Potential Role of Exposure to Poultry Products and By-products for Human H5N1 infections
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1 Potential Role of Exposure to Poultry Products and By-products for Human HN infections David E. Swayne, A. Lipatov, Y.K. Kwon, M. Jackwood & J. Beck USDA/Agricultural Research Service Southeast Poultry Research Laboratory Athens, Georgia Introduction HPAIV systemic infection with virus in meat, bone, organs and inside eggs Natural infections - imported raw duck meat from China into S. Korea (00) and Japan (00) Experimental studies: chickens, ducks, Japanese quail, geese Viruses: Eurasian-African HN viruses, N. Amer. HN, Eurasian HN Virus demonstrated by: Virus isolation RRT-PCR IHC
2 Virus Titers in HN HPAIV Infected Meat and Eggs Meat: HN HPAIV Species Clinical Features Titer (EID0/g) Chicken Dead.-8.0 Sick.0-. Non-sick.9-. Duck Sick - Non-sick.8 Eggs: 98 HN HPAIV from USA Tumpey et al., J Virol :-, 00 Swayne & Beck, Av Dis 9:8-8, 00 Swayne, Int J Food Micro 08:8-, 00 Thomas & Swayne, J Food Prot 0:-80, 00 Das et al., Av Dis :0-8, 008 No.+/total (log0 EID0/ml) d d d d Egg Shell 0/ 0/(.) /(.) - Albumen 0/ /(.) /(.9) - Yolk 0/ 0/(.8) /(.) - Swayne & Suarez, Develop. Biol. 0:-, 008 Pasteurization to Inactivate HPAIV at Levels Present in Eggs from Infected Hens Pasteurization at industry standards for temperature and times for salmonella inactivation were effective at inactivation of HPAIV in liquid products Questions as to dried egg white moisture content was not controlled in original test Recent work with.-8% moisture in dried product indicated pasteurization was effective at industry standards ex.. days at.ºc Swayne & Beck, Av. Pathol. ():-8, 00 Swayne & Thomas, Avian Influenza, Wylie-Blackwell, pp. 9-0, 008
3 Methodology to Demonstrate Kill of HPAIV in Meat Cooking 0ºC, last detection < s treatment, calculated endpoint.s Virus Titer (Log 0 EID 0 /gram) Temperature Curve Korea/0 - Thigh Korea/0 - Breast PA/8 - Thigh PA/8 - Breast Virus Isolation Limits Temperature ( o C) Time (s) Swayne, Int. J. Food Microbiol, 8-, 00 Thomas & Swayne, J. Food Protect. 0:-80, 00 Thomas & Swayne, J. Food Protect, :-, 008 Vaccination to Prevent HPAIV in Meat Challenge with A/chicken/Korea/ES/0 (HN) wks after vaccination Virus isolation from meat (Log 0 EID 0 /gm) Group Breast Thigh Virus Dose/Bird (Log 0 EID 0 ) Fowlpox-AIV-H vaccine - A - ND B Inactivated vaccine - - ND Sham. ND.8 A - = negative on virus isolation, ND = not done 0 SPF WL fed the meat 9 of 0 died Swayne & Beck, Avian Dis. 9():8-8, 00 Inactivated AI vaccine in domestic ducks prevented dk/vn/0 (HN) HPAIV in meat, blood and viscera (Beato et al. Vaccine, epub Feb 8, 00)
4 Vaccination to Prevent HPAIV in Eggs Single killed H AI or sham vaccine Challenge with A/Pennsylvania/0/8 (HN) HPAIV SHAM No.+/total (log0/ml) d d d d Egg Shell 0/ 0/(.) /(.) - Albumen 0/ /(.) /(.9) - Yolk 0/ 0/(.8) /(.) - AI Vax No.+/Total (log0/ml) d d d -d Egg Shell 0/ /(.) /0(.) 0/ Albumin 0/ 0/ 0/0 0/ Yolk 0/ 0/ 0/0 0/ Swayne & Suarez, Develop. Biol. 0:-, 008 Pig Transmission Model: Experimental Design IN Day after arrival: Day 0: Collection of blood Infection with and nasal swabs 0 EID 0 of virus DPI Euthanasia of two pigs and collection of organ samples DPI Termination of pigs, blood and organ collection IG Daily weight monitoring; Collection of nasal and rectal swabs on days,,,, 9, and Control IN infection - Ck/Indonesia/0 (.) - Vietnam/0/0 () - WS/Mongolia/0 (.) -MDK/Vietnam/0 (.) -Swine/NC/0 (HN) & HN (positive controls) IG infection (liquid) - Vietnam/0/0 - MDK/Vietnam/0 Chicken meat feeding - WS/Mongolia/0 -weeks-old a Land Race x Large White cross male castrated piglets were used in the experiments. (Lipatov et al., PLOS Pathogens :e0000, 008)
5 Titer (log 0 EID 0 /g) HN HPAIV & HN/HN SIV: IN Inoculation Titer (log0 EID0/ml) Ck/Indo/0 Day Day Day Day VN/0 than SIV Virus shedding and organ titers on DPI WS/Mong/0 MDk/VN/0 Ck/Indo/0 VN/0 WS/Mong/0 MDk/VN/0 Sw/NC/0 Sw/IN/88 Sw/NC/0 Nasal turbinate Tonsils Trachea Lungs Sw/IN/88 Mild weight loss: WS/Mong/0, Ck/Indo/0 HN and HN SIV WS/Mong/0, Ck/Indo/0 and VN/0 in nasal swabs, mainly days & Titers of HN viruses lower HN replicated respiratory tract Lung titers of HN viruses were lower than SIV, HPAIV similar SIV WS/Mong/0 virus was isolated from trachea and tonsils, and MDk/VN/0 virus was isolated from nasal turbinate Histology: HN - mild to moderate bronchiolitis and multifocal alveolitis. SIV, severe tracheobronchitis and bronchointerstitial pneumonia (Lipatov et al., PLOS Pathogens :e0000, 008) HN HPAIV: IG inoculation Liquid media (0 EID 0 WS/Mong./0 & VN/0/0), viruses no evidence of infection Feeding WS/0 infected meat (00g, 0 0 EID 0 ) Transmission by consumption of infected meat Respiratory infection, no evidence of digestive infection No lesions No weight loss No changes in food consumption No behavioral abnormalities No virus rectal swabs or GI tissues (Lipatov et al., PLOS Pathogens :e0000, 008)
6 Ferret model: Experimental Model ferrets euthanatized DPI Organs for histological and virological examination Daily monitoring of temperature; Measuring of body weight and collection of nasal washes and rectal swabs on days,,,, 9 and -0-weeks-old female ferrets, groups of were inoculated IN (0 EID 0 /0. ml) or IG (0 EID 0 / ml) Terminated DPI Serum collected IN infection IG infection (liquid) Chicken meat feeding IG infection (grind) Ck/Indonesia/0 VN/0/0 WS/Mongolia/0 MDk/Vietnam/0 Ck/Indonesia/0 VN/0/0 WS/Mongolia/0 MDk/Vietnam/0 VN/0/0 WS/Mongolia/0 MDk/Vietnam/0 VN/0/0 (Lipatov et al, JID in press) Ferret Model: Results IG virus in liquid: no infection IN virus in liquid: Ck/Indonesia/0 & MDk/Vietnam/0 - mild or asymptomatic respiratory infection WS/Mongolia/0 Severe respiratory inf. (nasal washes, turbinates & lung) & inf. of olfactory bulb Vietnam/0 systemic disease including respiratory and digestive systems (including liver & pancreas) Meat consumption: MDk/Vietnam/0 mild respiratory infection WS/Mongolia/0 infection & mild respiratory disease: dpe virus in tonsil and nasal turbinates, dpe virus negative (Lipatov et al, JID in press)
7 VN/0 Virus Exposure Route on Infection Virus in nasal washes Virus in rectal swabs Titer (log0 TCID0/ml 0 VN/0 IN VN/0 IG VN/0 Meat VN/0 Meat IG Titer (log 0 TCID0/ml) 0 VN/0 IN VN/0 IG VN/0 Meat VN/0 Meat IG Days after infection Days after infection DPI virus titer - = no detection + = = > = >. System IN IG Meat Meat-IG Tonsil Respiratory Nasal Turb Lung Digestive SI LI Liver Systemic Spleen Brain Olf bulb (Lipatov et al, JID in press) Summary HPAIV is contained in infected poultry meat and eggs Cooking and pasteurization of infected meat inactivated HPAIV while freezing (-0C) is unlikely to produce predictable inactivation In the pig model, consumption of WS/0 HN infected meat caused respiratory infection via tonsil and pharynx exposure In ferret model, consumption of MDk/0 and WS/0 caused non-fatal respiratory infection while VN/0 caused lethal infection with initiation of infection in both respiratory and digestive tracts
8 Summary Based on animal models, the potential for human infection would require consumption of raw HPAIV infected meat at higher doses of virus than through respiratory exposure, and intestinal component maybe virus strain dependent Mitigation steps: ) cooking or pasteurization, ) vaccination of poultry to prevent viremia and organ/muscle infection Thank You For You Attention! 8
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