Hepatitis E virus in animals, food and where else? · Reported number of cases of hepatitis E virus infection by year of notification and year of surveillance commencement, 22 European

Hepatitis E virus in animals, food and where else?

Nicole Pavio Anses Laboratoire de santé animale, UMR 1161 INRA,ENVA, Anses, France

[email protected]

Hepatitis E virus infection in human

Liver enzyme X 10 Anti-HEV IgM + HEV RNA +

• 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

• Hepatocellular carcinoma (Amougou Atsama, 2017; Borentain P et al 2017)

HEV1 to HEV4

Every year : 20 million hepatitis E infections

3 million symptomatic cases

56 600 hepatitis E-related deaths.

HEV 2018 June 4-5 2018

Water, spoiled foods

Genotypes 1, 2

Genotypes 3, 4

Endemic Regions

Tropical, subtropical

Authochtonous sporadic or grouped cases

Europe, Japan

Genotypes 3, 4

Virale hepatitis E epidemiology

Animal reservoirs

Food

HEV 2018 June 4-5 2018

HEV Phylogeny: HEPEVIRIDAE family

Cutthroat Trout

A

Orthohepevirus

Piscihepevirus

D B

C1

C

3ra 7

5-6

2

1 4

3

C2

1 2

3

4

Doceul et al 2016

Orthohepevirus A 7 genotypes 1 & 2 human 3-4 & 7 zoonotic (including Rabbit HEV-3ra)

HEV interspecies transmissions Zoonotic transmissions

HEV-3ra

HEV-7

HEV-3 -4

HEV 2018 June 4-5 2018

USA (Feagins et al., 2007) 11% RNA+; 2/3 infectious

The Netherlands (Bouwknegt et al., 2007) 6.5%

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 contaminated with HEV : grocery stores, slaughterhouse

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%

Pork sausages UK (Berto et al 2012) 10%

Germany (Szabo et al 2015) 20%

Sources/vehicules of HEV3 and HEV4

HEV 2018 June 4-5 2018

Spain and France (Clemente-Casares et al, EID 2003) Switzerland (Masclaux et al 2013) Italy (Iaconelli et al 2015)

Waste Water Treatment Plant

(Rutjes SA, EID 2009) Surface water in non endemic region (The Netherlands)

Strawberries (Fields) Berry fruit

(Brassard J, et al, 2012) (Maunula et al, 2013)

Swine Slurry

Corbicula Japonica, Japan (Li et al Am J Trop Med Hyg. 2007) Outbreak on Cruise Ship (Oyster HEV-3) (Said et al 2009) UK Mussels up to 92% HEV RNA (Crossan et al EID 2012) Bioaccumulation in Oyster (Grodski et al 2014)

Shell fish

Herb and Spices (Loisy-Hamon 2015)

Green leafy salads (Kokkinos et al 2012)

Slaughterhouse Personnel (hands) Surfaces, ustensiles (knife, hook) Effluents Transportation vehicules

Di Bartolo et al, 2012 Nantel-Fortier et al, 2016

Animal reservoirs

Food

Sources/vehicules of HEV3 and HEV4

HEV 2018 June 4-5 2018

Reported number of cases of hepatitis E virus infection by year of notification and year of surveillance

commencement, 22 European Union/European Economic Area (EU/EEA) countries, 2005–2015a

HEV 2018 June 4-5 2018

HEV Epidemiology in France

• National Seroprevalence : 22,4% • 3 hyperendemic regions • HEV RNA Prevalence blood donnors:

1/2848 -1/744 • Test in selected blood products (for solid

organ recipients) • Estimations: 60,000 acute cases/year 500 hospitalization 20 deaths

Mansuy et al, Hepatology 2016

Van Cauteren et al, 2018

Rose et al CIMID 2011, Bouquet et al 2011

Sample size

Number

positive

Prevalence

estimate

%

95% IC

HEV serology

Individual pig-level

Farm level

6565

186

1069

137

31

65

24 – 38

57 – 74

HEV virology (livers)

Individual pig-level

Farm level

3715

186

128

43

4

24

2 – 6

17 – 31

HEV prevalence in French swine (serology and RNA)

HEV 2018 June 4-5 2018

Hepatitis E Virus dynamic in pigs

anti-HEV IgG

anti-HEV IgM

HEV fecal excretion

0 2 4 6 8 10 12 14 16 18 20 22 24

Age in weeks

Maternal Anti-HEV antibodies

HEV Infection

ALT

Cholesterol Triglycerides

AST

Theoretical HEV infection in natural setting

HEV experimental infection in pigs

HEV 2018 June 4-5 2018

Days post-infection Days post-infection

Lo

g(g

e H

EV

) / g f

ece

s

Lo

g(g

e H

EV

) / g f

ece

s

Inoculated pigs Contact pigs

Co-infection with Hepatitis E and Porcin Reproductive and Respiratory Syndrom viruses

HEV + PRRSV : HEV chronic infection Increased risk of HEV in other organs (muscles)

Salines et al 2015

Co-infection with Hepatitis E and Porcin Reproductive and Respiratory Syndrom viruses

Salines et al submitted

Viremia kinetic (RNA copies/mL serum) HEV RNA in muscle at 49 dpi (RNA

copies/g) HEV RNA in

feces at 49 dpi

(RNA copies/g)

HEV RNA in

liver at 49 dpi

(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

Group 1

Inoculated nd nd nd 3.56.103 1.43.104 1.91.104 1.70.103 nd nd nd 1.49.107 1.46.106

Inoculated nd nd 4.22.104 3.85.104 7.70.104 2.14.104 4.02.103 nd nd nd 5.85.107 1.87.106

Inoculated nd nd 1.33.104 2.92.104 7.49.104 5.21.104 2.15.104 nd nd 1.43.104 6.40.107 1.02.106

Contact nd nd nd nd 2.12.104 2.80.104 9.44.103 nd nd nd 4.87.107 8.72.105

Contact nd nd nd nd nd nd nd nd nd nd 3.49.105 nd


Group 2

Inoculated nd nd 4.51.103 2.07.104 2.24.104 nd nd 4.33.103 nd 6.23.103 1.45.105 1.56.103

Inoculated nd nd 2.16.104 2.43.104 2.05.104 1.24.104 nd nd 2.18.104 2.70.103 6.80.106 2.42.106

Inoculated nd nd 6.31.104 1.69.104 5.55.104 2.80.104 nd 8.14.103 3.55.103 5.40.103 4.86.106 3.59.105

Contact nd nd nd nd nd 2.64.104 7.33.103 5.91.104 6.75.105 6.42.103 5.55.107 1.12.106

Contact nd nd nd nd nd 2.65.104 6.82.103 2.12.104 2.28.103 3.72.103 5.24.106 1.69.106

Contact nd nd nd nd 1.76.104 6.64.104 2.94.103 6.92.103 nd nd 3.07.108 3.44.106

Group 3

Inoculated nd nd nd 4.40.103 2.08.104 2.11.104 1.22.104 nd nd nd 2.08.108 3.24.106

Inoculated nd nd nd nd 8.68.103 1.72.104 1.92.104 nd 1.59.104 6.17.103 5.75.107 2.04.104

Inoculated nd nd 4.10.104 1.95.104 1.46.104 1.00.104 1.15.103 nd 1.09.106 1.62.104 5.61.107 9.63.105


Contact nd nd nd nd nd nd nd 5.32.103 nd nd 1.03.108 5.87.105


HEV-PRRSV

During intercurrent infection (PRRSV): HEV is detected in pork meat muscles

Biceps femoris (Ham)

Psoas Major (Filet Mignon)

• Risk assessement in Public Health: Inactivation procedure: heating time/temperature Drying, curing • Recommendation: deep cooking of pork meat • Survey at slaughterhouse: test faecal samples to detect pig at risk of HEV positive meat

• Up to 106/g = possible transmission Food consumption

HEV 2018 June 4-5 2018

71°C during 20 min Barnaud et al 2012

Hepatitis E Virus in Muscle at Slaughterhouse

Sample size positive samples

Prevalence estimate (%)

95% Confidence 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

1

0

2

4

6

8

10

12

14

[10^3 - 10^4] [10^4 - 10^5] [10^5 - 10^6] [10^7 - 10^8] [10^8 - 10^9]

Nu

mb

er o

f liv

ers

copies of HEV RNA per gram Feurer et al 2017

Different muscle: gluteus medius or semi-membranosus No indication PRRSV status

Perspectives: • Larger sampling • HEV, PCV2 and PRRSV status • Biceps or Psoas

HEV 2018 June 4-5 2018

Berto et al.2013 EID

Food category number of samples

analyzed (394 total)

RNA Quantification (Min-Max

in copies of HEV RNA/g)*

Prevalence and

[ 95% CI]

Figatelli and fitone 140 1.7 102-6.9 105 30 [23, 38]

Dried salted liver 30 6.9 105 3 [0, 1]

Quenelle and quenelle paste 55 2.6 102-2.83 105 25 [15, 37]

Dried or fresh liver sausages 169 1 102 -2.3 106 0.29 [22, 36]

Pavio et al 2014 EID

Frequent Hepatitis E Virus Contamination in Food Containing Raw Pork Liver, France

Phylogeny of HEV sequences present in raw pork liver foods.

Pavio et al EID 2014

All HEV genotype 3 High sequence identity : 99% (n=16) Food/ human sequences (France, UK, Spain) Food/pig sequences (France)

Swine Human (France) Human (other) Food

Food/pig

Food/pig

Food/ human SPAIN

Food/ human UK

Food/ human FRANCE

HEV 2018 June 4-5 2018

Renou et al EID 2014

ORF-2 ORF-1

Unequivocal food borne transmission of HEV

Direct foodborne transmission of HEV

HEV 2018 June 4-5 2018

Guillois et al CID 2015

Direct foodborne transmission of HEV

HEV 2018 June 4-5 2018

Kamar et al 2012

75 % foodborne origin

HEV 2018 June 4-5 2018

Van Cauteren et al, 2018

• ready-to-eat pork products as a major source for autochthonous hepatitis E (control-case study)

Pork liver>Wild boar>Pork>Liver sausage or liver pâté>Boiled sausage>Raw vegetables

Faber et al, 2018

Thanks to

CHU Purpan CNR VHE Jacques Izopet Jean-Michel Mansuy Florence Abravanel

Santé Publique France Elisabeth Couturier

Anciens CNR VHE Elisabeth Nicand Sophie Tessé Anne-Marie Roque -Afonso

ANSES Ploufragan Nicolas Rose Matthieu Andraud Morgane Salines Virginie Dorenlor Florent Eono Frederic Paboeuf

Institut du Porc Carole Feurer

HEV Team UMR 1161 Virologie Laboratoire de santé animale Virginie Doceul Elodie Barnaud Gaël Stéphant Antonin Demange Eugénie Bagdassarian Léa Meyer Sergiu Raileanu Marie Pellerin Stephan Zientara Jennifer Richardson

HEV 2018 June 4-5 2018

Documents

Hepatitis E virus in animals, food and where else? · Reported number of cases of hepatitis E virus infection by year of notification and year of surveillance commencement, 22 European