PARTNER PROJECT AGREEMENT #G-2101p
BETWEEN
‘THE INTERNATIONAL SCIENCE AND TECHNOLOGY CENTER,
‘THE NATIONAL CENTER FOR DISEASE CONTROL AND PUBLIC HEALTH.
AND
US Department of Health & Human Services / US Centers for Discase Control and
Prevention
Emerging zoonotic pathogens ia Georgian batsArtiste 16- Annexes
[As specified in Article 1.2, the Annexes are an integral par of the Agreement. They are:
Annex Work Plan
‘Annex Il General Conditions
‘Annex Ill Formats for Progress and Cost Reports
‘Amex IV Disclaimer
Article 17- Entry i
(o Force ofthe Agreement
‘The Agreement shall enter into force on February 1,2014.
Prepared in Moscow in the English language
For the Center| For the Recipient For the Parner
y >
Fr
c ye
David Cleave ‘Amiran Gamkelidee Eduardo Gomez
Acting Executive Director General Director Biosecurity Coordinator
iste [National Center for Disease Control US Department of Healthc
and Publie Health Human Services/US Centers for
DNA A, Od. d00Y
(ate) (date) (date)ANNEX I
Work Plan
1. Summary Projeet Information
1. Project Title
Emerging zoonotic pathogens in Georgian bats
2. Project Manager
Name: Lela Urshadze
Tie: S Positions _Viologit, Senor special
‘Streetaddress 9 MAvaiani a
Giys Toil Region: Tol
wars O17 Country: Georgia
Tels so9ssonnasisa Fax
E-mail llocde@gnaitcom
Lead Istntion
‘Short references NDC
Fell name: __ National Center fr Disease Convol and Pulic Health of Georgia (NCDCEPH)
Street adress 9 MA
‘Gye Toil ‘Regions Toil
wip: 0177 Country: Georgia
Name of Signature Authority: Arian Gamireidae
MD, PHD Positions _Oiretor Gener
99532251 1758 Far: 999552251 1755
agambasidzeaincdszs
Goveramental Agency: Ministry of eath
3.2. Other Participating Institutions
NoneSs
4. Foreign CollboratorsPartners
4. Colaboraors
‘wstttion: Center fo Disease Control and Prevention
‘iret address: 1600 Clifton RANE ie
Gigs Atlenata Region/State: Georgia
Zar: 30080 ‘Country: USA
Person: Dr Andres Velasco Vilaa
Titer Microbiologie Position: Lead, Genomics and Diagnostics Development
Unig Rabies Team, Poxvrus and Rabies Branch
Tes 404-639-1055, Fax: 404-639-1060
E-mail: AVelascoVilla I @ede gov
42, Parners
Institutions U.S. Centers for Disco Conrol and Proveton
‘erect address! 1600 Clifton Road NE, Maisop AS
‘Gly: Alles RegiowState: GA
Zar: 30329 CCounary: USA
Signatwre Authoriy: — Edvardo Gomez
Titles PAD Position; Biosecurity Coordinate
Tels > 1000) 659.7356 Fer
i909 235 0295
Emaik _ duligedegov
Project Coordinator: David all
Title PAD Position: BTEP Program Manager
Ta: = 1608) 69-7356 Rex: _ +1408) 235 0293
Email doul@edegov
5 Project Duration
18 mond
6. Project Location aed tauipment
Institution Location, Facies ad Equipment
[Lending tstittion [Tels Gera, National Cntr for Disease Conrol and Publi Heats
(CPHRL. building BSL3, BSL? FaciesSs
jad verical efoctrophoress equipment
lce1 documentation and analysis system, Genetic Analysis System CEQ-8000 Beckma
lcoulier, Sequencer - ABI 3130XL; WGS - Illumina MiSeq pluform; PCI
[Workstations
LL Specific information
1. Introduction and Overview
‘Bats as Important Reservoirs of Zoonotic Diseases
Bats are increasingly recognized as reservoirs of emerging zoonatic pathogens. The importance of bats as &
reservoir of infectious viral agents potentially transmissible to humans and other animals has become more
evident with the passage of time. For example, West Caucasian bat virus (WCBV) was isolted from an
insectivorous bat (Minioptorus schretberst) in the Russian part of the Great Caucasian Ridge ~300 kim away
From the Georgian border, This virus, and related viruses Tkoma and Lleida Viruses, is curently the mast
divergent member of Lyssavirus genus. Commercially avilable rabies biologics do not provide protection
against WCBV. The virus is pathogenic for laboratory animals (soch as mice, and hamsters), but it
Significance for publi health is unknown, uiven tt rabies diagnosis in Georgia, similar to ote counties of
Easter Europe, is typically clinical inthe majority of cases, and is not supplemented by virus idutiication.
‘The National Centers for Disease Control snd Publi Heath (NCDC&PH) in collocation with the U.S. CDC
conducted hat surveillance in Five regions of Georgia during lune, 2012. Eight species of bats were identified
mong captured 236 bts in toa. Samples from those bas are stored at NCDC&PH and in September 2013 will
be examined by direct Muorescent antigen (DFA) testing 10 detect Lyssaviuses. During the survey
approximately 40% of collected bats were Rhinolophus spp.. Accordingly, there is the possibility of existence
newly emergent CoV in Georgian bas
‘The data obiained from the above mentioned survey triggered initiation of this project. The objectives of the
project willbe: characterization of DFA postive Lyssaviruses by molecular methodologies; study prevalence of
‘coronaviruses in Georgian bats; comparison of human and bat Lyssaviruses.
Lyssa viruses
Rabies virus is genus of ssavirus, and is listed as « priority pathogen by the Georgian ministries of Health
and Agriculture. Even with Iinitations of the existing national surveillance system, 40-100 animal eases, and 6-
12 human cases of rabies are documented in the country every year. No revent rabies virus (RABV) isolates
have been characterized thoroughly atthe molecular of antigen Level, and information on their circulation
Patterns, host ranges and distribution is lacking.
In frame of the project it is plumed to cary out the following: PCR confirmation and sequencing of
Lyssaviuses fiom DFA positive samples; collection of fecal material and rectal swabs fiom bats for further
coronavirus investigation by molecular methodologies: comparison of sequence data of Lyssaviuses from
humo and ba samples.
Coronaviruses
‘Among the diferent viral families hosted by the bats, much attention has been pad tothe coronaviruses whichGs
In view of this, there have boen numerous studies in recent years with the goal of finding new species of
‘coronavinses in bal in order to monitor the word situation, to study the possible origin ofhuman SARS virus,
and to predict posible new coronavirus outbreaks in humans.
(On dhe other hand, the local surveillance of the emerging coronsvirus will prepare the Cauesss region forthe
possible outbreak of the SARS-related CoV and a novel beta (P)-CoV which eaus fatal respiratory diseases
‘The 2002 severe acute respiratory syndrome (SARS) epidemic in China resulted in 8,422 laboatory-confirmed
suman cases with 916 faites, The causative agent was a nevly emergent coronavirus (CoV) that may have
originated from a similar vis, SARS-rclated CoV, discovered in Chinese horseshoe bats (Rhinalophus spp.)
‘The recent implication of a novel beta (B)-CoV as tho cause of fatal respiratory disease in dhe Middle East,
emphasizes the importanes of surveillance for CoVs (MERS-COy) that may potentially jump species from bats
into humans. During the first bat survey Georgia ~40% of collected bas were Rhinolophus spp. accordingly,
‘hore exists tho possibilty of indontifying newly emergent CoV in Georgian bats. These date will serve as
preliminary data to support second survey consisting ofthe collection of fecal material and rectal swabs from
bats for further coronavirus investigation by molecular methodologies. The potential risk of a new group of
bat coronaviruses a a reservoir for human infections is suspected.
Benefits of Work
Inmplementation ofthis study will hep in the future to identify the infection source and pathogens transmission
path ffom the hats to humans in compliance with different epidemiological tools Under the project diagnostic
infrastructure is established that wil lad tothe early detection and prediction of emerging and reemering
isease in Georgia
‘The accomplishment ofthe project wll provide Georgian researches involved In the project withthe possibility
jence and establish international connections for performing fundamental, as well as applied
‘studies within the framework oF international seience and technology programs in the field of biology and
medicine,
‘The proposed project infends to research the emerging infectious diseases, specifically Iyssavius and
coronavirus, and knowledge of the antigenic and genetic propetis of Iyssaviruses will be important for
selecting futher vaccine candidates
In addition, although the significance of bats as resrvoits of zoonotic diseases in the Caucasus region is
unknown, we do know that West Caucasian bat vcuseireultes in Miniopreras schrlbers! bats inthe area, The
“sume bat species was recently implicated a reservoir of a nove floviras in Spain Aer sscond bet sampling.
“which willbe provide under this projet, will be collected different tissues frm the bats, fo Farther harvesting
for several emerging pathogens. By inferences from other parts of Europe and world, bats from Caucasus are
‘exported to maintain circulation of important zoonotic.
2. Expected Results and Their Application
The proposed study will be the frst investigation of Issavirus snd coronavinss conducted in Georgia.
Implementation ofthis study will help in the tare to identify the infection source and pathogens transmission
path from the bats to humans in compliance with diferent epidemiological tools
‘The project will allow detection of novel emerging and re-emerging pathogens, like coronavirus, a8 well as 8
beter understand of thee host range, pathobiology, distribution and circulation pattems, and th existing and
Potential threat for veterinary and human public health, as well as forthe bat populations, which perform an.
‘essential egological role.
Project will supply the laboratory with reagents necessry for providing reliable results that are important for
: 10 outbreaks offarther use for vaccine development and for the implementation of focal prevention and contol strategies to
prevent emerging infections diseases in humans.
'No recent lyssa and corona viruses have been investigated, characterized atthe molecular level and information
on their circulation pattems, host ranges and distribution is lacking. Improvements in laboratory capacity and
SAF training can accomplish this goal.
Phylogenetic analysis of lyssavirus strains isolated in the fame ofthe project will provide ws with findings for
understanding Iysavirus evolution
Antigenic and genetic properties of Iyssaviruses may be important for selecting futher vaccine candidates,
‘nd immunoglobulin preparations.
‘The ki of the project isto establish laboratory diagnostic inftastrucure that will lead tothe early detection
and prediction ofemerging and reemerging disease in Georgia
‘The incidence of rabies post exposure prophylaxis (PEP) in Gcorga is highest in Europe region, 876/100,000.
Expected that West Caucasian bat virus (WCB), circulate at Georgia, Commercially available eabies biologics
dd not provide protection against WCBV.
Implementation ofthis study wil help in the future 0 identify the infetion source and pathogens transmission
pat from the bas to humans in compliance with differen epidemiological tools
53. Meeting ISTC Goals and Objectives
The development of the project will provide Georgian researches that ate involved in the project with the
possbiliy to gain experience and establish international connections for performing fundamental, as well as
‘plied, studies within the framework of international science and technology programs in te fed of biology
and medicine. I also will allow the establishment and organization of prodvctiveintermational activities a the
‘National Center for Disease Control and Publie Heath.
4. Scope of Actvilee
Task 1
‘Task description and main milestones Participating Isttutions
| Bat surveillance for taking different types ofamples __[J-NCDC
‘Deseription of deliverables
| 1 Psdeting equipment and capacities for bat survey, working on the protocols ~ develop protocela based on]
fexperience of CDC, USA, Planing of survey
2 [Collecting bats ftom 2 regions of Georgia: Imereti and Kakhet and teking Wasues, for Lyssavia
jinestigation brains and blood ,for corona virus collection archival focal material and rectal swabs|
Handling ringing, and sampling, dissection by other tissues lang, kidney, liver, spleen, intestine for
|urther investigation on different vases
3 [Bat Samples arrangement and siorage for Further investigationTask 2
‘Task description and malo milestones
Strengthening of laboratory capacity for detection and]
[charactorization of Rabies etiological agents
Description of deliverables
1 Purchasing oF supplies and reagents for project needs, Development and implementation of standard
loperating protocols forthe detection and gentle characterization of etiological agents of rabies
2 [Screening of bats brain samples forthe detection of Lyssavis antigen by Direct Auoreseent antibody text
(DFA)
3 [Genetic characterization of all Lyssaviras positive samples by end point Reverse Teansripion-PCR (RT|
JPCR) coupled to nucloside sequence and phylogenetic analysis of the partial and complete N-geve|
sequences,
4 |Reconstruction ofthe evolutionary and dissemination dynamic history of recent and historical ysseviuses]
ound in his study
Task 3
“Task description and main milestones
laboratory PCR diaguostie for Corona virus ENCDC
Description of deliverables
1. [Ordering reagents for coronaviras PCR diagnostic, Implementation ofa pan-coronavius RT-PCR assay in
fats fecal and organ samples
2. [Coronavirus positive specimens by RY_PCR will be Further genetic characterization by sequencing and)
phylogenetic reconsructions.
5. [Combination of all obtained laboratory results with epidemiological data will be used to construct some
oredictive models which will be used co enhance disease surveillance stategies and beter implement
Jrevention and eontol activites
Tash~