AI surveillance in Vietnam and Laos: Report from Hokkaido University. Yoshihiro Sakoda, DVM, PhD
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1 Dec. 13, th OIE Regional Expert Group Meeting for Implementation for the program on Surveillance of Wild and domestic Birds along Migratory Flyways under the OIE/JTF Project for strengthening HPAI control in Asia AI surveillance in Vietnam and Laos: Report from Hokkaido University Yoshihiro Sakoda, DVM, PhD OIE Reference Laboratory for HPAI and LPAI Graduate School of Veterinary Medicine Hokkaido University, Sapporo, Japan
2 Characterization of AI viruses isolated from domestic birds in Vietnam Nam Dinh Dong Thap Ca Mau Bac Lieu
3 Screening test and virus isolation Sampling Egg inoculation 1-2hr Positive sample 2 days Reverse transcription loop-mediated isothermal amplification (RT-LAMP) Yoshida et al., J. Vet. Med. Sci.,
4 AI viruses isolated from domestic poultry in Vietnam Year Month Region Place Subtype of isolates (number of isolates) 2009 April Bac Lieu Farm H4N6 (7) H9N2 (18) LBM H3N2 (1) H3N8 (1) H9N2 (7) H11N3 (3) H11N9 (1) Slaughterhouse H9N2 (1) 2010 March Bac Lieu LBM H9N6 (1) 2010 October Bac Lieu LBM H6N2 (24) H6N6 (1) 2011 February Ca Mau LBM H5N1 (1) H6N2 (7) H11N5 (3) H11N9 (2) 2011 September Nam Dinh LBM H3N8 (1) H4N2 (1) H5N1 (1) H6N6 (3) 2011 October Ca Mau LBM H3N6 (1) H3N8 (7) H4N6 (4) H5N1 (15) H6N2 (39) H6N9 (2) 2012 February Dong Thap Farm H7N1 (1) H9N2 (1) H10N7 (3) LBM H4N6 (1) H5N1 (4) H6N2 (1) H7N1 (1) H9N2 (12) H11N3 (2) 2012 April Nam Dinh Farm H3N2 (4) H3N6 (3) H4N6 (4) H6N6 (1) H11N9 (1) LBM H3N2 (7) H3N6 (6) H5N1 (10) H6N6 (5) H9N2 (4) 2012 August Nam Dinh LBM H3N8 (7) H5N1 (14) H5N2 (1) H9N2 (5) H9N8 (1) 2012 October Dong Thap Farm H3N8 (2) H11N9 (3) LBM H3N8 (1) H4N6 (1) H5N1 (2) H6N2 (4) H11N6 (1)
5 AI viruses isolated from domestic poultry in Vietnam Year Month Region Place Subtype of isolates (number of isolates) 2009 April Bac Lieu Farm H4N6 (7) H9N2 (18) LBM H3N2 (1) H3N8 (1) H9N2 (7) H11N3 (3) H11N9 (1) Slaughterhouse H9N2 (1) 2010 March Bac Lieu LBM H9N6 (1) 2010 October Bac Lieu LBM H6N2 (24) H6N6 (1) 2011 February Ca Mau LBM H5N1 (1) H6N2 (7) H11N5 (3) H11N9 (2) 2011 September Nam Dinh LBM H3N8 (1) H4N2 (1) H5N1 (1) H6N6 (3) 2011 October Ca Mau LBM H3N6 (1) H3N8 (7) H4N6 (4) H5N1 (15) H6N2 (39) H6N9 (2) 2012 February Dong Thap Farm H7N1 (1) H9N2 (1) H10N7 (3) LBM H4N6 (1) H5N1 (4) H6N2 (1) H7N1 (1) H9N2 (12) H11N3 (2) 2012 April Nam Dinh Farm H3N2 (4) H3N6 (3) H4N6 (4) H6N6 (1) H11N9 (1) LBM H3N2 (7) H3N6 (6) H5N1 (10) H6N6 (5) H9N2 (4) 2012 August Nam Dinh LBM H3N8 (7) H5N1 (14) H5N2 (1) H9N2 (5) H9N8 (1) 2012 October Dong Thap Farm H3N8 (2) H11N9 (3) LBM H3N8 (1) H4N6 (1) H5N1 (2) H6N2 (4) H11N6 (1)
6 HA cleavage site of H5N1 isolates Isolates in North Vietnam R E R R R K R / G L F Isolates in South Vietnam E E R R K K R / G L F E G R R K K R / G L F Highly pathogenic avian influenza virus-like motif
7 Pathogenicity of H5 viruses in chickens Strain Intravenous pathogenicity Index (IVPI) North Dk/VN/OIE-2533/2011 (H5N1) Dk/VN/OIE-2202/2012 (H5N1) Ck/VN/OIE-2215/2012 (H5N2) South MDk/VN/OIE-3314/2011 (H5N1) MDk/VN/OIE-3509/2011 (H5N1) MDk/VN/OIE-0043/2012 (H5N1) Dk/VN/OIE-1287/2012 (H5N1)
8 Phylogenetic tree of the HA genes of H5 influenza viruses Blue : North Vietnam Red : South Vietnam Duck/Vietnam/OIE-2211/2012 Chicken/Vietnam/OIE-2215/2012 (H5N2) Duck/Vietnam/OIE-2236/2012 Duck/Vietnam/OIE-2202/2012 Duck/Vietnam/OIE-2212/2012 Environment/Chang Sha/1/2009 Whooper swan/hokkaido/4/2011 Hubei/1/2010 Duck/Vietnam/OIE-2533/2011 Duck/Vietnam/OIE-1287/2012 Oriental magpie robin/hong Kong/9298/2009 Common magpie/hong Kong/5052/2007 Wooper swan/hokkaido/2/2008 Environment/Guizhou/7/2009 Hunan/2/2009 Peregrine falcon/hong Kong/810/2009 Chicken/Vietnam/NCVD-20/2007 Vietnam/HN31388M1/2007 Environment/Guizhou/4/2009 Vietnam/HN31432M/2008 Indonesia/5/2005 Indonesia/7/2005 Chicken/Yamaguchi/7/2004 Bangladesh/207095/2008 Chicken/India/AS-NIV15983/2008 Whooper swan/mongolia/3/2005 Vietnam/1203/2004 Cambodia/R /2007 Cambodia/S / Egypt/3300-NAMRU3/2008 Chicken/Egypt/490N3-CLEVB/ Duck/Vietnam/OIE-0062/2012 Muscovy Duck/Vietnam/OIE-559/ Duck/Vietnam/OIE-3038/ Duck/Vietnam/OIE-3039/2012 Muscovy duck/vietnam/oie-0043/ Muscovy duck/vietnam/oie-3314/2011 Muscovy duck/vietnam/oie-3509/2011 Duck/Vietnam/OIE-3002/ Muscovy duck/vietnam/oie-3312/2011 Chicken/Liaoning/A-1/2007 Chicken/Shandong/A-10/2006 Chicken/Shanxi/10/2006 ChickenVietnam/NCVD-03/2008 C A B Clade
9 Phylogenetic tree of the HA genes of H5 influenza viruses Blue : North Vietnam Red : South Vietnam Duck/Vietnam/OIE-2211/2012 Chicken/Vietnam/OIE-2215/2012 (H5N2) Duck/Vietnam/OIE-2236/2012 Duck/Vietnam/OIE-2202/2012 Duck/Vietnam/OIE-2212/2012 Environment/Chang Sha/1/2009 Whooper swan/hokkaido/4/2011 Hubei/1/2010 Duck/Vietnam/OIE-2533/2011 Duck/Vietnam/OIE-1287/2012 Oriental magpie robin/hong Kong/9298/2009 Common magpie/hong Kong/5052/2007 Wooper swan/hokkaido/2/2008 Environment/Guizhou/7/2009 Hunan/2/2009 Peregrine falcon/hong Kong/810/2009 Chicken/Vietnam/NCVD-20/2007 Vietnam/HN31388M1/2007 Environment/Guizhou/4/2009 Vietnam/HN31432M/2008 Indonesia/5/2005 Indonesia/7/2005 Chicken/Yamaguchi/7/2004 Bangladesh/207095/2008 Chicken/India/AS-NIV15983/2008 Whooper swan/mongolia/3/2005 Vietnam/1203/2004 Cambodia/R /2007 Cambodia/S / Egypt/3300-NAMRU3/2008 Chicken/Egypt/490N3-CLEVB/ Duck/Vietnam/OIE-0062/2012 Muscovy Duck/Vietnam/OIE-559/ Duck/Vietnam/OIE-3038/ Duck/Vietnam/OIE-3039/2012 Muscovy duck/vietnam/oie-0043/ Muscovy duck/vietnam/oie-3314/2011 Muscovy duck/vietnam/oie-3509/2011 Duck/Vietnam/OIE-3002/ Muscovy duck/vietnam/oie-3312/2011 Chicken/Liaoning/A-1/2007 Chicken/Shandong/A-10/2006 Chicken/Shanxi/10/2006 ChickenVietnam/NCVD-03/2008 C A B Clade
10 Phylogenetic tree of the NA genes of N2 influenza viruses Blue : North Vietnam Red : South Vietnam Duck/Vietnam/OIE-3141/2011 (H6N2) Duck/Vietnam/OIE-3304/2011 (H6N2) Chicken/Korea/LPM03/2004 (H3N2) Duck/Hong Kong/Y439/17 (H9N2) Duck/Hokkaido/5/1977 (H3N2) Turkey/Massachusetts/3740/1965 (H6N2) Turkey/Wisconsin/1/1966 (H9N2) Chicken/Hebei/1/2002 (H7N2) Duck/Hong Kong/960/1980 (H6N2) Chicken/Iran/SS8/2011 (H9N2) Chicken/Dubai/463/2003 (H9N2) Quail/Hong Kong/G1/17 (H9N2) 98 Chicken/Yokohama/aq55/2001 (H9N2) Chicken/Vietnam/OIE-0056/2012 (H9N2) 98 Chicken/Hong Kong/G9/17 (H9N2) Chicken/Beijing/1/14 (H9N2) Duck/Hong Kong/Y280/17 (H9N2) Duck/Hebei/0908/2009 (H5N2) Duck/Jiangsu/m234/2012 (H5N2) Duck/Eastern China/1111/2011 (H5N2) Duck/Vietnam/OIE-1212/2012 (H3N2) 98 Duck/Hunan/491/2005 (H6N2) Duck/Hunan/573/2002 (H6N2) Duck/Eastern China/35/2006 (H3N2) Mallard/Korea/SH38-45/2010 (H13N2) Duck/Vietnam/OIE-2530/2011 (H4N2) Duck/Vietnam/OIE-2313/2009 (H9N2) Mallard/Jiangxi/8264/2004 (H6N2) Duck/Shantou/339/2000 (H6N2) Duck/Hunan/908/2005 (H6N2) Chicken/Vietnam/OIE-2215/2012 (H5N2) Chicken/Vietnam/OIE-1611/2012 (H9N2) Chicken/Tibet/LZ01/2010 (H5N2) Chicken/Hebei/1102/2010 (H5N2) Chicken/Guangdong/CYH12/2011 (H9N2) Chicken/Vietnam/OIE-2468/2012 (H9N2) Eastern China JX8264 -like ST339 -like Y439 Ck/Bei -like 0.02
11 BLAST search of each gene segment of A/chicken/Vietnam/OIE-2215/2012 (H5N2) Gene segment Region Name of strain with highest homology Homology (%) Accession No. PB A/chicken/Vietnam/OIE-1611/2012 (H9N2) AB PB A/chicken/Vietnam/OIE-1611/2012 (H9N2) 95.9 AB PA A/duck/Vietnam/OIE-2202/2012 (H5N1) AB HA Full genome A/duck/Vietnam/OIE-2202/2012 (H5N1).9 AB NP A/duck/Vietnam/OIE-2202/2012 (H5N1).9 AB NA A/chicken/Vietnam/OIE-1611/2012 (H9N2).8 AB M A/duck/Vietnam/OIE-2202/2012 (H5N1) AB NS A/chicken/Vietnam/OIE-2468/2012 (H9N2) 98.9 AB766243
12 AI viruses isolated from domestic poultry in Vietnam Year Month Region Place Subtype of isolates (number of isolates) 2009 April Bac Lieu Farm H4N6 (7) H9N2 (18) LBM H3N2 (1) H3N8 (1) H9N2 (7) H11N3 (3) H11N9 (1) Slaughterhouse H9N2 (1) 2010 March Bac Lieu LBM H9N6 (1) 2010 October Bac Lieu LBM H6N2 (24) H6N6 (1) 2011 February Ca Mau LBM H5N1 (1) H6N2 (7) H11N5 (3) H11N9 (2) 2011 September Nam Dinh LBM H3N8 (1) H4N2 (1) H5N1 (1) H6N6 (3) 2011 October Ca Mau LBM H3N6 (1) H3N8 (7) H4N6 (4) H5N1 (15) H6N2 (39) H6N9 (2) 2012 February Dong Thap Farm H7N1 (1) H9N2 (1) H10N7 (3) LBM H4N6 (1) H5N1 (4) H6N2 (1) H7N1 (1) H9N2 (12) H11N3 (2) 2012 April Nam Dinh Farm H3N2 (4) H3N6 (3) H4N6 (4) H6N6 (1) H11N9 (1) LBM H3N2 (7) H3N6 (6) H5N1 (10) H6N6 (5) H9N2 (4) 2012 August Nam Dinh LBM H3N8 (7) H5N1 (14) H5N2 (1) H9N2 (5) H9N8 (1) 2012 October Dong Thap Farm H3N8 (2) H11N9 (3) LBM H3N8 (1) H4N6 (1) H5N1 (2) H6N2 (4) H11N6 (1)
13 Phylogenetic tree of the HA genes of H9 influenza viruses Blue : North Vietnam Red : South Vietnam Chicken/Vietnam/OIE-2742/2012 Duck/Vietnam/OIE-2749/2012 (H9N8) Chicken/Vietnam/OIE-2468/2012 Swine/Guangdong/wxl/2004 Duck/Hong Kong/Y280/17 Chicken/Vietnam/OIE-1611/2012 Chicken/Anhui/WBQ/2011 Chicken/Wenshan/412/2009 Chicken/Vietnam/OIE-0056/2012 Duck/Vietnam/OIE-0061/2012 Chicken/Hong Kong/G9/17 Duck/Hong Kong/W213/17 Swine/Shandong/nc/2005 Hong Kong/2108/2003 Chicken/Israel/182/2008 Chicken/Pakistan/2/19 Hong Kong/33982/ Quail/Hong Kong/G1/17 Hong Kong/1074/17 Duck/Vietnam/OIE-2328/2009 Duck/Vietnam/OIE-2595/2009 Duck/Vietnam/OIE-2313/2009 Duck/Vietnam/OIE-2334/2010 Chicken/Korea/S22/2004 Duck/Korea/S23/2004 Swine/Korea/S452/2004 Chicken/Korea/029/19 Chicken/Korea/MS96/16 Mallard/England/ /2006 Turkey/Germany/EK224/15 Duck/Hong Kong/Y439/17 Ostrich/S.Africa/ /15 Duck/Hokkaido/294/2006 Duck/Hokkaido/W2/ Turkey/Wisconsin/1/1966 Y280 G1 Korean Eurasia North America
14 AI viruses isolated from domestic poultry in Vietnam 年月地域場所分離されたウイルスの亜型 ( 数 ) 2009 April Bac Lieu Farm H4N6 (7) H9N2 (18) LBM H3N2 (1) H3N8 (1) H9N2 (7) H11N3 (3) H11N9 (1) Slaughterhouse H9N2 (1) 2010 March Bac Lieu LBM H9N6 (1) 2010 October Bac Lieu LBM H6N2 (24) H6N6 (1) 2011 February Ca Mau LBM H5N1 (1) H6N2 (7) H11N5 (3) H11N9 (2) 2011 September Nam Dinh LBM H3N8 (1) H4N2 (1) H5N1 (1) H6N6 (3) 2011 October Ca Mau LBM H3N6 (1) H3N8 (7) H4N6 (4) H5N1 (15) H6N2 (39) H6N9 (2) 2012 February Dong Thap Farm H7N1 (1) H9N2 (1) H10N7 (3) LBM H4N6 (1) H5N1 (4) H6N2 (1) H7N1 (1) H9N2 (12) H11N3 (2) 2012 April Nam Dinh Farm H3N2 (4) H3N6 (3) H4N6 (4) H6N6 (1) H11N9 (1) LBM H3N2 (7) H3N6 (6) H5N1 (10) H6N6 (5) H9N2 (4) 2012 August Nam Dinh LBM H3N8 (7) H5N1 (14) H5N2 (1) H9N2 (5) H9N8 (1) 2012 October Dong Thap Farm H3N8 (2) H11N9 (3) LBM H3N8 (1) H4N6 (1) H5N1 (2) H6N2 (4) H11N6 (1)
15 Phylogenetic tree of the HA genes of H6 influenza viruses Blue : North Vietnam Red : South Vietnam Duck/Vietnam/OIE-527/2011 (H6N2) Duck/Vietnam/OIE-3301/2011 (H6N9) Duck/Vietnam/OIE-3225/2011 (H6N2) Duck/Vietnam/OIE-0033/2012 (H6N2) Duck/Vietnam/OIE-404/2011 (H6N2) Duck/Vietnam/OIE-3344/2011 (H6N2) Duck/Vietnam/OIE-4217/2010 (H6N2) Duck/Vietnam/OIE-4429/2010 (H6N2) Chicken/Eastern China/49/2010 (H6N6) Duck/Vietnam/OIE-2562/2011 (H6N6) Duck/Vietnam/OIE-1213/2012 (H6N6) Duck/Vietnam/OIE-3061/2012 (H6N2) Duck/Vietnam/OIE-3479/2012 (H6N2) 97 Duck/Hunan/908/2005 (H6N2) Duck/Fujian/1695/2005 (H6N2) Wild Duck/Shantou/2853/2003 (H6N2) Muscovy duck/fujian/fz01/2008 (H6N6) 57 Quail/Shantou/6825/2005 (H6N2) Teal/Hong Kong/W312/17 (H6N1) Eurasian wigeon/sweden/1/2004 (H6N2) Mallard/Minesota/2007 (H6N2) Duck/Hokkaido/7/19 (H6N2) Muscovy duck/vietnam/g33/2007 (H6N2) Duck/Hunan/573/2002 (H6N2) Duck/Hokkaido/262/2004 (H6N1) 67 Duck/Hokkaido/31/2010 (H6N2) Aquatic bird/korea/cn17/2009 (H6N2) Duck/Eastern China/1/2008 (H6N1) Duck/Hong Kong/d134/1977 (H6N2) Duck/Hong Kong/960/1980(H6N2) Gull/Tottori/105/1980 (H6N3) Duck/Hong Kong/1037-1/18 (H6N2) Duck/Shantou/339/2000 (H6N2) Chicken/Guangdong/C273/2011 (H6N2) Shearwater/Australia/1/1972 (H6N5) Turkey/Canada/1965 (H6N2) Turkey/Massachusetts/3740/1965 (H6N2) Group Ⅱ W312 Europe Group Ⅲ Early Group Ⅰ Eurasia North America
16 Summary 1. H5N1 and H5N2 HPAI viruses were isolated from apparently healthy ducks and chickens in south and north Vietnam. 2. LPAI viruses (H3, H4, H6, H7, H9, and H11 subtypes) were also isolated. 3. Genetic reassortment frequently occurred between HPAI and LPAI viruses. 4. Based on the results of this surveillance study, novel action plan should be drawn up for the control of avian influenza.
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