Detection of HEV in Estonia. Anna Ivanova, Irina Golovljova, Valentina Tefanova

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1 Detection of HEV in Estonia Anna Ivanova, Irina Golovljova, Valentina Tefanova

2 National Institute for Health Development Department of Virology Zoonotic pathogens Tick-borne pathogens: TBEV, Borrelia sp., Rickettsia sp., Babesia sp., Ehrlichia sp. Rodent-borne pathogens: Hantaviruses Hepatitis viruses HAV, HBV, HCV, HEV

3 Hepatitis E virus (HEV) Family: Hepeviridae Genus: Hepevirus HEV genome Small, non-enveloped virus ssrna (+) Hepatitis E: An emerging disease M. Pérez-Gracia et al. Infection, Genetics and Evolution, Volume 22, 2014, 40-59

4 Geographical distribution of hepatitis E virus genotypes. S. Mirazo, et al Hepat Med. 2014; 6:

5 Transmission routes of HEV Hepatitis E: An emerging disease M. Pérez-Gracia, et al. Infection, Genetics and Evolution, Volume 22, 2014, 40-59

6 Aims To assess HEV seroprevalence level in different human populations in Estonia To assess the presence of HEV infection in Estonian domestic pig and wild boar populations

7 Materials Human serum samples Retrospective samples Samples collected for the current study Ambulatory pt. without any known infectious desease (AP; 2003) 151 Acute nona-nonc ( ) 762 Health care workers (2004) 163 Hemodialysis pat. ( ) 176 IDU ( ) 205 Veterinarians 158 Pig farm workers 67 Hunters 144 Samples from domestic pigs and wild boars Serum samples from adult pigs ( 1,5 y) 380 Fecal samples from pigs (2 4 months) 449 Meat juice from wild boars 471

8 Location of 17 farms where samples were collected 1 - Serum samples of pig farm workers 2 - Serum samples of domestic pigs 3 Fecal samples from pigs

9 Collection of serum samples from hunters and wild boars 21/68 7/48 7/4 3/53 15/16 1/37 9/13 3/23 30/46 2/30 10/41 13/30 1/28 4/34

10 Methods Serological assays Human samples: ELISA: Confirmatory IB test: recomwell IgG and IgM (Mikrogen GmbH, Germany) recomline HEV IgG/IgM (Mikrogen GmbH, Germany) Pig serum and wild boar meet juice samples ELISA: PrioCHECK HEV porcine kit (Prionics AG, Switzerland) Detection of viral RNA qpcr for samples screening (Jothikumar et al., 2006) Samples positive in qpcr amplified in the ORF2 region (1011 nt)

11 % of HEV IgG/IgM positive samples Detection of antibodies against HEV in Estonian human population * IgM IgG * * * * 2 0 Pig farm workers Ambulatory Healthy Hemodialysis Hunters Healthcare patients workers IDU nona-nonc Veterinarians

12 % of HEV IgG positive samples Anti-HEV IgG seroprevalence in different age groups 10 * * >61

13 HEV antibodies and HEV RNA in domestic pigs and wild boars Hiiumaa F12 (50%) Saaremaa 13% 25% F5 (100%) Läänemaa F11 (58,3%) 4% F10 (80,6%) F pig farm (seroprevalence %) 13% 30% F3 (100%) F2 (71,4%) F1 (27,3%) 26% Pärnumaa Harjumaa 19% Raplamaa Seroprevalence in wild boar HEV RNA in domestic swine 11% Järvamaa 33% Viljandimaa Lääne- Virumaa Valgamaa F17 (38,5%) Jõgevamaa 23% 4% Tartumaa 7% F6 (35,7%) F7 (50%) Ida-Virumaa 12% Põlvamaa 7% Võrumaa 21% HEV RNA in wild boar F14 (60%) F16 (75%) F15 (85%) F13 (61,2%)

14 Phylogenetic tree based of Estonia HEV sequences on the partial ORF2 HEV sequences from wild boar HEV sequences from domestic pigs JF DQ KF GENOTYPE FJ e wild boar, Germany AB e wild boar, Japan JX e human, Russia HM e domestic pig, Hungary HQ domestic pig, Russia Est343 Est91 99 Est187 Est346 Est90 Est112 F9 4-5 F II F F F F F F AB human, Japan KC f domestic swine, Spain KC f human, Belgium EU f human, France HM f domestic swine, Sweden 99 F F HQ domestic pig, Russia HQ domestic pig, Russia Est442 F F F F F8 4-8 F F F F F F F IV JQ c human, France KJ i human, France KF b human, Brazil AF a human, USA AY j domestic pig, Canada AB GENOTYPE M74506 GENOTYPE 2 I III GENOTYPE 3

15 Geographical distribution of HEV lineages F2 F3 F6 Harjumaa Lääne- Virumaa Ida-Virumaa Hiiumaa Saaremaa F11 F8 Läänemaa Järvamaa Raplamaa Jõgevamaa Pärnumaa Viljandimaa Tartumaa Põlvamaa Valgamaa Võrumaa F4 F14 F16 F9 Lineage 1 Lineage 2 Lineage 3 Lineage 4 Sequences from wild boars Sequences from domestic pigs

16 Conclusions HEV circulates in Estonia Domestic pigs and wild boars are possible reservoirs and source of human HEV infection Seroprevalence in pig farm workers appears to be highest among the analysed groups All Estonian HEV strains from animals belong to genotype 3, but form 4 separate lineages

17 Acknowledgments Arvo Viltrop, Brian Lassen, Tiiu Saar, Department of Basic Veterinary Science and Population Medicine, Estonian University of Life Science Jevgenia Epstein, Kuulo Kutsar, Natalia Kerbo Public Health Board of Estonia The study was financially supported by the European Regional Development Fund, Programme TerVe, Projects ZoonRisk and EKZE