Archives of Virology

, Volume 163, Issue 5, pp 1187–1193 | Cite as

Experimental infection of Marmota monax with a novel hepatitis A virus

  • Jie-mei Yu
  • Li-li Li
  • Guang-cheng Xie
  • Cui-yuan Zhang
  • Yuan-yun Ao
  • Zhao-jun Duan
Original Article


To establish an animal model for the newly identified Marmota Himalayana hepatovirus, MHHAV, so as to develop a better understanding of the infection of hepatitis A viruses. Five experimental woodchucks (Marmota monax) were inoculated intravenously with the purified MHHAV from wild woodchuck feces. One animal injected with PBS was defined as a control. Feces and blood were routinely collected. After the animals were subjected to necropsy, different tissues were collected. The presence of viral RNA and negative sense viral RNA was analyzed in all the samples and histopathological and in situ hybridization analysis was performed for the tissues. MHHAV infection caused fever but no severe symptoms or death. Virus was shed in feces beginning at 2 dpi, and MHHAV RNA persisted in feces for ~2 months, with a biphasic increase, and in blood for ~30 days. Viral RNA was detected in all the tissues, with high levels in the liver and spleen. Negative-strand viral RNA was detected only in the liver. Furthermore, the animals showed histological signs of hepatitis at 45 dpi. MHHAV can infect M. monax and is associated with hepatic disease. Therefore, this animal can be used as a model of HAV pathogenesis and to evaluate antiviral and anticancer therapeutics.



The study was supported by the National Natural Science Foundation of China (Grant No. 81702007 and 81290345).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Six healthy male woodchucks (M. monax) (age, 1–3 years; weight, 1.8–2.4 kg) were purchased from the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Science and Peking Union Medical College.

All experiments involving MHHAV-infected animals were performed in biosafety level 2 containment at the Experimental Animal Center of the China CDC, in accordance with institutional guidelines. The animal study protocol was approved by the China CDC Animal Welfare Committee.

Supplementary material

705_2018_3715_MOESM1_ESM.tif (826 kb)
Supplemental Fig. 1. Electron micrographs of immune-complexed MHHAV. The virus particles shed in feces of the infected animals were identical to those shed by wild woodchucks, and both virus particles and empty capsids were present in feces (TIFF 825 kb)
705_2018_3715_MOESM2_ESM.tif (678 kb)
Supplemental Fig. 2. In situ hybridization of MHHAV RNA in woodchuck liver. (A) Liver of the control animal. (B) Liver of an MHHAV-infected animal. Left, middle, and right; fluorescence, bright-field, and superimposed images, respectively. Bright green dots indicate probe bound to MHHAV genomic RNA (TIFF 678 kb)


  1. 1.
    Amado LA, Marchevsky RS, de Paula VS, Hooper C, Freire Mda S, Gaspar AM, Pinto MA (2010) Experimental hepatitis A virus (HAV) infection in cynomolgus monkeys (Macaca fascicularis): evidence of active extrahepatic site of HAV replication. Int J Exp Pathol 91:87–97CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Anthony SJ, St Leger JA, Liang E, Hicks AL, Sanchez-Leon MD, Jain K, Lefkowitch JH, Navarrete-Macias I, Knowles N, Goldstein T, Pugliares K, Ip HS, Rowles T, Lipkin WI (2015) Discovery of a novel hepatovirus (Phopivirus of Seals) related to human hepatitis A virus. MBio 6:e01180CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Asher LV, Binn LN, Mensing TL, Marchwicki RH, Vassell RA, Young GD (1995) Pathogenesis of hepatitis A in orally inoculated owl monkeys (Aotus trivirgatus). J Med Virol 47:260–268CrossRefPubMedGoogle Scholar
  4. 4.
    Balayan MS (1992) Natural hosts of hepatitis A virus. Vaccine 10(Suppl 1):S27–S31CrossRefPubMedGoogle Scholar
  5. 5.
    Bishop NE, Hugo DL, Borovec SV, Anderson DA (1994) Rapid and efficient purification of hepatitis A virus from cell culture. J Virol Methods 47:203–216CrossRefPubMedGoogle Scholar
  6. 6.
    Deinhardt F (1992) Prevention of viral hepatitis A: past, present and future. Vaccine 10(Suppl 1):S10–S14CrossRefPubMedGoogle Scholar
  7. 7.
    Dienstag JL, Feinstone SM, Purcell RH, Hoofnagle JH, Barker LF, London WT, Popper H, Peterson JM, Kapikian AZ (1975) Experimental infection of chimpanzees with hepatitis A virus. J Infect Dis 132:532–545CrossRefPubMedGoogle Scholar
  8. 8.
    Drexler JF, Corman VM, Lukashev AN, van den Brand JM, Gmyl AP, Brunink S, Rasche A, Seggewibeta N, Feng H, Leijten LM, Vallo P, Kuiken T, Dotzauer A, Ulrich RG, Lemon SM, Drosten C (2015) Evolutionary origins of hepatitis A virus in small mammals. Proc Natl Acad Sci USA 112:15190–15195CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Feigelstock D, Thompson P, Mattoo P, Zhang Y, Kaplan GG (1998) The human homolog of HAVcr-1 codes for a hepatitis A virus cellular receptor. J Virol 72:6621–6628PubMedPubMedCentralGoogle Scholar
  10. 10.
    Feng Z, Hensley L, McKnight KL, Hu F, Madden V, Ping L, Jeong SH, Walker C, Lanford RE, Lemon SM (2013) A pathogenic picornavirus acquires an envelope by hijacking cellular membranes. Nature 496:367–371CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Glikson MGE, Oren R, Tur-Kaspa R, Shouval D (1992) Relapsing hepatitis A. Review of 14 cases and literature survey. Medicine (Baltimore) 71:14–23CrossRefGoogle Scholar
  12. 12.
    Gluud LL, Gluud C (2009) Meta-analyses on viral hepatitis. Infect Dis Clin N Am 23:315–330CrossRefGoogle Scholar
  13. 13.
    Hirai-Yuki A, Hensley L, McGivern DR, Gonzalez-Lopez O, Das A, Feng H, Sun L, Wilson JE, Hu F, Feng Z, Lovell W, Misumi I, Ting JP, Montgomery S, Cullen J, Whitmire JK, Lemon SM (2016) MAVS-dependent host species range and pathogenicity of human hepatitis A virus. Science 353:1541–1545CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Hornei B, Kammerer R, Moubayed P, Frings W, Gauss-Muller V, Dotzauer A (2001) Experimental hepatitis A virus infection in guinea pigs. J Med Virol 64:402–409CrossRefPubMedGoogle Scholar
  15. 15.
    Jiang YJ, Liao GY, Zhao W, Sun MB, Qian Y, Bian CX, Jiang SD (2004) Detection of infectious hepatitis A virus by integrated cell culture/strand-specific reverse transcriptase-polymerase chain reaction. J Appl Microbiol 97:1105–1112CrossRefPubMedGoogle Scholar
  16. 16.
    Krawczynski KK, Bradley DW, Murphy BL, Ebert JW, Anderson TE, Doto IL, Nowoslawski A, Duermeyer W, Maynard JE (1981) Pathogenetic aspects of hepatitis A virus infection in enterally inoculated marmosets. Am J Clin Pathol 76:698–706CrossRefPubMedGoogle Scholar
  17. 17.
    Lanford RE, Feng Z, Chavez D, Guerra B, Brasky KM, Zhou Y, Yamane D, Perelson AS, Walker CM, Lemon SM (2011) Acute hepatitis A virus infection is associated with a limited type I interferon response and persistence of intrahepatic viral RNA. Proc Natl Acad Sci USA 108:11223–11228CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Lemon SM, Binn LN, Marchwicki R, Murphy PC, Ping LH, Jansen RW, Asher LV, Stapleton JT, Taylor DG, LeDuc JW (1990) In vivo replication and reversion to wild type of a neutralization-resistant antigenic variant of hepatitis A virus. J Infect Dis 161:7–13CrossRefPubMedGoogle Scholar
  19. 19.
    Major MEDH, Mihalik K, Puig M, Rice CM, Neumann AU, Feinstone SM (2004) Hepatitis C virus kinetics and host responses associated with disease and outcome of infection in chimpanzees. Hepatology 39:1709–1720CrossRefPubMedGoogle Scholar
  20. 20.
    Martin A, Lemon SM (2006) Hepatitis A virus: from discovery to vaccines. Hepatology 43:S164–S172CrossRefPubMedGoogle Scholar
  21. 21.
    Mathiesen LR, Drucker J, Lorenz D, Wagner JA, Gerety RJ, Purcell RH (1978) Localization of hepatitis A antigen in marmoset organs during acute infection with hepatitis A virus. J Infect Dis 138:369–377CrossRefPubMedGoogle Scholar
  22. 22.
    Mathiesen LR, Moller AM, Purcell RH, London WT, Feinstone SM (1980) Hepatitis A virus in the liver and intestine of marmosets after oral inoculation. Infect Immun 28:45–48PubMedPubMedCentralGoogle Scholar
  23. 23.
    Munne MS, Canero Velasco MC, Moreiro R, Vladimirsky S, Otegui L, Castro R, Brajterman L, Soto S, Mutti J, Nucifora S, Lara E, Sosa A, Godoy P, Ciocca M, Cuarterolo M, Quarleri JF, Gonzalez JE (2006) Duration of viremia and fecal shedding of the virus in hepatitis A infected children. Acta Gastroenterol Latinoam 36:182–189PubMedGoogle Scholar
  24. 24.
    Pinto MA, Marchevsky RS, Baptista ML, de Lima MA, Pelajo-Machado M, Vitral CL, Kubelka CF, Pissurno JW, Franca MS, Schatzmayr HG, Gaspar AM (2002) Experimental hepatitis A virus (HAV) infection in Callithrix jacchus: early detection of HAV antigen and viral fate. Exp Toxicol Pathol 53:413–420CrossRefPubMedGoogle Scholar
  25. 25.
    Schulman AN, Dienstag JL, Jackson DR, Hoofnagle JH, Gerety RJ, Purcell RH, Barker LF (1976) Hepatitis A antigen particles in liver, bile, and stool of chimpanzees. J Infect Dis 134:80–84CrossRefPubMedGoogle Scholar
  26. 26.
    Shevtsova ZV, Lapin BA, Doroshenko NV, Krilova RI, Korzaja LI, Lomovskaya IB, Dzhelieva ZN, Zairov GK, Stakhanova VM, Belova EG et al (1988) Spontaneous and experimental hepatitis A in Old World monkeys. J Med Primatol 17:177–194PubMedGoogle Scholar
  27. 27.
    Stapleton JT, Jansen R, Lemon SM (1985) Neutralizing antibody to hepatitis A virus in immune serum globulin and in the sera of human recipients of immune serum globulin. Gastroenterology 89:637–642CrossRefPubMedGoogle Scholar
  28. 28.
    Summers J, Smolec JM, Snyder R (1978) A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci USA 75:4533–4537CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Thimme RBJ, Spangenberg HC, Wieland S, Pemberton J, Steiger C, Govindarajan S, Purcell RH, Chisari FV (2002) Viral and immunological determinants of hepatitis C virus clearance, persistence, and disease. Proc Natl Acad Sci 99:15661–15668CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Wong DC, Shih JW, Purcell RH, Gerin JL, London WT (1982) Natural and experimental infection of woodchucks with woodchuck hepatitis virus, as measured by new, specific assays for woodchuck surface antigen and antibody. J Clin Microbiol 15:484–490PubMedPubMedCentralGoogle Scholar
  31. 31.
    Yu JM, Li LL, Zhang CY, Lu S, Ao YY, Gao HC, Xie ZP, Xie GC, Sun XM, Pang LL, Xu JG, Lipkin WI, Duan ZJ (2016) A novel hepatovirus identified in wild woodchuck Marmota himalayana. Sci Rep 6:22361CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  • Jie-mei Yu
    • 1
  • Li-li Li
    • 1
  • Guang-cheng Xie
    • 1
  • Cui-yuan Zhang
    • 1
  • Yuan-yun Ao
    • 1
  • Zhao-jun Duan
    • 1
  1. 1.National Institute for Viral Disease Control and Prevention, CDC ChinaBeijingChina

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