and where else? Nicole Pavio Anses Laboratoire de sant animale, UMR - PowerPoint PPT Presentation

Hepatitis E virus in animals, food and where else? Nicole Pavio Anses Laboratoire de santé animale, UMR 1161 INRA,ENVA, Anses, France nicole.pavio@anses.fr

Hepatitis E virus infection in human HEV1 to HEV4 Every year : 20 million hepatitis E infections 3 million symptomatic cases 56 600 hepatitis E-related deaths. • Incubation time 40 days (2-9 weeks) • Acute Hepatitis enteric transmission (similar to hepatitis A) • Fulminate Hepatitis 1%-4%. Pregnant women: 15%-25% • Chronic Hepatitis Immuno-suppression (solid organ transplant recipient) RNA+> 6 Mo • Neurologic syndroms Guillain-Barré, Parsonage-Turner (CSF+ chronic infection) • Extra-hepatic Kidney dysfunction, pancreatisis  Liver enzyme X 10 • Hepatocellular carcinoma (Amougou Atsama, 2017; Borentain P et al 2017)  Anti-HEV IgM +  HEV RNA + IAFP Europe April 24-26 2019

Virale hepatitis E epidemiology Endemic Regions Authochtonous sporadic or grouped cases Tropical, subtropical Europe, Japan HEV Genotypes 3, 4 Water, spoiled foods Food Animal reservoirs HEV Genotypes 1, 2 IAFP Europe April 24-26 2019 HEV Genotypes 3, 4

HEV Phylogeny: HEPEVIRIDAE family Zoonotic transmissions Orthohepevirus 1 4 HEV-3 -4 2 3ra 7 5-6 3 4 A 1 3 B 2 D C HEV-3ra C2 C1 Piscihepevirus HEV-7 Cutthroat Trout IAFP Europe April 24-26 2019 Doceul et al 2016

Animal reservoirs of HEV Anti-HEV antibodies Anti-HEV antibodies and HEV RNA HEV RNA IAFP Europe April 24-26 2019

Sources/vehicules of HEV3 and HEV4  Swine Slurry  Slaughterhouse  Personnel (hands)  Surfaces, ustensiles (knife, hook) Di Bartolo et al, 2012  Effluents Nantel-Fortier et al, 2016  Transportation vehicules  Waste Water Treatment Plant Spain and France (Clemente-Casares et al, EID 2003) Switzerland (Masclaux et al 2013) Italy (Iaconelli et al 2015)  Surface water in non endemic region (The Netherlands) (Rutjes SA, EID 2009)  Strawberries (Fields) (Brassard J, et al, 2012) Berry fruit (Maunula et al, 2013)  Shell fish  Corbicula Japonica, Japan (Li et al Am J Trop Med Hyg. 2007)  Food Outbreak on Cruise Ship (Oyster HEV-3) (Said et al 2009)  UK Mussels up to 92% HEV RNA (Crossan et al EID 2012) Animal  Bioaccumulation in Oyster (Grodski et al 2014) reservoirs  Green leafy salads (Kokkinos et al 2012)  Herb and Spices (Loisy-Hamon 2015) IAFP Europe April 24-26 2019

Sources/vehicules of HEV3 and HEV4  Pork Liver contaminated with HEV : grocery stores, slaughterhouse USA (Feagins et al., 2007) 11% RNA+; 2/3 infectious The Netherlands (Bouwknegt et al., 2007) 6.5% (Boxman, et al 2019) 12,7% Japan (Yazaki et al., 2003) 1.9% Korea (Jung et al., 2007) 10.8% India (Kulkarni et al., 2008) 0.83% France (Rose et al 2011) 4% Germany (Wenzel JJ, 2011) 4% Italy (Di Bartolo et al 2012) 6% Canada (Wilhelm et al 2014) 3,5%  Pork Liver sausages contaminated with HEV France (Pavio et al 2014) 30% Germany (Szabo et al et al 2015) 22% Italy (Di Bartolo et al 2014) 24 % Switzerland (Moor et al 2018) 19% The Netherland (Boxman, et al 2019) 70%  Pork sausages UK (Berto et al 2012) 10% Germany (Szabo et al 2015) 20% Switzerland (Moor et al 2018) 6% IAFP Europe April 24-26 2019

HEV experimental infection in pigs Hepatitis E Virus dynamic in pigs Theoretical HEV infection in natural setting HEV fecal excretion anti-HEV IgG Maternal Anti-HEV antibodies AST ALT anti-HEV IgM HEV Infection Cholesterol Triglycerides 16 0 2 10 12 14 18 24 4 6 8 20 22 Age in weeks IAFP Europe April 24-26 2019 Rogée et al 2015

Hepatitis E Virus dynamic in pig herds Salines in revision IAFP Europe April 24-26 2019

Co-infection with Hepatitis E and Porcin Reproductive and Respiratory Syndrom viruses Inoculated pigs Contact pigs Log(ge HEV) / g feces Log(ge HEV) / g feces Days post-infection Days post-infection  HEV + PRRSV : HEV chronic infection  Increased risk of HEV in other organs (muscles) Salines et al 2015 IAFP Europe April 24-26 2019

Co-infection with Hepatitis E and Porcin Reproductive and Respiratory Syndrom viruses HEV RNA in muscle at 49 dpi (RNA HEV RNA in HEV RNA in Viremia kinetic (RNA copies/mL serum) copies/g) feces at 49 dpi liver at 49 dpi (RNA copies/g) (RNA copies/g) dpi 7 14 21 28 35 42 49 FB PM DP Control nd nd nd nd nd nd nd nd nd nd nd nd Control nd nd nd nd nd nd nd nd nd nd nd nd 3.56.10 3 1.43.10 4 1.91.10 4 1.70.10 3 1.49.10 7 1.46.10 6 Inoculated nd nd nd nd nd nd 4.22.10 4 3.85.10 4 7.70.10 4 2.14.10 4 4.02.10 3 5.85.10 7 1.87.10 6 Inoculated nd nd nd nd nd 1.33.10 4 2.92.10 4 7.49.10 4 5.21.10 4 2.15.10 4 1.43.10 4 6.40.10 7 1.02.10 6 Inoculated nd nd nd nd Group 1 2.12.10 4 2.80.10 4 9.44.10 3 4.87.10 7 8.72.10 5 Contact nd nd nd nd nd nd nd 3.49.10 5 Contact nd nd nd nd nd nd nd nd nd nd nd 1.20.10 6 Contact nd nd nd nd nd nd nd nd nd nd nd 4.51.10 3 2.07.10 4 2.24.10 4 4.33.10 3 6.23.10 3 1.45.10 5 1.56.10 3 Inoculated nd nd nd nd nd 2.16.10 4 2.43.10 4 2.05.10 4 1.24.10 4 2.18.10 4 2.70.10 3 6.80.10 6 2.42.10 6 Inoculated nd nd nd nd 6.31.10 4 1.69.10 4 5.55.10 4 2.80.10 4 8.14.10 3 3.55.10 3 5.40.10 3 4.86.10 6 3.59.10 5 Inoculated nd nd nd Group 2 2.64.10 4 7.33.10 3 5.91.10 4 6.75.10 5 6.42.10 3 5.55.10 7 1.12.10 6 Contact nd nd nd nd nd 2.65.10 4 6.82.10 3 2.12.10 4 2.28.10 3 3.72.10 3 5.24.10 6 1.69.10 6 Contact nd nd nd nd nd 1.76.10 4 6.64.10 4 2.94.10 3 6.92.10 3 3.07.10 8 3.44.10 6 Contact nd nd nd nd nd nd 4.40.10 3 2.08.10 4 2.11.10 4 1.22.10 4 2.08.10 8 3.24.10 6 Inoculated nd nd nd nd nd nd 8.68.10 3 1.72.10 4 1.92.10 4 1.59.10 4 6.17.10 3 5.75.10 7 2.04.10 4 Inoculated nd nd nd nd nd 4.10.10 4 1.95.10 4 1.46.10 4 1.00.10 4 1.15.10 3 1.09.10 6 1.62.10 4 5.61.10 7 9.63.10 5 Inoculated nd nd nd Group 3 7.39.10 5 Contact nd nd nd nd nd nd nd nd nd nd nd 5.32.10 3 1.03.10 8 5.87.10 5 Contact nd nd nd nd nd nd nd nd nd 8.29.10 5 Contact nd nd nd nd nd nd nd nd nd nd nd Salines et al 2018

HEV-PRRSV During intercurrent infection (PRRSV): HEV is detected in pork meat muscles Psoas Major (Filet Mignon) • Up to 10 6 /g = possible transmission Food consumption Biceps femoris (Ham) • Risk assessement in Public Health: Inactivation procedure: heating time/temperature Drying, curing • Recommendation: deep cooking of pork meat Barnaud et al 2012 71°C during 20 min • Survey at slaughterhouse: test faecal samples to detect pig at risk of HEV positive meat IAFP Europe April 24-26 2019

Hepatitis E Virus in Muscle at Slaughterhouse positive Prevalence 95% Confidence Sample size samples estimate (%) interval HEV serology 1034 629 60 57.8-63.7 HEV RNA (muscles) 1034 0 0 0-0.003 HEV RNA (liver) 1034 29 2.8 1.9-3.9 14  Different muscle: gluteus medius or semi-membranosus 12  No indication PRRSV status Number of livers 10 8 Perspectives: 6 • Larger sampling 4 • HEV, PCV2 and PRRSV status 2 • Biceps or Psoas 0 [10^3 - 10^4] [10^4 - 10^5] [10^5 - 10^6] [10^7 - 10^8] [10^8 - 10^9] copies of HEV RNA per gram IAFP Europe April 24-26 2019 Feurer et al 2017

HEV RNA +: 33/36 batches of (non- heated) liquid products 7/24 spray dried powder products IAFP Europe April 24-26 2019

Frequent Hepatitis E Virus Contamination in Food Containing Raw Pork Liver, France Food category number of samples RNA Quantification (Min-Max Prevalence and analyzed (394 total) in copies of HEV RNA/g)* [ 95% CI] Figatelli and fitone 140 1.7 10 2 -6.9 10 5 30 [23, 38] Dried salted liver 30 6.9 10 5 3 [0, 1] 2.6 10 2 -2.83 10 5 Quenelle and quenelle paste 55 25 [15, 37] 1 10 2 -2.3 10 6 Dried or fresh liver sausages 169 29 [22, 36] Pavio et al 2014 EID Berto et al.2013 EID

Phylogeny of HEV sequences present in raw pork liver foods. Food/ human SPAIN Food/ human FRANCE All HEV genotype 3 Food/pig High sequence identity : 99% (n=16) Food/ human sequences (France, UK, Spain) Food/pig sequences (France) Food/pig Swine Human (France) Human (other) Food Food/ human UK Pavio et al EID 2014 IAFP Europe April 24-26 2019

Direct foodborne transmission of HEV IAFP Europe April 24-26 2019 Guillois et al CID 2015

Foodborne transmission of HEV IAFP Europe April 24-26 2019 Yapa et al 2016

Foodborne transmission of HEV Case-control study among hepatitis E cases (n=270) aged 18 years or older notified between January 2012 and January 2014 and matching controls (n=1,159). => Consumption of undercooked pork liver, pork, wild boar meat, frankfurters, liver sausage and raw vegetables; contact with waste water (occupational). Pork liver>Wild boar>Pork>Liver sausage or liver pâté>Boiled sausage>Raw vegetables IAFP Europe April 24-26 2019 Faber et al 2018

Foodborne transmission of HEV 75 % foodborne origin of HEV Van Cauteren et al, 2018 A nationwide case-control study June 2015 and October 2017 (n=597). Traditional Dutch dry raw sausages of pork muscle meat, called , “ cervelaat ”, “ snijworst ”, and “boerenmetworst”were reported by 72% of the patients, and 46% of controls (OR 3.0; 95% CI 2.2 – 4.1) Direct contact with pigs and working with a septic tank were strong risk factors (aOR 3.1; 95%CI 1.3 – 7.3 and aOR 6.9; 95%CI 1.2 – 40.8, respectively), Kamar et al 2012 IAFP Europe April 24-26 2019 Tulen et al 2019

IAFP Europe April 24-26 2019