Swine influenza surveillance among pigs in Nan Province
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1 Swine influenza surveillance among pigs in Nan Province National Institute of Animal Health Department of Livestock Development Nan province 30 Aug 2011
2 การเฝ าระว งโรคไข หว ดใหญ ส กรในส กร จ งหว ดน าน จ งหว ด น าน 30 ส งหาคม 2554 ส จ รา ปาจร ยานนท สพ.บ. Ph.D. สถาบ นส ขภาพส ตว แห งชาต กรมปศ ส ตว
3 Outline Introduction Materials & Methods Results Gaps and needs Acknowledgements
4 Swine Influenza First isolated from pigs by Shope in 1930 Worldwide distribution High prevalence in North American and European swine farms All pathogenic SIV belong to type A influenza viruses
5 Influenza virus Hemagglutinin (HA, H: 1-16) Neuraminidase (NA, N: 1-9) Wild waterfowl serve as global reservoir because they can be infected with virus of all 16 HA and 9 NA subtypes SIV subtype defined by these two glycoproteins, the common subtype: H1N1, H3N2 and H1N2 other subtypes: H1N7, H3N1, H4N6 and H9N2
6 Influenza virus
7 Swine influenza virus Enveloped RNA virus belongs to the family Orthomyxoviridae 8 RNA segmented genomes, genetic exchange and reassortment can generate during mixed infections Infectious to human zoonotic agent Pigs as mixing vessel hosts human and avian flu can undergo genetic reassortment and cause new pandemic human influenza viruses
8 Swine influenza virus Virus attaches to respiratory tract and causes destruction of the airways of the lung, resulting in viral pneumonia
9 Influenza virus-induced pneumonia in pig
10 EM : virus particles in infected MDCK cell line Virus particles
11 Deadly virus : Spanish flu (H1N1) >500,000 dead in US, mil.ww : Asian flu (H2N2) 70,000 dead in US China->US : Hong Kong flu (H3N2) 34,000 dead in US 17: Hong Kong flu (H5N1) 19: Hong Kong flu (H9N2) 2003: Hong Kong flu (H5N1) 2003: Avian flu (H7N7) Netherlands 2003: Hong Kong flu (H9N2) Vietnam, Thailand, Cambodia (H5N1)
12 โรคไข หว ดใหญ เม กซ โก H1N Confirmed death and infections Confirmed infections Unconfirmed infections
13 Clinical signs of SIV High fever (> 104 O F) Clear nasal discharge Go off feed Become inactive, huddle, pile together Cough and/or sneeze Have difficulty breathing
14 Clinical signs of SIV Usually last 5-10 days unless complicated by secondary infections Pasteurella multocida Haemophilus parasuis Streptococcus pleuropneumoniae Mycoplasma hyopneumoniae
15 Swine influenza virus Primary pathogen causes respiratory disease or involves in PRDC The most common isolates involve in PRDC PRRS Mycoplasma hyopneumoniae SIV SIV causes an economic impact on pig production, adds 2 more weeks for fattening pigs to reach the market weight (Esterday, 2002)
16 Swine influenza in Thailand Only few reports regarding to SIV infection in Thailand Kupradinan et al. (11), SIVs were isolated from pigs with influenza-like illness in 1988, the isolates were SIV H1N1
17 H1 Phylogenetic tree California isolate Human case of classical swine H1 Thai isolates Japan isolates A/Sw/Korea/CY08/07(H1N2) A/Sw/Korea/PZ4/06(H1N2) A/Sw/Korea/Hongsong2/04(H1N2) A/Sw/Korea/S11/05(H1N2) A/Sw/MN/16419/01(H1N2) A/Sw/OH/511445/07(H1N1) A/Sw/Ohio/891/01(H1N2) A/Sw/MN/23124-S/01(H1N2) 98 A/Sw/Illinois/100085A/01 (H1N2)) A/Sw/Indiana/9K035/ (H1N2) A/Sw/Indiana/P12439/00 (H1N2) 100 A/Turkey/MO/24093/(H1N2) A/Sw/Guangxi/13/06(H1N2) A/Sw/North Carolina/93523/01 A/California/04/09(H1N1) (H1N2) A/Sw/North Carolina/98225/01(H1N2) 91 A/Sw/Minnesota/55551/00 (H1N2) A/duck/NC/91347/01(H1N2) A/Sw/CO/17871/01(H1N2) 100 A/Sw/Korea/CAS08/05(H1N1) A/Sw/Korea/CAN01/04(H1N1) A/Sw/Iowa/930/01(H1N2) A/Sw/MO/1877/01(H1N2) 97 A/Sw/Korea/CY02/02(H1N2) A/Sw/MN/34893/01(H1N2) 100 A/Sw/Wisconsin/464/98(H1N1) A/Sw/Wisconsin/136/97(H1N1) A/Sw/Wisconsin/238/97(H1N1) 100 A/Sw/Zhejiang/1/04(H1N2) A/Sw/Shanghai/1/07(H1N2) A/Sw/Guangdong/2/01(H1N1) 100 A/Sw/Shanghai/1/05(H1N1) A/Sw/MN/17138/01(H1N2) A/Sw/Wisconsin/457/98(H1N1) 100 A/Sw/Wisconsin/163/97(H1N1) A/Philippines/344/04(H1N2) A/Sw/Indiana/1726/88(H1N1) 97 A/Sw/Ratchaburi/NIAH1481/00(H1N1) A/Sw/Ratchaburi/NIAH550/03(H1N1) A/Sw/Alberta/56626/03(H1N1) A/Sw/Ontario/57561/03(H1N1) A/Sw/Taiwan/CO935/04(H1N2) A/Sw/Nebraska/1/92(H1N1) A/Sw/Ontario/11112/04(H1N1) 94 A/Sw/St-Hyacinthe/106/91(H1N1) A/Sw/Ontario/23866/04(H1N1) 100 A/Sw/Ontario/53518/03(H1N1) A/Sw/Saraburi/NIAH13021/05(H1N2) A/Sw/Chachoengsao/NIAH587/05(H1N1) A/Sw/Chonburi/NIAH589/05(H1N1) 98 A/Thailand/271/05(H1N1) 98 A/Sw/Chonburi/NIAH9469/04(H1N1) 100 A/Sw/Chonburi/NIAH977/04 (H1N1) 100 A/Sw/Miyazaki/1/06 (H1N2) 100 A/Sw/Miyagi/5/2003(H1N2) A/Sw/Saitama/96 (H1N2) 100 A/Sw/England/283902/93 (H1N1) A/Sw/England/117316/86 A/Sw/Ehime/1/80 (H1N2) A/Sw/Hokkaido/2/81 (H1N1) A/Sw/Kentucky/1/76(H1N1) A/Sw/New Jersey/11/76(H1N1) A/Sw/Tennessee/25/77(H1N1) A/Sw/Minnesota/24/75(H1N1) A/Sw/Kyoto/3/79 (H1N1) A/Sw/Niigata/1/77 (H1N1) A/Sw/Iowa/4/76(H1N1) A/Sw/Wisconsin/2/66(H1N1) 100 A/Sw/Wisconsin/1/57(H1N1) A/Sw/Wisconsin/1/61(H1N1) A/Sw/Illinois/63(H1N1) Classical Swine lineage Human lineage 0.02 Avian lineage
18 N1 Phylogenetic tree Classical Swine lineage California isolate Human case of avian like swine N1 Thai isolates Human lineage A/Sw/Cotes darmor/1488/ (H1N1) A/Sw/Ille et Vilaine/1455/ (H1N1) A/Sw/Cotes darmor/1121/00 (H1N1) A/Sw/Spain/50047/03(H1N1) A/Sw/England/195852/92 (H1N1) A/Sw/Belgium/74/85(H1N1)) A/Sw/Italy/1492/97 (H1N1) A/Sw/Italy/1513-8/98 (H1N1) A/Sw/Italy/1509-6/97 (H1N1) A/Sw/Italy/1424-4/95 (H1N1) A/Sw/Ratchaburi/NIAH550/03(H1N1) A/Sw/Ratchaburi/NIAH1481/00 (H1N1) A/Sw/Chachoengsao/NIAH587/05 (H1N1) A/Sw/Chonburi/NIAH589/05 (H1N1) A/Thailand/271/05(H1N1) A/Sw/Chonburi/NIAH9469/04 (H1 N1) A/Sw/Chonburi/NIAH977/04 (H1N1) A/California/04/09 (H1N1) A/Sw/Spain/51915/03(H1N1) A/Sw/Spain/53207/04(H1N1) A/Sw/Cotes darmor/1482/ (H1N1) A/ Sw/Cotes darmor/1515/ (H1N1) A/Sw/Virginia/670/87(H1N1) A/Sw/Virginia/671/87(H1N1) A/Sw/Lot/2979/82(H1N1) A/Sw/Belgium/1/83(H1N1) A/Sw/Finistere/3616/84(H1N1) A/turkey/France/87075/87(H1N1) Eurasian swine lineage Avian lineage 98 H5N1 viruses
19 Outline Introduction Materials & Methods Results Gaps and needs Acknowledgements
20 Collection of samples Nov Feb 2011 Nasal swabs 880 Serum 880 Farm size: small farm < 30 medium farm large farm > 300
21 Nasal swab collection Collect samples from early sick period especilly during febrile of illness Samples should be collected on polyester, not cotton swabs Swabbing the nasal passage for mucus obtained Suspend the swab in a cold transport medium and keep cold If the samples can not be process within 24 hours after collection, the samples should be storage at -70 O C
22 Transport media Keep at 4 O C: Medium + 0.5% BSA For longer time keeping, -20 O C is recommended Keep cold during tranportation Inform the responsible person in the lab before sending the samples
23
24 Diagnosis Virus isolation and identification Embryonated egg inoculation Tissue cultures: MDCK PCR & RRT-PCR (typing/subtyping) IHC for formalin-fixed paraffin-embedded tissue Specimens: nasal/tracheal/bronchial swab, lung grossly visible bronchi and bronchiole
25 Virus isolation Eggs inoculation Tissue culture (Two passages if no HA activity in the first passage)
26 Tissue culture: MDCK
27 RT-PCR
28 RT-PCR : detection and subtyping of the influenza virus M H1 H3 N1 N2 M M H1 H3 N1 N2 M PA M M = Marker NS NP 1500 bp 1150 bp 722 bp 1000 bp 500 bp 1082 bp 722 bp H1N1 H3N2
29 Real time PCR
30 Antibody detection HI (subtype specific) may have some cross-reaction ELISA: commercially available for SIV H1N1 & H3N2 and type A antibodies detection
31 Antibody detection: HI test
32
33
34 Outline Introduction Materials & Methods Results Gaps and needs Acknowledgements
35 Percentage of SIV subtype seropositive Percentage of SIV subtype seropositive by HI test All subtypes H1N1 550/46 H1N H3N2 59/47 6 H3N2 Nongwa 4 n = All subtypes H1N1 550/46 H1N H3N2 59/47 H3N2 Nongwa subtypes
36 Percentage of SI virus detection Percentage of SI virus detection 100% 90% 80% 70% 60% 50% 40% 30% VI PCR RRT-PCR 20% 10% 0% 0 VI 0 PCR 0 RRT-PCR methods
37 % seropositive Percentage of SIV seropositive among difference farm size Large Medium Small Large Medium Small Farm size
38 % seropositive Percentage of SIV seropositive among ages (18/545) (3/49) (20/66) (9/42) (21/96) (18/545) (3/49) (20/66) (9/42) (21/96) (4/18) >30 (27/64) (4/18) >30 (27/64) n = 880 months (+/total)
39 % seropositive Comparison of seropositive percentage of swine influenza between HI test and ELISA HI test ELISA All subtypes H1N1 H3N2 0.1 Subtypes
40 Outline Introduction Materials & Methods Results Gaps and needs Acknowledgements
41 Gaps and Needs Limitation of understanding of - Viral epidemiology - Transmission pigs humans - Vaccine strain selection Surveillance is crucial Lab network Sharing informations
42 Outline Introduction Materials & Methods Results Gaps and needs Acknowledgements
43 Acknowledgements Veterinarians and technicians from Nan Provincial Livestock Health Office Pig production companies Farmer from Nan Province Colleagues from Bureau of Epidemiology, MoPH US CDC Staff from NIAH
44 Thank you for your attention
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