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Hepatitis E virus infection in wild rabbit (Oryctolagus cuniculus) in Italy and in the UK: a serological, molecular, and pathological study

  • Francesca Parisi
  • Maurizio Mazzei
  • Ranieri Verin
  • Mario Forzan
  • Guido Rocchigiani
  • Charlotte Roper
  • Giacomo Bertelloni
  • Alessandro PoliEmail author
Original Article
  • 88 Downloads

Abstract

A novel animal strain of HEV demonstrating zoonotic potential rabbit HEV (rHEV) has been recently identified in farmed rabbits in China. To investigate the presence of rHEV in wild rabbit population, serum and tissue samples were taken from 65 rabbits, 35 Italian and 30 from UK. Sera were submitted to a double antigen sandwich ELISA, whereas hepatic tissues and other organs to molecular and pathological investigations. Sixteen serum samples (24.6%) scored positive for anti-HEV antibodies, and six samples (9%) of hepatic tissue were positive for HEV RT-PCR, while the other organs were negative. Sequencing and phylogenetic analysis of HEV RNA-positive samples indicated that while two Italian rabbits were infected with strains closely related to wild boar and swine strains (HEV-3), the other four (2 Italian and 2 English) were clustered within rHEV clade. Megalocytosis and multifocal areas of hepatocytes degeneration and necrosis with a pericentrilobular pattern were observed in rHEV-infected rabbits. In those infected by the strain analogous to HEV-3, the lesions were mainly localized in the periportal areas and were less severe. In both cases, inflammatory infiltrates were predominantly composed of CD3-positive lymphocytes and a reduced number of macrophages. By immunohistochemistry, only liver samples taken from HEV RNA-positive rabbits scored positive for viral antigen. Our results indicated that HEV infection is present in rabbit population with different clades and is endemic in the Italian and English wild rabbit population, suggesting the possibility that this species may be infected with rHEV or swine HEV-3 strains.

Keywords

Hepatitis E virus Histopathology Serology PCR Phylogenetic analysis Wild rabbit (Oryctolagus cuniculus) 

Notes

References

  1. Birke L, Cormier SA, You D, Stout RW, Clement C, Johnson M, Thompson H (2014) Hepatitis E antibodies in laboratory rabbits from 2 US vendors. Emerg Infect Dis 20:693–696.  https://doi.org/10.3201/eid2004.131229 CrossRefPubMedPubMedCentralGoogle Scholar
  2. Burt SA, Veltman J, Hakze-van der Honing R, Schmitt H, van der Poel WH (2016) Hepatitis E virus in farmed rabbits, wild rabbits and petting farm rabbits in the Netherlands. Food Environ Virol 8:227–229.  https://doi.org/10.1007/s12560-016-9239-3 CrossRefPubMedPubMedCentralGoogle Scholar
  3. Caruso C, Modesto P, Prato R, Scaglione FE, De Marco L, Bollo E, Acutis PL, Masoero L, Peletto S (2015) Hepatitis E virus: first description in a pet house rabbit. A new transmission route for human? Transbound Emerg Dis 62:229–232.  https://doi.org/10.1111/tbed.12348 CrossRefPubMedGoogle Scholar
  4. Cossaboom CM, Cordoba L, Dryman BA, Meng XJ (2011) Hepatitis E virus in rabbits, Virginia. USA Emerg Infect Dis 17:2047–2049.  https://doi.org/10.3201/eid1711.110428. CrossRefPubMedGoogle Scholar
  5. Di Bartolo I, De Sabato L, Marata A, Martinelli N, Magistrali CF, Monini M, Ponterio E, Ostanello F, Ruggeri FM (2016) Serological survey of hepatitis E virus infection in farmed and pet rabbits in Italy. Arch Virol 161:1343–1346.  https://doi.org/10.1007/s00705-016-2778-y CrossRefPubMedGoogle Scholar
  6. Geng Y, Zhang H, Huang W, T JH, Geng K, Li Z, Wang Y (2014) Persistent hepatitis e virus genotype 4 infection in a child with acute lymphoblastic leukemia. Hepat Mon 14:e15618.  https://doi.org/10.5812/hepatmon.15618 CrossRefPubMedPubMedCentralGoogle Scholar
  7. Giordani MT, Fabris P, Brunetti E, Goblirsch S, Romano L (2013) Hepatitis E and lymphocytic leukemia in Man, Italy. Emerg Infect Dis 19:2054–2056.  https://doi.org/10.3201/eid1912.130521 CrossRefPubMedPubMedCentralGoogle Scholar
  8. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41:95–98Google Scholar
  9. Hammerschmidt F, Schwaiger K, Dahnert L, Vina-Rodriguez A, Hoper D, Gareis M, Groschup MH, Eiden M (2017) Hepatitis E virus in wild rabbits and European brown hares in Germany. Zoonoses Public Health 64:612–622.  https://doi.org/10.1111/zph.12355 CrossRefPubMedGoogle Scholar
  10. Han J, Lei Y, Liu L, Liu P, Xia J, Zhang Y, Zeng H, Wang L, Wang L, Zhuang H (2014) SPF rabbits infected with rabbit hepatitis E virus isolate experimentally showing the chronicity of hepatitis. PLoS One 9:e99861.  https://doi.org/10.1371/journal.pone.0099861 CrossRefPubMedPubMedCentralGoogle Scholar
  11. Izopet J, Dubois M, Bertagnoli S, Lhomme S, Marchandeau S, Boucher S, Kamar N, Abravanel F, Guerin JL (2012) Hepatitis E virus strains in rabbits and evidence of a closely related strain in humans, France. Emerg Infect Dis 18:1274–1281.  https://doi.org/10.3201/eid1808.120057. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Krain LJ, Nelson KE, Labrique AB (2014) Host immune status and response to hepatitis E virus infection. Clin Microbiol Rev 27(1):139–165.  https://doi.org/10.1128/CMR.00062-13 CrossRefPubMedPubMedCentralGoogle Scholar
  13. Liu P, Li L, Wang L, Bu Q, Fu H, Han J, Zhu Y, Lu F, Zhuang H (2012) Phylogenetic analysis of 626 hepatitis E virus (HEV) isolates from humans and animals in China (1986-2011) showing genotype diversity and zoonotic transmission infection, genetics and evolution. Infect Genet Evol 12:428–434.  https://doi.org/10.1016/j.meegid.2012.01.017. CrossRefPubMedGoogle Scholar
  14. Ma H, Zheng L, Liu Y, Zhao C, Harrison TJ, Ma Y, Sun S, Zhang J, Wang Y (2010) Experimental infection of rabbits with rabbit and genotypes 1 and 4 hepatitis E viruses. PLoS One 5:e9160.  https://doi.org/10.1371/journal.pone.0009160 CrossRefPubMedPubMedCentralGoogle Scholar
  15. Macfarlane SB (1997) Conducting a descriptive survey: 2. Choosing a sampling strategy. Trop Doc 27:14–21.  https://doi.org/10.1177/004947559702700108 CrossRefGoogle Scholar
  16. Mao J, Zhao Y, She R, Cao B, Xiao P, Wu Q, Guo Z, Ma L, Soomro MH (2014) Detection and localization of rabbit hepatitis e virus and antigen in systemic tissues from experimentally intraperitoneally infected rabbits. PLoS One 9:e88607.  https://doi.org/10.1371/journal.pone.0088607 CrossRefPubMedPubMedCentralGoogle Scholar
  17. Mazzei M, Forzan M, Pizzurro F, Picciolli F, Bandecchi P, Poli A (2015a) Detection of hepatitis E virus antibodies in domestic and wild animal species in Central Italy. Clin Microbiol 4:227.  https://doi.org/10.4172/2327-5073.1000227 CrossRefGoogle Scholar
  18. Mazzei M, Nardini R, Verin R, Forzan M, Poli A, Tolari F (2015b) Serologic and molecular survey for hepatitis E virus in wild boar (Sus scrofa) in Central Italy. New Microbes New Infect 4(7):41–47.  https://doi.org/10.1016/j.nmni.2015.05.008. CrossRefGoogle Scholar
  19. Meng XJ, Halbur PG, Shapiro MS, Govindarajan S, Bruna JD, Mushahwar IK, Purcell RH, Emerson SU (1998) Genetic and experimental evidence for cross-species infection by swine hepatitis E virus. J Virol 72:9714–9721PubMedPubMedCentralGoogle Scholar
  20. Morozov VA, Morozov AV, Rotem A, Barkai U, Bornstein S, Denner J (2015) Extended microbiological characterization of Gottingen minipigs in the context of xenotransplantation: detection and vertical transmission of hepatitis E virus. PLoS One 10:e0139893CrossRefGoogle Scholar
  21. Nardini R, Verin R, Mazzei M, Forzan M, Poli A (2014) Hepatitis E virus-related liver alterations and viral antigen localization in European wild boar (Sus scrofa). Eur J Wildl Res 60:835–838.  https://doi.org/10.1007/s10344-014-0839-7 CrossRefGoogle Scholar
  22. Risalde MA, Rivero-Juarez A, Romero-Palomo F, Frias M, Lopez-Lopez P, Cano-Terriza D, Garcia-Bocanegra I, Jimenez-Ruiz S, Camacho A, Machuca I, Gomez-Villamandos JC, Rivero A (2017) Persistence of hepatitis E virus in the liver of non-viremic naturally infected wild boar. PLoS One 12:e0186858.  https://doi.org/10.1371/journal.pone.0186858 CrossRefPubMedPubMedCentralGoogle Scholar
  23. Schielke A, Ibrahim V, Czogiel I, Faber M, Schrader C, Dremsek P, Ulrich RG, Johne R (2015) Hepatitis E virus antibody prevalence in hunters from a district in Central Germany, 2013: a cross-sectional study providing evidence for the benefit of protective gloves during disembowelling of wild boars. BMC Infec Dis 15:440.  https://doi.org/10.1186/s12879-015-1199-y CrossRefGoogle Scholar
  24. Schlosser J, Eiden M, Vina-Rodriguez A, Fast C, Dremsek P, Lange E, Ulrich RG, Groschup MH (2014) Natural and experimental hepatitis E virus genotype 3-infection in European wild boar is transmissible to domestic pigs. Vet Res 45:121.  https://doi.org/10.1186/s13567-014-0121-8 CrossRefPubMedPubMedCentralGoogle Scholar
  25. Smith DB, Simmonds P, Jameel S, Emerson SU, Harrison TJ, Meng XJ, Okamoto H, Van der Poel WH, Purdy MA (2015) Consensus proposals for classification of the family Hepeviridae. J Gen Virol 96:1191–1192.  https://doi.org/10.1099/vir.0.000115 CrossRefPubMedGoogle Scholar
  26. Spahr C, Knauf-Witzens T, Vahlenkamp T, Ulrich RG, Johne R (2018) Hepatitis E virus and related viruses in wild, domestic and zoo animals: a review. Zoonoses Public Health 65:11–29.  https://doi.org/10.1111/zph.12405 CrossRefPubMedGoogle Scholar
  27. Sridhar S, Teng JLL, Chiu T-H, Lau SKP, Woo PCY (2017) Hepatitis E virus genotypes and evolution: emergence of camel hepatitis E variants. Int J Mol Sci 18:869CrossRefGoogle Scholar
  28. Van der Poel WH (2014) Food and environmental routes of hepatitis E virus transmission. Curr Opin Virol 4:91–96.  https://doi.org/10.1016/j.coviro.2014.01.006 CrossRefPubMedGoogle Scholar
  29. Villafuerte R, Castro F, Ramirez E, Cotilla I, Parra F, Delibes-Mateos M, Recuerda P, Rouco C (2017) Large-scale assessment of myxomatosis prevalence in European wild rabbits (Oryctolagus cuniculus) 60years after first outbreak in Spain. Res Vet Sci 114:281–286.  https://doi.org/10.1016/j.rvsc.2017.05.014 CrossRefPubMedGoogle Scholar
  30. Wang S, Cheng X, Dai X, Dong C, Xu M, Liang J, Dong M, Purdy MA, Meng J (2013) Rabbit and human hepatitis E virus strains belong to a single serotype. Virus Res 176:101–106.  https://doi.org/10.1016/j.virusres.2013.05.013 CrossRefPubMedGoogle Scholar
  31. Zhao C, Ma Z, Harrison TJ, Feng R, Zhang C, Qiao Z, Fan J, Ma H, Li M, Song A, Wang Y (2009) A novel genotype of hepatitis E virus prevalent among farmed rabbits in China. J Med Virol 81:1371–1379.  https://doi.org/10.1002/jmv.21536 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Francesca Parisi
    • 1
  • Maurizio Mazzei
    • 1
  • Ranieri Verin
    • 2
  • Mario Forzan
    • 1
  • Guido Rocchigiani
    • 2
  • Charlotte Roper
    • 2
  • Giacomo Bertelloni
    • 1
  • Alessandro Poli
    • 1
    Email author
  1. 1.Department of Veterinary SciencesUniversity of PisaPisaItaly
  2. 2.Department of Veterinary Pathology and Public HealthUniversity of LiverpoolLiverpoolUK

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