory viruses use common and unique strategies to mechanistically remodel the intracellular environment to enhance virus replication, regulate disease severity and promote virus transmission.Using SARS-CoV and H1 N1 2009 and a comparative systems biology approach with H5N1 avian influenza virus we will identify unique and common signaling circuitry that is essential for promoting severe disease profiles in the lung. Role: Co-Investigator Supplement to OMIC Pilot Award (Pl: Kawaoka) 6/1/14-5/31/16 Univ. of Wisconsin/NIH/NIAID Epigenetic Regulation of Interferon-Stimulated Genes Following MERS-CoV Infection The overriding hypothesis of this supplemental application is that MERS-CoV and H5N1 manipulate host epigenetic programs to specifically down-regulate certain classes of ISGs, which likely antagonize virus replication efficiency in vitro.The goal is to develop systems biology datasets and unbiased modeling algorithms to deconvolute the complex pathogen-host interactions that regulate severe disease outcomes following infection and identify common host pathways/genes that can be exploited for therapeutic control. 3 Page 56 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Role: Project Pl U19-Al100625 (Pl: Barie) 8/05/12-07/31/17 NIH/NIAID Systems lmmunogenetics of Biodefense Pathogens in the Collaborative Cross Specific Aims:ln this proposal, we are utilizing the Collaborative Cross (CC), a novel panel of reproducible, recombinant inbred (RI) mouse lines to identify genes and gene interactions, which regulate the induction, kinetics, and magnitude of the innate, inflammatory and adaptive arms of the immune response following virus infection.Specifically, we will develop novel modeling algorithms to predict and validate the causal relationships between natural genetic variation and host signaling networks, immune cell recruitment, and immune function. Role: Investigator and Co-Education Director Supplement to OMIC (Pl: Kawaoka) 6/1/16-5/31/17 Univ. of Wisconsin/NIH/NIAID Systems Virology for MERS-CoV in vivo The goal is to develop systems biology datasets and unbiased modeling algorithms to deconvolute the complex pathoQen-host interactions that regulate severe disease outcomes followinQ infection and identify common host pathways/genes that can be exploited for therapeutic control. These studies will build on our current data set by collecting data sets for MERS-CoV in vivo. Role: Project Pl (b)(4) The overall goal of this project is to test (b)(4) protease inhibitor/interferon cocktails in comparison to and with nucleoside analog compounds to determine the best course of treatment for patients infected with highly pathogenic human coronaviruses. Not Assigned (Pl: Barie) 08/01/17-06/30/18 Emory/NIH Elucidating the potential of nucleoside analog, EIDD-1931, as a broad-spectrum antiviral against highly pathogenic human coronavirus strains To define the activity, potency and mechanism of action of EIDD-1931 against highly pathogenic human coronaviruses for development as potential therapeutic. Role: Investigator 4 Page 57 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME POSITION TITLE Emil Ann Ha an Research Scientist -=;..;.;.;.;"'-'--.;;..;.;..;'-'-'-="'-'------------------1 eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION DEGREE MM/YYYY FIELD OF STUDY Hiram College BA 05/2008 Biol. Biomed. Humanities Columbia Univ. Mailman Sch. Public Health MPH 05/2013 Epidemiology Columbia Univ. Mailman Sch. Public Health CPH 08/2013 Public Health A. Personal Statement I have a background in laboratory science, veterinary science, epidemiology, and human behavioral health. My main focus in the current proposed R01 work is on the human behavioral work in Aim 2. My experience in understanding the implications of laboratory testing, in conducting and analyzing quantitative and qualitative human behavioral risk assessment and mixed-methods data analysis are exactly the tools required to conduct this work. As assistant to the Senior Behavioral Risk Scientist on the USAID-EPT-PREDICT project I have regularly applied behavioral analytical skills to research data from 28 countries. I have also conducted my own focused work in Bangladesh, analyzing the results of 2 years of survey work on zoonotic viral spillover risk - directly applicable to the current proposal. B. Positions and Honors Positions and Employment 2006-2007 Researcher, Hiram College, Hiram College, Cellular and Molecular Lab, Hiram, OH 2007 NSF REU Research Intern, University of Akron, Polymer Department, Akron, OH 2007-2008 Teaching Assistant, Hiram College, Organic Chemistry Department, Hiram, OH 2008-2012 RA, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 2011-2012 Team Manager, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 2013-2015 2015-2016 2016-2018 RA, EcoHealth Alliance, New York, NY Research Coordinator, EcoHealth Alliance, New York, NY Research Scientist and PREDICT Bangladesh Country Liaison, EcoHealth Alliance, NY Other Experience and Professional Memberships 2013- EcoHealth, reviewer 2016- WHO Bulletin, reviewer 2018- PLOS Neglected Tropical Diseases, reviewer Honors 2008 Biology Departmental Honors, Hiram College C. Contribution to Science 1. Human behavioral risk analysis. I have helped design, coordinate training for, and conduct on-theground human behavioral work for the USAID-EPT-PREDICT project. This work focuses on the risk of zoonotic spillover of novel viruses from wildlife to people at high-risk interfaces in developing countries, and is therefore directly applicable to the research proposed for this R01. I have conducted field survey work in Page 58 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter China and Bangladesh and have published two papers directly from this work. I am currently drafting 4 manuscripts concerning behavioral risk discoveries in multiple countries. a) Wang N, Li S, Yang X, Huang H, Zhang Y, Guo H, Luo C, Miller M, Zhu G, Chmura AA, Hagan E, Zhou J, Zhang Y, Wang L, Daszak P, Shi Z (2018). Serological evidence of bat SARS-related coronavirus infection in humans, China. Virologica Sinica, 33(1), 104-107. b) Miller M, Hagan E (2017). Integrated biological-behavioural surveillance in pandemic-threat warning systems. Bulletin of the World Health Organization, 95(1 ), 62. 2. Research collaboration in developing countries. I have worked to provide support in analyzing and reporting novel serological, quantitative, and qualitative findings from in-country staff in developing countries. This has taken the form of training local anthropologists, physicians, clinicians, and social scientists in ethical data collection, methods of quantitative and qualitative data analysis, usage of the R statistical software, usage of the MAXODA qualitative analysis software, and understanding the norms of scientific journal article preparation and submission. My scientific training and interdisciplinary professional experiences will be useful in the current proposed work through coordinating survey and sample collection work in China. a) Miller M, Hagan E (2017). Integrated biological-behavioural surveillance in pandemic-threat warning systems. Bulletin of the World Health Organization, 95(1 ), 62. b) Wang N, Li S, Yang X, Huang H, Zhang Y, Guo H, Luo C, Miller M, Zhu G, Chmura AA, Hagan E, Zhou J, Zhang Y, Wang L, Daszak P, Shi Z (2018). Serological evidence of bat SARS-related coronavirus infection in humans, China. Virologica Sinica, 33(1), 104-107. D. Additional Information: Research Support and/or Scholastic Performance Ongoing Research Support R01 Al110964 Daszak (Pl) 06/01/14-05/31/19 NIAID: Understanding the Risk of Bat Coronavirus Emergence Bat ecological, human risk behavioral and virological studies to understand the risk of bat coronavirus emergence Role: Research Scientist USAID EPT PREDICT-2 Mazet (Pl) 10/01/14 - 09/30/19 Conducting surveillance for novel pathogens in wildlife, livestock and people; characterizing human risk behavior; modeling risk of novel disease emergence; identifying mitigation strategies The goal of this project is to assist focal countries in monitoring viruses with pandemic potential, as well as the behaviors, practices, and conditions that are associated with viral evolution, spillover, amplification, and spread. Role: Research Scientist Completed Research Support USAID EPT PREDICT-1 Mazet (Pl) 10/01/09 - 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses 2 Page 59 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter This project preceded PREDICT-2, described above. $75 million award to identify hotspots of emerging diseases of pandemic potential and to help guide surveillance activities and disease control and prevention strategies across several countries. Role: Research Scientist 3 Page 60 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Guangjian Zhu eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTl11JTION AND LOCATION East China Normal University, Shanghai, China Hainan Normal University, Haikou, China East China Normal University, Shanghai, China A. Personal Statement POSITION TITLE Co-Investigator DEGREE BS MS PHO MMMV FIELD OF STUDY 07/03 Biology Science 07/03 Ecology 6/12 Biochemistry/Mo!. Biol. Throughout my graduate studies and work with East China Normal University, I have carried out molecular biology and field ecological research focused on bat genetics and viral diversity. I have co-authored multiple publications in the field of viral genetics and bat ecology under the mentorship of Ors. Daszak (EcoHealth Alliance) and Shi (Wuhan Inst. Virol.). For the past 9 years I have been the in-country coordinator for all EcoHealth Alliance work in China on USAID- EPT-PREDICT, as well as for the previous NIAID R01 on bat Co Vs. During this time I have been responsible for the identification of high-risk interfaces between wildlife and people, where close contact might allow for zoonotic pathogen spillover. I have also led wildlife surveys which involved bat and rodent capture and sampling for viral discovery. Through this work I have led site-selection and wild and domestic animal sampling in Guangxi, Yunnan, Guangdong and Shanghai, and have compiled archived and current samples from birds in Shanghai Chongming Reserve for H7N9 avian influenza analyses. Under the USAID PREDICT program I collected over 1,000 bat samples which have been tested for coronaviruses and other viral families at the Wuhan Institute o1 Virology. Under our previous NIAID award (R01Al110964), I am responsible for developing and leading a wildlife team to sample bats, rodents, and other small mammals in the live animal markets of southern China. I will continue these efforts in our renewal proposal as we focus our efforts on centers of CoV diversity in southern China. Through my graduate and professional work I have developed expertise in collecting high-quality, non-destructive samples from wildlife as well as expertise in molecular diagnostics. B. Positions and Honors Positions and Employment 2007- Assistant Researcher, Guangdong Entomological Institute, China Other Experience and Professional Memberships Honors 2009 Biology Prize of the 2009 lg Nobel Prize (Tan et al. 2009, PLoS One) C. Selected peer-reviewed publications most relevant to the current application Ge XY, Li JL, Yang X-L, Chmura M, Zhu G, Epstein JH, Mazet JK, Hu B, Zhang W, Peng C, Zhang YJ, Luo CM, Tan B, Wang N, Zhu Y, Crameri G, Zhang SY, Wang LF, Oaszak P, Shi Z (2013). Isolation and characterization of a bat SARS-like Coronavirus that uses the ACE2 receptor. Nature 503: 535-538. Page 61 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Zhu G, Han N, Hong T, Tan M, Yu D, Zhang L (2008). Echolocation Call, Roost and ND 1 Sequence Analysis of New Record of Nyctalus plancyi (Chiroptera: Vespertilionidae) on Hainan Island. Zoological Research, 29(4): 447-451. Zhu G, Li D, Ye J, Hong T, Zhang L (2008). New Record of la io in Hainan Island, its Echolocation Pulses and ND1 Analysis. Chinese Journal of Zoology, 43(5): 69-75. Sun Y, Yu D, Zhu G, Liu X, Zhang SY, Chen J (2009). Isolation and characterization of 11 microsatellite loci in Scotophilus kuhlii (Lesser Asiatic Yellow House Bat). Conservation Genetics, 10: 1857-1859. Mao X, Zhu G, Zhang SY, Rossiter SJ (2010). Pleistocene climatic cycling drives intra-specific diversification in the intermediate horseshoe bat (Rhinolophus affinis) in Southern China. Molecular Ecology, 19(13): 2754-2769. Hua P, Zhang L, Zhu G, Jones G, Zhang SY, Rossiter SJ (2011 ). Hierarchical polygyny in multiparous lesser flat-headed bats. Molecular Ecology, 20(17): 3669-3680. Additional recent publications of importance to the field (in chronological oder) Mazet JAK, Wei Q, Zhao G, Cummings DAT, Desmond JS, Rosenthal J, King CH, Cao W, Chmura AA, Hagan EA, Zhang S, Xiao X, Xu J, Shi Z, Feng F, Liu X, Pan W, Zhu G, Zuo G, Daszak P (2015). Joint China-Us Call for Employing a Transdisciplinary Approach to Emerging Infectious Diseases. EcoHealth 12(4 ): 555-559. Hu B, Chmura AA, Li J, Zhu G, Desmond JS, Zhang YJ, Zhang JS, Epstein JH, Daszak P, Shi Z (2014). Detection of Diverse Novel Astroviruses from Small Mammals in China. Journal of General Virology 95: 2442-2449. Zhu G, Wang R, Xuan F, Daszak P, Anthony SJ, Zhang SY, Zhang L, He G (2013). Characterization of Recombinant H9n2 Influenza Viruses Isolated from Wild Ducks in China. Veterinary Microbiology 166(4): 327-336. Zhu G, Chmura AA, Zhang L (2011 ). Morphology, echolocation calls and diet of Scotophilus kuhlii (Chiroptera: Vespertilionidae) on Hainan Island, south China. Acta Chiropterologica, 14(1): 175-181. Ma J, Jones G, Zhu G, Metzner W (2010). Echolocation behaviours of the Japanese pipistrelle bat Pipistrellus abramus during foraging flight. Acta Theriologica, 55(4): 315-332. Tan M, Jones G, Zhu G, Ye J, Hong T, Zhou S, Zhang S, Zhang L (2009). Fellatio by fruit bats prolongs copulation time. PLOS One, 4(10), e7595. Zhang L, Zhu G, Jones G, Zhang SY (2009). Conservation of bats in China: problems and recommendations. ORYX, 43(2): 179-182. Zhu G, Tang Z, Liang B, Zhang X (2007). Diet and Roost Site of Cynopterus sphinx in Winter in Haikou. Chinese Journal of Zoology, 42(4): 22-27. D. Research Support Ongoing Research Support USAID EPT PREDICT-1 Biosketches Mazet (Pl) 1 0/01 /09 - 09/30/14 2 Page 62

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Amount: $18 million subcontract on a $75 million award Role: Lead Field Scientist 1 R01 Al110964 Daszak (Pl) 06/01 /14 - 05/31 /19 NIAID: Understanding the Risk of Bat Coronavirus Emergence Bat ecological, human risk behavioral and virological studies to understand the risk of bat coronavirus emergence Amount: $2.5 million Role: Lead Field Scientist Completed Research Support USAID EPT PREDICT-1 Mazet (Pl) 10/01/09 - 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Amount: $18 million subcontract on a $75 million award Role: Lead Field Scientist 1 R01Al079231 Daszak (Pl) 09/18/08-08/31/13 NIAID Non-Biodefense Emerging Infectious Diseases Risk of viral emergence from bats. To model hotspots for bat viral diversity, identify & characterize new bat viruses & understand their pathology Role: Research Scientist 3 Page 63 Biosketches

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Contact PD/Pl: DASZAK, PETER NAME Linfa Wang eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION East China Normal University, Shanghai University of California, Davis A.PERSONAL STATEMENT Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH POSITION TITLE Co-Investigator DEGREE BS PHD MMNY FIELD OF STUDY 01/1982 Biology 07/2086 Biochemistry My 20+ years of research focused on designing and applying novel testing platforms to discover zoonotic pathogens has direct applicability to the current proposal. I am trained as a biochemist and molecular biologist, and have been working in the field of virology and infectious diseases for more than 20 years, playing a key role in identification of animal links with several high profile zoonotic agents, including Hendra virus in Australia, Nipah virus in Malaysia and SARS virus in China. During this time, I've directed largescale laboratory diagnostic studies based on serological and PCR surveys of wildlife, domestic animals and people; and worked with multidisciplinary lab, field and modeling teams, including those at EcoHealth Alliance, to interpret the results. In my current role as director of the Program in Emerging Infectious Diseases at Duke-NUS Graduate Medical School, I have initiated major projects to continue this work, and to analyze bat genomics and basic bat biology to better understand bat-unique biological features such as longevity and co-existence with pathogens with no or minimal clinical disease. This work has led to a number of patented techniques, as well as novel reagents that I have made available to collaborators and the greater scientific community. Over the years, I have established an extensive collaborative network with scientists all around the world, covering research and surveillance work into infections of human, animal and wildlife in a truly One Health approach, including many of the collaborators on the current proposal. Li W, Shi Z, Yu M, Ren W, Smith C, Epstein JH, Wang H, Crameri G, Hu Z, Zhang H, Zhang J, McEachern J, Field H, Daszak P, Eaton BT, Zhang S, Wang LF (2005) Bats are natural reseNoir of SARS-like coronaviruses. Science 31 0: 676-679. Ge XY, Li JL, Yang X-L, Chmura AA, Zhu G, Epstein JH, Mazet JK, Hu 8, Zhang W, Peng C, Zhang YJ, Luo CM, Tan B, Wang N, Zhu Y, Crameri G, Zhang SY, Wang LF, Daszak P, Shi Z (2013). Isolation and characterization of a bat SARS-like Coronavirus that uses the ACE2 receptor. Nature 503: 535-538. Zhang G, Cowled C, Shi Z, Huang Z, Bishop-Lilly KA, Fang X, Wynne JW, Xiong Z, Baker ML, Zhao W, Tachedjian M, Zhu Y, Zhou P, Jiang X, Ng J, Yang L, Wu L, Xiao J, Feng Y, Chen Y, Sun X, Zhang Y, Marsh GA, Crameri G, Broder CC, Frey KG, Wang LF Wang J (2013) Comparative Analysis of Bat Genomes Provides Insight into the Evolution of Flight and Immunity. Science 339: 456-60. Zhou P, Fan H, Lan T, Yang XL, Shi WF, Zhang W, Zhu Y, Zhang YW, Xie QM, Mani S, Zheng XS, Li B, Li JM, Guo H, Pei GO, An XP, Chen JW, Zhou L, Mai KJ, Wu ZX, Li D, Anderson DE, Zhang LB, Li SY, Mi 20, He TT, Cong F, Guo PJ, Huang R, Luo Y, Liu XL, Chen J, Huang Y, Sun Q, Zhang XL, Wang VY, Xing SZ, Chen YS, Sun Y, Li J, Daszak P, Wang LF, Shi ZL, Tong YG, Ma JY (2018). Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin. Nature 556: 255-258 Patents: A protease deficient Bacillus subtilis mutant strain US patent No. 5,585,253; Bacillus subtilis expression and secretion system. US patent No. 7,238,560; Footrot antigens, vaccines and diagnostic assays. Page 64 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Australian Patent No. 38377/93; A novel epitope tagging system for protein surveillance and purification. Australian Patent No. PM? 419/94; Assay for the Parallel Detection of Biological Material Based on PCR. PCT/SG2013/000455; A Chimeric Animal Comprising Stably Transplanted Bat Cells. IMC/P/10031/00/SG B. POSITIONS AND HONORS Positions and Employment 1986 - 89 1990 1990 - 92 1992 - 96 1996 - 04 2004 - 08 2008 - 15 2012 - Post-doctoral Fellow, Department of Biochemistry, University of California, Davis, USA Senior Research Officer, Centre for Molecular Biology and Medicine, Monash University, Australia Research Scientist, CSIRO Australian Animal Health Laboratory (AAHL), Geelong, Australia Senior Research Scientist, CSIRO AAHL, Geelong, Australia Principal Research Scientist, CSIRO AAHL, Geelong, Australia Senior Principal Research Scientist, CSIRO AAHL, Geelong, Australia OGE Science Leader, CSIRO AAHL, Geelong, Australia Professor & Director, Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore Other Experience and Professional Memberships 1996 - 2003 2005- 2006 - 2006 - 7 2008- 2009- 2010 - 2012 - 2012 - 2012 - 2013 - 2017 - Editorial Board, Asia Pacific J. Mot. Biol. Biotech. WHO SARS Scientific Research Advisory Committee Honorary Professor, Wuhan Institute of Virology, Chinese Academy of Sciences Editorial Board, Chinese J. Virol.; Zoonoses & Pub/. Hlth. NH & MRC Grant Review Panel Chair, ICTV Study Group, Paramyxoviridae Honorary Professor, University of Melbourne, Australia Editorial Board, Frontiers Virol. Editor-in-Chief, Viral. J. Board of Directors, Singapore Eye Research Institute Executive Committee, Australasian Society of Virology WHO International Health Regulations, Roster of Experts World Economic Forum, Global Health Threat Advisory Board Selected Awards/Honors: 2006 CSIRO Award for Excellence in Partnership 2007 Finalist, Eureka Prize for Scientific Research 2008 CSIRO CEO Science Leader Award 2010 Elected fellow of the Australian Academy of Technological Sciences and Engineering 2011 Gardner Lecture Award, European Society of Clinical Virologist 2013 CSIRO Chairman's Medal for Research 2014 Winner, Eureka Prize for Infectious Disease Research 2014 Finalist, Prime Minister's Science Award, Australia C. CONTRIBUTION TO SCIENCE 1. Application of both molecular and serological platforms to pathogen discovery My work at CSIRO AAHL, and now at Duke-NUS has focused on the development and use of PCR and serological assays to identify novel pathogens in wildlife, livestock and people, often under outbreak conditions. This includes the discovery of bats as a reservoir for SARS-CoV, using novel serological assays and PCR techniques I developed. 2 Page 65 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter a. Bossart KN, McEacherna JA, Hickey AC, Choudhry V, Dimitrov DS, Eaton BT, Wang LF (2007) Neutralization assays for differential henipavirus serology using Bio-Plex Protein Array Systems. Journal of Virological Methods, 142: 29-40. b. Thalmann CM, Cummins DM, Yu M, Lunt R, Pritchard LI, Hansson E, Crameri S, Hyatt A, Wang LF (2010) Broome virus, a new fusogenic Orthoreovirus species isolated from an Australian fruit bat. Virology 402:26-40. c. Cui J, Tachedjian G, Tachedjian M, Holmes EC, Zhang SY, Wang LF (2012) Identification of diverse groups of endogenous gammaretroviruses in mega- and microbats. Journal of General Virology 93:2037-2045. d. Wang J, Selleck P, Yu M, Ha W, Rootes C, Gales R, Wise T, Crameri S, Chen H, Broz I, Hyatt A, Woods R, Meehan B, McCullough S, Wang LF (2014) Novel Phlebovirus with Zoonotic Potential Isolated from Ticks, Australia. Emerging Infectious Diseases 20:1040-1043. 2. Identification of bats as major reservoir of emerging zoonotic viruses I have used surveillance in wildlife, livestock and humans, coupled with experimental infections under BSL2, -3, and -4, and laboratory assays to identify evidence that bats are the reservoir for a series of emerging viruses in people, including Hendra virus, Nipah virus, SARS-CoV, and others. This work has been one of the foundations for current interest in bats in emerging infectious disease research. a. Eaton BT, Broder CC, Middleton D, and Wang LF, (2006). Hendra and Nipah viruses: different and dangerous. Nature Reviews Microbiology, 4: 23-35. b. Chua KB, Crameri C, Hyatt A, Yu M, Tompang MR, Rosli J, McEachern J, Crameri S, Kumarasamy V, Eaton BT, Wang LF (2007). A previously unknown reovirus of bat origin is associated with an acute respiratory disease in humans. Proceedings of the National Academy of Sciences, 27: 11424-11429. c. Mahalingam S, Herrero LJ, Playford G, Spann K, Herring B, Rolph R, Middleton D, McCall B, Field H, Wang LF (2012) Hendra virus: an emerging paramyxovirus in Australia. Lancet Infectious Diseases 12: 799-807. d. Clayton BA, Middleton D, Arkinstall R, Frazer L, Wang LF, Marsh GA (2016) The Nature of Exposure Drives Transmission of Nipah Viruses from Malaysia and Bangladesh in Ferrets. PLOS Neglected Tropical Diseases, 10(6): e0004775. 3. Establishment of bats as a new mammalian model system to study virus-host interaction and evolutionary biology Working with collaborators around the world, my lab has amassed an unprecedented collection of serological, tissue and other samples from bat surveillance programs. I have used these to develop and disseminate primary and immortalized bat cell lines, and a host of reagents which my team and collaborators are using to test hypotheses about why bats are able to host so many distinct viruses. Current projects include bat genomics and proteomics; examining the bat MHC, using gene knockout technology to identify links between flight, viral resistance, and longevity. a. Wynne JW, Shiell BJ, Marsh G, Boyd V, Monaghan P, Zhou P, Klein R, Todd S, Mok L, Green D, Tachedjian M, Baker M, Matthews D, Wang LF (2014). Proteomics informed by transcriptomics reveals Hendra virus sensitizes bat cells to TRAIL mediated apoptosis. Genome Biology 15: 532. b. Zhou P, Tachedjian M, Wynne JW, Boyd V, Cui J, Smith I, Cowled C, Ng JH, Mok L, Michalski WP, Mendenhall lH, Tachedjian G, Wang LF, Baker ML (2016). Contraction of the type I IFN locus and unusual constitutive expression of IFN-a in bats. Proceedings of the National Academy of Sciences, 113: 2696-2701. 3 Page 66 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter c. Xie J, Li Y, Shen X, Goh G, Zhu Y, Cui J, Wang LF, Shi Z, Zhou P (2018). Dampened STINGDependent Interferon Activation in Bats. Cell Host and Microbe, 23(3):297-301. d. Yong KSM, Ng JHJ, Her Z, Hey YY, Tan SY, Tan WWS, lrac SE, Liu M, Chan XY, Gunawan M, Foo RJH, Low DHW, Mendenhall IH, Chionh YT, Dutertre CA, Chen Q, Wang LF (2018). Batmouse bone marrow chimera: a novel animal model for dissecting the uniqueness of the bat immune system. Science Reports, 8(1) :4726. D.RESEARCHSUPPORT Ongoing research support NRF2012NRF-CRP001-056 Wang (Pl) 01 /11/13-31/10/18 National Research Foundation (NRF, Singapore) Learning from bats: from genomics to controlling viral infection and combating cancer Using bats as model to study immunology, inflammation and other cellular/molecular mechanisms which are responsible for the unique biological features of bats, such as longevity and infection with no or less diseases. Role: Leading Pl Al212961 NIH Crump (Pl) Investigating Febrile Deaths in Tanzania (INDITe) 01/02/16-31/01/21 To identify actionable patient management and health system interventions that could avert fatal outcomes among patients with severe febrile illness in low-resource areas. Role: Co-Pl (b) (4) Development of multiple serological platforms for differentiation of Zika and dengue virus infections Using multiple multiplex serological platforms to develop antibody tests which can differentiation infections of Zika virus from Dengue virus and other closely related flaviviruses. Role: Pl Completed (b) (4) Establishment of serological diagnostic capability for highly virulent zoonotic viral infections in Singapore Using most advanced technological platforms to enhance the capability in diagnosing and responding to future zoonotic disease outbreaks in Singapore. Role: Pl (b) (4) Understanding the host pathogen relationships of Hendra Virus in bats, horses and humans Examines why bats can be infected with Hendra Virus with no apparent symptoms, yet the virus causes severe disease in other mammals including humans. We hope this information can be used to design new drugs or vaccines to Hendra Virus. Role: Co-Pl 4 Page 67 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter (b) (4) Improving the management of an emerging viral disease In Australia: determination of the mechanisms of neuroinvasion by Hendra Virus and their control, leading to optimization of post-exposure therapy following contact with Hendra Virus Using a recently established mouse infection model, this study aims to elucidate the mechanism of Hendra Virus neuroinvasion and to optimize the post-exposure therapy strategies. Role: Chief lnvestigaor (b)(4) New targets in antiviral therapies Development of novel antiviral strategies based on the interruption of nuclear localization process of key virus proteins in the families of Paramyxoviridae and Rhabdoviridae. Role: Co-Pl Al077995 NIH/NIAID Broder (Pl) Vaccines and therapeutics for Nipah and Hendra Virus 01/06/07-31/05/13 Establish virus infection, lethal dose, and detection parameters of Nipah virus in a ferret model. 2. Evaluate the protective efficacy of recombinant sG as a subunit vaccine for Nipah virus in the ferret. 3. Determine the passive protective efficacy of neutralizing, anti-G, fully-human monoclonal antibody therapy for Nipah virus infection in the cat and ferret. 4. Determine the solution structure of Nipah sG and in complex with its receptor ephrinB2. Role: Co-Pl 5 Page 68 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Lili Ren eRA COMMONS USER NAME (credential, e.g., agency login) (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Norman Bethune Univ. Med. Sci., China Medical College, Jilin university Ctr. Disease Control & Prevention, China A. Personal Statement POSITION TITLE Co-Investigator DEGREE MM/YY BS 07/1998 MS 07/2002 Ph.D 07/2005 FIELD OF STUDY Clinical medicine Pathology, pathophysiol. Immunology I have expertise in the pathogenesis and evolution of respiratory viruses. Using systems biological concepts and technical systems, my group has identified several new emerging respiratory virus epidemics in China, clarifying new causative viral agents of severe pneumonia, providing insights into the pathogenic mechanisms of viruses, and improving clinical pathogen diagnostics. My group has also conducted a longitudinal study to analyze evolution of known common respiratory viruses since 2005 - the largest and the longest continuous study of its type in China. My multiple pathogen detection system approach has been transferred to more than 300 national sentinel sites. In addition, my group has established several platforms to identify the serological epidemiology of common respiratory viruses including human CoVs. This will be a critical reference for the proposed R01 work, and have implications on the design of our serological screening platform. I have also established a collaborative group with clinicians in China to study the ecology of emerging viral pathogens. My extensive experience in this type of study will greatly assist in my role overseeing clinical sample collection, and screening by PCR and serological methods in southern China. B. Positions and Honors. Positions and Employment 2005-2010 Research assistant, Institute of Pathogen Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, China 2010-2016, Research scientist, Institute of Pathogen Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, China 2016- Senior Scientist, Institute of Pathogen Biology, Chinese Academy of Medical Sciences/Peking Union Medical College, China Other Experience and Professional Memberships 2016-2019 2014-2020 2016-2021 2017- Biosketches Member, Expert Committee on Biosafety Assessment of the National Health and Family Planning Commission Member, Youth Committee of the Chinese Medical Association Medical Virology Branch Member, China Research Hospital, Space Microbiology and Infection Committee Branch Editorial board, Chinese Journal of Experimental & Clinical Virology Page 69

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Contact PD/Pl: DASZAK, PETER Honors 2008 - 10 2015 2016 2017 Program Director/Principal Investigator: Daszak, Peter Outstanding Young Talents of New Century (NCET-07-0506), granted by Ministry of Education, China. Principal Investigator. 2nd prize of the advanced science and technology progress award, second author, granted by Ministry of Education, China. 1st prize of the advanced science and technology progress award, second author, granted by Ministry of Education, China. Excellent teacher of Peking Union Medical College (PUMC), China C. Selected peer-reviewed publications most relevant to the current application *=Co-corresponding or first authors Ren L*, Richard G*, Wang Z*, Xiang Z*, Wang Y*, Zhou H, Li J, Xiao Y, Yang Q, Zhang J, Chen L, Wang W, Li Y, Li T, Meng X, Zhang Y, Guy V, Chen J, Jin Q, Wang J (2009) Prevalence of human respiratory viruses in adults with acute respiratory tract infections in Beijing, 2005-2007. Clinical Microbiology and Infection, 15(12): 1146-1153. Ren L, Gonzalez R, Xu X, Li J, Zhang J, Vernet G, Paranhos-Baccala G, Jin Q, Wang J (2009) WU polyomavirus in fecal specimens of children with acute gastroenteritis, China. Emerging Infectious Diseases, 15(1 ): 134-135. Ren L, Gonzalez R, Xiao Y, Xu X, Chen L, Vernet G, Paranhos-Baccala G, Jin Q, Wang J (2009) Saffold cardiovirus in children with acute gastroenteritis, Beijing, China. Emerging Infectious Diseases 15(9): 1509-1511. Ren L, Gonzalez R, Xu J, Xiao Y, Li Y, Zhou H, Li J, Yang Q, Zhang J, Chen L, Wang W, Vernet G, Paranhos-Baccala G, Wang Z, Wang J (2011 ). Prevalence of human coronaviruses in adults with acute respiratory tract infections in Beijing, China. Journal of Medical Virology 83(2): 291-297. Yang J*, Yang F*, Ren L*, Xiong Z, Wu Z, Dong J, Sun L, Zhang T, Hu Y, Du J, Wang J, Jin Q (2011). Unbiased parallel detection of viral pathogens in clinical samples by use of a metagenomic approach. Journal of Clinical Microbiology, 49(10): 3463-3469. Guo L*, Zhang X*, Ren L*, Yu X*, Chen L*, Zhou H, Gao X, Teng Z, Li J, Hu J, Wu C, Xiao X, Zhu Y, Wang Q, Pang X, Jin Q, Wu F, Wang J (2014). Human antibody responses to avian influenza A(H7N9) virus, 2013. Emerging Infectious Diseases, 20(2): 192-200. Ren L*, Yu X*, Zhao B*, Wu F, Jin Q, Zhang X, Wang J (2014). Infection with possible precursor of avian influenza A(H7N9) virus in a Child, China, 2013, Emerging Infectious Diseases, 20(8): 1362-1365. Ren L*, Zhang Y*, Li J, Xiao Y, Zhang J, Wang Y, Chen L, Paranhos-Baccala G, Wang J (2015). Genetic drift of human coronavirus OC43 spike gene during adaptive evolution. Scientific Reports, 5:11451. doi .org/1 0 .1 038/srep 11451 Zhang Y, Li J, Xiao Y, Zhang J, Wang Y, Chen L, Paranhos-Baccala G, Ren L*, Wang J* (2015). Genotype shift in human coronavirus OC43 and emergence of a novel genotype by natural recombination. Journal of Infection, 70(6): 641-650. Yan F, Xiao Y, Li M, Zhang H, Zhang R, Zhou H, Shen H, Wang J, Li W*, Ren L* (2017). Metagenomic analysis identified human rhinovirus B91 infection in an adult suffering from severe pneumonia. American Journal of Respiratory and Critical Care Medicine, 195(11 ):1535-1536. 2 Page 70 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Ren L*, Yang D*, Ren X*, Li M, Mu X, Wang Q, Cao J, Hu K, Yan C, Fan H, Li X, Chen Y, Wang R, An F, An S, Luo M, Wang Y, Xiao Y, Xiang Z, Xiao Y, Li L, Huang F, Jin Q, Gao Z, Wang J (2017). Genotyping of human rhinovirus in adult patients with acute respiratory virus infections identified predominant infections of genotype A21 . Scientific Reports, 7:41601 . Ren L*, Zhang R*, Rao J, Xiao Y, Zhang Z, Yang B, Cao D, Zhong H, Ning P, Shang H, Li M, Gao Z, Wang J (2018). Transcriptionally Active Lung Microbiome and Its Association with Bacterial Biomass and Host Inflammatory Status. mSystems, 3:e00199-18. D. Research Support Ongoing Research Support 1. 2017ZX10103004, Key project of infectious diseases 01 /01/2017-12/31/2020 Viral etiology and spectrum of respiratory tract infections and the mutations characteristics The goal of this project is to identify the etiology of community acquired pneumonia in China and the epidemic and mutations of the important respiratory viruses. Role: Pl Completed Research Support 1. 2012ZX10004-206 Key project of infectious diseases 01/01/2012-12/31/2015 Viral etiology and spectrum of respiratory tract infections and the mutations characteristics The goal of this project is to investigate the etiology of acute respiratory tract infections in China and the seroepidemiological of important respiratory viruses. Role: Pl 2. 2009ZX10004-206 Key project of infectious diseases 01/01/2009-12/31 /2010 Viral etiology and spectrum of respiratory tract infections and the mutations characteristics The goal of this project is to establish the surveillance network and to investigate the etiology of acute respiratory tract infections in China. Role: Pl 3 Page 71 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter NAME Li Guo eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Bengbu Medical College, China Anhui Medical University, China BIOGRAPHICAL SKETCH POSITION TITLE Co-Investigator DEGREE B.S. M.S. Chinese Center for Disease Control & Prevention M.D. A. Personal Statement MM/YY FIELD OF STUDY 07/1997 Clinical Medicine 07/2002 Microbiology 07/2006 Immunology I have direct expertise in the proposed work described in this R01. I have been working on the etiology and immunology of respiratory viruses since 2003. I have evaluated cross-reactivity of NP among HCoVs and developed a competitive ELISA (cELISA) for detecting anti-N lgG antibodies against HCoV -229E, -OC43, -NL63, and -HKU1. These data indicate differential transmission of HCoVs in the Chinese population and that anti-N lgG may serves as an index for susceptibility to HCoV infections. In addition, I have evaluated human antibody responses to A(H7N9) influenza virus, differential seroprevalence of human bocavirus species 1-4 in China, and found bocavirus in children with respiratory tract infections. B. Positions and Honors. Positions and Employment 1997 - 99 2002- 09 2009- 15 2015 - Teaching Assistant, Bengbu Medical College Assistant Professor, Chinese Center for Disease Control and Prevention Associate Professor, Institute of Pathogen Biology (IPB), Chinese Academy of Medical Science (CAMS) /Peking Union Medical College. Professor, Institute of Pathogen Biology (IPB), Chinese Academy of Medical Science (CAMS) /Peking Union Medical College C. Selected peer-reviewed publications • = Co-corresponding or first author Guo L, Wang D, Zhou H, Wu C, Gao X, Xiao Y, Ren L, Paranhos-Baccala G, Shu Y, Jin Q, Wang J (2016). Cross-reactivity between avian influenza A (H7N9) virus and divergent H7 subtypic and heterosubtypic influenza A viruses. Scientific Reports, 6, 22045. Gao X, Zhou H, Wu C, Xiao Y, Ren L, Paranhos-Baccala G, Guo L *, Wang J* (2015). Antibody against nucleocapsid protein predicts susceptibility to human coronavirus infection. Journal of Infection 71 (5): 599-602. Chen z•, Wang J*, Bao L*, Guo L*, Zhang W, Xue Y, Zhou H, Xiao Y, Wang J, Wu F, Deng Y, Qin C, Jin Q (2015). Human monoclonal antibodies targeting the haemagglutinin glycoprotein can neutralize H7N9 influenza virus. Nature Communications, 6:6714. Page 72 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Yang J*, Zhang T*, Guo L *, Hu YF, Li JL, Su HX, Xiao Y, Ren XW, Dong J, Sun LL, Xiao Y, Li Li, Yang F, Wang JW, Yuan H, Jin Q (2014). Mutations of Novel Influenza A(H10N8) Virus in Chicken Eggs and MOCK Cells. Emergining Infectious Diseases, 20(9) :1541-1543. Zhou Z, Gao X, Wang Y, Zhou H, Wu C, Paranhos-Baccala G, Vernet G, Guo L*, Wang J* (2014). Conserved B-Cell Epitopes among Human Bocavirus Species Indicate Potential Diagnostic Targets. PLOS One, 9(1): e86960. Guo L, Zhang X, Ren L, Yu X, Chen L, Zhou H, Gao X, Teng Z, Li J, Hu J, Wu C, Xiao X, Zhu Y, Wang Q, Pang X, Jin Q, Wu F, Wang J (2014). Human antibody responses to avian influenza A(H7N9) virus. Emerging Infectious Diseases, 20(2): 192-200. Guo L, Wu C, Zhou H, Wu C, Paranhos-Baccala G, Vernet G, Jin 0, Wang J, Hung T (2013). Identification of a nonstructural DNA-binding protein (DBP) as an antigen with diagnostic potential for human adenovirus. PLOS One, 8(3): e56708. Guo L, Wang Y, Zhou H, Wu C, Song J, Li J, Paranhos-Baccala G, Vernet G, Wang J, Hung T (2012). Differential seroprevalence of human bocavirus species 1-4 in Beijing, China. PLOS One, 7(6): e39644. Guo L, Gonzalez R, Zhou H, Wu C, Vernet G, Wang Z, Wang J (2012). Detection of three human adenovirus species in adults with acute respiratory infection in China. European Journal of Clinical Microbiology and Infectious Disease, 31(6): 1051-1058. Guo L, Gonzalez R, Xie Z, Zhou H, Liu C, Wu C, Paranhos-Baccala G, Vernet G, Shen K, Jin Q, Wang J (2011). Bocavirus in children with respiratory tract infections. Emerging Infectious Diseases, 17(9): 1775-1777. Wang Y, Gonzalez R, Zhou H, Li J, Li Y, Paranhos-Baccala G, Vernet G, Guo L*, Wang J* (2011). Detection of human bocavirus 3 in China. European Journal of Clinical Microbiology and Infectious Disease, 30(6): 799-805. Guo L, Gonzalez R, Wang W, Vernet G, Paranhos-Baccala G, Wang J (2010). Complete genome sequence of human astrovirus genotype 6. Virology Journal, 7: 29. Guo L, Xu X, Song J, Wang W, Wang J, Hung T (2010). Molecular Characterization of Astrovirus Infection in Children with Diarrhea in Beijing, 2005-2007. Journal of Medical Virology, 82(3): 415-423. Guo L, Zhou H, Wang M, Song J, Han B, Shu Y, Ren L, Si H, Ou J, Zhao Z, Wang J, Hung T (2009). A recombinant adenovirus prime-virus-like particle boost regimen elicits effective and specific immunities against norovirus in mice. Vaccine, 27(38): 5233-5238. Guo L, Song J, Xu X, Ren L, Li J, Zhou H, Wang M, Ou J, Wang J, Hung T (2009). Genetic analysis of norovirus in children affected by acute gastroenteritis in Beijing, 2004-2007. Journal of Clinical Virology 44(1): 94-98. Guo L, Wang J, Zhou H, Si H, Wang M, Song J, Han B, Shu Y, Ren L, Ou J, Hung T (2008) Intranasal administration of a recombinant adenovirus expressing the norovirus capsid protein stimulates specific humeral, mucosal, and cellular immune responses in mice. Vaccine, 26(4): 460-468. D. Research Support Ongoing Research Support 2018ZX10734404-006 Biosketches 01/01 /2018-12/31 /2020 2 Page 73

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Key technologies for the identification and identification of important respiratory viruses and establishment of reference libraries Role: Co-Pl Completed Research Support (b) (4) (b)(4) Study on the immunoprotection of recombinant adenovirus vaccine against Norovirus by using virus-like particles as a control Role: Pl 3 Page 74 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Peng Zhou eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION College of Life Science, Henan University, China Wuhan Inst. Virol., Chinese Acad. Sci., China Australian Animal Health Laboratory, CSIRO Sinqapore Duke-NUS Medical School, Sinqapore A. Personal Statement POSITION TITLE Co-Investigator DEGREE BS PHO Postdoc Postdoc MM/YY FIELD OF STUDY 07/04 Bioengineering 01/11 Virology 07/14 Viral Immunology 02/16 Viral lmmunoloqy My virological expertise is directly related to the proposed work in this R01 renewal, including next generation diagnostic tool development for monitoring bat virus spillover, bat pathogen discovery and bat viral immunology. I have worked on bat virology since 2004, and participated in the work that led to the discovery of SARS-like coronaviruses in Mainland China. I worked on bat viral immunology to explore reasons why bats can coexist with high viral diversity and viral loads. My findings show that in bats, a constitutively expressed interferon and a dampened STING-dependent interferon production pathway exists, which may explain why bats can control viral replication and tolerate viral diseases. My most recent work on Swine Acute Diarrhea Syndrome (SADS) characterized the spillover of CoV from bats to swine causing a large-scale pandemic. I have worked closely with Dr. Linfa Wang from Singapore Duke-NUS medical school, on developing next-generation viral nucleotide, serological and isolation tools for coronavirus from bats and other animals. I have also collaborated with, and published papers with Dr. Daszak and other staff at EcoHealth Alliance. I will be in charge of diagnostics, genomics, and virus isolation in this project. B. Positions and Honors. Positions and Employment 2016- Principle Investigator, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China Other Experience and Professional Memberships 2009 - 1 o Joint PhD study in Australia Animal Health Laboratory, CSIRO Honors 2009 Scholarship for China-Australia Joint PhD study, China Scholarship Council. 2017 Natural Science Award (the first rank) of Hubei province, China. 2018 National Science Fund for Excellent Young Scholars, China. c_ Selected peer-reviewed most relevant to the current application • = Co-corresponding or first authors 1. Zhou P, Fan H, Lan T, Yang XL, Shi WF, Zhang W, Zhu Y, Zhang YW, Xie QM, Mani S, Zheng XS, Li B, Li JM, Guo H, Pei GO, An XP, Chen JW, Zhou L, Mai KJ, Wu ZX, Li D, Anderson D, Zhang LB, Li SY, Mi ZQ, He TT, Cong F, Guo PJ, Huang R, Luo Y, Liu XL, Chen J, Huang Y, Sun Q, Zhang XLL, Wang YY, Xing SZ, Chen YS, Sun Y, Li J, Daszak P, Wang LF, Shi ZL, Tong YG, Ma JY (2018) Fatal swine acute diarrhea syndrome Page 75 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter caused by an HKU-2 related coronavirus of bat origin. Nature, 556: 255-258. doi.org/10.1038/s41586-018-0010-9 2. Xie J, Li Y, She X, Goh G, Zhu Y, Cui J, Wang LF, Shi Z, Zhou P (2018) Dampened STING-dependent interferon activation in bats. Cell Host and Microbes 23(3): 297-301 . doi.org/10.1016/j.chom .2018.01 .006 3. Zhou P, Tachedjian M, Wynne JW, Boyd V, Cui J, Smith I, Cowled C, Ng JH, Mok L, Michalski WP, Mendenhall lH, Tachedjian G, Wang LF, Baker ML (2016). Contraction of the type I IFN locus and unusual constitutive expression of IFN-a in bats. Proceedings of the National Academy of Sciences, 113(10): 2696-701. doi.org/10.1073/pnas.1518240113 4. Wu L*, Zhou P*, Ge XY, Wang LF, Baker ML, Shi Z (2013). Deep RNA Sequencing Reveals Complex Transcriptional Landscape of a Bat Adenovirus. Journal of Virology, 87(1): 503-511. doi.org/10.1128/JVl.02332-12 5. Zhou P, Li H, Wang H, Wang LF, Shi Z (2012). Bat severe acute respiratory syndrome-like coronavirus ORF3b homologues display different interferon antagonist activities. Journal of General Virology, 93: 275-281. doi.org/10.1099/vir.0.033589-0 6. Zhou P, Cowled C, Todd S, Crameri G, Virtue ER, Marsh GA, Shi ZL, Wang LF, and Baker ML (2011). Type Ill Interferons in pteropid bats: differential expression patterns provide evidence for distinct roles in antiviral immunity. Journal of Immunology, 186(5): 3138-3147. doi.org/ 10.4049/jimmunol.1003115 7. Zhou P, Han Z, Wang LF, Shi Z (2009). lmmunogenicity difference between the SARS coronavirus and the bat SARS-like coronavirus spike (S) proteins. Biochemal Biophysical Research Communications, 387(2): 326-329. doi.org/10.1016/j.bbrc.2009.07.025 D. Research Support Ongoing Research Support Combating the next SARS- or MEAS-like emerging infectious disease outbreak by improving active surveillance Role: Pl Interferon responses in SARS-Like Coronavirus infected Bat cells Role: Pl Bat virology Role: Pl Completed Research Support Immune responses an transcnptome ana ys1s of bat adenovirus infected bat cells Role: Co-Pl 2 Page 76 Biosketches (b)(4l (b)(4) (b) (4) (b) (4~

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Ben Hu eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Huazhong Agricultural University, China Huazhong Agricultural University, China Wuhan Inst. Virology, Chinese Acad. Sci. A. Personal Statement POSITION TITLE Co-Investigator DEGREE BS MS PHO MMNY FIELD OF STUDY 2007 Veterinary medicine 2010 Preventive vet. med. 2015 Microbiology I obtained my PhD degree in 2015 and have been working as a Research Assistant in Dr. Zhengli Shi's laboratory for over 3 years. My research is focused on the discovery and characterization of viruses in small mammals, especially in bats and rodents. My work forms the basis for some of the key findings built on in the current R01 proposal, in particular: 1) the identification of diverse bat SARSr-CoVs at cave sites in Yunnan, China; 2) the potential recombination origin of SARS-CoV; and 3) characterization of spillover risk for bat SARSr-CoVs. I have also reported genetically diverse novel astroviruses in bats, rodents, and shrews in China. I have collaborated with EcoHealth Alliance scientists on multiple research projects for the past 3 years. B. Positions and Honors. Positions and Employment 2015- Assistant Researcher, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, China Honors C. Selected peer-reviewed publications most relevant to the current application Note: * = Corresponding Author 1. Ge X, Li J, Yang X, Chmura AA, Zhu G, Epstein JH, Mazet JK, Hu B, Zhang w, Peng C, Zhang YJ, Luo CM, Tan B, Wang N, Zhu Y, Crameri G, Zhang SY, Wang LF, Daszak p•, Shi Z (2013). Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor. Nature, 503: 535-538. doi.org/10.1038/nature12711 2. Hu B, Chmura AA, Li J, Zhu G, Desmond JS, Zhang Y, Zhang W, Epstein JH, Daszak P, Shi Z (2014). Detection of diverse novel astroviruses from small mammals in China. Journal of General Virology 95, 2442-2449. doi.org/10.1099/vir.0.067686-0 3. Hu B, Ge X, Wang LF, Shi Z (2015). Bat origin of human coronaviruses. Virology Journal, 12(1 ): 221. doi.org/10.1186/s 12985-015-0422-1 4. Ge XY, Wang N, Zhang W, Hu B, Li B, Zhang YZ, Zhou JH, Luo CM, Yang XL, Wu LJ, Wang B, Zhang Y, Li ZX, Shi Z (2016). Coexistence of multiple coronaviruses in several bat colonies in an abandoned mineshaft. Virologica Sinica, 31 (1): 31-40. doi.org/10.1007/s 12250-016-3713-9 Page 77 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 5. Wang MN, Zhang W, Gao YT, Hu B, Ge XY, Yang XL, Zhang YZ, Shi Z (2016). Longitudinal surveillance of SARS-like coronaviruses in bats by quantitative real-time PCR. Virologica Sinica, 31 (1 ): 78-80. doi.org/ 10.1007 /s 12250-015-3703-3 6. Yang XL, Hu B, Wang B, Wang MN, Zhang Q, Zhang W, Wu LJ, Ge XY, Zhang YZ, Daszak P, Wang LF, Shi Z (2016). Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus. Journal of Virology, 90(6): 3253-3256. doi.org/ 10.1128/JVl.02582-15 7. Waruhiu C, Ommeh S, Obanda V, Agwanda B, Gakuya F, Ge XY, Yang XL, Wu LJ, Zohaib A, Hu B, Shi Z (2017). Molecular detection of viruses in Kenyan bats and discovery of novel astroviruses, caliciviruses, and rotaviruses. Virologica Sinica, 32(2): 101-114. doi.org/ 10.1007 /s 12250-016-3930-2 8. Hu B, Zeng LP, Yang XL, Ge XY, Zhang W, Li B, Xie JZ, Shen XR, Zhang Y2, Wang N, Luo DS, Zheng XS, Wang MN, Daszak P, Wang LF, Cui J, Shi Z (2017). Discovery of A Rich Gene Pool of Bat SARS-related Coronaviruses Provides New Insights into the Origin of SARS Coronavirus. PLOS Pathogens, 13(11): e1006698. 9. Luo CM, Wang N, Yang XL, Liu HZ, Zhang W, Li B, Hu B, Peng C, Geng QB, Zhu GJ, Li F, Shi Z (2018). Discovery of Novel Bat Coronaviruses in South China That Use the Same Receptor as Middle East Respiratory Syndrome Coronavirus. Journal of Virology, 92(13): e00116-18. 10. Luo Y, Li B, Jiang RD, Hu BJ, Luo DS, Zhu GJ, Hu B, Liu HZ, Zhang YZ, Yang XL, Shi ZL (2018). Longitudinal Surveillance of Betacoronaviruses in Fruit Bats in Yunnan Province, China during 2009-2016. Virologica Sinica, 33(1):87-95. doi.org/ 10.1007/s12250-018-0017-2 D. Research Support Ongoing Research Support 31800142 National Natural Science Foundation of China Hu (Pl) 01/01/2019-12/31/2021 Pathogenicity studies of two novel bat SARSr-CoVs on transgenic mice expressing human ACE2 Role: Pl R01 Al110964 Daszak (Pl) 06/01 /14-05/31 /19 Understanding Risk of Bat Coronaviruses The goal of this study is to analyze the risk of coronavirus spillover from bats to humans in Southern China Role: Research Scientist Emerging Pandemic Threat Program, USAID Mazet (Pl) 10/01 /14-09/30/19 PREDICT 2 The goal of this project is to create and implement a global virus surveillance system in animals and humans and analyze spillover risk. Role: Laboratory Scientist Completed Research Support USAID EPT PREDICT-1 Mazet (Pl) 10/01/09 - 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Amount: $18 million subcontract on a $75 million award Role: Laboratory Scientist 2 Biosketches Page 78

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter BIOGRAPHICAL SKETCH NAME Aleksei A. Chmura eRA COMMONS USER NAME (b)(6) EDUCATION/TRAINING INSTITUTION AND LOCATION Columbia University School of Life Sciences, Kinaston University (UK) A. Personal Statement POSITION TITLE Research Scientist DEGREE MMNY BS 06/2004 PhD 08/2018 FIELD OF STUDY Biology Bioloav Or. Chmura has an interdisciplinary background in ecology, wildlife biology, virology, and extensive on-the-ground experience conducting wildlife sampling in China. Fluent in spoken and written Mandarin, for the past decade, Or. Chmura has acted as a key coordinator among EcoHealth Alliance headquarters staff, and laboratory and field teams in China. Dr. Chmura's personal research involves the wildlife origins of SARS-CoV, wildlife paramyxovirus diversity and evolution, and human-wildlife contact behavior in southern China. His work has been funded by USAID EPT/PREDICT since 2009. As part of his doctoral work, he spent over a year in the Wuhan Institute of Virology laboratory in China under the direction of Dr. Zheng Ii Shi and Dr. Peter Daszak. B. Positions and Honors. Positions and Employment 2001-2004, Volunteer Curator, Dept. of Mammalogy, American Museum of Natural History, USA 2001-2005, Program Assistant, Ctr. Environmental Research and Conservation, Columbia University, USA 2002-2005, Instructor, Columbia University Tropical Field Ecology Programs, USA/Oomician Republic/Brazil 2005-Present, Program Coordinator, EcoHealth Alliance, USA 2006-Present, Managing Editor, EcoHealth, New York, USA Other Experience and Professional Memberships 2000-2005 2002-present 2005-present 2009-present The Explorers Club American Museum of Natural History International Association for Ecology and Health Society for Applied Microbiology C. Selected peer-reviewed publications most relevant to the current application Monagin C, Ning L, Schneider B, Chmura AA, Epstein JH, Wu D, Paccha B, Ke C, Oaszak P, Rabinowitz P (2018) Serologic and behavioral risk survey of workers with wildlife contact in China. PLOS ONE, 13(4): e0194647. Wang N, Li, S, Yang X, Huang H, Zhang Y, Guo H, Luo C, Miller M, Zhu G, Chmura AA, Hagan E, Zhou J, Zhang Y, Wang L, Daszak P, Shi Z (2018) Serological evidence of bat SARS-related coronavirus infection in humans, China. Virologica Sinica, 33(1): 104-107. Zeng L, Ge X, Peng C, Yang X, Tan B, Gao Y, Chen J, Chmura AA, Daszak P, Shi Z (2016) Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response. Journal of Virology, 90(14): 6573-6582. Mazet JAK, Wei Q, Zhao G, Cummings DAT, Desmond JS, Rosenthal J, King CH, Cao W, Chmura AA, Hagan EA, Zhang S, Xiao X, Xu J, Shi Z, Feng F, Liu X, Pan W, Zhu G, Zuo G, Daszak P (2015). Joint China-Us Call for Employing a Transdisciplinary Approach to Emerging Infectious Diseases. EcoHealth 12(4): 555-559. Page 79 Biosketches

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Hu B, Chmura AA, Li J, Zhu G, Desmond JS, Zhang YJ, Zhang JS, Epstein JH, Daszak P, Shi Z (2014). Detection of Diverse Novel Astroviruses from Small Mammals in China. Journal of General Virology 95: 2442-2449. Ge XY, Li JL, Yang X-L, Chmura AA, Zhu G, Epstein JH, Mazet JK, Hu B, Zhang W, Peng C, Zhang YJ, Luo CM, Tan B, Wang N, Zhu Y, Crameri G, Zhang SY, Wang LF, Daszak P, Shi Z (2013). Isolation and characterization of a bat SARS-like Coronavirus that uses the ACE2 receptor. Nature 503: 535-538. Zhu G, Chmura AA, Zhang L (2011 ). Morphology, echolocation calls and diet of Scotophi/us kuhlii (Chiroptera: Vespertilionidae) on Hai nan Island, south China. Acta Chiropterologica, 14( 1 ): 175-181. Kilpatrick AM, Chmura AA, Gibbons OW, Fleischer RC, Marra PP, Daszak P (2006). Predicting the global spread of H5N 1 avian influenza. PNAS 103: 19368-19373. D. Research Support Ongoing Research Support R01 Al110964 Understanding Risk of Bat Coronaviruses Daszak (Pl) 06/01 /14-05/31 /19 The goal of this study is to analyze the risk of coronavirus spillover from bats to humans in Southern China Role: Research Scientist Emerging Pandemic Threat Program, USAID Mazet (Pl) 10/01 /14-09/30/19 PREDICT 2 The goal of this project is to create and implement a global virus surveillance system in animals and humans and analyze spillover risk. Role: Program Coordinator Completed Research Support USAID EPT PREDICT-1 Mazet (Pl) 10/01/09- 09/30/14 Modeling hotspots for disease emergence and conducting surveillance in wildlife in hotspots for new emerging zoonoses Role: Program Coordinator 2 R01TW005869 Daszak (Pl) 09/01/08 - 08/31/13 NIH Ecology of Infectious Diseases (Fogarty International Center) The Ecology, Emergence and Pandemic Potential of Nipah virus in Bangladesh To conduct mathematical modeling and fieldwork to understand the dynamics of Nipah virus in Bangladesh Role: Research Scientist NSF DEB-1257513 Daszak ( Pl) 08/15/12-07 /31/13 US-China Ecology and Evolution of Infectious Diseases Collaborative Workshop; Kunming, China 2012 Role: Program Coordinator 1 R01 AI079231 Daszak (Pl) 09/18/08 - 08/31 /13 NIAID Non-Biodefense Emerging Infectious Diseases Risk of viral emergence from bats. To model hotspots for bat viral diversity, identify & characterize new bat viruses & understand their pathology Role: Research Scientist 2 Page 80 Biosketches

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 1 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project 0 Subaward/Consortium Enter name of Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2019 A. Senior/Key Person Prefix First Name• Last Name• Suffix Project Role* End Date•: 05-31-2020 Budget Period: 1 Base Calendar Academic Summer Requested Fringe 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 Funds Requested ($)* 1 . Dr . PETER Middle Name DASZAK PD/Pl Salary ($) Months Months Months Salarv (_$)* Benefits ($0,),*------~~~~ (b) (4). (b) (6) . 2..:. ~.r: ....... ~~?i~ ....................... ~ .................... <?.l.i\l~I ................................... <?.°.~.1.~~.El~.ti~~t<:>r __ ............................ . ~.:. ~.r: ........ ~.~il~~i .................... Y. ...................... t='r~.~~!~.C.() ................................ <?.°.~.1.~- -El~.ti~.~t<:>r ............................. .. . ~.·-~.r: ........ ~°.~.rr.! ................................................ ~?.5.5. .......................................... <?.°.~i~~-El~.ti~_~t<:>r .............................. . 5 . Ms. Hongying Li Research Scientist 6. Dr. Alice Latinne Research Scientist ................... ···························································· ·······••·••························· ............................................................................ . 7 . Ms. Emily A Hagan Research Scientist .................................................................................................... ···············"•··· .. ···· .............................................................. . 8 . Dr. Aleksei A Chmura Research Scientist Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 223,713.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ........................................................................................................................................................ , ................................................................................................................................................. , ................................................................... . Graduate Students ............................. Undergraduate_ Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-8) (Funds Requested) Tracking Number: GRANT12743073 Page 81 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) 0.00 223,713.00 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 1 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2019 End Date*: 05-31-2020 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Budget Period: 1 Total Equipment Total Travel Cost Funds Requested ($)* 0.00 Funds Requested ($)* 9,440.00 29,958.00 39,398.00 Funds Requested ($)* Number of Participants/Trainees Total Participant Trainee Support Costs 0.00 RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Page 82 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 1 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2019 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2020 Budget Period: 1 Funds Requested ($)* 20,850.00 79,750.00 190,649.00 Total Other Direct Costs 291,249.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . EcoHealth Alliance Indirect Cost 2 . EcoHealth Alliance Indirect Cost on 3 Subawards (IPB, UNG, WIV) 3 . University of North Carolina at Chapel Hill Indirect Cost 4 . IPB and WIV Subawards (2) Indirect Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 554,360.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 32.74 363,710.00 119,079.00 32.74 75,000.00 24,555.00 55.5 50,000.00 27,750.00 8.0 140,649.00 11,252.00 Total Indirect Costs 182,636.00 I. Total Direct and Indirect Costs Funds Requested($)* Total Direct and Indirect Institutional Costs (G + H) 736,996.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: EHA_NIAID_COV_BUDGET_JUSTIFICATION_FINAL.pdf (Only attach one file.) 736,996.00 RESEARCH & RELATED Budget [F-K} (Funds Requested) Page 83 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 2 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project 0 Subaward/Consortium Enter name of Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2020 A. Senior/Key Person Prefix First Name• Last Name• Suffix Project Role* End Date•: 05-31-2021 Budget Period: 2 Base Calendar Academic Summer Requested Fringe 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 Funds Requested ($)* 1 . Dr . PETER Middle Name DASZAK PD/Pl Salary ($) Months Months Months Salarv ~,_•_B_e_n_e_f,_'ts__,(,$":,)_* -----~~~~~ (b) (4), (b) (6) . 2..:. ~.r: ....... ~~?i~ ....................... ~ .................... <?.l.i\l~I ................................... <?.°.~.1.~~.El~.ti~~t<:>r __ ........................... . ~.:. ~.r: ........ ~.~il~~i .................... Y. ...................... t='r~.~~!~.C.() ................................ <?.°.~.1.~- -El~.ti~.~t<:>r ............................. . . ~.·-~.r: ........ ~°.~.rr.! ................................................ ~?.5.5. .......................................... <?.°.~i~~-El~.ti~_~t<:>r ............................. . 5 . Ms. Hongying Li Research Scientist 6. Dr. Alice Latinne Research Scientist ................... ···························································· ·······••·••························· ........................................................................... . 7 . Ms. Emily A Hagan Research Scientist ····················--· ............................................................................. ···············"•····················--·········· ...................................... . 8 . Dr. Aleksei A Chmura Research Scientist Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 223,713.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ............................................................................................................................................................................................................................................................................................................................................................................... Graduate Students ............................. Undergraduate_ Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-8) (Funds Requested) Tracking Number: GRANT12743073 Page 84 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) 0.00 223,713.00 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 2 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2020 End Date*: 05-31-2021 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Budget Period: 2 Total Equipment Total Travel Cost Funds Requested ($)* 0.00 Funds Requested ($)* 9,440.00 29,958.00 39,398.00 Funds Requested ($)* Number of Participants/Trainees Total Participant Trainee Support Costs 0.00 RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Page 85 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 2 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2020 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2021 Budget Period: 2 Funds Requested ($)* 14,850.00 6,000.00 79,750.00 190,649.00 Total Other Direct Costs 291,249.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . EcoHealth Alliance Indirect Cost 2 . University of North Carolina at Chapel Hill Indirect Cost 3. IPB and WIV Subawards (2) Indirect Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 554,360.00 Indirect Cost Rate (%) Indirect Cost Base ($) 32.74 363,710.00 55.5 50,000.00 Funds Requested ($)* 119,079.00 27,750.00 8.0 140,649.00 11,252.00 -------------1 Total Indirect Costs 158,081.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 712,441.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: EHA_NIAID_COV_BUDGET_JUSTIFICATION_FINAL.pdf (Only attach one file.) 712,441.00 RESEARCH & RELATED Budget (F-K} (Funds Requested) Page 86 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 3 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project 0 Subaward/Consortium Enter name of Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2021 End Date•: 05-31-2022 Budget Period: 3 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* 1 . Dr . PETER DASZAK PD/Pl (b) (4). (b) (6) . 2..:. ~.r: ....... ~~?i~ ....................... ~ .................... <?.l.i\l~I ................................... <?.°.~.1.~~.El~.ti~~t<:>r __ ........................... . ~.:. ~.r: ........ ~.~il~~i .................... Y. ...................... t='r~.~~!~.C.() ................................ <?.°.~.1.~- -El~.ti~.~t<:>r ............................. . . ~.·-~.r: ........ ~°.~.rr.! ................................................ ~?.5.5. .......................................... <?.°.~i~~-El~.ti~_~t<:>r ............................. . 5 . Ms. Hongying Li Research Scientist 6. Dr. Alice Latinne Research Scientist ................... ···························································· ·······••·••························· ........................................................................... . 7 . Ms. Emily A Hagan Research Scientist ····················--· ............................................................................. ···············"•····················--·········· ...................................... . 8 . Dr. Aleksei A Chmura Research Scientist Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 223,713.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ............................................................................................................................................................................................................................................................................................................................................................................... Graduate Students ............................. Undergraduate_ Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-8) (Funds Requested) Tracking Number: GRANT12743073 Page 87 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) 0.00 223,713.00 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 3 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2021 End Date*: 05-31-2022 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Budget Period: 3 Total Equipment Total Travel Cost Funds Requested ($)* 0.00 Funds Requested ($)* 9,440.00 29,958.00 39,398.00 Funds Requested ($)* Number of Participants/Trainees Total Participant Trainee Support Costs 0.00 RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Page 88 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 3 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2021 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2022 Budget Period: 3 Funds Requested ($)* 14,850.00 6,000.00 79,750.00 190,649.00 Total Other Direct Costs 291,249.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . EcoHealth Alliance Indirect Cost 2 . University of North Carolina at Chapel Hill Indirect Cost 3. IPB and WIV Subawards (2) Indirect Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 554,360.00 Indirect Cost Rate (%) Indirect Cost Base ($) 32.74 363,710.00 55.5 50,000.00 Funds Requested ($)* 119,079.00 27,750.00 8.0 140,649.00 11,252.00 -------------1 Total Indirect Costs 158,081.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 712,441.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: EHA_NIAID_COV_BUDGET_JUSTIFICATION_FINAL.pdf (Only attach one file.) 712,441.00 RESEARCH & RELATED Budget (F-K} (Funds Requested) Page 89 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 4 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project 0 Subaward/Consortium Enter name of Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2022 End Date•: 05-31-2023 Budget Period: 4 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* 1 . Dr . PETER DASZAK PD/Pl (b) (4). (b) (6) . 2..:. ~.r: ....... ~~?i~ ....................... ~ .................... <?.l.i\l~I ................................... <?.°.~.1.~~.El~.ti~~t<:>r __ ............................ . ~.:. ~.r: ........ ~.~il~~i .................... Y. ...................... t='r~.~~!~.C.() ................................ <?.°.~.1.~- -El~.ti~.~t<:>r .............................. . . ~.·-~.r: ........ ~°.~.rr.! ................................................ ~?.5.5. .......................................... <?.°.~i~~-El~.ti~_~t<:>r .............................. . 5 . Ms. Hongying Li Research Scientist 6. Dr. Alice Latinne Research Scientist ................... ···························································· ·······••·••························· ............................................................................ . 7 . Ms. Emily A Hagan Research Scientist ····················--· ............................................................................. ···············"•····················--·········· ....................................... . 8 . Dr. Aleksei A Chmura Research Scientist Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 223,713.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ............................................................................................................................................................................................................................................................................................................................................................................... Graduate Students ............................. Undergraduate_ Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-8) (Funds Requested) Tracking Number: GRANT12743073 Page 90 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) 0.00 223,713.00 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 4 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2022 End Date*: 05-31-2023 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Budget Period: 4 Total Equipment Total Travel Cost Funds Requested ($)* 0.00 Funds Requested ($)* 9,440.00 29,958.00 39,398.00 Funds Requested ($)* Number of Participants/Trainees Total Participant Trainee Support Costs 0.00 RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Page 91 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 4 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2022 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2023 Budget Period: 4 Funds Requested ($)* 14,850.00 6,000.00 79,750.00 190,649.00 Total Other Direct Costs 291,249.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . EcoHealth Alliance Indirect Cost 2 . University of North Carolina at Chapel Hill Indirect Cost 3. IPB and WIV Subawards (2) Indirect Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 554,360.00 Indirect Cost Rate (%) Indirect Cost Base ($) 32.74 363,710.00 55.5 50,000.00 Funds Requested ($)* 119,079.00 27,750.00 8.0 140,649.00 11,252.00 -------------1 Total Indirect Costs 158,081.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 712,441.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: EHA_NIAID_COV_BUDGET_JUSTIFICATION_FINAL.pdf (Only attach one file.) 712,441.00 RESEARCH & RELATED Budget (F-K} (Funds Requested) Page 92 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 5 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project 0 Subaward/Consortium Enter name of Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2023 End Date•: 05-31-2024 Budget Period: 5 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* 1 . Dr . PETER DASZAK PD/Pl (b) (4), (b) (6) . 2..:. ~.r: ....... ~~?i~ ....................... ~ .................... <?.l.i\l~I ................................... <?.°.~.1.~~.El~.ti~~t<:>r __ ........................... . ~.:. ~.r: ........ ~.~il~~i .................... Y. ...................... t='r~.~~!~.C.() ................................ <?.°.~.1.~- -El~.ti~.~t<:>r ............................. . . ~.·-~.r: ........ ~°.~.rr.! ................................................ ~?.5.5. .......................................... <?.°.~i~~-El~.ti~_~t<:>r ............................. . 5 . Ms. Hongying Li Research Scientist 6. Dr. Alice Latinne Research Scientist ................... ···························································· ·······••·••························· ........................................................................... . 7 . Ms. Emily A Hagan Research Scientist ····················--· ............................................................................. ···············"•····················--·········· ...................................... . 8 . Dr. Aleksei A Chmura Research Scientist Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 223,713.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ............................................................................................................................................................................................................................................................................................................................................................................... Graduate Students ............................. Undergraduate_ Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-8) (Funds Requested) Tracking Number: GRANT12743073 Page 93 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) 0.00 223,713.00 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 5 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2023 End Date*: 05-31-2024 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs E. Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Budget Period: 5 Total Equipment Total Travel Cost Funds Requested ($)* 0.00 Funds Requested ($)* 9,440.00 29,958.00 39,398.00 Funds Requested ($)* Number of Participants/Trainees Total Participant Trainee Support Costs 0.00 RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Page 94 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 5 ORGANIZATIONAL DUNS*: 0770900660000 Budget Type*: • Project O Subaward/Consortium Organization: ECOHEAL TH ALLIANCE, INC. Start Date*: 06-01-2023 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2024 Budget Period: 5 Funds Requested ($)* 14,850.00 6,000.00 79,750.00 190,649.00 Total Other Direct Costs 291,249.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . EcoHealth Alliance Indirect Cost 2 . University of North Carolina at Chapel Hill Indirect Cost 3. IPB and WIV Subawards (2) Indirect Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 554,360.00 Indirect Cost Rate (%) Indirect Cost Base ($) 32.74 363,710.00 55.5 50,000.00 Funds Requested ($)* 119,079.00 27,750.00 8.0 140,649.00 11,252.00 -------------1 Total Indirect Costs 158,081.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 712,441.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: EHA_NIAID_COV_BUDGET_JUSTIFICATION_FINAL.pdf (Only attach one file.) 712,441.00 RESEARCH & RELATED Budget (F-K} (Funds Requested) Page 95 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Program DirectOf/Principal Investigator: Daszak, Peter ECOHEAL TH ALLIANCE BUDGET JUSTIFICATION A. Senior/Key personnel: The PD/Pl, Dr. Peter Daszak, will commit, H4 ),(b)(1per year in each year of this budget. He will be primarily responsible for overseeing the project, general management, communication and collaboration with subaward partners, as well as contributing to data analysis and manuscript writing. Co-Investigator, Dr. Kevin Olival. will commit (bH4 >, (b)(6) per year in each year of this budget. Dr. Olival will lead the design and implementation of the bat sampling fieldwork (Aim 1 ); facilitate overall project management; and train and oversee field teams. Dr. Olival will also oversee modeling and analyses under Aims 1 & 3, participate in regular conference calls, and help write manuscripts and reports. Co-Investigator, Dr. Leilani Francisco, will commit Cb) (4), (b) (6) per year in each year of this budget. Dr. Francisco will lead the implementation of the community and clinic-based surveillance (Aim 2), including adherence to study design, sampling methodology, and ethics in human subjects research; data collection instrument development; data management, cleaning, and analysis; and, findings dissemination. Co-Investigator, Dr. Noam Ross, will commit (b)( 4 ),~ per year in each year of this budget. Dr. Ross will lead modeling work and assist in with data analyses and manuscript writing. He will also advise on data management, statistical approaches, and computational work. B. Other Personnel Research Scientist, Ms. Hongying Li, will commit > (4), (b) (6) per year in each year of this budget. Ms. Li will coordinate the field and laboratory activities in China, maintaining the financial administration, results reporting, and data management, as well as work closely with Dr. Lili Ren at the Institute of Pathogen Biology to refine protocols, oversee field data collection, and perform data analysis for human study. Research Scientist, Dr. Alice Latinne, will commit (b) (4), (b) (6) per year in each year of this budget. Dr. Latinne will assist in with phylogenetic and phylogeographic analyses and manuscript writing. She will also advise on data management and field activities. Research Scientist. Dr. Aleksei Chmura, will commit (b) < 4 >, ~per year in each year of this budget. Dr. Chmura will coordinate regular calls, reports, maintainEccfF1ealth Alliance and subaward budgets and both project and financial reporting, draft subcontracts, and set-up project databases, advise field activities, assist with statistical analysis, and manuscript writing. Research Scientist. Ms. Emily Hagan, will commit > (4), (b) C per year in each year of this budget. Ms. Hagan will assist with the development of human data collection instruments, testing, and implementation; advise on data storage, data analyses, and manuscript writing. She will also provide training for field teams conducting human subjects research. Fringe benefits for Year 1 are calculated for EcoHealth Alliance's federally approved rate of 31.5% of base salary and is included in all subsequent years. C. Equipment No Equipment costing more than $5,000 will be purchased D. Travel Domestic Travel $9,440 is requested annually for Years 1 through 5 for the PD/Pl, 3 Co-Investigators, and 1 Research Scientist to attend and present on research results at the annual American Society for Tropical Medicine and Hygiene and the American Public Health Association meetings. 2 night and 3 day travel to Washington, DC is Page 96 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program DirectOf/Principal Investigator: Daszak, Peter calculated as follows: $205 for hotels ($251 x 2 nights x 5 people x 2 trips= $5,020); $76 for meals and incidentals ($76 x 2.5 days x 5 people x 2 trips = $1,900); and $252 for round-trip train ($252 x 5 x 2 = $2,520). International Travel $11,998 is requested annually in Years 1 to 5. This will support round-trip flights from New York to Beijing and Wuhan for the field annual meetings for 3 Senior/Key Personnel and 1 for the PD/Pl (Daszak) at $1,055 each. Five nights and six days of hotels, meals, and incidentals for 3 Senior/Key Personal and 1 PD/Pl are calculated at $1,944.50 per year: hotels at $258 per night (x 5 nights and 4 personnel= $5,160) and meals and incidentals at $119 per day (x 5.5 days and 4 personnel = $2,856). $17,960 is requested annually in Years 1 to 5 for EHA Research Scientists (Ms. Li and Ms. Hagan) who will travel to China for two field training and supervising visits per year for duration of 21 days each. Support for this request, annually, is $17,960 and is calculated as follows: 2 round trip flights= $4,400; hotel $258 x 20 nights x twice a year= $4,732; meals and incidentals at $119 per day x 20.5 days x twice a year= $3,570 E. Participant/Trainee Support Costs There are no participant/trainee support costs. F. Other Direct Costs Materials & Supplies We request $7,000 in Year 1 for sample collection materials to be shipped to China including bat catching equipment ($1,000); PPE ($2,000); and 1 liquid nitrogen dry shipper ($1,000) for Wuhan Institute of Virology in China to be used by Dr. Guanjian Zhu for field work. In Years 2 through 5, field and human sampling will be completely underway; we request support for PPE ($2,000) and other sample collection materials ($2,000) in each of these years. Publication Costs We request $6,000 per year for only Years 2 to 5 for publication fees required to publish research findings in peer-reviewed journals such as Nature, Public Library of Science, and other journals Subawards/Consortium/Contractual Costs We are requesting consortium/contractual support for our three partners: Wuhan Institute of Virology (WIV), Institute of Pathogen Biology (IPB), and University of North Carolina (UNC). We have fully detailed these direct and indirect costs in their respective sub-award budgets. Computers, Software, Reference Materials and Dataset Acquisition We request support of $6,000 to permit two Research Scientists to purchase 1 laptop each (2 x $3,000 including insurance and software). We also request $1,000 per year in each year to cover software and reference materials, and an additional $1,000 per year in each year for acquisition of datasets. Shipping We will be shipping the materials and supplies detailed above to our subaward institutions in China (IPB and WIV). Shipping box and all taxes are estimated at $1,667 per shipment. We estimate 3 shipments of supplies and materials will be sent every year through the duration of this project. Consultants Dr. Linfa Wang, Co-Investigator/Consultant We request H4>. (bH per year for each year of the project for a consultancy for Dr. Linfa Wang who will focus on PCR development, serological testing strategy and virus characterization, and will also participate in regular meetings with collaborators. Dr. Wang has more than 20 years of research experience in designing and applying novel testing platforms to discover zoonotic pathogens. 2 Page 97 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program DirectOf/Principal Investigator: Daszak, Peter Dr. Guangjian Zhu, Co-Investigator/Consultant In total, we request $368,000 for the consultancy of Dr. Guangjian Zhu from Year 1 to Year 5 of the project including: $204,390 for field personnel, $124,750 for field travel; $33,548 for field supplies and materials, and $5,255 for other costs. Detailed expenses are calculated as the follows: Personnel ($204,390) Dr. Guangjian Zhu Co-Investigator/Consultant will oversee all field sampling activities in China by coordinating with local partners and stakeholders to lead the specimen collection and bats population monitoring at selected surveillance sites. Dr. Zhu is a zoologist and ecologist specializing in bats surveillance in southern and western China and has been leading EcoHealth Alliance's field surveillance work in China for more than 15 years. We request H4>.CbH6J annual stipend for Zhu, who will commi (b)(4), (b)(6) per year to this project in years 1-3 and increase time to (b)(4), (b)(6) per year for in years 4-5, since he will allocate more time to collaborating on peer-reviewed publications in the last two years of the project. Research Assistant (TBD) will assist the Co-Pl and Field Coordinator (Zhu) for project data management, reporting, and administration. We request H4),(b)(6) p.a. salary for this Research Assistant who will dedicate 2 months p.a. on this project from Years 1-5. Field Assistants (2 in each province, TBD) will assist all field surveillance activities including specimen collection and data entry and management. The assistants will commit a total of 50 days per year to this project from years 1-5. We request H4),(b)(6) per year to support each assistant for the field surveillance work. Travel ($124,750) Inter-Province Travel. We request 1) $1,200 per year for all five years of this project to cover 3- per-year round-trip flights/trains each from Shanghai, to Yunnan, Guangdong, Guizhou, and Guangxi for Dr. Zhu to meet with collaborating institutions, train field teams, and ensure sample collection, storage, and shipments. Each round-trip flight is estimated at $400, in total $6,000 for 5 years; 2) $2,400 per year for all five years of this project to cover 2-per-year round-trip flights/trains for 2 field assists traveling to the field sites in Yunnan, Guangdong, Guizhou, and Guangxi for sampling work. Each round-trip flight is estimated at $400, in total $12,000 for 5 years. Field Transportation. Field work will take place for 50 days per year for 5 years, the expenses of local transportation include 1) car rental at the rate of $79/car/day, with 1 car for 50 days, in total of $3,950 per year, and $19,750 for 5 years; 2) Gas and toll fee at the rate of $32/car/day, with 1 car for 50 days, in total of $1,600 per year, and $8,000 for 5 years. Meal and Lodging. We request 1) $6,400 to cover the expense of meals for 4 field team members in the field for 50 days per year, at the rate of $32/person/day, totaling $32,000 in 5 years; 2) $9,400 for lodging expenses of 4 field team members in the field for 50 days at the rate of $47/person/night. totaling $247,000 in 5 years. Supplies and Materials ($33,548) Biological sampling supplies ($25,165) We request $25,165_to purchase supplies for biological sampling during the 5 years of the project, including 1) puritan calcium alginate swabs $8,800 (5,000 IND); 2) viral sample collection tubes $6,875 (15,000 IND); 3) heparinized glass hematocrit tubes $190 (~4,000IND): 4) mist nets for bats trapping $2,200 (~500IND); 5) cloth bags for bats trapping $2,400 (~1,000IND); 6) Viral Transport Media $4,700 {~7,000 ml). Personal Protection Equipment ($4,336): We request 1) $3,440 for 3M N95 respirators (~1,600IND) for field work across Year 1-5; 2) $470 for eye protection glasses (~100 IND) for 3 Page 98 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter the use in field across Year 1-5; 3) $426 for nitrile gloves (~3,000IND) for sampling work for Year 1-5. Cold Chain Maintenance ($4,047): We request $4,047 to purchase 3 liquid nitrogen dry shippers for preserve biological samples in the field before transported an ultra-low temperature freezer. The expense is calculated at the rate of $1,349 each, with 1 purchased per year from Year 1-3, totaling $4,047. Equipment ($0) No equipment over $5,000 will be purchased. Other Costs ($6,399) We request 1) a total of $1,275 for specimen transportation or delivery from the field to partners' labs from Year 1-5, at the rate of $85/delivery with 1,000 tubes, with three times per year; and 2) a total of $3,980 for rabies and tetanus vaccination 4 field team members from Year 1-5, at the rate of $199/yea ripe rson. H. Indirect Costs We are requesting the EcoHealth Alliance federally approved indirect cost rate of 32.74% on all applicable direct costs. Indirect is taken only on the first $25,000 for each consortium/contractual agreement in each year. As there are 3 (Wuhan Institute of Virology, Institute of Pathogen Biology, and University of North Carolina), a total of $24,555 ($8,185 x 3) is requested as indirect costs on consortium/contractual/subaward agreements. This is not included as part of direct cost calculations and is only requested for year 1. In years 2-5 no indirect will be taken on consortium/contractual agrrement subcontracts. 4 Page 99 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other 2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Section K, Total Costs and Fee (I + J) Tracking Number: GRANT12743073 Totals($) 0 47,200.00 149,790.00 0.00 0.00 0.00 0.00 0.00 0 80,250.00 24,000.00 398,750.00 0.00 953,245.00 0.00 0.00 0.00 0.00 0.00 Page 100 1,118,565.00 0.00 1,118,565.00 0.00 196,990.00 0.00 1,456,245.00 2,771,800.00 814,960.00 3,586,760.00 0.00 3,586,760.00 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 1 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2019 A. Senior/Key Person End Date•: 05-31-2020 Budget Period: 1 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 Prefix First Name• Middle Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Name Salary ($) Months Months Months Salary ($)* Benefits ($)* :t: ~?:::::: ;!r :::::::::::::::::::::. ~:::::::::::::::::::::: ;;t: ........... :::::·::::::: ::::: ~r m~;;:ir~;:;~:::::: :::·::::::::::: ::::::r) ( 4 ). ~: :::::::::::::::::: :::::::::::::::::::: :1:::::::::::::::::::::::::::::(b:):( 4 :):. (b:):(:6) Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person I Cb> <4>, Cb><~ B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••-- .. •••••••••--••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical .............................................................................. , (b) (4), (b) <4 1 Total Number Other Personnel Total Other Personnel (b) (4), (b) (6) -----! Total Salary, Wages and Fringe Benefits (A+B) RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 101 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 1 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2019 End Date*: 05-31-2020 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs Budget Period: 1 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* Total Travel Cost 0.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 102 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 1 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2019 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2020 Budget Period: 1 Funds Requested ($)* 15,960.00 Total Other Direct Costs 15,960.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . All Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 50,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 55.5 50,000.00 _______ 27,750.00 ___, Total Indirect Costs 27,750.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) n,750.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_UNC_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 77,750.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 103 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 2 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2020 A. Senior/Key Person End Date•: 05-31-2021 Budget Period: 2 Prefix First Name• Middle Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Name Salary ($) Months Months Months Salary ($)* Benefits ($)* 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 Funds Requested ($)* :t: ~?::::::: ;;r:::::::::::::::::::::. ~:::::::::::::::::::::: ;;t: ....................................... ~.;:.:.~~.;;.::;;:;~ .............................. :I ___________________________ (b_)_< 4 _)_. (b-)-(~6) Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person I (b) (4). (b) <~ B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••--••••••••n••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical · · 1 ··--· · · .. Laboratory 'f echn.idari · ·· ..... ····· ····· · ... ·1 (b) <4>. (b) <~ ·· .... · · · · ··· ... · ····· ····· .. · ····· ···· ................................... ··· · ..... ·1 (b) (4). (b) (6) (b) (4). (b) (6) 1 Total Number Other Personnel RESEARCH & RELATED Budget {A-B) (Funds Requested) Tracking Number: GRANT12743073 Page 104 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 2 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2020 End Date*: 05-31-2021 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs Budget Period: 2 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* Total Travel Cost 0.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 105 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 2 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2020 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2021 Budget Period: 2 Funds Requested ($)* 15,960.00 Total Other Direct Costs 15,960.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . All Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 50,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 55.5 50,000.00 _______ 27,750.00 ___, Total Indirect Costs 27,750.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) n,750.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_UNC_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 77,750.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 106 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 3 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2021 A. Senior/Key Person Prefix First Name• Last Name• Suffix Project Role* End Date•: 05-31-2022 Budget Period: 3 Base Calendar Academic Summer Requested Fringe 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 Middle Funds Requested ($)* Name Salary ($) Months Months Months Salarv ~,_•_B_e_n_e_f,_'ts__,(,$":,)_* -----~~~~~ (b) (4), (b) (6) .1 .... 1?.r:...... f:<1IP.h. ................... ?.................. 8.a.r~~ ........................................ <?.°.~.1.~~.E:~.~i~<1t<:>r .............................. .. . 2..:.1?.r: ...... t'.l.mY ................................................ ?.irr,. 7 ................................... <?°.~.1.~~.E:~.ti~<1t<:>r ............................... ------------------------------ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: B. Other Personnel Number of Project Role* Personnel* Post Doctoral Associates Total Senior/Key Person I Cb> (4), Cb> (6) Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• .. •••••••••n••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical ............................................... ·······. ······· ......... . 1 Laboratory Technician 1 Total Number Other Personnel RESEARCH & RELATED Budget {A-B) (Funds Requested) Tracking Number: GRANT12743073 I Cb) < 4 >, Cb><~ . . . . . ..... . ..... ..... .. . ..................................................... ·1 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Page 107 -----1 (b) (4), (b) (~ (b) (4), (b) (6) Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 3 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2021 End Date*: 05-31-2022 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs Budget Period: 3 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* Total Travel Cost 0.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 108 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 3 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2021 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2022 Budget Period: 3 Funds Requested ($)* 15,960.00 Total Other Direct Costs 15,960.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . All Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 50,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 55.5 50,000.00 _______ 27,750.00 ___, Total Indirect Costs 27,750.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) n,750.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_UNC_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 77,750.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 109 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 4 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2022 A. Senior/Key Person End Date•: 05-31-2023 Budget Period: 4 Prefix First Name• Middle Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Name Salary ($) Months Months Months Salary ($)* Benefits ($)* 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 Funds Requested ($)* :t: ~?::::::: ;;r:::::::::::::::::::::. ~:::::::::::::::::::::: ;;t: ....................................... ~.;:.:.~~.;;.::;;:;~ .............................. :I ___________________________ (b_)_< 4 _>_. (b_)_~_6) Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person r (b) (4). (b) <~ B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••--••••••••n••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical · · 1 ··--· · · .. Laboratory 'f echn.idari · ·· ..... ····· ····· · ........ i(b> <4>. Cb><~ ·· .... · · · · ··· ... · ····· ····· .. · ····· ···· ................................... ··· · ..... ·1 (b) (4), (b) (6) (b) (4), (b) (6) 1 Total Number Other Personnel RESEARCH & RELATED Budget {A-B) (Funds Requested) Tracking Number: GRANT12743073 Page 110 Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 4 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2022 End Date*: 05-31-2023 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs Budget Period: 4 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* Total Travel Cost 0.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 111 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 4 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2022 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2023 Budget Period: 4 Funds Requested ($)* 15,960.00 Total Other Direct Costs 15,960.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . All Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 50,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 55.5 50,000.00 _______ 27,750.00 ___, Total Indirect Costs 27,750.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) n,750.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_UNC_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 77,750.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 112 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 5 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2023 A. Senior/Key Person End Date•: 05-31-2024 Budget Period: 5 Prefix First Name• Middle Last Name• Suffix Project Role* Base Calendar Academic Summer Requested Fringe Name Salary ($) Months Months Months Salary ($)* Benefits ($)* 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 Funds Requested ($)* :t: ~?::::::: ;;r:::::::::::::::::::::.~:::::::::::::::::::::: ;;t: ................................. ~.;:.:.~~.;;.::;;:;~ __ ..................... :l ___________________________ (b_)(_ 4 _)._(b_)_(6) Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person 20,819.00 B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••-- .. •••••••••--••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical · ·1 ··--· ·.. La6ora1oi-y'fechn.idari · ·· ..... ····· ····· · ... --··1Cb>< 4>.Cb><~ ·· .... ·· · · ··· ... · ········· --········· ...................................... · ..... --1 Cb><4>.Cb><6) --------'---------------.1-----------------~----------------, (b)(4).(b)(6) 1 Total Number Other Personnel Total Other Personnel Total Salary, Wages and Fringe Benefits (A+B) RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 113 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 5 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2023 End Date*: 05-31-2024 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) 2. Foreign Travel Costs Budget Period: 5 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* Total Travel Cost 0.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 114 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 5 ORGANIZATIONAL DUNS*: 6081952770000 Budget Type*: O Project • Subaward/Consortium Organization: The University of North Carolina at Chapel Hill Start Date*: 06-01-2023 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2024 Budget Period: 5 Funds Requested ($)* 15,960.00 Total Other Direct Costs 15,960.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . All Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 50,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 55.5 50,000.00 _______ 27,750.00 ___, Total Indirect Costs 27,750.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) n,750.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_UNC_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 77,750.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 115 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL BUDGET JUSTIFICATION, SUBAWARD A. Senior/Key Personnel Ralph Barie, PhD Co-Investigator CbH4>. CbH6J. Dr. Barie is a known expert in coronavirus cross species transmission and pathogenesis and has studied this group of viruses for over 30 years. His group developed the first reverse genetic systems for epidemic and zoonotic SARS-like coronaviruses and they have studied the ability of these viruses to replicate efficiently in various primary human airway epithelial cell cultures as well as other key primary cell types. His group has also studied the sensitivity of these viruses to be controlled by existing vaccines and therapeutics both in vitro and in vivo. Dr. Barie will lead the studies at the University of North Carolina at Chapel Hill. He will design research strategies, interpret findings and review research outcomes with Dr. Sims and Mr. Yount. At a regular basis, Dr. Barie will report the results of the teams research to Dr. Daszak and Dr. Shi and together, they will use this information to identify additional research priorities and design downstream studies. Ors. Daszak, Shi and Barie have published together in the past and participated on research project applications. Dr. Barie recently spent several days in Wuhan, China, where he discussed research strategies and collaborations with Dr. Daszak and Dr. Shi. He will work closely with Dr. Sims and Mr. Yount to prepare timely reports, share research and discuss future research directions with the group. Amy Sims, PhD Co-Investigator (b)(4).(b)(6)_ Dr. Sims has over 22 years of research studying coronavirus molecular biology, replication and pathogenesis. She has published over 50 papers including seminal papers on characterizing host response patterns of primary human lung airway epithelial cells and other cell types after infection with SARS-CoV, MERS-CoV, influenza and various SARS-like bat coronaviruses. She is not only well versed in the preparation, cultivation and maintenance of primary human lung cells but also proficient at studying virus infection outcomes, in the presence and absence of antiviral therapeutics. In consultation with Dr. Barie, Dr. Sims will design experiments, perform infections and characterize epidemic and bat SARS-like coronavirus replication in human cells. She will compile data and share these results with the research team. Dr. Sims will also interface and work closely with Mr. Yount, who will assist in these studies, including infections, cell preparations and characterizing virus growth efficiency in these cultures. Dr. Sims has over 15 years of experience working in a BSL3 laboratory and oversees the management of these facilities. She has select agent clearance. B. Other Personnel Mr. Boyd Yount, Laboratory Technician CbH4).(b)(6) Mr. Yount has published over 50 papers on coronaviruses and developed the first reverse genetic platforms for SARS-CoV, MERS-CoV and various SARS-like bat corona viruses. He will work closely with Ors. Barics, Shi and Daszak to design and recover select bat SARS-like coronaviruses for downstream studies in the Barie and Shi laboratories, including characterizing virus phenotypes in primary cells as well as cells expressing various human, civet and bat ACE2 receptors. He will prepare virus stocks, Mr. Yount will work closely with Ors. Barie and Sims to design and implement experiments in the BSL3 laboratory, prepare reports and research outcomes during the course of the program. Mr. Yount has over 15 years of experience in a BSL3 setting and is well versed in all the techniques used in this proposal. He has select agent clearance. Fringe Benefits. Benefits are for faculty, staff and postdoctoral research associates are calculated as follows: Faculty and Staff- 24.519% Social Security and retirement and $6,104 for health insurance, Supplies and Reagents. $15,960 C. Equipment No equipment over $5,000 will be purchased. D. Travel No travel will be requested for this subaward. Travel to EcoHealth Alliance and other collaborators will be covered from other UNC funding. Page 116 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter F. Other Direct Costs Materials and Supplies. A variety of culture media and serum ($3,000), primary cell procurement ($2500), recombinant enzymes ($1500) and antibodies ($1500), synthetic DNAs ($2,500) and an assortment of miscellaneous supplies (e.g., gloves, chemicals, plasticware, etc.)($2460) are needed during the course of the program to recover recombinant viruses and maintain cells in culture, perform virus growth curves and identify virus tropisms by immunohistochemistry. In addition, personnel protective equipment (PPE), portal breathing apparati (PAPR), globes and protective clothing are used in the BSL3 setting ($2500). H. Indirect Costs In an agreement with DHHS dated 11/23/2016 the indirect cost rate for The University of North Carolina is 55.5% of MTDC, excluding equipment and tuition. 2 Page 117 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other 2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Section K, Total Costs and Fee (I + J) Tracking Number: GRANT12743073 Totals($) 5 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0 79,800.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Page 118 104,095.00 66,105.00 170,200.00 0.00 0.00 0.00 79,800.00 250,000.00 138,750.00 388,750.00 0.00 388,750.00 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 1 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology Start Date*: 06-01-2019 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2020 Budget Period: 1 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .. :.1?.r:...... i'.'.~.e.~.9.!i ........................................... S.~.i....... .. ..................... <?°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) . 2..:.1?.r: ...... f=>~.~9 .............................................. i'.'.~<:>.U. .................................... <?°.~.1.~~.E:~.tigat<:>r ............................... . 3..: .. [).r: ........ 13.~.~ .................................................... !:il! .............................................. <?.°.~.1.~- -E:~.~i~.~t<:>r ................................ ,_ ____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel Number of Project Role* Personnel' Post Doctoral Associates File Name: Total Senior/Key Person 17,667.00 Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ················-·····································-····························-·················· ······································•···-··································•························································································· ................................................................................................ . Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel 0.00 --------- Total Salary, Wages and Fringe Benefits (A+B) 17,667.00 RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 119 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 1 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2019 End Date*: 05-31-2020 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 1 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 120 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 1 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2019 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2020 Budget Period: 1 Funds Requested ($)* 48,668.00 Total Other Direct Costs 48,668.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,649.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,648.00 _______ 5,652.00 ___, Total Indirect Costs 5,652.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 76,301.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_WIV_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 76,301.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 121 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 2 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology Start Date*: 06-01-2020 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2021 Budget Period: 2 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salarv (_$)* Benefits ($0,),*------~=~~ .1 .. :.1?.r:...... i'.'.~.e.~.9.!i ........................................... S.~.i....... .. ..................... <?°.~.1.~~.E:~.~i~~t<:>r .............................. .. (b) (4), (b) (6) . 2..:.1?.r: ...... f=>~.~9 .............................................. i'.'.~<:>.U. .................................... <?°.~.1.~~.E:~.tigat<:>r .............................. . 3..: .. [).r: ........ 13.~.~ .................................................... !:il! .............................................. <?.°.~.1.~- -E:~.~i~.~t<:>r ............................... _____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel Number of Project Role* Personnel' Post Doctoral Associates File Name: Total Senior/Key Person 17,667.00 Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ················-·····································-····························-·················· ······································•······································•················································································· ... ·-··· ................................................................................................ . Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel 0.00 --------- Total Salary, Wages and Fringe Benefits (A+B) 17,667.00 RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 122 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 2 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2020 End Date*: 05-31-2021 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 2 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 123 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 2 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2020 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2021 Budget Period: 2 Funds Requested ($)* 48,668.00 Total Other Direct Costs 48,668.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,649.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,648.00 _______ 5,652.00 ___, Total Indirect Costs 5,652.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 76,301.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_WIV_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 76,301.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 124 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040-0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 3 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology Start Date*: 06-01-2021 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2022 Budget Period: 3 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .. :.1?.r:...... i'.'.~.e.~.9.!i ........................................... S.~.i....... .. ..................... <?°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) . 2..:.1?.r: ...... f=>~.~9 .............................................. i'.'.~<:>.U. .................................... <?°.~.1.~~.E:~.tigat<:>r ............................... . 3..: .. [).r: ........ 13.~.~ .................................................... !:il! .............................................. <?.°.~.1.~- -E:~.~i~.~t<:>r ............................... . Tot a I Funds Requested for all Senior Key Persons in the attached file ------------------------------ Additional Senior Key Persons: B. Other Personnel Number of Project Role* Personnel' Post Doctoral Associates File Name: Total Senior/Key Person 17,667.00 Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ················-·····································-····························-·················· ······································•···-··································•························································································· ................................................................................................ . Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel 0.00 --------- Total Salary, Wages and Fringe Benefits (A+B) 17,667.00 RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 125 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 3 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2021 End Date*: 05-31-2022 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 3 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 126 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 3 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2021 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2022 Budget Period: 3 Funds Requested ($)* 48,668.00 Total Other Direct Costs 48,668.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,649.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,648.00 _______ 5,652.00 ___, Total Indirect Costs 5,652.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 76,301.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_WIV_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 76,301.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 127 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 4 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology Start Date*: 06-01-2022 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2023 Budget Period: 4 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .. :.1?.r:...... i'.'.~.e.~.9.!i ........................................... S.~.i....... .. ..................... <?°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) . 2..:.1?.r: ...... f=>~.~9 .............................................. i'.'.~<:>.U. .................................... <?°.~.1.~~.E:~.tigat<:>r ............................... . 3..: .. [).r: ........ 13.~.~ .................................................... !:il! .............................................. <?.°.~.1.~- -E:~.~i~.~t<:>r ................................ _____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel Number of Project Role* Personnel' Post Doctoral Associates File Name: Total Senior/Key Person 17,667.00 Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ················-·····································-····························-·················· ······································•···-··································•························································································· ................................................................................................ . Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel 0.00 --------- Total Salary, Wages and Fringe Benefits (A+B) 17,667.00 RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 128 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 4 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2022 End Date*: 05-31-2023 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 4 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 129 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 4 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2022 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2023 Budget Period: 4 Funds Requested ($)* 48,668.00 Total Other Direct Costs 48,668.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,649.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,648.00 _______ 5,652.00 ___, Total Indirect Costs 5,652.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 76,301.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_WIV_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 76,301.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 130 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 5 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Wuhan Institute of Virology Start Date*: 06-01-2023 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2024 Budget Period: 5 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .. :.1?.r:...... i'.'.~.e.~.9.!i ........................................... S.~.i....... .. ..................... <?°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) . 2..:.1?.r: ...... f=>~.~9 .............................................. i'.'.~<:>.U. .................................... <?°.~.1.~~.E:~.tigat<:>r ............................... . 3..: .. [).r: ........ 13.~.~ .................................................... !:il! .............................................. <?.°.~.1.~- -E:~.~i~.~t<:>r ................................ ------------------------------ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: B. Other Personnel Number of Project Role* Personnel' Post Doctoral Associates File Name: Total Senior/Key Person 17,667.00 Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ················-·····································-····························-·················· ······································•···-··································•························································································· ................................................................................................ . Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel 0.00 --------- Total Salary, Wages and Fringe Benefits (A+B) 17,667.00 RESEARCH & RELATED Budget {A-B) (Funds Requested) Page 131 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 5 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2023 End Date*: 05-31-2024 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 5 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 132 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 5 ORGANIZATIONAL DUNS*: 5290274740000 Budget Type*: O Project • Subaward/Consortium Organization: Wuhan Institute of Virology Start Date*: 06-01-2023 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2024 Budget Period: 5 Funds Requested ($)* 48,668.00 Total Other Direct Costs 48,668.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,649.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,648.00 _______ 5,652.00 ___, Total Indirect Costs 5,652.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 76,301.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_WIV_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 76,301.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 133 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Program DirectOf/Principal Investigator: Daszak, Peter WUHAN INSTITUTE OF VIROLOGY BUDGET JUSTIFICATION, SUBAWARD A. Senior/Key Personnel: Dr. Zhengli Shi, Co-Investigator. Senior Research Scientist at Wuhan Institute of Virology (WIV) Chinese Academy of Sciences, will commit CbH4),(b)(6l per year H 4),(b)( to this project to oversee the laboratory implementation at WIV. At a regular basis, Dr. Shi will meet with other Co-Pis to refine study protocols, report back results, and prepare publications. Dr. Shi has been working on the discovery and characterization of novel viruses from bats and other wildlife since 2004. This included the discovery that Chinese horseshoe bats are the natural reservoir of SARSr-CoVs and the likely origin of SARS-CoV. Her lab at WIV isolated SARSrCoVs from bats sharing high homology with human SARS-CoV and demonstrated their interspecies transmission risk, largely confirming bats as the source of SARs. She will lead her team to carry out then systematic studies on the epidemiology, genetic evolution, interspecies infection mechanism and pathogenesis of a series of bat-borne CoVs on this R01 renewal proposal. Dr. Peng Zhou. Co-Investigator. Research Scientist at Wuhan Institute of Virology (WIV) Chinese Academy of Sciences, will commit will commit Cb) (4), (b)(6) per year {b)(4), (b)(6) to this project to be in charge of the diagnostics, genomics, and virus isolation work at WIV. Dr. Zhou have been working on bat virology since 2004, who will contribute his expertise in next generation diagnostic tool development for monitoring bat virus spillover, bat pathogen discovery, and bat viral immunology to this R01 renewal proposal. Dr. Ben Hu, Co-Investigator. Research Associate and coordinator of Shi's laboratory at WIV, will commit Cb>< 4 >,~ CbH4>.CbH per year H4),(b)(6) to this project to directly supervisor the lab technicians, and coordinate with our collaborators in this R01 proposal for communication, reporting, and or organizing meetings. Dr. Hu has been working on discovery and characterization of viruses in small mammals in bats and rodents since 2010 at WIV, and had extensive experience managing international collaborative projects. B. Equipment No equipment over $5,000 will be purchased. C. Travel We are requesting $4,314 per year for all years for Dr. Shi to travel to the United States to meet with EcoHealth Alliance (Daszak, Francesco, Oliva!, Ross) and University of North Carolina at Chapel Hill (Barie, Sims) collaborators. Travel is calculated at one round trip airfare from Wuhan to New York City ($1,000), ninenight hotel in New York City ($288 per night), and 1 0 days per diem at $76 per day except for first and last day, which have a reduced per diem of $57. D. Other Direct Costs We are requesting support for laboratory experiments and related testing costs with a minimum base of 2,000 samples from 1,000 animals per year. RNA Extractions We will be running RNA Extractions for 1,000 bats per year (two samples per bat: rectal and blood) in each year of the project. This will cost $6,214 per year (QIAamp ViralRNA Mini Kit with Axygen Pipette Tips and Filter Tubes at $3.11 per sample). RT-PCR Costs for 1-Step RT-PCR assays for Coronavirus conducted on 2,000 samples per year for each year of the project total $6,358 and are detailed as follows: Superscript Ill one step kit ($2.31 per sample); Platinum Tag DNA Polymerase ($0.25 per sample); nuclease-free water ($0.07 per sample); and Axygen Pipette Tips and Filter Tubes ($0.54 per sample). DNA Sequencing In each year of the project, DNA Sequencing will be performed on 1,500 samples at a cost of $4.34 per reaction. We request a total of $6,503 per year in each year. Page 134 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter In vitro Infection Experiment We are requesting support for in vitro infection experiments using pseudoviruses carrying the spike proteins (wild type or mutants) or live viruses in cell lines of different origins, binding affinity assays between the spike proteins (wild type or mutants) and different cellular receptor molecules, and humanized mouse experiments. In each year of the project, we request $1,040 for Lipofectamine3000 transfection reagent; $3,612 for GIBCO Fetal Bovine Serum, $517 for GIBCO antibiotic antimycotic, and 2,601 for GIBCO medium that will be used in the in vitro infection experiments that will be used for in vitro infection experiment, as well as $6,000 for cell lines, in total of $8,639 per year. Luciferase lmmunoprecipitation System (LIPS) Assay We are requesting $18,642 to support the in each year of the project to develop LIPS assay for bat CoV antibody detection, with detailed cost as follows: $9,827 for Protein A/G Ultralink Resin; $434 for Monoclonal ANTI-FLAG(R) M2 antibody; $5,636 for Ren ii la Luciferase Assay System; $2,168 for Merck-Millipore MSBVN1850 MultiScreen HTS; $578 for Axygen Polypropylene PCR Tube Strips. The Enzyme-Linked lmmunosorbent Assay (ELISA) We also request $2,312 to support the serological testing of 1,000 bat serum samples per year with ELISA plates, at the cost of $2.31 per sample. E. Indirect Costs We are requesting an extremely low indirect cost of 8% on all direct costs. 2 Page 135 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other 2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Section K, Total Costs and Fee (I + J) Tracking Number: GRANT12743073 Totals($) 0 0.00 21,570.00 0.00 0.00 0.00 0.00 0.00 0 243,340.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Page 136 88,335.00 0.00 88,335.00 0.00 21,570.00 0.00 243,340.00 353,245.00 28,260.00 381,505.00 0.00 381,505.00 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 1 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Institute of Pathogen Biology Start Date*: 06-01-2019 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2020 Budget Period: 1 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .... 1?.r: ....... ~.iii ..................................................... Ren ..................................... <?.°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4), (b) (6) 2 . Dr. Li Guo Co-Investigator .................................................................................................................................................................................................. .._ ____________________________ --! Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: B. Other Personnel Number of Project Role* Personnel* Post Doctoral Associates Total Senior/Key Person I Cb> (4), Cb> c~ Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••••••••••••••••••• •••••••••--•••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-B} (Funds Requested) Tracking Number: GRANT12743073 Page 137 Total Other Personnel Total Salary, Wages and Fringe Benefits (A~B) 0.00 I Cb> (4). Cb> c'1 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 1 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2019 End Date*: 05-31-2020 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 1 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 138 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 1 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2019 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2020 Budget Period: 1 Funds Requested ($)* 48,686.00 Total Other Direct Costs 48,686.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,000.00 _______ 5,600.00 ___, Total Indirect Costs 5,600.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 75,600.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_1PB_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 75,600.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 139 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 2 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Institute of Pathogen Biology Start Date*: 06-01-2020 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2021 Budget Period: 2 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .... 1?.r: ....... ~.iii ..................................................... Ren ..................................... <?.°.~.1.~~.E:~.~i~~t<:>r .............................. .. (b) (4), (b) (6) 2 . Dr. Li Guo Co-Investigator ................................................................................................................................................................................................. _____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person I Cb> <4>, Cb><~ B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••--••••••••n••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel Total Salary, Wages and Fringe Benefits (A~B) 0.00 I (b) (4), (b) (6) ~----------------------------------------------------------~· RESEARCH & RELATED Budget {A-B} (Funds Requested) Page 140 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 2 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2020 End Date*: 05-31-2021 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 2 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 141 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 2 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2020 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2021 Budget Period: 2 Funds Requested ($)* 48,686.00 Total Other Direct Costs 48,686.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,000.00 _______ 5,600.00 ___, Total Indirect Costs 5,600.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 75,600.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_1PB_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 75,600.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 142 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 3 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Institute of Pathogen Biology Start Date*: 06-01-2021 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2022 Budget Period: 3 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .... 1?.r: ....... ~.iii ..................................................... Ren ..................................... <?.°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) 2 . Dr. Li Guo Co-Investigator .................................................................................................................................................................................................. _____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person I (b) (4). (b) (6) B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••-- .. •••••••••--••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel Total Salary, Wages and Fringe Benefits (A~B) 0.00 I (b) (4). (b) (6) ~-----------------------------------------------------------~· RESEARCH & RELATED Budget {A-B} (Funds Requested) Page 143 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 3 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2021 End Date*: 05-31-2022 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 3 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 144 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 3 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2021 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2022 Budget Period: 3 Funds Requested ($)* 48,686.00 Total Other Direct Costs 48,686.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,000.00 _______ 5,600.00 ___, Total Indirect Costs 5,600.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 75,600.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_1PB_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 75,600.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 145 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 4 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Institute of Pathogen Biology Start Date*: 06-01-2022 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2023 Budget Period: 4 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salarv (_$)* Benefits ($0,),*------~=~~ .1 .... 1?.r: ....... ~.iii ..................................................... Ren ..................................... <?.°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4), (b) (6) 2 . Dr. Li Guo Co-Investigator .................................................................................................................................................................................................. ------------------------------ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: B. Other Personnel Number of Project Role* Personnel* Post Doctoral Associates Total Senior/Key Person r (b) (4). (b) <~ Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• •••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••----•••••••••--••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel RESEARCH & RELATED Budget {A-B} (Funds Requested) Tracking Number: GRANT12743073 Page 146 Total Other Personnel Total Salary, Wages and Fringe Benefits (A~B) 0.00 I (b) (4), (b) <~ Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 4 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2022 End Date*: 05-31-2023 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 4 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 147 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 4 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2022 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2023 Budget Period: 4 Funds Requested ($)* 48,686.00 Total Other Direct Costs 48,686.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,000.00 _______ 5,600.00 ___, Total Indirect Costs 5,600.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 75,600.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_1PB_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 75,600.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 148 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER 0MB Numbe·: 4040·0001 Expiration Date: 10/31/2019 RESEARCH & RELATED BUDGET - SECTION A & B, Budget Period 5 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Enter name of Organization: Institute of Pathogen Biology Start Date*: 06-01-2023 A. Senior/Key Person Prefix First Name• Middle Name Last Name• Suffix Project Role* End Date•: 05-31-2024 Budget Period: 5 Base Calendar Academic Summer Requested Fringe Funds Requested ($)* Salary ($) Months Months Months Salary ($)* Benefits ($)* .1 .... 1?.r: ....... ~.iii ..................................................... Ren ..................................... <?.°.~.1.~~.E:~.~i~~t<:>r ................................ . (b) (4). (b) (6) 2 . Dr. Li Guo Co-Investigator .................................................................................................................................................................................................. _____________________________ _ Total Funds Requested for all Senior Key Persons in the attached file Additional Senior Key Persons: File Name: Total Senior/Key Person I (b) (4). (b) <~ B. Other Personnel Number of Project Role* Personnel* Calendar Months Academic Months Summer Months Requested Salary ($)* Fringe Benefits* Funds Requested ($)* Post Doctoral Associates ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••n••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••--••••••••n••••••••••••••••••••••••••••••••• ••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••••• Graduate Students ............................. Undergraduate.Students ................................................................................................................................................................................................................................................................................................ . Secretarial/Clerical 0 Total Number Other Personnel Total Other Personnel Total Salary, Wages and Fringe Benefits (A~B) 0.00 I (b) (4). (b) (6) ~----------------------------------------------------------~ RESEARCH & RELATED Budget {A-B} (Funds Requested) Page 149 Tracking Number: GRANT12743073 Funding Opportunity Number: PA· 18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - SECTION C, D, & E, Budget Period 5 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2023 End Date*: 05-31-2024 C. Equipment Description List items and dollar amount for each item exceeding $5,000 Equipment Item Total funds requested for all equipment listed in the attached file Additional Equipment: File Name: D. Travel 1. Domestic Travel Costs ( Incl. Canada, Mexico, and U.S. Possessions) Budget Period: 5 Funds Requested ($)* Total Equipment 0.00 Funds Requested ($)* 2. Foreign Travel Costs 4,314.00 Total Travel Cost 4,314.00 E. Participant/Trainee Support Costs Funds Requested ($)* 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other: Number of Participants/Trainees RESEARCH & RELATED Budget {C-E) (Funds Requested) Tracking Number: GRANT12743073 Total Participant Trainee Support Costs 0.00 Page 150 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET- SECTIONS F-K, Budget Period 5 ORGANIZATIONAL DUNS*: 5281563570000 Budget Type*: O Project • Subaward/Consortium Organization: Institute of Pathogen Biology Start Date*: 06-01-2023 F. Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations End Date*: 05-31-2024 Budget Period: 5 Funds Requested ($)* 48,686.00 Total Other Direct Costs 48,686.00 G. Direct Costs Funds Requested ($)* H. Indirect Costs Indirect Cost Type 1 . Direct Costs Cognizant Federal Agency (Agency Name, POC Name, and POC Phone Number) Total Direct Costs (A thru F) 70,000.00 Indirect Cost Rate (%) Indirect Cost Base ($) Funds Requested ($)* 8.0 70,000.00 _______ 5,600.00 ___, Total Indirect Costs 5,600.00 I. Total Direct and Indirect Costs Funds Requested ($)* Total Direct and Indirect Institutional Costs (G + H) 75,600.00 IJ. Fee Funds Requested m·1 K. Total Costs and Fee Funds Requested($)* L. Budget Justification* File Name: NIAID_COV_2019_1PB_BUDGET_JUSTIFICATION.pdf (Only attach one file.) 75,600.00 RESEARCH & RELATED Budget (F-K) (Funds Requested) Page 151 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 . Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Program DirectOf/Principal Investigator: Daszak, Peter WUHAN INSTITUTE OF VIROLOGY BUDGET JUSTIFICATION, SUBAWARD A. Senior/Key Personnel: Dr. Lili Ren. PhD Co-Investigator CbH4),(b)( . Dr. Ren is an expert in the pathogenesis and evolution of respiratory viruses. She will refine study protocols, coordinate research, oversee implementation of all activities, analyze data, lead regular meetings with other Co-Investigators and Other Senior/Key Personnel as well as draft papers. Dr. Li Guo, PhD Co-Investigator and Senior Research Technician Cb> <4>, Cb> (6) per year to perform all laboratory work. She has been working on etiology and immunology research on respiratory viruses since 2003 and has evaluated the cross-reactivities of N among HCoVs and developed a competitive ELISA (cELISA) for detecting anti-N lgG antibodies against HCoV -229E, -OC43, -NL63, and -HKU1. B. Other Personnel No other Personnel will be required for this subaward. All Institute of Pathogen Biology salaries include the US benefits, so benefits are not calculated separately. C. Equipment No equipment over $5,000 will be purchased. D. Travel We are requesting $4,314 per year for all years for Dr. Ren or Dr. Guo to travel to the United States to meet with EcoHealth Alliance (Daszak, Francesco, Oliva!, Ross) and University of North Carolina at Chapel Hill (Barie, Sims) collaborators. Travel is calculated at one round trip airfare from Beijing to New York City ($1,000), nine-night hotel in New York City ($288 per night), and 10 days per diem at $76 per day except for first and last day, which have a reduced per diem of $57. F. Other Direct Costs We are requesting support for laboratory experiments and related costs with a minimum base of 1,000 samples expected per year. RNA Extractions We will be running 1,000 RNA Extractions per year in each year of the project. This will cost $10,450 per year for QIAamp Viral RNA Mini Kit with an additional $2,023 per year for Axygen Pipette Tips and Filter Tubes at $1.08 per sample. 1-STEP RT-PCR Costs for 1-Step RT-PCR assays for Coronavirus conducted on 1,000 samples per year for each year of the project total $6,358 and are detailed as follows: Superscript Ill one step kit ($4.62 per sample); Platinum Tag DNA Polymerase ($0.51 per sample); nuclease-free water ($0.15 per sample); and Axygen Pipette Tips and Filter Tubes ($1.08 per sample). DNA Sequencing In each year of the project, DNA Sequencing will be performed on 3,200 samples at a cost of $2.62 per reaction. We request a total of $2,601 per year in year. Cell Culture We request a total of $4,913 in year one; $6,647 in year two (upon expectation of 35% increase in positive samples); and $8,671 (a 30% increase) per year for the remaining years (3-5) of the project to cover costs for cell culturing. This will require GIBCO Fetal Bovine Serum, antibiotic antimycotic. growth medium. and cell culture plates and flasks. Flask costs are estimated at $1.45 per plate/flask. Page 152 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Protein Expression and Purification For protein expression and purification, we request a total of $6,590 in year 1, $6,705 in year 2, and $5,260 in each year of years 3-5. Details of protein expression and purification costs include histidine and sepharose tagged protein purification fast flow columns ($116 each), Q sepharose fast flow media ($116 each), protease inhibitor ($289 each), eStain Protein staining kits ($145 each), protein G sepharose 4 fast flow ($145 each), Q and SP sepharose fast flow ($723 each), and bacterial culture plates ($116 each). It is expected that more samples will be processed in year 1, so costs are estimated to reduce by 20% in years 2-5 Serological Tests We request support for serology assays. None will be conducted in year one until we have samples. Costs are estimated for years 2-5 with $9,220 in year 2 and due to additional lgM and lgG secondary antibodies costs and increase in estimate to $9,436 in years 3-5. Cost estimates include lgM and lgG secondary antibodies at $72.25; mouse and rabbit lgG antibodies at $434; mouse and rabbit lgM antibodies at $434; and ELISA plates at $2.90 each. Lab Supplies Funding is requested to support laboratory supplies including three (3) -80°C freezers ($4,340 each); reagents including agarose, sodium chloride, yeast extract, phosphate buffer, Tris and other biochemical reagents (average of $307 per year); centrifuge tube costs are estimated to increase in years 2-5 (average of $665 per year); disposable personnel protective equipment (PPE), portal breathing apparati (PAPR), globes and protective clothing to be used in BSL settings are estimated at $723 per year; cell lines will be required in years 1 and 2 ($867 each); and in vitro culture (lipofectamine2000) will be required at cost of $434 in year 1 and $2,168 per year in years 2-5. H. Indirect Costs We are requesting an extremely low indirect cost of 8% on all direct costs. 2 Page 153 Budget Justification Attachment

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Contact PD/Pl: DASZAK, PETER RESEARCH & RELATED BUDGET - Cumulative Budget Section A, Senior/Key Person Section B, Other Personnel Total Number Other Personnel Total Salary, Wages and Fringe Benefits (A+B) Section C, Equipment Section D, Travel 1. Domestic 2. Foreign Section E, Participant/Trainee Support Costs 1. Tuition/Fees/Health Insurance 2. Stipends 3. Travel 4. Subsistence 5. Other 6. Number of Participants/Trainees Section F, Other Direct Costs 1. Materials and Supplies 2. Publication Costs 3. Consultant Services 4. ADP/Computer Services 5. Subawards/Consortium/Contractual Costs 6. Equipment or Facility Rental/User Fees 7. Alterations and Renovations 8. Other 1 9. Other 2 10. Other 3 Section G, Direct Costs (A thru F) Section H, Indirect Costs Section I, Total Direct and Indirect Costs (G + H) Section J, Fee Section K, Total Costs and Fee (I + J) Tracking Number: GRANT12743073 Totals($) 0 0.00 21,570.00 0.00 0.00 0.00 0.00 0.00 0 243,430.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 Page 154 85,000.00 0.00 85,000.00 0.00 21,570.00 0.00 243,430.00 350,000.00 28,000.00 378,000.00 0.00 378,000.00 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Total Direct Costs less Consortium F&A NIH policy (NOT-OD-05-004) allows applicants to exclude consortium/contractual F&A costs when determining if an application falls at or beneath any applicable direct cost limit. When a direct cost limit is specified in an FOA, the followina table can be used to determine if vour aoolication falls within that limit. Category Budget Period 1 Budget Period 2 Budget Period 3 Budget Period 4 Budget Period 5 TOTALS Total Direct Costs less Consortium F&A 515,358 515,358 515,358 515,358 515,3581 2,576,790 Tracking Number: GRANT12743073 Page 155 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER PHS 398 Cover Page Supplement 1. Vertebrate Animals Section Are vertebrate animals euthanized? O Yes e No If "Yes" to euthanasia Is the method consistent with American Veterinary Medical Association (AVMA) guidelines? O Yes 0 No If "No" to AVMA guidelines, describe method and provide scientific justification 2. *Program Income Section *Is program income anticipated during the periods for which the grant support is requested? O Yes e No 0MB Number. 0925-0001 Expiration Date: 03/3112020 If you checked "yes" above (indicating that program income is anticipated), then use the format below to reflect the amount and source(s). Otherwise, leave this section blank. *Budget Period *Anticipated Amount($) *Source(s) Tracking Number: GRANT12743073 Page 156 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER PHS 398 Cover Page Supplement 3. Human Embryonic Stem Cells Section *Does the proposed project involve human embryonic stem cells? O Yes e No If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell line(s) from the following list: http://grants.nih.gov/stem_cells/registry/current.htm. Or, if a specific stem cell line cannot be referenced at this time, check the box indicating that one from the registry will be used: Specific stem cell line cannot be referenced al this lime. One from the registry will be used. Cell Line(s) (Example: 0004): 4. Inventions and Patents Section (Renewal applications) *Inventions and Patents: O Yes • No If the answer is "Yes" then please answer the following: *Previously Reported: Yes 0 No 5. Change of Investigator/Change of Institution Section □ Change of Project Director/Principal Investigator Name of former Project Director/Principal Investigator Prefix: *First Name: Middle Name: *Last Name: Suffix: 0 Change of Grantee Institution *Name of former institution: Tracking Number: GRANT12743073 Page 157 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Introduction 1. Introduction to Application (for Resubmission and Revision application$) Research Plan Section 2. Specific Aims 3. Research Strategy* 4. Progress Report Publication List Other Research Plan Section 5. Vertebrate Animals 6. Select Agent Research 7. Multiple PD/Pl Leadership Plan 8. Consortium/Contractual Arrangements 9. Letters of Support 10. Resource Sharing Plan(s) 11. Authentication of Key Biological and/or Chemical Resources Appendix 12. Appendix Tracking Number: GRANT12743073 PHS 398 Research Plan NIAID_COV _2019_SPECIFIC_AIMS_final.pdf NIAID_COV _2019_RESEARCH_STRATEGY _final.pdf 0MB Numbe~ 0925-0001 Expiration Date: 03/31/2020 NIAi D _ GOV _2019_PROGRESS_REPORT _PUBLI CATI ON_LI ST .pdf NIAID_COV _2019_ VERTEBRATE_ANIMALS_Final.pdf NIAID_COV _2019_SELECT _AGENTS_Final.pdf NIAi D _ COV _2019_ CONSORTIUM_ CONTRACTUAL_Final.pdf NIAID_COV _2019_LOS_Final.pdf N IAI D_ GOV _2019_RESOU RCE_SHARI NG_PLAN_Final .pdf NIAID_COV _2019_AUTHENTICATION_OF _KEY _BIO_RSCS.pdf Page 158 Funding Opportunity Number: PA-18-484. Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER SPECIFIC AIMS Zoonotic coronaviruses are a significant threat to global health, as demonstrated with the emergence of Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) in 2002, and the continuing spread of Middle East Respiratory Syndrome (MERS-CoV). The wildlife reservoirs of SARS-CoV were identified by our group as bat species, and since then we have sequenced dozens of novel SARS-related CoV (SARSr-CoV) strains. Our previous R01 work demonstrates that bats in southern China harbor an extraordinary diversity of SARSr-CoVs, some of which are able to use human ACE2 to enter into human cells, can infect humanized mouse models to cause SARS-like illness, and evade available therapies or vaccines. We found that the bat hosts of SARSr-CoVs appear to no longer be traded in wildlife markets, and that people living close to bat habitats are the primary risk groups for spillover. At one of these sites, we found diverse SARSr-CoVs containing every genetic element of the wild-type SARS-CoV genome, and serological evidence of human exposure among people living nearby. Thus, there is significant potential for future spillover of SARSr-CoVs, and of public health impacts. Yet salient questions remain: Are there specific bat communities and sites that harbor CoV strains with higher risk for bat-to-human spillover? Which human behaviors drive risk of bat SARSr-CoVexposure that could lead to infection? Does human exposure to these viruses cause SARSlike or other illness? Can we characterize viral strain diversity, bat traits and human behaviors to assess risk of potential future CoV spillover? The proposed work in this renewal R01 builds on these findings to address these issues by conducting: 1) focused sampling of bats in southern China to identify viral strains with high predicted risk of spillover; 2) community-based, and clinic-based syndromic, sampling of people to identify spillover, and assess behavioral risk factors and evidence of illness; and 3) conduct in vitro and in vivo viral characterization and analyze epidemiological data to identify hotspots of future CoV spillover risk. This work will follow 3 specific aims: Aim 1: Characterize the diversity and distribution of high spillover-risk SARSr-CoVs in bats in southern China. We will conduct targeted bat sampling at sites where we predict that undiscovered high risk SARSrCoV strains exist. Bat sampling will be targeted geographically and by host species to test predictions about evolutionary diversity of SARSr-CoV. We will analyze RdRp and S protein sequences to test their capacity for spillover to people in Aim 3. Aim 2: Community- and clinic-based surveillance to capture SARSr-CoV spillover, routes of exposure and potential public health consequences. We will conduct focused, targeted human surveys and sampling to identify key risk factors for SARSr-CoV spillover and evidence of illness. To maximize our opportunity of capturing human exposure to bat CoVs, we will conduct community-based surveillance in regions with high SARSr-CoV prevalence and diversity, and individuals having contact with bats. We will assess bat-CoV seropositive status against a small number of questions about human-wildlife contact and exposure. We will conduct clinic-based syndromic surveillance close to these sites to identify patients presenting with influenzalike illness and severe acute respiratory illness, assess their exposure to bats via a questionnaire, and test samples for PCR- and serological evidence of SARSr-CoV infection. We will conduct follow-up sampling to capture patients who had not yet seroconverted at the time of clinic visit. Aim 3: In vitro and in vivo characterization of SARSr-CoV spillover risk, coupled with spatial and phylogenetic analyses to identify the regions and viruses of public health concern. We will characterize the propensity of novel SARSr-CoVs to infect people in vitro using primary human airway epithelial cells and in vivo using the transgenic hACE2 mouse model. We will use mAb and vaccine treatments to test our hypothesis that SARSr-CoVs with 10-25% divergence in S protein sequences from SARS-CoV are likely able to infect human cells, and to evade mAb therapeutics and vaccines. We will then map the geographic distribution of their bat hosts and other ecological risk factors to identify the key 'hotspots' of risk for future spillover. Overall, our SARSr-CoV program serves as a model platform to integrate virologic, molecular and ecologic factors contributing to CoV emergence while informing high impact strategies to intervene and prevent future pandemics. This includes providing critical reagents, therapeutic interventions and recombinant viruses for future SARSr-CoV pandemic and public health preparedness. Page 159 Specific Aims

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Contact PD/Pl: DASZAK, PETER 1. RESEARCH STRATEGY A. Significance: Severe Acute Respiratory Syndrome coronavirus (SARS-CoV) emerged in China threatening public health and global economies ( 1, 2). Like most other emerging pathogens (3), it originated in animal reservoir hosts, initially thought to be carnivores (4), and later shown by our group to be bats (5). Bats harbor a high diversity of 13- coronaviruses, including those related to Middle Eastern Respiratory Syndrome coronavirus (MERS-CoV) (6-9) and the newly emerged Swine Acute Diarrhea Syndrome coronavirus (SADS-CoV) ( 10), and may be the progenitor hosts of all Coronaviridae (5, 11-15). SARS-CoV uses the angiotensin-converting enzyme 2 (ACE2) receptor to gain entry to human cells ( 16). In 2012, we isolated and characterized two bat SARS-related coronaviruses (SARSr-CoVs) in China that use the ACE2 receptor and are closely related to SARS-CoV ( 17). Since then, under an R01 awarded in 2014, we have discovered >50 bat SARSr-CoVs in southern China. Some of these strains can bind to and infect human cells, cause SARS-like clinical signs in a humanized mouse model, and evade therapeutic and vaccine candidates against SARS-CoV ( 18). The Rhinolophus spp. bat hosts of these viruses are abundant across southern China, where hunting and consumption of wildlife is common and human population growth high, and where we have now identified serological evidence of exposure to SARSr-CoVs and other bat Co Vs ( 19). Thus, there is significant potential for future spillover of SARSr-CoVs, and of their subsequent spread. Yet salient questions remain: Are there specific bat communities and sites that harbor Co V strains with higher risk for bat-to-human spillover? Which human behaviors drive risk of bat SAR Sr-Co V exposure that could lead to infection? Does human exposure to these viruses cause SARS-like or other illness? Can we characterize viral strain diversity, bat traits and human behaviors to assess risk of potential future Co V spillover? This R01 renewal proposal aims to address these critical issues by conducting: 1) focused sampling of bats in southern China to identify viral strains with high predicted risk of spillover; 2) community-based, and clinic-based syndromic, sampling of people to identify spillover, and assess behavioral risk factors and evidence of illness; and 3) conduct in vitro and in vivo viral characterization and analyze epidemiological data to identify hotspots of future CoV spillover risk. B. Progress report: R01 Al110964, Daszak Pl, Project Period: 06/01/2014 -05/31/2019 The aims of our previous R01 were to: 1) Assess bat SARSr-CoV spillover potential at high risk human-wildlife interfaces, e.g. the wildlife trade, as reported for the 2003 outbreak (4, 20); 2) Analyze how viral diversity and phylogeny relates to host range and risk of emergence; and 3) Use binding assays, cell culture and mouse models to test the propensity of different SARSr-CoVs to infect humans. We made significant discoveries leading to 18 published peer-reviewed papers (18, 19, 21-33}, including two papers in Nature ( 10, 34), and a review in Cell (35) (see Progress Report Publication List). These findings include: Diversity and distribution of bat 13-and SARSr-CoVs in Southern China. We sampled and PCR-screened >16,000 individual bats from 6 families (16 genera) in southern China, finding A --- ......... ... ,.. - ---- ..... ·-.~-· ...... ~;•--- ...... --- .. _ .... _ 9 species positive (5,730 individuals screened) for SARSr-CoVs (Table 1, Fig. 1 ). We identified 178 novel '3-CoVs, of which 172 were novel (52 novel SARSr-CoVs). This includes members of a new '3- CoV clade, "lineage E" (26), and diverse HKU3- related CoVs (179 sequences) within a 'sister' clade to the SARS-CoV lineage . ..:.. • :;. :- ~ Fig. 1 (left): Bat sampling at 47 sites in China under ,.... · .. , ... ,._ ~ our previous R01. Yellow= sampling effort, red= CoV +ve bats. Fig. 2 (right): Map of bat species found positive for SARSr-CoVs in our previous R01, highlighting S. China (particularly Yunnan Province) as a center of diversity for SARSr-CoV reservoir host species. We found 6.7% mean PCR prevalence of SARSr-CoVs across bat hosts, with a small number of Rhinolophus spp. horseshoe bats having significantly higher PCR prevalence than other species sampled (Table 1). These bats are widely distributed, diverse, abundant, and roost and feed close to people and livestock, suggesting high potential for future SARSr-CoVs spillover. Distribution data for SARSr-CoV bat hosts suggest viral strain diversity is likely highest in southern China, particularly Yunnan Province (Fig. 2). 1 Page 160 Research Strategy

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Contact PD/Pl: DASZAK, PETER Table 1: Species found PCR-positive for Bat Species Individuals # SARSr-CoV SARSr-CoV SARSr-CoVs in our R01, with sample sizes tested positive mean prev. prev. range and prevalence estimates. Rhinolophus sinicus 1,328 113 8.5% 7.1-10.1% We identified one cave system (the "Jinning Cave") in Yunnan Province that R. macrotis 70 3 4.3% 0.9 · 12% harbors Rhinolophus spp. bats with diverse SARSr-CoVs, including some R. ferrumequinum 406 12 3.0% 1.5-5.1% with S proteins able to use human R. spp. 331 10 3.0% 1.5-5.5% ACE2 as entry receptors. Bats in this R. affinis 792 7 0.9% 0.4-1.8% cave carried SARSr-CoVs with all R. pusillus 1,023 8 0.8% 0.3-1.5% unique genetic elements of the SARS-CoV outbreak virus, suggesting Aselliscus stoliczkanus 269 2 0.7% 0.1-2.7% that this site may be a potential public Hipposideros pratti 323 2 0.6% 0.1 -2.2% health risk (29). H. armiger 1,188 0.1% 0.0-0.5% We used a novel phylogeographic analysis, Maximum Clade Credibility (MCC) tree, to reconstruct the geographic areas of evolutionary origin for 13-CoVs that we sequenced. Results suggest that: 1) Guangdong Province is the ancestral center of diversity of 13-CoVs (data not shown); 2) Guizhou is the likely origin of the HKU3-related clade (lineage 1); and 3) Guangdong and Guangxi are the likely ancestral origins of the SARS-CoV outbreak sequences (lineage 2) YN GO l GD VN (Fig. 3). Despite our intensive sampling at some sites, around half of the 20 Rhinolophus spp. we identified were captured at sample sizes below the minimum required to detect SARSr-CoVs at prevalences we found (n=11O, power 80%), and 5 others were SARSr-CoV negative in our study. To estimate sampling gaps, we used a viral 'mark-recapture' approach we previously published (36, 37). Results suggest we are approaching saturation of CoV strain discovery at some sites, whereas other sites contain rich pools of SARSr-CoVs that remain undiscovered (Fig. 4). In the current proposal, we have used these analyses to estimate geographic and species-specific sampling targets to more effectively identify new strains and CoV lineages needed to support experimental infection studies and risk assessment. Fig.3 (left): MCC phylogeny of lineage B 13-CoVs, including SARSr-CoVs (black box). Lineage 1 includes HKU3-related Co Vs, lineage 2 includes SARS-CoV unaaga, outbreak strains and close relatives (red box). Branches colored according to (HKU3l province of inferred ancestral origin (Guangdong Cororuiv1ru•5 •mplinobyProv1 ""' SARSr-CoVs ~ := 1,_---========..J GD, Yunnan YN, Guizhou, GZ). f,~ Fig. 4 (right): Estimates of SARSr-CoV strain diversity in the bats we sampled (strain j.., e·· . - defined as >10% sequence divergence in RdRp gene). GD and YN harbor highest CoV I"' diversity, but discovery has not yet saturated. We estimate proposed additional sampling of j __ _ 5,000 bats will identify >80% of remaining 13-CoV strains in bat hosts from these regions. o 20(0 '<"I> 60CO 90()) Numoer oc bans tes'6d In vitro & in vivo characterization of SARSr-CoV potentiaf for human infection -Gwngdang-o.ang,,--v ... ,,,." We conducted in vitro and in vivo experiments to characterize the pathogenic potential of novel SARSr-CoVs. We isolated three SARSr-CoVs from bat feces: WIV1, WIV16 and Rs4874, with S protein sequences that diverged from SARS-CoV by 3% to 7% ( 17, 22, 29). We conducted full-length genome sequencing of 12 other novel SARSr-CoVs from the Jinning Cave, some highly similar to outbreak SARS-CoV in the most variable genes: N-terminal domain and receptor binding domain (RBD) of the S gene, ORF8 and ORF3 (29). Using our reverse genetics system, we constructed chimeric viruses with SARSr-CoV WIV1 backbone and the S gene of different variants, including WIV1-Rs4231 Sand WIV1-Rs7327S. All 3 SARSr-CoV isolates and the two chimeric viruses replicated efficiently in Vero E6 cells and in Hela cells expressing hACE2, but not in Hela cells that don't express ACE2 ( 17, 22, 29) (Fig. Sa). In collaboration with Ralph Barie (UNC), we used the SARS-CoV reverse genetics system (38) to generate a chimeric virus with a mouse-adapted SARS-CoV 2 Page 161 Research Strategy

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Contact PD/Pl: DASZAK, PETER backbone expressing SHC014 S protein with 10% sequence divergence from SARS-CoV S. This chimera replicated in primary human airway epithelium, using the human ACE2 receptor to enter into cells ( 18) (Fig. 5b). Thus, SARSr-CoVs with diverse variants of SL-CoV S protein without deletions in their RBD can use human ACE2 as receptor for cell entry. - Hela-hACE2 _.,_w,vi -► Rs487• 1 _..,_ WIVJ,R.s.423) I 4 -► WIVl•R.57327 ! Fig. 5a (left): RT-PCR shows that bat SARSr-CoVs WIV1, Rs4874, and chimeras WIV1-Rs4231S, WIV1- Rs7327S grow in Hela cells expressing human ACE2. Fig. 5b (right): Viral replication of SARS-CoV Urbani (black) and SARS-SHC014S (green) primary ,2 z• 36 ,a air-liquid interface human airway epithelial cell u .. Timepostinf•clioo(h) cultures at an MOI of 0.01. How pos-tinf•«ion We infected transgenic mice expressing hACE2 with10 5 pfu of full-length recombinant WIV1 and three chimeric viruses (WIV1 backbone with SHC014S, WIV16S and Rs4231 S). hACE2 transgenic mice challenged with rWIV1 -SHC014S experienced ~20% body weight loss by 6dpi; rWIV1 and rWIV-4231 S produced less body weight loss, and rWIV1-WIV16S led to no body weight loss (Fig. 6a). At 2 and 4 dpi, viral loads in lung tissues of mice challenged with all three chimeras reached> 106 genome copies/g, significantly higher than rWIV1 infection (Fig. 6b). This demonstrates that pathogenicity of SARSr-CoVs in humanized mice differs with divergent S proteins, confirming the value of this model in assessing novel SARSr-CoV pathogenicity. E -¥° 1.1 10 ~ I -~~~*~~f::~ ...... Control ... t:11 ,l;' 1.0,- .... WIV-1 ,; 8 ~ -+- SHCOl4 ~ -- WIV-16 ~ 6 <ti 0.9 11D \VIV-1 El3 SHC014 la;al WIV•l6 !DD 4231 Fig. 6: In vivo infection of SARSr-CoVs in hACE2 transgenic mice. 6a {left) Body weight change after infection; 6b {right) Viral load in lung tissues. ~ 0.8 §> 07 i -+- 4231 ~ 4 ~ Infection of rWIV1-SHC014S j 2 caused mild SARS-like clinical " 0.6 8. 0 2 3 ◄ s , 0 'I- .. .. ;,,.~ signs in the transgenic hACE2 0P1 mouse model that weren't reduced by immune-therapeutic monoclonals that attenuate SARS-CoV pathogenecity. Vaccination against SARS-CoV did not reduce severity of clinical signs in mice subsequently infected with rSARSSHC014S ( 18). We found 2/4 broad human mAbs against SARS-CoV RBD cross-neutralized WIV1, but none could efficiently neutralize SHC014 which is less similar to SARS-CoV in the RBD (39). We repeated this virus characterization approach with chimeras using HKU3r-CoV S proteins that are ~25% divergent from SARSCoV S, and found that they are unable to use the ACE2 receptor. Additionally, we were unable to culture HKU3r-CoVs in Vero E6 cells, or human cell lines. The ability of HKU3r-CoVs to infect people, and their receptor binding target, remain unknown. This work has three implications for our R01 renewal: 1) some SARSr-CoVs currently circulating in bats in southern China are likely able to infect and replicate within people; 2) clinical outcomes of infection may include SARS-like illness that is currently not treatable with mAb nor preventable with experimental vaccines; 3) SARSr-CoV ability to bind human ACE2 is lost with S protein divergence between 10% (SHC014) and 25% (HKU3r-CoVs). Although no viruses within this range have so far been described, these strains likely use hACE2 but could escape existing vaccines and immunotherapeutics and represent significant public health threats. In our R01 renewal proposal, we will actively seek to identify viruses with this level of S protein divergence, characterize their binding targets in vitro, and their capacity to produce SARS-like disease that evades immunotherapy and vaccination in vivo. Discovery of a novel bat-origin a-CoV associated with pig die-offs Coronaviruses have a well-described propensity to jump the species barrier and cause new outbreaks ( 40). In 2016-17, we analyzed fecal samples from pigs at 5 farms in Guangdong Province (GD) affected by a fatal diarrheal disease. We discovered an a-CoV closely related to HKU2, and used PCR, serological and pathological data, followed by infection experiments to demonstrate that this novel virus, Swine Acute Diarrheal Syndrome coronavirus (SADS-CoV), caused the death of more than 20,000 pigs at these farms ( 10). We 3 Page 162 Research Strategy

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Contact PD/Pl: DASZAK, PETER identified SADSr-CoVs in Rhinofophus spp. bats in GD, and analyzed >30 full-length genomes to provide phylogenetic evidence that SADS-CoV originated in these bats (Fig. 7). MF094A1~VF.-....A .. M~o-M4U M,D·.C•V "- e A MFtMIPIA0$-CoY F...,...C tlll 11 ........ ~f..CoYF,mOA Mf1lM6$}' 6"1)8(~ 1•1'°6 .P, MF'-Ol4&U SAOS,-CoV ,au,o..,. i=~~=, .... IMOM$8SaA0$f.l;dl/~ .... SAo.sctu.-. NC002&1S1~GOl'OMwut.2¥ .I. NC0281521Canw1~..,...camet~11•1no1s 'ft 1(¥01374'7.1 22t£.....,.ed bat~in.lS 90ln 8t<Y22Slf,.1 .... hC0061)1JM..m,on~,.N.t3Jt NC 032t!l7 1 Nlfli3,~ bl! OCll'Cll'1wiNa 9"111IIIKYhl,13.9,a ..,_ ~0181111Rousettutt.1~HKU10 ... NC0298U 1 BtRf~H..e20t3 .-., AF353611.1 PEDl/1tW1CV1n A KJ16al521 PEOV at•,1111tAK,M .. JN),lltZl:'19..lfl'l!OVltill"OfS 'fl!III Fig. 7: Bayesian phylogenetic tree of the full-length genome sequences of SADS-CoV (red), bat SADSr-CoVs (blue), and related acoronaviruses. Host species represented by symbol. SADS-CoV replicates in Vero cells ( 10, 41, 42), but its capacity to replicate in human cell lines, and its zoonotic potential remains unknown. We developed a novel Luciferase lmmunoprecipitation _ _,, • .,..,., Systems (LIPS) antibody assay for SADS-CoV and found no evidence of spillover to pig farm workers at affected farms (0/33 people seropositive) (10). In the current proposed work we will include SADS-CoV diagnostic reagents in our serological panel to opportunistically screen human samples for JX647847 I PEDVM11111001 1lll (.196()178 l flEDV._VNJFPtOU tr.t)ll'VmAN'VietnlnA ICF272t20 1 PEDV .._ USNC010,1d:tl21U3 A039997& I FEDV..., OH861 1111: (F7'111,751 PEDV..,...CHNNl0,l.6/201J A KR061'58..1PEDVVWIIMl',tf7ZW20091'li ICIJa36«3atPEDVIICli6lt~2000A evidence of spillover into people exposed to bats in southern China Mapping bat viral emergence risk We analyzed host and viral data for all known mammalian viruses and used a generalized additive models to correct for underlying sampling and reporting biases (34). This approach allowed us to predict the relative number of yet-to-be-described or 'missing' viruses that a species likely harbors. For China, there are distinct .. hotspots of unknown bat viral diversity in Yunnan Province (Fig. 8) . . , Fig. 8: Spatial distribution of predicted 'missing' or as-yet undiscovered viruses, from (34). Yellow= highest diversity, red triangle= Jinning Cave, Yunnan (29). In a separate paper, we found that bat host diversity and climatic variability are l correlates of viral diversity within bats, and that human population density, bushmeat /"'~l hunting, and livestock production are correlates of the risk of transmission for viruses that spillover (26). The risk of spillover and spread differ spatially, suggesting that locations where bat viruses are most diverse may not be the most strategic sites for public health intervention (21). Work in the current proposal will improve on both approaches to identify hotspots of CoV emergence risk, by using data from the high-risk locations and interfaces identified in our previous R01, including better characterization of SARSr-CoV diversity in bats, and the potential of these viruses to cause infection. Human risk behavior. the wildlife trade, and evidence of bat SARSr-Co V spillover. Qualitative Study: Our previous R01 hypothesis was that SARSr-CoV spillover would most likely occur through the trade in bats for food, via the same market chains that to the emergence of SARS (20). To test this, we conducted an exploratory study using standardized one-on-one semi-structured ethnographic interviews and observational data in southern China among 88 people involved in trading wild bats, to assess local social and cultural norms and individual attitudes underlying contact with bats (publication in prep.). Our results suggest that in the 11 years since the emergence of SARS, there have been substantial changes to the wildlife trade: 1) Former wildlife markets are now predominantly selling captive-bred species {poultry, livestock, farmed wildlife); and 2) few bats are now sold through markets. We identified other risk factors for spillover, including people living near to bat roosts, and those visiting bat caves for hunting or recreation. Human Questionnaire & Sero-surveillance: We used qualitative study findings to develop a human behavioral risk questionnaire on the type and frequency of animal contact, wildlife observed in daily life, and unusual illnesses reported over the past 12 months. We conducted a cross-sectional study among populations that live near bat caves or roosts where we had detected bat SARSr-CoVs. Study participants provided biological samples, and bats were concurrently captured and sampled. Questionnaires and biological samples Yunnan •• • • Research Strategy • G1,,111ngdong •• A {oropharynx swab, serum, plasma) were collected from 1,585 participants from 7 sites in Yunnan, Guangxi, and Guangdong provinces (Fig. 9) . Fig. 9: Concurrent sampling in bats and target human population in communities in Yunnan, Guangxi, and Guangdong provinces. Pie-charts indicate sampling effort (bat sampling = blue, Human questionnaire and sampling = purple, Ethnographic interview = yellow, bat Co V seropositive = red) 4 Page 163

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Contact PD/Pl: DASZAK, PETER Serological Evidence of Exposure: We developed serological assays for HKU9 CoVs (~). SARSr-CoV Rp3 W), HKU1 O CoV (a), and MERS-CoV (13) and used ELISA and Western blot to test serum samples collected in 2016/17. We found 7 individuals (7/733, 0.95%) living within a 6 km radius of the Jinning Cave, and 6/209 people (2.87%) at one site, with evidence of exposure to bat SARSr-CoVs (Table 2; Fig. 8). Site # Bat CoV + (%) SARSr-CoV HKU10 + HKU9 + MERS-CoV+ Table 2 (left): tested Rp3 + (%) (%) (%) (%) ELISA and Jinning, Yunnan Mengla, Yunnan Jinghong, Yunnan Lufeng, Yunnan Guangdong Guangxi 209 6 (2.87) 6 (2.87) Western blot 168 1 (0.6) 1 (0.6) confirmed 212 testing of 144 human sera for 420 antibodies to 4 412 2(0.48) 2(0.48) batCoVs. Fig. 10: Western blot reactivity of human sera to SARSr-CoV Rp3 NP. We found evidence among human populations in Guangxi Province of people with prior exposure to the bat (J.,-CoV HKU1 O (2/412, 0.48%). This is of potential public health interest because HKU10 is known to be able to jump host species within bats, and therefore may have high propensity for emergence (43). Behavioral Risk Factors: Questionnaire response and demographic data suggest specific risk factors included type of occupation, keeping of pets and visiting wet markets. Seropositive individuals were mostly farmers/peasants (8/9), living in Yunnan _ JNO<l (0 70) _ JN162(0.5J} INI 11 (0.51) INO-tl (0.4?) - $ARSpal><Oot(J 19) _ Jl'l0)9 (0.l8) _ Jl<Oi9(0H) _ JN036 (0 40) == _ JN0I0 (0.36) WHU7(Ml) province (7/9), 41-60 yrs old (7/9), with domestic contact with rodents (6/9), and are male (6/9). However, these characteristics were common and occurred in ~40% of survey respondents. Although these results are preliminary and don't provide detailed information on routes of exposure, they suggest that further refining use serological tests coupled with qualitative and questionnaire data will identify likely routes of exposure to novel Co Vs in China. In Aim 2 of this R01 renewal proposal, we identify strategies to better target at-risk people, and conduct focused questionnaires and serosurveys to produce statistically significant findings. Analysis of self-reported illness: We analyzed data on self-reported symptoms of fever with cough and shortness of breath or difficulty breathing (severe acute respiratory illness - SARI), and fever with muscle aches, cough, or sore throat (influenza-like illness - ILi) from study participants. We used a least absolute shrinkage and selection operator (LASSO) regression to identify associations between ILi and/or SARI symptoms and contact with animals in the last year. Salient predictors or combination of predictors were, in descending order of odds ratio: 1) having eaten raw or undercooked carnivores; 2) having slaughtered poultry and being a eai.nraNl)(UlldO<COOl<edcamlvores (s•0.74,n•17) I O resident of Guangxi province; 3) having had contact ··••ohtor ... po,.,U,y•pm,;n-.Gu•noXl <••0- 7S:n= 55) 0° with cats or bats in the house·, and 4) being a male pe1cats (s=064.n=183} In house bats (S. 077; n• 201) 0 who has raised poultry (Fig. 11 ). handled l>ve poullry <• = 0!!8. n = 743) 0 ra!S«lcattleOf buffalo (s • 062: n• 137) 0 raised poullrygeoder male (s • O 75: n = 459) I 0 sll,red ,.aterw,11> animals for wasning:g.-, female (s • 0.63: n ~ 95) e I a90•1to60 (a•0.61,n•766) e I ocx:op;,llon ether (s = 0.6; n • 182) e 1 1 1.5 2 O<l<ls Ratios (log-oods scale) 2. APPROACH Fig. 11: Predictors of self-reported ILi and/or SARI in prior 12 months (s = bootstrap support; n = number +ve out of 1,585 respondents). Orange circles= odds ratios > 1 (positively associated with the outcome); purple= odds ratios <1 (negatively associated with the outcome). Rationale and Innovation : Our previous R01 work demonstrates that bats in southern China harbor an extraordinary diversity of SARSr-CoVs, some of which are able to use human ACE2 to enter into human cells, can infect humanized mouse models to cause SARS-like illness, and evade available therapies or vaccines. We found that the bat hosts of SARSr-CoVs appear to no longer be traded in wildlife markets, and that people living close to bat habitats are the primary risk groups for spillover. At one of these habitats, we found diverse SARSr-CoVs containing every genetic element of the wild-type SARS-CoV genome, and serological evidence of human exposure among people living nearby. The proposed work in this renewal R01 builds on these 5 Page 164 Research Strategy

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Contact PD/Pl: DASZAK, PETER findings:. In Aim 1, we will conduct targeted bat sampling at sites where we predict that undiscovered high risk SARSr-CoV strains exist. Bat sampling will be targeted geographically and by host species to test predictions about evolutionary diversity of SARSr-CoV. We will analyze RdRp and S protein sequences to test their capacity for spillover to people in Aim 3. In Aim 2, will conduct focused, targeted human surveys and sampling to identify key risk factors for SARSr-CoV spillover and evidence of illness. To maximize our opportunity of capturing human exposure to bat CoVs, we will conduct community-based surveillance in regions with high SARSr-CoV prevalence and diversity, and individuals having contact with bats. We will assess bat-CoV seropositive status against a small number of questions about human-wildlife contact and exposure. We will conduct clinic-based syndromic surveillance close to these sites to identify patients presenting with influenza-like illness and severe acute respiratory illness, assess their exposure to bats via a questionnaire. and test samples for PCR- and serological evidence of SARSr-CoV infection. We will conduct follow-up sampling to capture patients who had not yet seroconverted at the time of clinic visit. In Aim 3, we will characterize the propensity of novel SARSr-CoVs to infect people in vitro using primary human airway epithelial cells and in vivo using the transgenic hACE2 mouse model. We will use mAband vaccine treatments to test our hypothesis that SARSr-CoVs with 10-25% divergence in S protein sequences from SARS-CoV are likely able to infect human cells, and to evade mAb therapeutics and vaccines. We will then map the geographic distribution of their bat hosts and other ecological risk factors to Identify the key 'hotspots' of risk for future spillover. Our SARSr-CoV program serves as a model platform to integrate virologic, molecular and ecologic factors contributing to CoV emergence while informing high impact strategies to intervene and prevent future pandemics. This includes providing critical reagents, therapeutic interventions and recombinant viruses for future SARSr-CoV pandemic and public health preparedness. Research team and management: We have reinforced our original collaboration between EcoHealth Alliance (EHA), a global leader in field investigations of emerging viruses from wildlife and modeling/analysis of viral risk, and Wuhan Institute of Virology, a global leader in bat viral investigations (Fig. 12). First, we have included senior behavioral risk scientists Co-I Francisco (EHA) and Ren (Inst. Pathogen Biol., Beijing) to oversee human survey and sampling work in Aim 2. Second, Prof. Linfa Wang (Duke-NUS), a world leader in @ EcoHealthA,,,ance • Cov ecte-en1n,g . cov sero10gy • v-.ra1hnh0Slene • Host re<:ep10t bind • s p,o,e,n se,q:uen • in vi/Jo and lo 1/M> ors.. oaau.Jc. Olivat. franci • • . Ta:ge""' cat~ . ·. • Host~ virus ctiaraeteflza il.81.ion @Ors.Shi.Zhou, == Wllnillllnlll!U•rt ·- .,..._ ............ • CoV . reve,se ge.r.ebc:& -· $ • Sm3ff .:i.nimol models of nity $t,11Vetllince viral pathogenes,,s • • S)'!'ldfotll.C surv~ilJanc& • Ptimery h4.Jmen cd CLtl • ~g=Ysequerteing • RecongtrudJon of •oon . EPidemiOICgy on. 8"rlc and Sims • Beha...,.1 nsk .-"fft u ors. fteo and Guo e,. !WJ, •u~!" •uu ' =:~~ \lti understanding the role of bats as hosts of emerging viruses, will act as a consultant by advising and assisting in the development of PCR and serological tests and virus characterization. Prof. Wang has developed a unique array of bat immunological reagents that enrich the serological arms of the proposal. Third, Prof Ralph Barie (UNG) will use his expertise in CoV characterization to conduct primary human epithelial airway cell infections to identify high risk strains that are poised for human emergence. He will oversee and participate in animal experiments in Aim 3. This expanded team will work on a more focused set of goals, based on the results of our previous R01. Pl Daszak has collaborated with all partners for between 3 and 15 yrs and will host monthly calls, annual in-person meetings, conduct quarterly adaptive management to refine research lines of work. Fig. 12: Interdisciplinary team & roles in the proposed R01 renewal work. Aim 1: Characterize the diversity and distribution of high spillover-risk SARSr-CoVs in bats in southern China 1.1 Rationale/Innovation: Our previous R01 work identified diverse SARSr-CoVs with high propensity for human infection ( 18, 19, 29). Characterization of these suggests SARSr-CoVs that are up to 10%, but not 25% different in the spike glycoprotein use human and bat ACE2 receptors for docking and entry. Uneven sampling gaps (e.g., no strains were found with 10-25% spike variation) prevent a thorough understanding of the transition point where the most divergent strains lose human ACE2 receptor usage. Our viral discovery curves (Fig. 4) suggest further sampling will reveal a rich diversity of as-yet-undiscovered SARSr-CoVs. In this aim we will use phylogenetic and viral discovery analysis to specifically target bat species and regions that are undersampled to allow sufficient power to identify and characterize missing SARSr-CoV strain diversity. Our 6 Page 165 Research Strategy

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Contact PD/Pl: DASZAK, PETER previous work also suggests that SARSr-CoVs with S proteins that are ~ 10% divergent from SARS-CoV resist neutralization by therapeutic mAbs and escape SARS-CoV vaccines (17, 18, 23), suggesting that. However, viruses with 10-25% divergence in S proteins may bind to human cell receptors, but completely evade therapeutic and vaccine effects, and could therefore be a higher risk for public health. We will sequence the S proteins of novel SARSr-CoVs to prioritize viruses for experimental work in Aim 3 to test this hypothesis. 1.2 General Approach: We will use sampling, testing, and CoV sequence data from our previous R01 to pinpoint sites and host species needing additional sampling. We will work in 4 provinces (Yunnan, Guangxi, Guizhou and Guangdong) that we have identified phylogenetically as having the highest diversity of as-yetundiscovered SARSr-CoVs and with competent natural hosts. Precise sampling site locations will be refined in Y1. We will target at least 5,000 individual bats over 5 years from 15 currently undersampled species of Rhinolophus bats, which we calculate will allow us to almost fully characterize the expected natural diversity of SARSr- and other ~-CoVs in the region. Bats will be captured, sampled, and released at the site. Specimens will be transported in liquid N2 to Wuhan Inst. Virology (WIV) for PCR screening, and positive samples selected for further molecular characterization and S Protein sequencing. EHA will lead the study design, field sampling, and data analysis for this Aim; and WIV will lead the testing and viral sequencing. 1.3 Sampling and testing of bats: 1.3.a Site selection & sample sizes: In Y1 we will use our bat host and viral trait modeling, phylogeographic analyses of RdRp and S Protein sequences, and geographic and host species-based viral discovery curve analyses to identify SARSr-CoV diversity hotspot regions for bat sampling. We will sample at 8 new sites in four provinces. We will use cave site data (44), and demographic information to identify two sites in each of Yunnan, Guangxi, Guangdong, and Guizhou where humans likely have contact with bats. In Yunnan, we will identify two unsampled caves close to, but distinct from, the Jinning cave (29). This will provide adequate coverage of lineage 1 and 2 SARSr-CoVs, including a rich source of new HKU3rCoVs, which have unknown potential for zoonotic spillover. Sampling will begin towards the end of Y1. We will use survey data from our previous R01 and host-specific viral accumulation curve data to target an additional 10 under-sampled Rhinolophus spp., 5 that were SARSr-CoV negative in our study, and a small number of related bat genera (including Hipposideros spp. and Asel/iscus spp.) we previously found PCR positive for SARSr-CoVs (Table 1). We will sample at least 5,000 bats from these 4 provinces (~1250 per province). Given ~5-12% prevalence of SARSr-CoVs in Rhinolophus spp. at our previous sites, this sample size would give us 425 (±175) positive individual bats, and ~125 novel strains. 1.3.b CoV screening, isolation: Viral RNA will be extracted from bat fecal pellets/anal swabs with High Pure Viral RNA Kit (Roche). RNA will be aliquoted, and stored at -BOC. One-step hemi-nested RT-PCR (lnvitrogen) will be used to detect the presence of CoV sequences using primers that target a 440-nt fragment in the RNAdependent RNA polymerase gene (RdRp) of all known a- and ~-CoVs (45). PCR products will be gel purified and sequenced with an ABI Prism 3730 DNA analyzer. We will attempt isolation on samples with diverse and interesting novel CoVs, using Vero E6 cells and bat primary cell culture. 1.3.c Sequencing S proteins: For all novel SARSr-CoV strains, we will sequence the complete S gene by amplifying overlapping fragments using degenerate primers as shown previously (17, 29). Full-length genomes of selected SARSr-CoV strains (representative across subclades) will be sequenced via high throughput sequencing method followed by genome walking. The sequencing libraries will be constructed using NEBNext Ultra II DNA Library Prep Kit for lllumina and sequenced on a MiSeq sequencer. PCR will be performed to fill gaps in the genome. The full length S gene sequences, including the amount of variation in the S receptor binding residues that bind the ACE2 receptor, will be used to select strains for Aim 3 experiments. 1.3.d Host ACE2 receptors: We will sequence host ACE2 receptors of different bat species or different bat populations from a single species (e.g. Rhinolophus sinicus) to identify relative importance of different hosts of high risk SARSr-CoVs and the intraspecific scale of host-CoV coevolution. Of particular interest is homology across 18 bat and human orthologue ACE2 contact interface residues that engage the SARS RBD as a potential indicator of SARSr-CoV cross species transmission potential and growth in human cells (46). 1.4 Analyses: 1.4.a Bat-CoV evolution and distribution: We will use Bayesian and Maximum Likelihood phylogenetic analyses of RdRp, Spike, and full genome sequence (when available) data to reconstruct the evolutionary history of the novel bat SARSr-CoVs we identify. We will rerun MCC analyses (Fig. 3) to 7 Page 166 Research Strategy

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Contact PD/Pl: DASZAK, PETER reconstruct 13-CoV evolutionary origins using our expanded dataset. Ecological niche models will be used to predict spatial distribution of PCR-positive species, identify target sites for additional bat sampling, and analyze overlap with people. We will use our viral discovery/accumulation curve approach (Fig. 4) to monitor progress towards discovery of >80% of predicted diversity of SARSr-CoVs (lineage 1 & 2) within species and sites, halting sampling when this target is reached, and freeing up resources for work other work (37, 47). 1.4.b Viral strain prioritization: Of the expected 100-200 novel SARSr-CoV strains, we will down-select to prioritize for further characterization based on S genes that are: i) different from SHC014, WIV1, SARS-CoV with diversity ranges of 10-25%; ii) have virus S RBD that could use human/bat receptors; iii) have recombinant chimeric spikes indicative of gene flow between clade I and II strains; iv) have bat ACE2 receptors that might select for spike RBDs that can use human receptors for entry (15/18 conserved residues in human/bat ACE2 molecules that bind SARSs-CoV S RBD domains are likely more efficient receptors than 3/18 conserved sites). Using structural models based on the SARS S glycoprotein, the extent and location of antigenic variation will be annotated onto the structure, further resolving the locations of highly evolving and conserved structural motifs/epitopes that function in protective immunity ( 48-51). 1.5 Potential problems/alternative approaches: Permission to sample bats in sites or provinces we select. We have a > 15-year track record of successful field work in southern China and have worked with local authorities and communities to ensure access. We have existing permissions in place which will be renewed at the start of this project. Due to the abundance of distinct caves in the region, if access to a site is withdrawn, we will rapidly identify a suitable, complementary site with similar bat species composition and abundance. We may not identify J3-CoVs in our sample bat species due to seasonality of viral shedding. The sampling regions we have selected are subtropical, and our previous data, and even published studies in temperate regions (52), do not suggest a strong pattern of seasonality in SARSr-CoV shedding. Nonetheless to account for this we will conduct sampling evenly on quarterly basis within each province. Aim 2: Community and clinic-based surveillance to capture SARSr-CoV spillover, routes of exposure and potential public health consequences. 2.1 Rationale: Our previous R01 study identified serological evidence of exposure to SARSr-CoVs in certain communities in S. China (Table 2, Fig. 10) ( 19). However, the low seroprevalence (0.6%-2. 7% at positive sites) suggests we need a larger sample size, and a focused, targeted study to maximize the likelihood of identifying seropositive cases. In Aim 2, we will use combined biological-behavioral risk surveillance in targeted populations within the community and clinical settings to 1) identify risk factors correlated with seropositivity (exposure to) and PCR positive status (infection with) bat SARSr-CoVs; and 2) assess possible health effects of SARSr-CoVs infection in people. Obtaining this information could be a significant step in understanding the likelihood of recent 'hidden' spillover events and their public health impacts, as well as the risk of future emergence of SARS-like diseases. It will also support the development of risk-mitigation strategies by public health authorities within China and other countries with bats that harbor these viruses (e.g. most of SE Asia). 2.2 General Approach/Innovation: We will use a dual study design to gain in depth understanding of exposure and risk factors for SARSr-CoV spillover (Fig. 13). We will conduct community-based surveillance, with more focused questionnaires and biological sampling to determine the seroprevalence of SARSr-CoVs in at-risk human populations, and to identify risk-factors for SARS-CoV spillover in these communities. We will conduct clinic-based syndromic surveillance and biological sampling at sites that include the community-based surveillance sites within their catchment. This will include follow-up sampling to capture seroconversion of recently infected people and the full course of symptoms. We will also use PCR to test for present of active SARSr-CoV replication. Both community-based and clinic-based syndromic surveillance programs are casecontrol studies designed with the sample sizes necessary to statistically quantify (with a power of 80%) risk factors and health impacts for SARSr-CoV spillover, linked to serological status and symptoms. 2.3 Target population & sample size: We will target sites in the same four provinces, and close to those for bat sampling, based on: 1) sites of likely divergence of SARSr-CoVs; 2) a high diversity of SARSr-CoVs within the S protein sequence divergence of 5-25%; and 3) high rates of human-wildlife interactions. Communitybased surveillance will be conducted at 2 sites in each of the 4 provinces, a total of 8 sites. From our previous work we anticipate that 10-30% of the community population will have had exposure to bats allowing us to 8 Page 167 Research Strategy

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Contact PD/Pl: DASZAK, PETER capture highly exposed and non-exposure individuals at each site. Individuals living or working around bat roosts, who hunt wildlife, work with wildlife or livestock farming, transportation, selling, or slaughtering wildlife in the surveyed areas will be targeted so that they make up ;;,:30% of the sampled population in each community. We will stratify sampling to ensure appropriate representation of sex, demographic, and socio-economic factors at each community site. We will initiate active clinic-based syndromic surveillance at 2 county-level hospitals and 1 provincial-level hospital in each of the 4 provinces, in total 12 hospital sites, all within the catchment areas for bat sampling, and which are used by people in our community-based surveillance. Patients ~ 12 years old presenting at the health facility who meet the syndromic and clinical case definitions for Sito S.ll<:llon PopulatJoo Data Collection Analysl.s SARI and ILi will be recruited into the study. We will enroll Community-based surveillance Clinic-basedsyndromicsurveillance a total of at least 2,750 individuals for clinical studies, :"~~~:8~~;':!.'i!a~~--~ which accounts for up to 40% loss from follow-up. Study -po:;entt,,,...«;og••·•~cu,..,.,. .. d;n... data will be pooled across sites, as clinical patients are --=~.~~~ 0 ~.:: 1 •.,.,,._, limited by the number of individuals presenting at - bkef'ydfV9fQMCe of SARSc-COVs - 8 hioh diwrsity of SARSr .COVs - high mes d human•wikllife inlenM:tlons -comrrunity populs:lon -~18yowsoroider Consent and EnrOllm•nt DAY 0 hospitals. For community-based surveillance, we will enroll 1,650 individuals per province, pooled across two sites for each province, allowing us to make province-level comparisons of differing effects. Estimating 5% overall - QueslOt'lf'l#e edrnlniS4r#IIOft - ~ blood eno serum 1atnpiK - qwsbomalro M:lminb:171tlO'I -whOl•bbc>d--,Mt\lm.-,plff - nasal 0t CW09Ml'Yngiea1 s.wabt ~ seroprevalence in these high-exposure populations, these sample sizes are sufficient to estimate effect sizes of behavioral risk factors of 2X or greater with 80% power. Fig. 13: Human survey and sampling study design overview 2.4 Data & sample collection: At both community and clinical settings, following enrollment with signed consent form, biological specimens (two whole blood samples, one max. 500 µL; two 500 µL serum samples) will be collected from all eligible participants, and a questionnaire will be administered. We will investigate five risk factors, so as to maximize the power of the analyses, all related to high risk wildlife exposure, based on continuing analysis of our previous work, and will include: 1) occupation; 2) observed or reported interactions with bats in/around house; 3) proximity to nearby bat roosts; 4) working or regular visit to animal markets; 5) self-reported ILi/SARi. An additional two nasal or oropharyngeal swabs will be collected from patients enrolled in the clinic-based syndromic study. With permission from each clinic, and consent from participants, we will review clinical records to collect data on medical history, clinical syndromes, and patient etiology. 2.5 Clinic enrollment and follow-up: We will recruit inpatients and outpatients after initial screening to meet the clinical case definition of 1) severe/acute respiratory illness (SARI/ ARI) of unknown etiology; or 2) Influenza-like illness (Ill) of unknown etiology. Once enrolled, biological samples will be collected and a questionnaire administered by trained hospital staff that speak appropriate local dialects. Samples will be taken concurrently when collecting samples for normative diagnostics. For inpatients, samples will be collected within 1 O days of reported onset of illness to increase the chance of PCR CoV detection (53). We will follow up 35 days after enrollment to collect an additional two 500 µL serum samples conduct a standardized questionnaire supplement to collect additional data on the course of symptoms in the interim period. 35 days gives adequate time for development of lgG, which occurs <28 days after onset of symptoms for SARS patients (54). 2.6: Laboratory analysis; 2.6.a Serological testing: In our previous R01, we expressed his-tagged nucleocapsid protein (NP) of SARSr-CoV Rp3 in E.coli and developed a SARSr-CoV specific ELISA for serosurveillance using the purified Rp3 NP. The specificity of this assay was evaluated using polyclonal antibodies against HKU1, OC43, 229E, NL63, MERS-CoV and EBOV and no significant cross-activity was detected ( 19). While this shows it is a specific test for Rp3, it suggests that if we can expand our serology tests to cover other bat Co Vs, we may identify many more seropositive individuals. In this renewal, we will therefore use two serological testing approaches. First, we will expand test all human sera collected from both community- and clinic- based sampling for a panel of bat Co Vs that will include SARS-CoV (outbreak strain), a range of lineage 2 SARSr-CoVs (including WIV1 and SHC014), lineage 1 HKU3r-CoVs, MERS-CoV, and the a-Co Vs SADS-CoV and HKU10. We previously found serological evidence of human exposure to HKU1 O ( 19), but HKU1 O is known to jump from one host bat species to another ( 43) and is 9 Page 168 Research Strategy

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Contact PD/Pl: DASZAK, PETER therefore likely to have infected people more widely. lncoporating serological testing for SHC014 is also likely to yield higher seroprevalence because it is readily divergent from SARS-CoV wildtype and therefore unlikely to have been picked up in our earlier testing. It is possible that non-neutralizing cross reactive epitopes exist that afford an accurate measure of cross reactivity between clade1 and clade 2 strains which would allow us to target exposure to strains of 15-25% divergence from SARS-CoV. Additionally, we will test samples for antibodies to common human Co Vs (HCoV NL63, OC43 - see potential pitfalls/solutions below). Secondly, we recognize that CoVs have a high propensity to recombine. To serologically target 'novel' recombinant virus exposure, we will conduct 1) ELISA screening with SARSr-CoV S or RBD; 2) confirm these results by Western blot; then 3) use NP based ELISA and LIPS assays with a diversity of SARSr-CoV NP. For NP ELISA, microtiter plates will be coated with 100 ng/well of recombinant batCoV NP and incubated with human sera in duplicates followed by detection with HRP labeled goat anti-human lgG antibodies. For confirmation, all ELISA positive samples will be subjected to Western blot at a dilution of 1:100 by using batCoV-NP as antigen. We will use an S protein-based ELISA to distinguish the lineage of SARSr-CoV. As new SARSr-CoVs are discovered, we will rapidly design specific Luciferase immunoprecipitation system (LIPS) assays targeting the S1 genes of bat-CoV strains for follow-up serological surveillance, as per our previous work ( 10). 2.6.b RT-PCR testing. Specimens will be screened using RT-PCR for the RdRp gene (See section 1.3.b for details). Positive samples will be subjected to full genome sequencing and RT-PCR amplification of the S glycoprotein gene. Samples from the clinic-based syndromic surveillance will also be tested using RT-PCR for Influenza A & B, HCoV NL63, OC43, HKU1, SARS-CoV & 229E as rule-outs, and SADS-CoV and HKU10 as an opportunistic survey for potential spillover these Co Vs as proposed above. 2.7 Epidemiological analysis: We will conduct a case-control study to identify risk factors for SARSr-CoVs spillover. "Cases" are defined as participants whose samples tested positive for SARSr-CoVs by serological tests. "Controls" will be selected from the pool of participants admitted to the studies but testing negative. We will use nearest neighbor matching to pair cases demographically with controls at a 1-to-3 ratio or greater. We will use generalized linear models to analyze correlation between serological/PCR status and risk factors including: Activities with likely exposure to 1) bats; 2) livestock; and 3) locations of residence and work. We will use the same procedure to determine how clinical presentation differs between SARSr-COVs-exposed and unexposed enrollees, in the time course of illness, severity of symptoms, and type of symptoms. 2.8 Potential problems/alternative approaches: Rarity of spillover events means it may be difficult to identify sufficient seropositives to statistically analyze risk behavior or illness. First, we are now targeting our community-based surveillance to subpopulations with high-levels of bat exposure, at sites selected for diverse and prevalent SARSr-CoVs, and are adding clinic-based syndromic surveillance of SARI and ILi cases in these same regions - both will increase likelihood of finding positive individuals. Second, our serology testing will include a panel of assays for a large diversity of lineage 1 and 2 SARSr-CoVs as well as SADS-CoV, HKU1 O and other bat-borne CoVs. Rhinolophus spp. bats host all of these (overall bat CoV PCR prevalence, 11.8%; ~-CoV, 3.4%; a-CoV, 9.1 %). Thus, using this broad serological panel to screen individuals in likely contact with these species increases the potential for detecting spillover with enough power for statistical analyses, and will shed light on behaviors that predispose to CoV spillover from bats. Third, we will include common human CoVs in our panel, so that even if low prevalence of bat CoVs is found, we will be able to conduct a valuable crosssectional study of the seroprevalence of human CoVs. Finally, we will be able to assess relative measures of human-wildlife contact from our survey work. We will analyze intensity of contact against other risk factors and clinical outcomes to provide useful proxy information for spillover risk. Patients visiting clinics may have cleared virus, but not yet developed lgG antibodies, reducing seropositive cases. Our 35 day follow-up sampling should avoid this because the maximum lag time between SARS infection and lgG development was ~28 days (53). We also expect that patients in rural communities will only visit clinics when symptoms have progressed, likely coinciding with late illness and onset of lgG. We will also have data from our community study, so won't be completely reliant on hospital data to identify PCR- or seropositives. Finally, the risk is outweighed by the potential public health importance of discovering active spillover of a new SARSr-CoV. Serological testing may not match known CoVs due to recombination events. We will use the threetiered serological testing system outline in 2.6.a to try to identify these 'novel' Co Vs, however, we will also remain flexible on interpretation of data to ensure we account for recombination. 10 Page 169 Research Strategy

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Contact PD/Pl: DASZAK, PETER Aim 3: In vitro and in vivo characterization of SARSr-CoV spillover risk, coupled with spatial and phylogenetic analyses to identify the regions and viruses of public health concern 3.1 Rationale/Innovation: In Aim 1, we aim to expand the known diversity of SARSr-CoVs by over 125 strains, targeting 10-25% S protein divergence that we predict infers high spillover risk and evasion of immune therapeutic and vaccine efficacy. In Aim 3, we will further characterize the zoonotic potential of a selected group of these novel SARSr-CoVs, using infectious clone technology, in vitro and in vivo infection experiments and analysis of HKU3r-CoV receptor binding to test the hypothesis that S protein% sequence divergence thresholds predict spillover potential ( 18, 55). We will analyze data from these viral characterization and infection experiments, coupled with bat host distribution, viral diversity and phylogeny, human survey of risk behaviors and illness, and human serology to assess spillover risk of SARSr-CoVs in different bat species across southern China. This will enable future development of public health interventions and enhanced surveillance to prevent the emergence of a novel SARSr-CoV. 3.2 General Approach: We will use S protein sequences to select a range of viral strains that cover the 10- 25% S protein divergence we predict as high public health potential and construct chimeric SARSr-CoVs using the WIV1 backbone and these S genes as done previously ( 12, 18, 38). We will rescue of full-length clones and assess infection of non-permissive cells expressing human, bat and civet ACE2 receptors, Vero cells, primary human airway epithelial cells, and CaCo cells for HKU3r-CoVs (which have not been cultured and may use intestinal epithelium in nature). We will conduct experimental infections in hACE2 transgenic mice to assess pathogenicity and clinical signs (18). Finally, using a panel of mAbs that neutralize SARS-CoV infection in vitro and in vivo, and vaccine against SARS-CoV S protein, we will examine the capacity of strains with divergent S protein sequences to evade therapeutics, revealing strains with high public health potential. We will also conduct limited experiments to analyze HKU3r-CoV receptor binding and assess spillover potentia. Using these results, and data from Aims 1 and 2, we will use spatial modeling techniques to identify geographic hotspots in southern China where bat species that harbor high risk SARSr-CoVs inhabit, where communities that have high exposure to bats exist, where serological or PCR evidence of spillover has been identified, and where underlying demographic or environmental trends suggest high risk of future emergence. 3.3 Virus characterization: 3.3.a Construction of chimeric SARSr-CoV viruses: Infectious clones with the S gene of novel SARSr-CoVs and the SARSr-CoV WIV1 genome backbone using the reverse genetic system developed in our previous R01 (24). The correct infectious BAC clones will be screened by BAC DNA digestion with appropriate restriction enzyme or PCR amplification. The chimeric viruses will be rescued in Vero cells and then verified by sequence analyses. Our research group is well versed in coronavirus reverse genetics. 3.3.b Cell entry analysis: Hela cells expressing human ACE2 are cultured on coverslips in 24-well plates incubated with the chimeric bat SARSr-CoVs with different spike proteins at a multiplicity of infection (MOI) = 1.0 for 1 h. The inoculum is removed and the cells are washed twice with PBS and supplemented with medium. Hela cells without ACE2 are used as negative control. Twenty-four hours after infection, cells are rinsed with PBS and fixed with 4% formaldehyde in PBS (pH7.4) at 4 ·c for 20 min. ACE2 expression is detected by using goat anti-human ACE2 immunoglobulin followed by FITC-labelled donkey anti-goat immunoglobulin. Virus replication is detected by using rabbit antibody against the nucleocapsid protein of bat SARSr-CoV followed by Cy3-conjugated mouse anti-rabbit lgG. In parallel with the immunofluorescence assay, plaque assay will be conducted to determine the viral titers and growth kinetics in the infected cells at different times post-infection. 3.3.b Primary human airway epithelial cell culture: Primary human ciliated airway epithelial cells (HAE) cultures from the lungs of transplant recipients represent highly differentiated human airway epithelium containing ciliated and non-ciliated epithelial and goblet cells, grown on an air-liquid interface for several weeks prior to use ( 18, 55, 56). We will prepare HAE cultures from three different patient codes in triplicate in collaboration with the tissue procurement facility at the Cystic Fibrosis Center at UNC. Cultures will be inoculated with chimeric bat SARSr-CoVs to assess efficient replication. At 72 hpi, cultures will be fixed for immunofluorescent staining using antisera to the SARS-CoV conserved nucleocapsid protein (N) (57, 58). SARSr-CoVs that differ significantly in S protein sequence (11-24%) from epidemic SARS-CoV yet replicate in vitro, will also be evaluated for sensitivity to neutralization in Vero cells by PRNTS0 assays using broadly SARS-CoV cross reactive human mAbs S227.14, S230.15, S215.17, and S109.8 ( 49, 55). As controls, the S 11 Page 170 Research Strategy

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Contact PD/Pl: DASZAK, PETER genes of novel SARSr-CoV will be inserted into VEE 3526 replicon vectors (VRP3526-S), packaged and used to vaccinate mice (59). Polyclonal sera will be harvested and tested for ability to cross neutralize SARS-CoV, GD03, WIV-1, SHC014, WIV-16, other novel SARSr-CoV and HKU3-SRBD by PRNT50 assay (55, 60, 61). Using PRNT50 titers with sera (n=4 each) among these viruses, antigenic cartography (62) will allow comparison of antigenic vs. phylogenetic distance, identifying the transition at which SARSr-CoV strains escape SARS-CoV based vaccines, informing immunotherapeutic and vaccine design strategies (63-65). 3.3.c Humanized mouse infection experiments: Briefly, in BSL3, n=5 10- to 20-week old hACE2 transgenic mice will be intranasally inoculated with 1 x 104 PFU of wildtype WIV-1 or chimeric bat SARSr-CoVs with different spike proteins, then monitored daily for weight loss, morbidity, and clinical signs of disease. Mice will be euthanized at 2, 4, and 6 dpi (endpoint of 14 dpi), organs harvested and virus quantified by SARS-CoV NP RT-PCR. After 7 days in formalin, tissues for histopathology will be removed from the BSL3 and stained with H&E, and for immunohistochemistry using polyclonal antibodies against the N protein. We will conduct limited evaluation of existing countermeasures using therapeutic monoclonal antibodies in vitro and in vivo. Existing SARSr-CoV mAbs will be diluted in DMEM starting at 1 :20, and serial 2-fold dilutions mixed with an equal volume of 50 PFU of chimeric bat SARSr-CoVs with different spike proteins, then incubated on Vero E6 cells at 37°C for 1 h, then changed to a semi-solid medium (DMEM containing 1 % methylcellulose and 2% FBS). Antibody neutralization titers will be evaluated by plaque quantification at 4-5dpi. hACE2 transgenic mice will be injected with SARS-CoV mAbs, and infected with chimeric bat SARSr-CoVs. Clinical signs and morbidity will be assessed and tissue pathology examined and compared with mice without treatment of mAbs to determine the therapeutic effect on SARSr-CoV infection, and protection of SARSr-CoV by wildtype SARS-S based vaccines assessed as described (56, 66). We will sequence full length genomes of high risk strains that are antigenically distinct and escape SARS cross neutralization, synthetically reconstruct a small subset (1-2) and evaluate the ability of nucleoside analogues to inhibit growth in HAE cultures and/or in vivo (55, 56). 3.3.d HKU3 clade cellular receptor: We will screen potential receptor molecules by pull-down analysis on membrane proteins interacting with the spike protein, initially using bat primary intestinal epithelial cell lines and lysates to extract protein, isolate membranes, and proteomically sequence intestinal proteins. The fusion protein of the HKU3 and HKU3r-CoV S proteins containing human lmmunoglobulin Fe fragment will be eukaryotically expressed and purified. SARSr-CoV Swill be incubated as bait protein with the membrane proteins extracted from Rhinolophus sinicus intestinal cells, to capture and precipitate membrane proteins that interact with the S protein. Mass Spectrometry will be performed to screen for the candidate receptor molecules and Co-lmmunoprecipitation assay to confirm binding of the SARSr-CoV S protein to the candidate receptor. Alternatively, retroviruses pseudotyped with the SARSr-CoV S protein will be constructed and used to infect cells trans-expressing the candidate receptor molecule. Luciferase activity will be measured to test whether the S protein can bind to the receptor. If successful, this work will allow future research to clone and study human HKU3 receptor ortholog's ability to function as a receptor for other clade 2 strains and will allow better assessment of risk of clade 2 SARSr-CoV spillover to humans. 3.4 Combined spatial risk 'hotspot' analyses: We will use data from 3.3 to identify rank SARSr-CoV strains most likely to infect people and evade therapeutic and vaccine modalities. We will use bat survey and zoological data ( 44) to build species distribution models ( 67) and predict the distribution of bat species that harbor low, medium and high risk viral strains. Stacking these modeled distributions for the ~20-30 Rhinolophus and related species that occur in the region will allow estimates of SARSr-CoV diversity for a given locality. We will use machine learning models (boosted regression trees) and spatial 'hotspot' mapping approaches to identify the ecological, socio-economic and other correlates of SARSr-CoV diversity and spillover (from serosurveys) (21, 68, 69). We will include data from our human behavioral surveys and sampling to give a direct measure of where risk of spillover to people is likely to be highest in the region. Potential problems/alternative approaches: We may not be able to glean further information about the capacity of HKU3r-CoVs to infect human cells, or bind to human cell surface receptors. If attempts at culture are unsuccessful, and efforts to identify the receptor too costly or time-consuming, we will cease this line of work. In that event, we will focus entirely on filling out the gaps in the 10-25% S protein sequence divergence from SARS-CoV, by working on a greater diversity of lineage 2 SARSr-CoVs. 12 Page 171 Research Strategy

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Progress report publication list: R01 Al110964, Daszak Pl, Project Period: 06/0112014- 05/31/2019 The following are peer-reviewed papers published from work funded by this NIAID R01 during the project period ( 1-18). Other manuscripts on behavioral risk, phylogenetic and viral risk characterization are in prep and awaiting submission prior to the end date of the grant. 1. B. Hu et al., Detection of diverse novel astroviruses from small mammals in China. Journal of General Virology 95, 2442-2449 (2014). PMID: 25034867 2. B. Hu, X. Y. Ge, L. F. Wang, Z. L. Shi, Bat origin of human coronaviruses. Virology journal 12, (2015). PMID: 26689940 3. V. D. Menachery et al., A SARS-like cluster of circulating bat coronaviruses shows potential for human emergence. Nature Medicine 21, 1508-+ (2015). PMID: 26552008 4. J. N. Mandi et al., Reservoir Host Immune Responses to Emerging Zoonotic Viruses. Ce/1160, 20-35 (2015). PMID: 25533784 5. M. N. Wang et al., Longitudinal surveillance of SARS-like coronaviruses in bats by quantitative real-time PCR. Virologica Sinica 31, 78-80 (2016). PMID: 26920711 6. L. Brierley, M. J. Vonhof, K. J. Olival, P. Daszak, K. E. Jones, Quantifying Global Drivers of Zoonotic Bat Viruses: A Process-Based Perspective. American Naturalist 187, E53-E64 (2016). PMID: 26807755 7. X. Y. Ge et al., Coexistence of multiple coronaviruses in several bat colonies in an abandoned mineshaft. Virologica Sinica 31, 31-40 (2016). PMID: 26920708 8. L.-P. Zeng et al., Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIV1 Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response. Journal of Virology 90, 6573-6582 (2016). PMID: 27170748 9. X. L. Yang et al., Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus. Journal of Virology 90, 3253- 3256 (2016). PMID: 26719272 10. K. J. Olival et al., Host and viral traits predict zoonotic spillover from mammals. Nature 546, 646-650 (2017). PMID: 28636590 11. L. P. Zeng et al., Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses. Science China-Life Sciences 60, 1399-1402 (2017). PMID: 29134417 12. X. L. Yang et al., Genetically Diverse Filoviruses in Rousettus and Eonycteris spp. Bats, China, 2009 and 2015. Emerging Infectious Diseases 23, 482-486 (2017). PMID: 28221123 13. B. Hu et al., Discovery of a rich gene pool of bat SARS-related coronaviruses provides new insights into the origin of SARS coronavirus. PLoS pathogens 13, (2017). PMID: 29190287 14. N. Wang et al., Serological Evidence of Bat SARS-Related Coronavirus Infection in Humans, China. Virologica Sinica, (2018). PMID: 29500691 15. C. M. Luo et al., Discovery of Novel Bat Coronaviruses in South China That Use the Same Receptor as Middle East Respiratory Syndrome Coronavirus. Journal of Virology 92, (2018). PMID: 29669833 16. Y. Luo et al., Longitudinal Surveillance of Betacoronaviruses in Fruit Bats in Yunnan Province, China During 2009-2016. Virologica Sinica 33, 87-95 (2018). PMID: 29500692 17. Z. Wu et al., Comparative analysis of rodent and small mammal viromes to better understand the wildlife origin of emerging infectious diseases. Microbiome 6, 178 (2018). PMID: 30285857 18. P. Zhou et al., Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin. Nature 556, 255-258 (2018). PMID: 29618817 Page 172 Progress Report Publication List

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Contact PD/Pl: DASZAK, PETER PHS Human Subjects and Clinical Trials Information Are Human Subjects Involved • Yes Is the Project Exempt from Federal regulations? O Yes Exemption Number 01 Other Requested Information Tracking Number: GRANT12743073 0 No e No 0MB Number: 0925-0001 and 0925--0002 Expiration Date: 03/31/2020 02 03 04 05 06 07 OB Page 173 Funding Opportunity Number: PA-18-484 Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Human Subject Studies Study# 1 Tracking Number: GRANT12743073 Study Title Clinical Trial? Understanding the Risk of Bat Coronavirus Emergence: Community and clinic-based No surveillance to capture SARSr-CoV spillover, routes of exposure and potential public health consequences Page 174 Funding Opportunity Number: PA-18-484 Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Section 1 - Basic Information (Study 1) 1 . 1 . Study Title * 0MB Number: 0925-0001 and 0925-0002 Expiration Date: 03/3112020 Understanding the Risk of Bat Coronavirus Emergence: Community and clinic-based surveillance to capture SARSr-CoV spillover, routes of exposure and potential public health consequences 1.2. Is this study exempt from Federal Regulations • 0 Yes e No 1.3. Exemption Number □ 1 0 2 O 3 O 4 05 06 07 08 1.4. Clinical Trial Questionnaire • 1 .4.a. Does the study involve human participants? 1.4.b. Are the participants prospectively assigned to an intervention? 1.4.c. Is the study designed to evaluate the effect of the intervention on the participants? 1.4.d. Is the effect that will be evaluated a health-related biomedical or behavioral outcome? 1.5. Provide the ClinicalTrials.gov Identifier (e.g. NCT87654321) for this trial, if applicable Tracking Number: GRANT12743073 • Yes o Yes 0 Yes O Yes 0 No e No e No e No Funding Opportunity Number: PA-18-484 Received Date: Page 175 201s-11-05T16:31 :22.000-05:oo

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Contact PD/Pl: DASZAK, PETER Section 2 - Study Population Characteristics (Study 1) 2.1. Conditions or Focus of Study 0 Emerging zoonotic disease from bat Coronavirus 2.2. Eligibility Criteria Participants to be enrolled in this study will be people living, working, or visiting the high-risk sites: 1) of likely divergence of SARSr-CoVs; 2) with a high diversity of SARSr-CoVs within the S protein sequence divergence of 5-25%; and 3) with high rates of human-wildlife interactions in four provinces of Yunnan, Guangxi, Guizhou, and Guangdong in China, who meet the inclusion criteria outlined below. Study sites are prioritized according to ecological and epidemiological conditions associated with a high risk for SARSr-CoVs spillover. Research participants will be enrolled in two settings of community and hospital or clinic. Community People living, working, or visiting targeted high-risk communities (as defined above) who have close contact with bats, we anticipate interviewing and collecting biological samples from individuals with a range of exposure to bats. Enrolled research participants will be asked to provide biological samples and complete a questionnaire that is designed to obtain information about living circumstances (e.g. distance between the living house and closest bat roost, observed bats in house), income or livelihood, experience with SARI and ILi-iike illness and involvement in activities with direct or indirect (e.g. via livestock) contact with bats. Additional inclusion criteria for community participants - Adults (18 years of age or older) who provide informed consent - Pregnant women will be considered eligible for inclusion Exclusion criteria for community participants - Adults (18 years of age or older) who are unable to provide informed consent, including individuals with physiologically or medically induced cognitive impairments - Individuals under 18 years of age - Prisoners Hospital or clinic Patients at clinics or hospitals presenting with clinically defined symptoms of severe/acute respiratory illness (SARI/ARI) and/ or influenza-like illness (ILi) with unknown origin. As with the community-based group, biological samples will be collected from the patients, and the patients or his/her designate will complete a questionnaire. We will follow up with these participants 35 days after enrollment to collect another biological sample to assess the development of lgG and collect additional data on the course of symptoms in the interim period. Additional inclusion criteria for hospital or clinic participants - Adults ( 18 years of age or greater) who provide informed consent - Children aged 12 years or older with an accompanying parent or guardian who is able to provide informed consent, with the assent of children 12 years or older also required - Pregnant women will be considered eligible for inclusion Exclusion criteria for hospital or clinic participants - Individuals over the age of 12 years who refuse to provide informed consent - Adults unable to provide informed consent, including individuals with physiologically or medically induced cognitive impairments - Children, aged 12-17, without an accompanying parent or guardian who is able to provide informed consent, or a child aged 12 to 17 who is unable or unwilling to provide assent - Children < 12 years of age - Prisoners 2.3. Age Limits Min Age: 12 Years Max Age: N/A (No limit) 2.4. Inclusion of Women, Minorities, and Children NIAID_COV _2019_Inclusion_of_Women_Minorities_Children_Final.pdf 2.5. Recruitment and Retention Plan NIAID_COV_2019_Recruitment_Retention_Final.pdf 2.6. Recruitment Status Not yet recruiting 2. 7. Study Timeline NIAID_COV _2019_Study_ Timeline_Final.pdf 2.8. Enrollment of First Subject 06/01/2020 Anticipated Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 Received Date: Page 176 201a-11-osT16:31 :22.000-os:oo

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter INCLUSION OF WOMEN AND MINORITIES: This proposal will enroll men and women as study participants without regard to ethnicity. • At community sites, exposure to bats in working and living environments will be the primary criteria for identifying participants in community. We will make every effort to have men and women equally represented in this study and no individuals will be excluded based on ethnicity. • At clinic sites, only patients who present at the health facility who meet the clinical case definition of 1) severe/acute respiratory illness (SARI/ARI) of unknown origin; or 2) Influenza-like illness (Ill) of unknown origin will be recruited for this study, and no patients will be excluded (or included) based on ethnicity or gender. INCLUSION OF CHILDREN: Children aged 12 years or older will be included in the clinical syndromic study. • Previous clinic-based studies have shown that children are one of the major populations who are affected by the severe/acute respiratory illness (SARI/ARI) and/or Influenza-like illness (Ill). • Children aged 12 years or older are post-primary school and are able to respond to the questionnaire on their own which increases the reliability of responses. • Our human research team at the Institute of Pathogen Biology are all well-trained and have extensive experience working with children at this age, as well as their parents, in a clinical setting since 2009. • Every effort will be made to protect the privacy, dignity, and well-being of children who participate in this study. • Inclusion of children in the study would increase the sample size to allow for the estimation of effect sizes of behavioral risk factors by two-fold (2X) or greater with 80% power. We will not include children in the community-based surveillance because children in target communities are mainly school children who have very limited exposures to bats or other wild animals under the scenarios of interest to the study, prolonged time spent in the forests or markets. Page 177 Inclusion of Women, Minorities, and Children

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter RECRUITMENT AND RETENTION PLAN In order to improve recruitment within target communities, introductory visits will be made by project staff to each of the selected sites. These visits will be advertised through word of mouth and a project description letter to town/city leaders that can be posted in a central community location. The letter will inform the community that a team will be coming on a particular day(s) to discuss health issues related to animal contact. The letter will not be advertised for recruitment purposes. It will only be used to inform the community of the research visits. The project description letter will be written in the local language with a Flesch-Kincaid readability score equivalent to a 7th grade and up level (post-primary in China), to assure that potential community participants understand the study purpose, eligibility, and inclusion guidelines. During community visits, discussions and meetings will be held firstly with local authorities and community leaders to introduce our project, and when appropriate and following approval from local authorities, the study team will post flyers to inform the community when the team will be coming back to speak about enrollment. This "town hall" style meeting will be completely voluntary, and, based on our experience, those interested would likely attend. Although local authorities may be present to introduce the study team members, they will not be involved in the recruitment and/or consent of the participants for the study. If research visits or recruitment are held at a workplace, subjects will be clearly informed during this recruitment process that their participation in the study is voluntary and will not impact their employment, nor will information discussed be shared with employers. With the local permission and accompanied will local authorities or community leaders, study team members will also engage in community 'walkabouts' during which they will discuss study details, as well as dates, times, and locations for enrollment and participation in the study. Participation in the study will be strictly voluntary and will require signed informed consent for all participants and signed assent for clinical participants aged 12-17 along with parent or guardian consent. Participants will be given a consent form prior to being asked to participate in this study. Our research staff will read the consent form to potential participants, and they will review the consent form with the research staff and be given time to ask questions. After reviewing the consent form, study staff will explain details of the study including: why they were selected, what the study procedures are and what will be expected from them, potential risks and benefits of their participation, that their participation is completely voluntary, and that they can withdraw their participation at any time. Responses will be kept strictly confidential. Measures will be taken to assure the privacy, dignity, and respect of each participant. During training of research staff, we will emphasize the importance of avoiding coercion and protecting the privacy of participants. Community-based recruitment: Participants from the community will be recruited through town hall meetings and community 'walkabouts' as described above. Meeting dates, times, and locations for enrollment and participation will be shared during these activities, and participants who wish to enroll can volunteer to participate at these times and locations. Clinic-based recruitment: Patients eligible for enrollment will be identified at intake areas or in the emergency room, ward, or intensive care unit of each participating clinic and hospital by clinic staff, according to standard operating procedures at collaborating sites. Employed staff at each location will identify potential participants meeting the clinical case definition of severe/acute respiratory illness (SARI/ARI) and/or influenza-like illness (Ill) with unknown origin. Patients will be screened for eligibility according to the inclusion/exclusion criteria based on available clinical information. For larger provincial-level hospitals, interval sampling will be implemented by selecting every Nth case at the site among those individuals who meet enrollment criteria. The interval will be determined by local implementing partners based on an evaluation of the expected number of cases presenting at the site within a given year in order to best meet study design and sample size criteria. In terms of retention, we will express our gratitude to subjects for their participation and discuss the importance of the follow-up data collection. Nonetheless, we expect to have an approximate 40% loss to follow up and have included this in our sample size calculations. Page 178 Recruitment and Retention Plan

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter STUDY TIMELINE Patients/participants will be asked to volunteer approximately 1 hour of their time for participation in the study, including providing biological samples and completing the questionnaire at each sampling time point. This will be an ongoing, five-year project (June 01, 2019 -- May 31, 2024). We anticipate to starting human subject enrollment on June 01, 2020, and completion of preliminary analyses is expected in 2024. Page 179 Study Timeline

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Contact PD/Pl: DASZAK, PETER Inclusion Enrollment Reports IER ID# Enrollment Location Type Enrollment Location Study 1, IER 1 Foreign Local community and hospital/clinic in Yunnan, Guangdong, Guangxi, Guizhou Provinces Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 Received Date: Page 180 201a-11-osT16:31 :22.000-os:oo

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Contact PD/Pl: DASZAK, PETER Inclusion Enrollment Report 1 Using an Existing Dataset or Resource* : O Yes • No Enrollment Location Type* : O Domestic • Foreign Enrollment Country(ies): CHN: CHINA Enrollment Location(s): Local community and hospital/clinic in Yunnan, Guangdong, Guangxi, Guizhou Provinces Comments: This is a renewal, the cumulative enrollment from the previous funding period 5R01Al110964-05 is 980 females and 616 males, in total 1,596 Asians. We don't plan to use the existing dataset. Planned Ethnic Categories Racial Categories Total Not Hispanic or Latino Hispanic or Latino Female Male Female Male American Indian/ 0 0 0 0 0 Alaska Native Asian 4675 4675 0 0 9350 Native Hawaiian or 0 0 0 0 0 Other Pacific Islander Black or African 0 0 0 0 0 American White 0 0 0 0 0 More than One Race 0 0 0 0 0 Total 4675 4675 0 0 9350 Cumulative (Actual) Ethnic Categories Unknown/Not Racial Categories Not Hispanic or Latino Hispanic or Latino Reported Ethnicity Total Unknown/ Unknown, Unknown/ Female Male Not Female Male Not Female Male Not Reported Reported Reported American Indian/ 0 0 0 0 0 0 0 0 0 0 Alaska Native Asian 0 0 0 0 0 0 0 0 0 0 Native Hawaiian or 0 0 0 0 0 0 0 0 0 0 Other Pacific Islander Black or African 0 0 0 0 0 0 0 0 0 0 American White 0 0 0 0 0 0 0 0 0 0 More than One Race 0 0 0 0 0 0 0 0 0 0 Unknown or 0 0 0 0 0 0 0 0 0 0 Not Reported Total 0 0 0 0 0 0 0 0 0 0 Page 181 Tracking Number: GRANT12743073 Funding Opportunity Number: PA-18-484 Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Section 3 - Protection and Monitoring Plans (Study 1) 3.1. Protection of Human Subjects NIAID _ COV _2019_Protection_Human_Subjects_Final.pdf 3.2. Is this a multi-site study that will use the same protocol to O Yes e No 0 N/A conduct non-exempt human subjects research at more than one domestic site? If yes, describe the single IRB plan 3.3. Data and Safety Monitoring Plan 3.4. Will a Data and Safety Monitoring Board be appointed for this study? 3.5. Overall structure of the study team Tracking Number: GRANT12743073 O Yes e No Funding Opportunity Number: PA-18-484 Received Date: Page 182 201a-11-osT16:31 :22.000-os:oo

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter PROTECTION OF HUMAN SUBJECTS: 1. Risks to Human Subjects 1.1 Human Subjects Involvement, Characteristics, and Design This project is a study of human exposure to animal coronaviruses in southern China. Subjects will be enrolled on a voluntary basis and informed consent will be obtained from all participants. Consenting participants will provide biological samples and complete a questionnaire. Subjects will be individuals: 1) who are highly exposed to bats in community settings, including through hunting, butchering, or general handling within the context of their living or working environment (~ 18 years old); and 2) patients admitted to hospitals and clinics presenting with disease symptoms of clinically-defined severe/acute respiratory illness (SARI/ARI) or Influenza-like illness (Ill) of unknown origin (~ 12 years old). The study population will be selected from the Yunnan, Guangxi, Guangdong, and Guizhou provinces of China. We will aim to enroll: 1) in 12 clinic sites across the four provinces, 2,750 individuals (accounting for an estimated 40% loss from follow-up); and 2) in 8 community sites, 1,650 individuals per each of the four provinces, pooled across two sites for each province for a total of 6,660 (1,650*4) participants, allowing us to make province-level comparisons of differing effects (one time data collection, no follow-up among community participants). The community and clinical sites are further defined in "Specific Aim 2: Using community-based and clinical biological-behavioral surveys to identify SARSr-CoV spillover, routes of exposure and public health consequences of human infection". There are no data to suggest a gender or ethnic bias for coronavirus exposure or infection, therefore subjects will be enrolled based on exposure criteria, and subjects will not be excluded based on ethnicity or gender. We will also stratify sampling to ensure representation of sex, demographic, and socio-economic factors in each community site. 1.2 Sources of Materials Samples to be collected and screened for coronaviruses include whole blood and nasal/oropharyngeal swabs. Samples will be collected and a questionnaire will be administered by trained medical personnel from the local CDC, hospitals, and clinics. In community sites, whole blood samples (only) will be collected once during Years 2-4 of the study, and samples will be screened for coronaviruses using developed ELISA at the Institute of Pathogen Biology and the Wuhan Institute of Virology. In clinic sites, both whole blood samples and nasal/oropharyngeal swabs will be collected at enrollment, and samples will be screened for coronaviruses using ELISA and consensus PCR (cPCR). Patients who test positive for coronavirus or antibodies to coronavirus will be followed up 35 days after enrollment, when additional blood samples will be collected for serological testing with ELISA. At the enrollment, a standardized questionnaire will be administered at both community and clinic sites to collect data on living circumstances (e.g. distance between the living house and closest bat roost, observed bats in house), income or livelihood, experience with SARI and Ill-like illness and involvement in activities with direct or indirect (e.g. via livestock) contact with bats. During the follow-up with clinic study participants, a standardized questionnaire supplement will be administered to collect additional data on the course of symptoms in the interim period. All electronic data will be password protected, and all hardcopy files and biological samples will be stored in secure storage facilities. All consent forms will be stored separately from any data in separated locked filing cabinets. 1.3 Potential Risks The potential risks to study participants resulting from study participation are minimal. The volume of blood being collected is within normal safety limits. The questionnaire will be designed to assess exposure risk, and may ask personal questions, but they will conducted in private and confidentially to protect privacy. There may be some stress to subjects who are informed that they have been exposed to an animal virus, but counseling will be available and options for medical care will be included in the discussion. Page 183 Protection of Human Subjects

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 2. Adequacy of Protection against Risks 2.1 Recruitment and Informed Consent Potential study participants at each site will be identified by well-trained in-country research team in partnership with local CDC staff (for community participants) and medical personnel (for clinic participants). The team will be thoroughly trained on communicating the research objectives, what is being asked of participants, any risks or benefits, and will be able to address any questions that potential subjects may have. Both written and oral descriptions of the study details will be provided in Chinese Mandarin (or orally via an interpreter in local dialects if necessary) as part of the informed consent process. Contact details of the collaborators at the local CDC or hospital and the study Pl will be provided to all subjects, and CDC or hospital personnel on the research team will be available onsite to answer questions from the study subjects. Test results will be communicated to each subject and counseling offered to minimize stress. 2.2 Protection against Risks After the informed consent process, the questionnaire will be conducted in private, ensuring that others cannot overhear responses. Individual sessions will be held in areas where there are no other individuals within a 10- foot distance. A barrier will be created so that no other individuals can view the participants during their interview. Depending on the location, this could be a private room, behind a building or fence, or behind a line of trees, obstructing view so that confidentiality may be maintained. The interview team will take care to pair interviewers and respondents by sex to the best of their ability to increase the level of comfort of the participant and the team will ensure the privacy and confidentiality of responses. Children will not be interviewed in the absence of a parent or guardian. This study will not involve greater than minimal risk, and every effort will be made to ensure the privacy, dignity, and well-being of children who participate in this study. 3. Potential Benefits to Subjects and Others There are potential benefits to the study subjects including receiving a physical exam/health check from a medical officer and the potential benefit of identifying a health hazard. At the conclusion of the study, we will deliver an educational workshop for high risk individuals (open to study subjects and non-study subjects) describing the health benefits of using PPE and hand-washing during animal handling activities throughout the day, as well as to share other prevention interventions that emerge from the research data. 4. The Importance of Knowledge to be Gained There are valuable potential benefits to the general public from the knowledge to be gained by this study, as it may identify sources of zoonotic coronaviruses in the market system or through hunting. Avoidance of these animals or extra care when handling them may substantially reduce the risk of CoVs (and other zoonotic pathogen) transmission. Page 184 Protection of Human Subjects

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Contact PD/Pl: DASZAK, PETER Section 4 - Protocol Synopsis {Study 1) 4.1. Brief Summary 4.2. Study Design 4.2.a. Narrative Study Description 4.2.b. Primary Purpose 4.2.c. Interventions J Type I Name ] Description 4.2.d. Study Phase Is this an NIH-defined Phase Ill Clinical Trial? O Yes 4.2.e. Intervention Model 4.2.f. Masking o Participant Yes o Care Provider 4.2.g. Allocation 4.3. Outcome Measures Type Name 4.4. Statistical Design and Power 4.5. Subject Participation Duration Time Frame 4.6. Will the study use an FDA-regulated intervention? 4.6.a. If yes, describe the availability of lnvestigational Product (IP) and lnvestigational New Drug (IND)/ lnvestigational Device Exemption (IDE) status 4.7. Dissemination Plan O Yes 0 No O No o Investigator o Outcomes Assessor Brief Description 0 No Funding Opportunity Number: PA-18-484 Received Date: Tracking Number: GRANT12743073 Page 185 201a-11-osT16:31 :22.000-os:oo

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Contact PD/Pl: DASZAK, PETER Delayed Onset Studies Delayed Study Title Onset Study# The form does not have any delayed onset studies Tracking Number: GRANT12743073 Page 186 Anticipated Clinical Justification Trial? Funding Opportunity Number: PA-18-484 Received Date: 2018-11-05T16:31 :22.000-05:00

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter VERTEBRATE ANIMALS: 1. Detailed description of animal use. Work with vertebrate animals will be conducted at Wuhan University at the School of Medicine in Wuhan, China and the University of North Carolina, Chapel Hill, USA. Capture and sampling techniques for all wild animals (bats) described in this study have been previously approved by multiple Institutional Animal Care and Use Committees (IACUCs) for projects led by EcoHealth Alliance. These institutions include: UC Davis IACUC (Mazet and Epstein; UC Davis 15898; current); and The Cummings School of Veterinary Medicine at Tufts University (Olival and Epstein, current), Animal Welfare Assurance (#A4059-01) on file with the Office of Laboratory Animal Welfare at the National Institutes of Health. We have prepared a draft IACUC application for this project, and will submit it within 1 month of the project's start date (to Tufts University) to minimize delays in beginning Year 1 field sampling. Experimental work using humanized mice will be conducted at the Center for Animal Experiment Biosafety 3 lab of Wuhan University at the School of Medicine in Wuhan, China and the University of North Carolina at Chapel Hill, the Institute for Pathogen Biology. The Wuhan laboratory is AAALAC accredited and has both an Institutional Biosafety Committee and an Institutional Animal Care and Use Committee. We will submit our protocols for IACUC approval should this proposal be funded. Conditions for animal use are described below. Both laboratories (Wuhan and UNC) have Internal Biosafety Committees and are accredited BSL-2 and BSL 3 laboratories. Animals will be housed in a BSL-3 facility and will be under the care of a full-time veterinarian. All experimental work using infectious material will be conducted under appropriate biosafety standards. Disposal of hazardous materials will be conducted according to the institutional biosafety regulations. Note: The majority of wild bats captured and sampled will be done using non-destructive, techniques. In a small number of instances (~ 2 bats per species for previously unsampled Rhinolophus sp.), where intestine and lung tissue is required to establish cell lines, animals will be humanely euthanized and a necropsy performed according to accepted protocols (see euthanasia section) Bat capture. Free-ranging bats will be captured using either a mist net or harp trap. The net system is manned by two people during the entire capture period, and bats are removed from the net as soon as they become entangled to minimize stress and prevent injury. In the Co-Pl's (Dr. Olival) experience, a maximum of 20-30 bats can be safely held and processed by a team of three people per trapping period. Duration of trapping will depend on the capture rate. Bats are placed into a pillowcase or small cloth bag and hung from a branch or post until samples are collected. Bats are held for a maximum of six hours (typically less than 3 hours), and released after sampling. Laboratory mice. Lab mice will be sourced commercially by the Wuhan Center for Animal Experiment at Wuhan University. Sample Collection: Bats: Bats will be manually restrained during sampling. Depending on the species and size of bat, swabs will be taken from the oropharynx, urogenital tract, and rectum. Fresh feces will be collected if available, in which case a rectal swab will not be collected. Blood will be collected from fruit bats either from the cephalic vein or from the radial artery or vein using a 25 gauge needle and 1 cc syringe. Blood Page 187 Vertebrate Animals

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter will be collected from bats weighing less than 100g according to published techniques (126). Rodents: Anesthesia for captive small rodents will be conducted using plastic tubes, with the animals transferred directly from the traps to the tubes containing a cotton swab soaked in ether, isoflurane, or methoxyflurane for anesthetic induction. For larger rodents, chemical restraint and anesthesia (ketamine alone, or ketamine combined with xylazine) will be applied either through the squeeze cages by syringe if applicable. Once anesthetized a small blood sample will be collected using a capillary tube placed into the retro-orbital sinus. Only trained technicians will perform retro-orbital bleeding and it will only be performed on anesthetized rodents. Femoral or jugular venipuncture may be used for larger rodents (e.g. rats). In all rodents, blood volumes of no more than 1 % of body weight will be withdrawn. (example 0.2 ml blood from a 20 gram rodent). Laboratory Mice. Humanized mice will be bred at the University of Wuhan and University of North Carolina at Chapel Hill. Mice will be inoculated with a specific dose (e.g. 1x106 TCID50) of virus through different routes (intranasally and intraperitoneally). Mouse body temperature will be monitored with implanted temperature sensing microchips (LifeChip Bio-thermo, Destron Fearing), and mice will be weighed daily. Animals will be observed daily for clinical signs of illness. Moribund mice will be euthanized, according to AVMA recommendations. Live animals will be euthanized at three weeks postinoculation and organs harvested. We will collect sera on days 10, 15 and 21 to test for neutralizing antibodies against bat CoVs. We will collect nasal washes, oral swabs, and rectal swabs, and urine every two days. These are minimally invasive procedures, and will be performed by experienced lab technicians under the supervision of a full-time veterinarian. 2. Justify use of animals, choice of species, numbers to be used. Species and number used in study: The purpose of this study is to conduct targeted, but extensive surveillance of bat populations in Southern China to detect coronaviruses that may pose a risk to the health of both humans and animals. The experimental work is designed to understand the ability of bat coronaviruses to bind to human receptors. In this renewal application, we propose a total bat sample size comparable to our initial R01 effort of -5000 animals. These animals will be sampled from -15-20 species collected across 4 provinces. Given -5-12% prevalence of SARSr-CoVs in Rhinolophus spp. at our previous sites during our initial R01, this sample size would give us 425 (±175) positive individual bats, and allow us to identify ~125 novel strains. Assuming a conservative prevalence rate, a sample size of n=110 individuals per species will allow us to detect SARSr-CoV using PCR with a power of 80%. Wild bats: We will sample a minimum of 110 individuals from ~15-20 different bat species from sites across four provinces in Southern China (Yunnan, Guangxi, Guizhou and Guangdong). Sampling will focus on species in the family Rhinolophidae, genus Rhinolophus., but will also include individuals in the related genera Hipposideros and Aselliscus. In every situation, sampling of wildlife will be conducted in the most humane manner while minimizing the impacts on individual animals and their wild populations. In all instances, the fewest number of animals will be sampled that will provide valid information and statistical inference for the pathogen and disease of interest and every effort will be made to minimize stress and discomfort for the animal. A small number of bats (maximum 2 per species) representing each of the species in this study may be euthanized in order to collect lung and intestinal tissue required for characterizing coronavirus receptors. Voucher specimens may also be collected at the discretion of the team leader for the accurate identification of species using molecular methodology. Humanized mice for experimental infection for Specific Aim 3: In order to understand whether select strains of bat-borne CoVs utilize receptors found in people have the potential to infect people, we will use Swiss albino mice (standard breed at Wuhan University) that have been genetically modified to have 2 Page 188 Vertebrate Animals

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter human receptors. We'll infect them with cultured bat coronaviruses and determine which organs become infected and whether these mice are capable of shedding infectious virus. Humanized mice will be genetically modified to carry human ACE2 gene will be used to evaluate pathogenesis of CoVs. We cannot anticipate exactly how many viruses we will find that are candidates for experimental models, however will likely identify approximately 5-6 bat SARSr-CoV strains that will be used for mouse infection experiments. We will use 15-20 adult mice per virus strain, and therefore will require a maximum of 120 mice over the study period. 3. Provide information on veterinary care. For wild caught animals, there is no specific veterinary care that is appropriate, nor will clinical veterinary facilities be available. Animals that are injured during the capture or sampling process will be assessed by an experienced team leader, and if the animal is determined to be unlikely to survive if released, it shall be euthanized humanely (see euthanasia section). Animals will be released within hours of capture. Laboratory mice will be housed in BSL-3 small animal facilities at the Center for Animal Experiment at Wuhan University and University of North Carolina at Chapel Hill, the Institute for Pathogen Biology. Experimental animals will be regularly monitored by experienced staff and a supervising veterinarian. The animal facility operates 24 hours a day and has full-time veterinarians on staff. All animals will be provided with food and water ad libitum and will otherwise receive standard care. 4. Procedures for ensuring animal comfort, lack of distress, pain, or injury: Wild bats will not be held longer than 6 hours during the sampling process. Co-Pl Olival has extensive experience in capture, anesthesia, and sampling wildlife, especially bats. In our team's experience, bats tolerate the described procedure well. Mist nets will be attended continuously during capture periods, and bats will be extracted from the net as soon as they become entangled. This will minimize stress and injury from entanglement. Bats will be placed individually in cotton bags and hung from tree branches while awaiting processing and during recovery. The bags are sufficiently porous as to allow for ventilation and are designed for bat capture. The enclosed environment seems to calm the bats, as they do not struggle once inside, but they hang quietly - this is a standard and accepted practice in the bat research world and best way to minimize stress to the animal. Animals will be monitored by a veterinarian or experienced field team member during all stages of capture, processing, and release. Animals will be kept in a cool place while in the pillowcases. The procedures used in this experiment (blood draw, nasal, oral, and rectal swabs) are minimally invasive. Mice that show signs of morbidity post-infection will be examined and euthanized according to AVMA standards (see below). 5. Euthanasia: In the event of injury to an animal that results in pain and suffering, and reasonable veterinary care is unavailable, the animal will be euthanized by a veterinarian or trained field team member using ketamine injected intramuscularly 37.5mg/kg and sodium pentobarbital injected intravenously at a dose of 1.0ml per 5kg injected intravenously. This protocol is in accordance with the AVMA euthanasia report (2007). Any animal that is euthanized using a chemical agent will be disposed such that it will not be permitted to enter the food supply either through markets or hunting. 3 Page 189 Vertebrate Animals

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter SELECT AGENT RESEARCH/BIOHAZARDS. No select agent research. Agents: SARS-related bat coronaviruses (SARSr-CoV), like WIV1, WIV16 and SHC014. These bat viruses are distantly related to the epidemic human SARS-CoV which emerged in 2003 and caused 8,000 cases and 800 deaths worldwide. While the epidemic human SARS-CoV is a BSL3 select agent, the SARSr-CoV are BSL3 pathogens in the US and not select agents. The proposal will use a SARSr-CoV molecule clone designated WIV1 during the course of these studies, which is NOT a select agent. This strain has not been shown to cause human disease or be transmissible between humans. All recombinant DNA work will use the bat SARSr-CoV WIV1 molecular clone. At the University of North Carolina (US Government select agent certified laboratory), some virus growth studies will be conducted in primary human airways, comparing wildtype SARS-CoV, WIV1 and various SARSr-CoV WIV1 chimeric virus growth kinetics. Wildtype SARS-CoV strain research will not be conducted at the Wuhan Institute of Virology. Registration status of all entities where select agent(s) will be used. Wildtype SARS-CoV is a select agent. UNG-Chapel Hill is currently registered with the CDC for select agent use, including SARS-CoV, as required by select agent regulations (42 CFR 73). The UNC SARS select agent laboratories are routinely inspected by the environmental health and safety department at UNC and by the CDC. Workers receive select agent and BSL3 training focused on SARS-CoV safety, procedures and protective clothing/PAPR training each year. Introduction and Background. SARS-CoV caused outbreaks with significant case fatality rates, and there are no vaccines available for this agent. SARS-CoV is classified as a BSL-3 select agent. Wildtype SARS-CoV is currently thought extinct in the wild. The work proposed in this application will involve two aspects: field work and laboratory work, focusing on distantly SARS-like bat coronaviruses (SARSrCoV). Fieldwork involves the highest risk of exposure to SARSr-related or other bat CoVs, while working in caves with high bat density overhead and the potential for fecal dust to be inhaled. There is also some risk of exposure to pathogens or physical injury while handling bats, civets, rodents or other animals, their blood samples or their excreta. The Co-Pis and field team have extensive experience and certification working with wildlife species and high-biosecurity pathogens (Nipah virus, ebolavirus, SARS), and great care will be taken in the field to limit the risk of accidental exposure to known or unknown animal pathogens. We have strict procedures for handling bats and working with samples from them as they are secured in the field and transported to the lab. Field team members handling animals will be trained to utilize personal protective equipment (PPE) and practice proper environmental disinfection and biosafety techniques. This includes wearing coveralls or dedicated clothing, nitrile gloves, eye protection, and a P95 or P100 respirator during bat handling and sampling. Fully Tyvek suits and HEPA-filtered Powered Air Purifying and Supplied Air Respirator Systems (PAPRs) will additionally be worn in cave systems where there is a higher risk of contact with aerosolized bat feces. All field clothing and equipment will be disinfected using Virkon disinfectant. All biological waste from field surveys will be disposed of in the appropriate container (sharps box or an autoclave bag) and will be autoclaved at local hospitals or university labs. All personnel will be vaccinated against rabies and have a neutralizing antibody titer, in accordance with WHO and CDC recommendations. Field teams will carry rabies boosters in the field and will receive a booster in the event of a potential rabies exposure. Field safety protocol: Our procedures to deal with bites, needle-sticks etc. are as follows: The wound is washed thoroughly with soap and water to clean away dirt and debris, then vigorously scrubbed with a sterile gauze bandage and benzalkonium chloride for 5 minutes. If bleeding, pressure is applied with a sterile bandage for until bleeding has stopped. If the wound continues to bleed, medical attention at the nearest hospital is sought. The bat from which the bite or exposure originated is identified, and the samples collected from it labeled on the data sheet that these were involved in an exposure. Our procedures require that the person potentially exposed reports to a major hospital within 24 hours to Page 190 Select Agent Research

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter have wound examined and receive a rabies post-exposure booster vaccines as per WHO/CDC protocols. The laboratory work is lower risk, as samples placed in lysis buffer will be non-infectious. Samples placed in viral transport medium and frozen will be stored at ultra-low temperatures (-86°C) until viral isolation is required. Serum will be heat inactivated at 56°C for 30 minutes prior to testing. Lab biosafety: The University of North Carolina at Chapel Hill, the Institute for Pathogen Biology, the Wuhan Institute of Virology, and the Wuhan University Center for Animal Experiment BSL-3 laboratories all have respective Internal Biosafety Committees and are accredited BSL-2 and BSL 3 laboratories. All experimental work using infectious material will be conducted under appropriate biosafety standards. Disposal of hazardous materials will be conducted according to the institutional biosafety regulations. Available Treatments: No approved treatments are related for SARS and the SARSr-related bat coronavirus infections. However, therapeutic antibodies and nucleoside analogues have been successfully used in SARS-infected rodents and primates, which could be approved for compassionate use in humans exposed to the SARSr-CoV. UNC Facilities where the select agent(s) will be used. SARS-CoV will be manipulated in research activities including establishment of viral replication curves, infection of rodent animal models and performance of plaque assays in laboratory spaces that meet operational and procedural criteria for BSL-3 activities as outlined in the CDC/NIH "Biosafety in Microbiological and Biomedical Laboratories", 5th edition, as well as BSL-3 criteria outlined in the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (April 2016). In addition, all mouse studies at UNG-Chapel Hill will be performed in an approved and registered BSL-3/ABSL-3 laboratory equipped with Techniplast Sealsafe™ HEPA-filtered animal housing for rodents. All animal protocols will be approved by the UNG-Chapel Hill IACUC. UNC BSL-3/ABSL-3/select agent laboratories are equipped with biosafety cabinets, incubators, centrifuges with containment features, cold storage units, an autoclave, sink, eyewash and life safety equipment, and mechanical system monitors and alarms to support effective isolation and containment of operations involving SARS-CoV and SARSr-CoV. The anterooms to the BSL-3 laboratories house PAPR charging stations, laboratory and safety supplies, and a changing area. For both the BSL-2 and BSL-3 select agent spaces, access to select agents is restricted by the door between the hallway and anteroom and the door between the anteroom and BSL-3 space, requiring a combination of swipe card and punch code for entry. All select agent materials (SARS-CoV virus and genome length RNA) are stored in locked freezers and incubators. UNC Procedures for monitoring possession, use and transfer of select agents. All personnel who will have access to select agent-regulated materials have been added to the Select Agent registration following security risk assessments prescribed by the CDC Select Agent Program. Personnel have completed training in all aspects of select agent compliance requirements and have adopted changes to standard operating procedures as applicable to assure that these requirements are met. Personnel will follow all procedures prescribed for accessing and securing the laboratory, documenting laboratory activities and materials used, and responding to incidents that could result in theft, loss, or release of select agent-regulated materials. Transfers of select agentregulated materials will be coordinated by the laboratory managers and Responsible Official in accordance with standard operating procedures, including obtaining appropriate permits for shipping select agent materials and observing all regulations for shipping, both under dangerous goods and select agent regulations. Transfer of select agent RNA in TRlzol from registered BSL-3 to registered BSL-2 space and cDNA from registered BSL-2 space to non-registered BSL-2 space is conducted according to current select agent rules, regulations, and guidelines, including the new inactivation policies released in 2017. 2 Page 191 Select Agent Research

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter UNC Biosafety, biocontainment, and security of the select agent(s). The Barie laboratories have been operational with BSL-3 core policies and procedures for ~15 years. Standard operating procedures at BSL-3 have been reviewed and approved by the UNC Chapel Hill Institutional Biosafety Committee and undergo both annual review and approval as well as updates as laboratory processes change or biosafety procedures evolve. The content of these documents has been formatted to conform to select agent regulations for the biosafety, security, and incident response plans. Additionally, lab-specific security risk assessments have been completed and recommendations implemented to ensure that security measures and procedures are sufficient to effectively minimize the possibility of unauthorized access to select agent-regulated materials. The UNC Chapel Hill facilities have undergone multiple CDC inspections and are currently in compliance with CDC requirements relating to SARS-CoV and select agent status. Our three-year renewal inspection occurred in June 2018 and we have been renewed for another three years. UNC Biocontainment resources. All BSL-3 laboratories are under negative pressure, with redundant systems to ensure that negative pressure is maintained. All BSL-3 facilities have autoclaves to decontaminate waste materials as well as approved protocols for treatment or inactivation of any materials leaving the laboratory. All personnel are extensively trained in basic virology and safety protocols before being approved for select agent work and undergo additional extensive training to work with SARS-CoV and related SARSr-CoV as a BSL-3 pathogen. In both laboratories, annual testing is performed to verify that biosafety cabinets, laboratory supply/exhaust systems (including alarms), and other laboratory equipment are functioning as designed. The laboratories are secured at all times, and only personnel who have successfully completed Select Agent clearance and laboratory specific training requirements are permitted to enter without an escort. P3CO Research. Recognizing the implementation of new gain of function research guidelines under P3CO, SARS-CoV and MERS-CoV are subject to these guidelines, and as such, reverse genetic studies are subject to review. Our group has considerable expertise in interfacing with the appropriate NIH P3CO institutional review boards to review, revise and finalize research designs that have the potential to modify pathogenesis or transmissibility in mammals. Importantly, we are not proposing to genetically manipulate SARS-CoV over the course of this proposal. However, we are proposing to genetically manipulate the full length bat SARSr-CoV WIV1 strain molecular clone during the course of the proposal, which is not a select agent, has not been shown to cause human infections, and has not been shown to be transmissible between humans. 3 Page 192 Select Agent Research

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter REFERENCES l. C. Drosten et al., Identification of a novel coronavirus in patients with severe acute respiratory syndrome. New England Journal of Medicine 348, 1967-1976 (2003). 2. T. Ksiazek, D. Erdman, C. Goldsmith, A novel coronavirus associated with severe acute respiratory syndrome. The New England Journal of Medicine 348, 1953-1966 (2003). 3. L. H. Taylor, S. M. Latham, M. E. Woolhouse, Risk factors for human disease emergence. Philosophical transactions of the Royal Society of London. Series B, Biological sciences 356, 983-989 (2001). 4. Y. Guan et al., Isolation and characterization of viruses related to the SARS coronavirus from animals in Southern China. Science 302, 276-278 (2003). 5. W. Li et al., Bats are natural reservoirs of SARS-like coronaviruses. Science 310, 676-679 (2005). 6. A. Annan et al., Human Betacoronavirus 2c EMC/2012-related Viruses in Bats, Ghana and Europe. Emerging infectious diseases 19, 456-459 (2013). 7. $. Wacharapluesadee et al., Identification of Group C Betacoronavirus from Bat guano fertilizer, Thailand. Emerging Infectious Diseases [Internet], (2013). 8. S. Anthony et al., Coronaviruses in bats from Mexico. Journal of General Virology 94, (2013). 9. Z. A. Memish et al., Middle East respiratory syndrome coronavirus in bats Saudi Arabia. Emerg Infect Dis 19, (2013). 10. P. Zhou et al., Fatal swine acute diarrhoea syndrome caused by an HKU2-related coronavirus of bat origin. Nature 556, 255-258 (2018). 11. J. F. Drexler et al., Amplification of Emerging Viruses in a Bat Colony. Emerging Infectious Diseases 17, 449-456 (2011). 12. J. Huynh et al., Evidence Supporting a Zoonotic Origin of Human Coronavirus Strain NL63. Journal of Virology 86, 12816-12825 (2012). 13. S. K. P. Lau et al., Ecoepidemiology and Complete Genome Comparison of Different Strains of Severe Acute Respiratory Syndrome-Related Rhinolophus Bat Coronavirus in China Reveal Bats as a Reservoir for Acute, Self-Limiting Infection That Allows Recombination Events. Journal of Virology 84, 2808-2819 (2010). 14. P. L. Quan et al., Identification of a Severe Acute Respiratory Syndrome Coronavirus-Like Virus in a Leaf-Nosed Bat in Nigeria. Mbio 1, (2010). 15. S. Tong et al., Detection of novel SARS-like and other coronaviruses in bats from Kenya. Emerg Infect Dis 15, 482-485 (2009). 16. W. H. Li et al., Angiotensin-converting enzyme 2 is a functional receptor for the SAR$ coronavirus. Nature 426, 450-454 (2003). 17. X. Y. Ge et al., Isolation and characterization of a bat SARS-like coronavirus that uses the ACE2 receptor. Nature 503, 535-538 (2013). 18. V. D. Menachery et al., A SA RS-like cluster of circulating bat coronaviruses shows potential for human emergence. Nature Medicine 21, 1508-+ (2015). 19. N. Wang et al., Serological Evidence of Bat SARS-Related Coronavirus Infection in Humans, China. Virologica Sinica, (2018). 20. R. H. Xu et al., Epidemiologic clues to SARS origin in China. Emerging Infectious Diseases 10, 1030-1037 (2004). 21. L. Brierley, M. J. Vonhof, K. J. Oliva I, P. Daszak, K. E. Jones, Quantifying Global Drivers of Zoonotic Bat Viruses: A Process-Based Perspective. American Naturalist 187, E53-E64 (2016). Page 193 References Cited

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 22. X. L. Yang et al., Isolation and Characterization of a Novel Bat Coronavirus Closely Related to the Direct Progenitor of Severe Acute Respiratory Syndrome Coronavirus. Journal of Virology 90, 3253-3256 (2016). 23. L. P. Zeng et al., Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses. Science China-Life Sciences 60, 1399-1402 (2017). 24. L.-P. Zeng et al., Bat Severe Acute Respiratory Syndrome-Like Coronavirus WIVl Encodes an Extra Accessory Protein, ORFX, Involved in Modulation of the Host Immune Response. Journal of Virology 90, 6573-6582 (2016). 25. X. L. Yang et al., Genetically Diverse Filoviruses in Rousettus and Eonycteris spp. Bats, China, 2009 and 2015. Emerging Infectious Diseases 23, 482-486 (2017). 26. X. Y. Ge et al., Coexistence of multiple coronaviruses in several bat colonies in an abandoned mineshaft. Virologica Sinica 31, 31-40 (2016). 27. B. Hu et al., Detection of diverse novel astroviruses from small mammals in China. Journal of General Virology 95, 2442-2449 (2014). 28. B. Hu, X. Y. Ge, L. F. Wang, Z. L. Shi, Bat origin of human coronaviruses. Virology journal 12, (2015}. 29. B. Hu et al., Discovery of a rich gene pool of bat SARS-related coronaviruses provides new insights into the origin of SARS coronavirus. PLoS pathogens 13, (2017). 30. C. M. Luo et al., Discovery of Novel Bat Coronaviruses in South China That Use the Same Receptor as Middle East Respiratory Syndrome Coronavirus. Journal of Virology 92, (2018). 31. Y. Luo et al., Longitudinal Surveillance of Betacoronaviruses in Fruit Bats in Yunnan Province, China During 2009-2016. Virologica Sinica 33, 87-95 (2018). 32. M. N. Wang et al., Longitudinal surveillance of SARS-like coronaviruses in bats by quantitative real-time PCR. Virologica Sinica 31, 78-80 (2016). 33. Z. Wu et al., Comparative analysis of rodent and small mammal viromes to better understand the wildlife origin of emerging infectious diseases. Microbiome 6, 178 (2018). 34. K. J. Olival et al., Host and viral traits predict zoonotic spillover from mammals. Nature 546, 646-650 (2017}. 35. J. N. Mandi et al., Reservoir Host Immune Responses to Emerging Zoonotic Viruses. Ce// 160, 20-35 (2015). 36. S. J. Anthony et al., A strategy to estimate unknown viral diversity in mammals. mBio 4, e00598-00513 (2013). 37. S. J. Anthony et al., Non-random patterns in viral diversity. Nature Communications 6: Article number 8147, (2015}. 38. B. Yount et al., Reverse genetics with a full-length infectious cDNA of severe acute respiratory syndrome coronavirus. Proceedings of the National Academy of Sciences of the United States of America 100, 12995-13000 (2003}. 39. Z. Lei-Ping et al., Cross-neutralization of SARS coronavirus-specific antibodies against bat SARS-like coronaviruses. SCIENCE CHINA Life Sciences 60, 1399 (2017). 40. V. D. Menachery, R. L. Graham, R. S. Barie, Jumping species-a mechanism for coronavirus persistence and survival. Current opinion in virology 23, 1-7 (2017). 41. L. Gong et al., A New Bat-HKU2-like Coronavirus in Swine, China, 2017. Emerging Infectious Diseases 23, 1607-1609 (2017). 42. Y. F. Pan et al., Discovery of a novel swine enteric alphacoronavirus (SeACoV) in southern China. Veterinary Microbiology 211, 15-21 (2017). 43. S. K. P. Lau et al., Recent transmission of a novel alphacoronavirus, bat coronavirus HKU10, from Leschenault's rousettes to pomona leaf-nosed bats: first evidence of interspecies transmission of coronavirus between bats of different suborders. Journal of virology 86, 11906-11918 (2012). 2 Page 194 References Cited

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 44. J. H. Luo et al., Bat conservation in China: should protection of subterranean habitats be a priority? Oryx 47, 526-531 (2013). 45. S. Watanabe et al., Bat coronaviruses and experimental infection of bats, the Philippines. Emerg Infect Dis 16, 1217-1223 (2010). 46. R. L. Graham, R. S. Barie, Recombination, Reservoirs, and the Modular Spike: Mechanisms of Coronavirus Cross-Species Transmission. Journal of Virology 84, 3134-3146 (2010). 47. D. Carroll et al., The Global Virome Project. Science 359, 872-874 {2018). 48. M. Rani et al., Increased antibody affinity confers broad in vitro protection against escape mutants of severe acute respiratory syndrome coronavirus. J Virol 86, 9113-9121 {2012). 49. B. Rockx et al., Escape from human monoclonal antibody neutralization affects in vitro and in vivo fitness of Severe Acute Respiratory Syndrome Coronavirus. Journal of Infectious Diseases 201, 946-955 (2010). SO. L. Wang et al., Importance of neutralizing monoclonal antibodies targeting multiple antigenic sites on MERS-CoV Spike to avoid neutralization escape. Journal of virology 92, e02002-02017 (2018). 51. J. Pallesen et al., lmmunogenicity and structures of a rationally designed prefusion MERS-CoV spike antigen. Proceedings of the National Academy of Sciences 114, E7348-E735 7 (2017). 52. J. F. Drexler et al., Genomic characterization of severe acute respiratory syndrome-related coronavirus in European bats and classification of coronaviruses based on partial RNA-dependent RNA polymerase gene sequences. J Virol 84, 11336-11349 (2010). 53. K. H. Chan et al., Detection of SARS coronavirus in patients with suspected SARS. Emerging infectious diseases 10, 294-299 (2004). 54. P. R. Hsueh, L. M. Huang, P. J. Chen, C. L. Kao, P. C. Yang, Chronological evolution of lgM, lgA, lgG and neutralisation antibodies after infection with SARS-associated coronavirus. Clinical Microbiology and Infection 10, 1062-1066 (2004). 55. V. D. Menachery et al., SA RS-like WIVl-CoV poised for human emergence. Proceedings of the National Academy of Sciences of the United States of America 113, 3048-3053 (2016). 56. T. P. Sheahan et al., Broad-spectrum antiviral GS-5734 inhibits both epidemic and zoonotic coronaviruses. Science translational medicine 9, eaal3653 (2017). 57. A. C. Sims et al., Severe acute respiratory syndrome coronavirus infection of human ciliated airway epithelia: role of ciliated cells in viral spread in the conducting airways of the lungs. Journal of virology 79, 15511-15524 (2005). 58. T. Scobey et al., Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus. Proceedings of the National Academy of Sciences of the United States of America 110, 16157-16162 (2013). 59. S. Agnihothram et al., Development of a Broadly Accessible Venezuelan Equine Encephalitis Virus Replicon Particle Vaccine Platform. Journal of virology 92, e00027-00018 (2018). 60. S. Agnihothram et al., A mouse model for Betacoronavirus subgroup 2c using a bat coronavirus strain HKUS variant. MBio 5, e00047-00014 (2014). 61. D. Deming et al., Vaccine efficacy in senescent mice challenged with recombinant SARS-CoV bearing epidemic and zoonotic spike variants. PLoS medicine 3, e525-e525 (2006). 62. J. M. Fonville et al., Antibody landscapes after influenza virus infection or vaccination. Science (New York, N. Y.) 346, 996-1000 (2014). 63. K. Debbink et al., Within-host evolution results in antigenically distinct Gll.4 noroviruses. J Virol 88, 7244-7255 (2014). 64. L. C. Lindesmith et al., Broad blockade antibody responses in human volunteers after immunization with a multivalent norovirus VLP candidate vaccine: immunological analyses from a phase I clinical trial. PLoS Med 12, e1001807 (2015). 3 Page 195 References Cited

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter 65. J. A. Swanstrom et al., Dengue Virus Envelope Dimer Epitope Monoclonal Antibodies Isolated from Dengue Patients Are Protective against Zika Virus. mBio 1, e01123-01116 (2016). 66. X.-C. Tang et al., Identification of human neutralizing antibodies against MERS-CoV and their role in virus adaptive evolution. Proceedings of the National Academy of Sciences of the United States of America 111, E2018-E2026 (2014). 67. P. Daszak et al., Interdisciplinary approaches to understanding disease emergence: The past, present, and future drivers of Nipah virus emergence. Proceedings of the National Academy of Sciences, (2012). 68. K. E. Jones et al., Global trends in emerging infectious diseases. Nature 451, 990-993 (2008). 69. T. Allen et al., Global hotspots and correlates of emerging zoonotic diseases. Nature Communications 8, 1124 (2017). 4 Page 196 References Cited

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter CONSORTIUM/CONTRACTUAL ARRANGEMENTS: This project is a multi-institutional collaboration led by EcoHealth Alliance, New York (Daszak, Pl), which will subcontract funds to three institutions: the Wuhan Institute of Virology (Dr. Shi), the University of North Carolina at Chapel Hill (Dr. Barie), and the Institute of Pathogen Biology (Dr. Ren). In addition, Dr. Linfa Wang from Duke-National University of Singapore (Duke NUS) will act as a senior consultant with no requested subcontract funds, and will primarily advise on the serological and molecular based diagnostic platforms. Dr. Daszak has over 15 years previous experience managing collaborative projects including two R01s on Nipah virus ecology and the current R01 on Coronavirus (Al110964) that involve multiple, separate foreign institutions; a 5-year NSF/NIH Ecology of Infectious Disease award on West Nile virus which involved multiple subcontracts, a NIAID R01 on bat viral discovery that involved multiple international contracts, and a multi-million dollar per year contract from USAID that involves 21 international partners. The applicant organization (Eco Health Alliance) is justified in taking the lead on this project because it specializes in understanding the ecological, and virological processes underlying zoonotic disease emergence. Dr. Daszak has conducted significant preliminary work on this issue including 15-years of research on the ecological and related factors of the emergence of SARS and 15- years of work in China. The subcontract institutions will work on specific issues and areas in which they have proven expertise. These areas are: • Human community surveillance, human clinical or hospital syndromic surveillance, CoV serology, full genome sequencing, epidemiology, and behavioral risk (Institute of Pathogen Biology, Dr. Ren) • CoV screening and serology of non-human samples, viral pathogenesis, serological testing protocol development, host receptor binding, S protein sequencing, in vitro and in vivo virus characterization (Wuhan Institute of Virology, Dr. Shi) • Small animal models of viral pathogenesis, primary human cell cultures, CoV reverse genetics, reconstruction of zoonotic CoV (University of North Carolina at Chapel Hill, Dr. Barie). Dr. Daszak has had inter-institutional contractual agreements with the Wuhan Institute of Virology for over 13 years. Ors. Shi and Daszak have collaborated together since 2002 and have been involved in running joint conferences, collaborating on papers, and shipping samples into and out of China. Ors. Barie, Shi, Wang, and Daszak have collaborated closely for over 10 years on Coronavirus and other emerging disease research. Ors. Shi and Ren have collaborated on viral discovery projects 8 years. Page 197 Consortium/Contractual Arrangements

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Contact PD/Pl: DASZAK, PETER WUHAN INSTITUTE OF VIROLOGY THE CHINESE ACADEMY OF SCIENCES Address:Xiaohongshan 44, Wuchang, Wuhan 430071, Hubei, P .A.China Tel:+86-27-87199162 Fax: +86-27-87199162 http://www.whiov.ac.cn Dr. Peter Daszak President Eco Health Alliance 460 W 34th St. 17th Floor NewYork,NY10001 USA Dear Dr. Daszak, 31 October 2018 The Wuhan Institute of Virology, Chinese Academy of Sciences, has an high interest in working with Eco Health Alliance and its scientists to identify and prevent the transmission of bat coronaviruses to human populations globally. In particular, the NIAID funded R01 renewal proposal entitled "Understanding the risk of bat coronavirus emergence" will provide an excellent opportunity to achieve these goals. The Wuhan Institute of Virology, Chinese Academy of Sciences, recognizes the mutual benefits to be gained through research cooperation and a successful partnership with EcoHealth Alliance in the field of identification and prevention of zoonotic disease transmission. It is vital to not only identify the diseases themselves, but also identify high-risk human populations and the actions that put them at risk for infection along with evaluating approaches to intervention and disease management. Understanding and preventing exposure and transmission of zoonotic diseases from wildlife to humans remains a high priority for prevention of pandemics. In our discussion with EcoHealth Alliance, we have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to epidemic disease outbreaks - particularly those of animal origin. To assist in this study, we will provide participating laboratories in China with human samples both new and archived and support research in bat coronaviruses. We at Wuhan Institute of Virology, Chinese Academy of Sciences, look forward to our continued collaborations with the EcoHealth Alliance team and working further on this worthwhile study. Sincerely, Dr.Y Dire Chin Xiao Wuh Letters of Support Page 198

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Contact PD/Pl: DASZAK, PETER Dr. Peter Daszak President EcoHealth Alliance 460 W 34th St 17th Floor New York, NY 10001 USA Dear Dr. Daszak, 01 November2018 The Institute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College (!PB, CAMS&PUMC) has an high interest in working with EcoHealth Alliance and its scientists to identify and prevent the transmission of bat corona viruses to human populations globally. In particular, the NIAID funded ROl proposal entitled "Understanding the risk of bat coronavirus emergence" will provide an excellent opportunity to achieve these goals. The IPB, CAMS&PUMC recognizes the mutual benefits to be gained through research cooperation and a successful partnership with EcoHealth Alliance and long term colleague Dr. Zhang Shu-Yi in the field of identification and prevention of zoonotic disease transmission. It is vital to not only identify the diseases themselves, but also identify high-risk human populations and the actions that put them at risk for infection along with evaluating approaches to intervention and disease management. Understanding and preventing exposure and transmission of zoonotic diseases from wildlife to humans remains a high priority for prevention of pandemics. In our discussion with EcoHealth Alliance, we have agreed to participate in activities that will strengthen the ability of China and other countries in the region to respond to epidemic disease outbreaks - particularly those of animal origin. To assist in this study, we will provide participating laboratories in China with human samples both new and archived and support research in bat coronaviruses. We at IPB, CAMS&PUMC look forward to our continued collaborations with the EcoHealth Alliance team and working further on this worthwhile study. Sincerely, !.\.I' Lili Ren 1/. \r·, • I~~ 1 ;;t Co-Investigator I-'.>,._ ff> 1 • • Institute of Pathogen Biology,'~· ~ J,\~ Chinese Academy of Medic 'rr Sc-iences & Pel<ing UniQn Medical College No.9 Dong Dan San Tiao, Do -g:Cheng pistr.ict '-~-,. Beijing 100730 P.R.China :-;-i., ,..:i ;I •. _ 7, ~( E-mail (b)(6) 11iJ lhJ°:rY. f ~ Ji.171 Page 199 Letters of Support

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Contact PD/Pl: DASZAK, PETER 1,11.l.l:'-.l,', 'l,\ H,)•JL ill· \;t,{lllAI P{:!¼i 1( llI Al.Tl! October 24, 2018 Peter Daszak, PhD EcoHealth Alliance 460 West 34th Street - 17th floor New York, NY 10001 "I HE UNIV!iRSITY of NORTH CAROLINA '11 CHAPEi.. HILL Reference: Program Announcement number PA-18-484, entitled NIH Research Project Grant (Parent R01 Clinical Trial Not Allowed), dated December 6, 2017 Dear Dr. Daszak, This letter confirms that the appropriate program and administrative personnel at The University of North Carolina at Chapel Hill (UNC-CH) have reviewed the above referenced program announcement and are committed to enter into a subcontract with the EcoHealth Alliance for the performance period of July 1, 2019 to June 30, 2024. The work to be performed by UNC-CH does not include animal and/or human research subjects. UNC-CH maintains an active and enforced conflict of interest policy meeting the requirements of 42 CFR Part 50, Subpart F and 45CFR Part 94. EcoHealth Alliance's Principal Investigator on this proposal is Dr. Peter Daszak. The UNC-CH budget, budget justification and scope of work are provided as separate enclosures to this letter. The estimated cost of the proposed subcontract will not exceed $388,750 and includes appropriate direct and indirect costs. Furthermore, by submission of this commitment letter UNC-CH and its Principal Investigator (Pl) certify (1) that the information submitted within the application is true, complete and accurate to the best of the UNC-CH's and Pl's knowledge; (2) that any false, fictitious, or fraudulent statements or claims may subject the UNC-CH and Pl to criminal, civil, or administrative penalties; and (3) that the Pl agrees to accept responsibility for the scientific conduct of the project and to provide the required progress reports if an award is made as a result of UNC-CH's application. If you have any questions, please contact the undersigned at 919-966-3895. Sincerely, ~Te(f£~rc: Vice Chancellor for Research Enclosed: Budget Budget Justification Scope of Work Letters of Support Ralph S Barie, PhD Professor, Epidemiology, Microbiology and Immunology Page 200

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Contact PD/Pl: DASZAK, PETER ~DukeNUS ~ Medical School 31 Oct, 2018 Dr. Peter Daszak EcoHealth Alliance 460 West 34th Street, Suite 1701 New York, NY 10001, USA Dear Peter, I am writing this letter in strong support of the proposed renewal of the NIH (R0l All 10964, Understanding the Risk of Bat Coronavirus Emergence) project led by EcoHealth Alliance. As you know, I have long experience with EIDs associated with bats, having worked with Hendra virus in Australia, Nipah virus in Malaysia, Singapore and Bangladesh, SARS related viruses in China, Reston ebolavirus in the Philippines. More recently, in collaboration with scientists in EcoHealth and China, I played an important leadership role in coordinating and directing the research which discovered abat HUK2-related coronavirus as the causative agent of a major swine acute diarrhea syndrome (SADS) outbreak in Southern China. Your current proposal complements and substantially expands this approach in promising novel and more powerful tools to understand how host immune dynamics and heterogeneity in immune response affect the timing, location, and severity of disease outbreaks in wildlife, and risk of spillover from wildlife to human populations. I very much looking forward to participating in this initiative and should EcoHealth Alliance be successful in its application for renewal, I agree to participate in activities associated with this project, including contributing expertise, helping identify, locate and interpret relevant data, and participating in partner meetings. This letter conveys my strong interest and commitment to making this initiative a success. I am excited to be part of the initiative to develop tools to better interrogate seroprevalence data and better predict zoonotic disease emergence. I'm confident that the approach will improve our understanding of the dynamics of infectious diseases and have wide application in public health, and I look forward to working with EcoHealth Alliance on this project Yours sincerely, Lin/a (Lin-Fa} WANG, PhD FTSE Professor & Director, Programme in Emerging Infectious Diseases 8 College Road, Singapore 169857 T 6516 7666 F 6221 7396 www.duke-nus.edu.sg A school of the National University of Singapore (RCB No: 200604346E) Letters of Support Page 201

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter RESOURCE SHARING PLAN Data Sharing Plan: Data will be available to the public and researchers without cost or registration, and be released under a CC0 license, as soon as related publications are in press. Data will be deposited for in long-term public scientific repositories - all sequence data will be made publicly available via GenBank, species location data via the Knowledge Network for Biodiversity, and other data will be deposited in appropriate topic-specific or general repositories. Computer code for modeling and statistical analysis will be made available on a code-hosting site (GitHub), and archived in the Zenodo repository under an open-source, unrestrictive MIT license. Limited human survey and clinical data will be released following anonymization and aggregation per IRB requirements. Publications will be released as open-access via deposition to PubMed commons. Viral isolates will remain at the Wuhan Institute of Virology initially. Isolates, reagents and any other products, should they be developed, will be made available to other NIH-funded researchers via applicable Wuhan Institute of Virology and EcoHealth Alliance Material Transfer Agreements and/or licensing agreements. Sharing Model Organisms: We do not anticipate the development of any model organisms from this study. Should any be developed, they will be made available to other NIH-funded researchers via applicable Wuhan Institute of Virology and EcoHealth Alliance Material Transfer Agreements and/or licensing agreements. Genomic Data Sharing: We anticipate obtaining genetic sequence data for 1 00s of novel coronavirus genotypes, including RNA-dependent RNA polymerase (RdRp) and Spike genes for all strains/genotypes. In addition, we will generate full viral genomes for a subset of the bat SARSr-CoVs that we identify. All sequence data will be deposited in the NIH genetic sequence database, GenBank. We will ensure that all meta-data associated with these sequences, including collection locality laUlong, species-level host identification, date of collection, and sequencing protocols will also submitted. The genotype data will be made publicly available no later than the date of initial publication or six months after the receipt of final sequencing data, whichever comes first. We anticipate sequence generation will occur over the 5 year proposed project period. Genome Wide Association Studies (GWAS): Not applicable. Page 202 Resource Sharing Plan(s)

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter Rigor and Authentication of Key Biological and/or Chemical Resources Our project aims to understand the risk of bat SARS related coronavirus (SARSr-CoV) disease emergence in people, and will use some non-standard biological and chemical resources that require validation and authentication. We will construct chimeric SARSr-CoVs using a WIV1 backbone and the S genes of selected SARSr-CoV strains, and assess capacity to infect hACE2, bACE2 and cACE2 Vero cells, Hela cells, primary human airway epithelial cells, and potentially CaCo cells for HKU3r-CoVs (which have not yet been cultured in human cell lines and may use intestinal epithelium in nature). We will then conduct experimental infections in hACE2 transgenic mice to assess pathogenicity and clinical signs. Each of these methods have been previously validated and published by our collaborative research team, as highlighted in the research proposal. Each laboratory-based research partner in our project (including University of North Carolina (UNC), Wuhan Institute of Virology (WIV), Institute of Pathogen Biology (IPB) at Chinese Academy of Medical Sciences, and Duke National University of Singapore (DukeNUS)) each have specialized strategies to oversee the authentication of key biological resources, reagents and chemical resources. EcoHealth Alliance will actively engage with each partner to ensure that the highest quality science, public accountability, and social responsibility in the conduct of science are maintained throughout. The overall goal is to ensure that the underlying scientific foundation of the project from conception to completion is scientifically sound. To ensure scientific rigor (e.g., determining group sizes, analyzing anticipated results, reducing bias, ensuring independent and blinded measurements, improving precision and reducing variability including or excluding research subjects, and managing missing data), EcoHealth Alliance will review scientific approaches and outcomes throughout the duration of the award. We will ensure that experimental designs will include considerations of sex as a "Relevant Biological Variables" in all studies involving human subjects or vertebrate animals. Unless otherwise specified and justified, all experiments will include male and females. UNC and WIV will implemented an "audit trail" that tracks animals used in experimental investigations (Aim 3) from parents, through birth, shipment, experimentation, results, QC and analyses, providing outside researchers the ability to track experiments from conception through publication. Cells. Early passage primary human airway epithelial cells are a key reagent for the proposed studies. Human lung cells are derived from donors of both sexes and from all ages and ethnic groups. Care is taken during cell isolation to only handle one human organ at a time. Similarly, primary cell populations are handled carefully, only one donor cell type from a single donor at a time to avoid any mixing. The cells are observed to exhibit well-described prototypical characteristics of human primary lung cells in cell type specific medias in culture. For quality control, the cells are cultured in antibiotic free media to test for bacterial and fungal microbial contamination and are subjected to mycoplasma testing. Once the epithelial cells are grown as polarized and differentiated monolayers, a representative sample is subject to quality control histological analysis of cell morphology and Short Terminal Repeat (STR) marker profiling by the UNC Lineberger Cancer Center's Tissue Culture Facility (TCF). Routine evaluations for mycoplasma contamination are routinely performed in the laboratory. • Certain experiments also employ immortal cell lines. Cell lines are obtained from the ATCC, or from the TCF. The TCF maintains cell lines, utilizing STR marker profiling and records of authentication are available. New cell lines not available directly from the TCF can be authenticated through the STR marker service provided by the TCF. Cell lines are routinely evaluated for mycoplasma contamination. • When receiving cell lines, lab members initially maintain isolation and keep them isolated from other authenticated cell lines until mycoplasma testing and STR marker profiling is performed. All cell lines must be authenticated before commencing experimental work with them. • Records are maintained for each of the cell lines regarding 1) the origin of the cell lines; 2) when they were resuscitated; 3) number of passages; 4) all test results; 5) any unique distinguishing growth behavior; and 6) any known genetic features. • Cells that have been passaged for 6 months after receipt or from resuscitation will be re-authenticated, or a new vial of the working stock will be thawed. • Lab members routinely examine cultured cell morphology by phase microscopy and monitor the growth characteristics in culture. New vials of the working stock are thawed if deviation from the baseline is Page 203 Authentication of Key Biological and/or Chemical Resources

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Contact PD/Pl: DASZAK, PETER Program Director/Principal Investigator: Daszak, Peter observed. • Mycoplasma contamination is re-checked whenever cells are extensively passaged to create new stocks. Animals (Mice) • Rodent genotyping for mouse strain genetic validation. Inbred mouse strains are an invaluable tool for biomedical research, and hACE2 transgenic mice represent a key aspect of Aim 3 to assess pathogenicity and clinical signs for SARSr-CoVs. To ensure that the genetic background of all mice used within this program is known and when applicable they are part of a known inbred strains, we will genotype each mouse strain used within this program on the appropriate MUGA platform (Morgan, AP et.al., G3 2016, Dec 18). The most recent iteration of this state of the art genotyping array contains over 140,000 markers and can be used to precisely determine the genetic background at the substrain level and the precise location (at <1 megabase resolution) of genomic regions derived from different mouse inbred strains. In this way, the identity and genomic integrity of all mice used within these studies will be ensured. As new diagnostic assays become available, we will assess their utility and cost effectiveness the different MUGA arrays and implement them as appropriate. • Furthermore, for each mutant mouse strain used within the project, positive diagnoses of the mutation will be assessed for each cohort of experimental animals with a diagnostic validated PCR assay or Sanger sequencing diagnostic to ensure proper results. Recombinant and Wildtype Viruses and Mutant Derivatives. • Recombinant and wildtype viruses contain unique marker mutations that allow for distinguishing strains and mutation profiles, using a combination of full genome sequencing, reverse transcription-polymerase chain reaction (RT-PCR) or RT-PCR restriction fragment length polymorphism analyses (RT-PCR RFLP). Our group has developed defined primer pairs to distinguish between SARS-CoV and SARS-related bat coronaviruses as well as MERS-CoV and MERS-related bat coronaviruses. All viruses will be validated and certified pure of contaminating viruses prior to use or shipment to other laboratories. 2 Page 204 Authentication of Key Biological and/or Chemical Resources

Understanding the Risk of Bat Coronavirus Emergence

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