CONTENTS. Brief introduction. Epidemiology. Diagnosis LOGO. D. Batchuluun, B. Batsuren, Ts. Badamsuren, Ts. Erdene-Ochir, J.

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1 УЛСЫН МАЛ ЭМНЭЛЭГ АРИУН ЦЭВРИЙН ТӨВ ЛАБОРАТОРИ STATE CENTRAL VETERINARY LABORATORY SCVL 26 September 2008 D. Batchuluun, B. Batsuren, Ts. Badamsuren, Ts. Erdene-Ochir, J. Bekh-Ochir CONTENTS Brief introduction Epidemiology Diagnosis

2 Brief introduction Causative agent Classification Prevalence 4 Clinical features Causative agent Influenza virus of type A, family Orthomyxoviridae Isolated first in 1956 /H7N7/ 2 subtypes 1:Equi-1-А/equine/Praga/56-H7N7 2:Equi-2-А/equine/Miami/63-H3N8 Infects all equine breeds and oddtoed ungulates /human/

3 Classification International infection bureau /OIE/ lists it in B classification diseases Prevalence Prevalent globally Clinical features Incubation period 4-10 days Running nose Fever Cough / 3-7 days /

4 Epidemiolgy 1 2 Globally In Mongolia Globally

5 In Japan, Australia Japan Kazakhstan, Kirgizstan Singapore China Mongolia Australia In Mongolia Enters through Khovd and Bayan-Ulgii aimag areas which border with PRC and Kazakhstan and reaches Sukhbaatar and Dornod. Tendency to repeat the pattern in years

6 Outbreak in 2007 In Bulgan, Uench, Myangat and Buyant soums of Khovd aimag Ulaankhus, Bulgan, Bugat and Tsengel soums of Bayan-Ulgii aimag Vaccination done Diagnosis 1 2 Diagnosis Confirmation

7 Diagnosis Nasal swab specimens 115 Serum specimens 281 Total specimens aimags Test sequencing Immunochromatography test to detect influenza of A type (Anijen-RoK) Virus culture Hemagglutination assay Hemagglutination inhibition testing (HAI) kit (A/H3/H7) Diagnose A type by reverse transcription polymerase chain reaction (RT-PCR)

8 Results Detect influenza A type by immunochromatography testing (Anijen -RoK) /Field specimen (-) Egg liquid (+)/ Virus culture Hemagglutination assay /after 4-5 passages titer was detected/ Hemagglutination inhibition testing / (HAIT) kit (A/H3/H7), H3 antibody with 1:64-1:512 titer Н7 antibody/ Reverse transcription polymerase chain reaction (RT- PCR) diagnosis of А type (+) А/H3 subtype infection Verification testing Verify diagnosis of А/H3 subtype infection For purposes of identifying subtype and origins nasal swabs specimen 10 serum specimen 10, total 20 specimens were sent for testing to Avian Influenza Reference Laboratory for Asia Pacific countries, Veterinary School, Japan s Hokkaido University

9 Results Confirmed that infection was caused by А/H3 subtype Detected influenza virus nucleotide sequencing on cleavage site HA Comparison of nucleotide sequencing for A/equine/Mongolia/1/08 (H3N8) Gene Closest origin Establishing sequencing (хос суурь/bp) Comparibility (%) PB2 A/equine/California/8560/2002(H3N8) PB1 A/equine/California/8560/2002(H3N8) PA A/equine/California/8560/2002(H3N8) HA A/equine/Bari/2005(H3N8) NP A/equine/California/8560/2002(H3N8) NA A/equine/California/8560/2002(H3N8) M A/equine/Wisconsin/1/03(H3N8) NS A/equine/California/8560/2002(H3N8)

10 Phylogenetic analysis of influenza virus of Н3 genes Phylogenetic analysis of Equine Influenza viruses in Mongolia in 2008 Eq/Mongolia/1/2008 (H3N8) Eq/Mongolia/2/2008 (H3N8) H3 HA gene ( ) Eq/Mongolia/3/2008 (H3N8) Eq/Mongolia/4/2008 (H3N8) Eq/Mongolia/5/2008 (H3N8) Eq/Mongolia/6/2008 (H3N8) Eq/Mongolia/7/2008 (H3N8) Eq/Mongolia/8/2008 (H3N3) Eq/Mongolia/9/2008 (H3N8) Eq/Ibaraki/1/2007 Eq/Kanazawa/2007 North Eq/Wisconsin/1/2003 American Eq/Kentucky/5/2002 Eq/Massachussetts/213/2003 Eq/Roma/2004 Eq/Argentina/3/1999 Eq/Argentina/4/1999 Eq/Kentucky/9/1995 Eq/Kentucky/1/1994 Eq/Kentucky/1/1998 Eq/Oklahoma/2000 Eq/La Plata/1/1993 Eq/Kentucky/1/1992 Eq/Kentucky/1277/1990 Eq/Saskatoon/1/1990 Eq/Italy/788/1991 Eq/Suffolk/1989 Eq/Yvelines/2136/1989 Eq/Berlin/3/1989 Eurasian Eq/Ibadan/6/1991 Eq/Sussex/1989 Eq/Newmarket/1993 Eq/Aboyne/1/2005 Eq/Lambourn/22778/1992 Eq/Berlin/1/1991 Eq/Hong Kong/1/1992 Eq/Avesta/1993 Eq/Algiers/1972 Eq/Tokyo/1971 Eq/Miami/1/1963 Eq/Uruguay/1/1963 Predivergent Eq/New Market/1976 Eq/NewMarket/D64/1979 Eq/Fontainebleau/1976 Eq/Romania/ Conclusion It was established that equine seasonal influenza of was caused by А H3N8 virus. Equine influenza virus isolated from Mongolia was similar to HA genes of equine influenza viruses of North America (A/equine/California/8560/2002(H3N8), A/equine/Bari/2005(H3N8) and Japan (Eq/Kanazawa/07, Eq/Ibaraki/07)

11 Acknowledgements Ministry of Food and Agriculture Veterinary Department Veterinary Division, Veterinary Laboratories of aimags and capital city Veterinary Units Biocombinate Avian Influenza Reference Laboratory for Asia Pacific Countries, Hokkaido University, Japan SCVL 2008

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