Food and Environmental Virology

, Volume 2, Issue 3, pp 176–182 | Cite as

Occurrence of Norovirus and Hepatitis A Virus in U.S. Oysters

  • Jacquelina W. Woods
  • William BurkhardtIII
Original Paper


Noroviruses (NoV) and hepatitis A virus (HAV) are the leading causes of non-bacterial gastroenteritis in shellfish consumers worldwide. This study determined the seasonal and geographical distribution of NoV (genogroups I and II) and HAV in live U.S. market oysters. Samples were analyzed to determine the occurrence and levels of NoV and HAV using RT-qPCR and conventional RT-PCR. NoV and HAV were detected in 3.9 and 4.4%, respectively. NoV genogroups I and II were detected, with genogroup II predominating. Sequencing identified genotypes II.4, II.3, and II.7. The GII.4 strain showed ≥98% similarity with 2006–2007 circulating strains, Minerva and Laurens. HAV sequences from the 5′ non-coding region (NCR) of the genome were from genotypes I, II, or III. The incidence of NoV in oysters harvested from Atlantic Coast states was higher than that in oysters from other regions and its occurrence was greatest during the cooler months (December to February). HAV was detected at a higher frequency in shellfish harvested from the Gulf Coast and also predominated during cooler months. The seasonal occurrence of viruses in this study corresponded to the reported incidence of shellfish-associated viral illnesses. This investigation provides an overview of the occurrence and distribution of NoV and HAV in U.S. market shellfish.


Norovirus Hepatitis A virus Market survey Virus detection 



We would like to thank the GCSL retail study team (Angelo DePaola, Jessica Jones, Kevin Calci, and Jeffrey Krantz); individuals at participating FDA-ORA laboratories (Emily Jacobs and Angela Swinford—FDA SRL, Khamphet Nabe and June Wetherington—FDA PRL-NW, Kuppuswamy Kasturi and Jose Obano—FDA NRL, and Donna Hill-Williams and Nelly Tran—FDA PRL-SW. We thank the ISSC for financial support and FDA personnel and individuals from the collecting states. We would also like to thank Susan McCarthy for review of this manuscript.


  1. Ando, T., Noel, J. S., & Fankhauser, R. L. (2000). Genetic classification of Norwalk-like viruses. Journal of Infectious Disease, 181, S336–S348.CrossRefGoogle Scholar
  2. Beuret, C., Kohler, D., Baumgartner, A., & Luthi, T. M. (2002). Norwalk-live virus sequences in mineral waters: one year monitoring of three brands. Applied and Environmental Microbiology, 68, 1925–1931.CrossRefPubMedGoogle Scholar
  3. Blanton, L. H., Adams, S. M., Beard, R. S., Wei, G., Bulens, S. N., Widdowson, M., et al. (2006). Molecular and epidemiologic trends of Calicivirus associated with outbreaks of acute gastroenteritis in the United States, 2000–2004. Journal of Infectious Disease, 183, 413–421.CrossRefGoogle Scholar
  4. Burkhardt III, W., Calci, K. R., Watkins, W. W., Rippey, S. R., & Chirtel, S. J. (2000). Inactivation of indicator microorganisms in estuarine waters. Water Research, 34, 2207–2214.CrossRefGoogle Scholar
  5. Burkhardt III, W., Woods, J. W., Nordstrom, J., & Hartman, G. (2006). A realtime RT-PCR protocol for the simultaneous detection of norovirus and enterovirus. Laboratory information bulletin #4369. Washington, DC: U.S. Food and Drug Administration.Google Scholar
  6. Centers for Disease Control and Prevention (CDC). (2006). Prevention of specific infectious diseases: hepatitis, viral, type A. In Travelers’ health: Yellow book. Health information for international travel, 2005–2006. Available at: Accessed March 2008.
  7. Center for Disease Control and Prevention (CDC). (2007). Norovirus activity—United States, 2006–2007. Morbidity and Mortality Weekly Report, 56, 842–846.Google Scholar
  8. Center for Disease Control and Prevention (CDC). (2009). Surveillance for foodborne outbreaks—United States. Morbidity and Mortality Weekly Report, 38, 609–615.Google Scholar
  9. Constantini, V., Loisy, F., Jones, L., LeGuyader, F. S., & Saif, L. J. (2006). Human and animal enteric caliciviruses in oysters from different coastal regions of the United States. Applied and Environmental Microbiology, 72, 1800–1809.CrossRefGoogle Scholar
  10. Costa-Mattioli, M., Di Napoli, A., Ferre, V., Billaudel, S., Perez-Bercoff, R., & Cristina, J. (2003). Genetic variability of hepatitis A virus. Journal of General Virology, 84, 3191–3201.CrossRefPubMedGoogle Scholar
  11. Depaola, A., Jones, J. L., Woods, J. W., Burkhardt III, W., Calci, K. R., Krantz, J. A., Bowers, J. C., Kastrui, K., Byars, R. H., Jacobs, E., Hill-Williams, D., & Nabe, K. (2010). Bacterial and viral pathogens in live oysters: U.S. markets survey 2007. Applied and Environmental Microbiology. doi: 10.1128/AEM.02590-09.
  12. Desenclos, J. C., Klontz, K. C., Wilder, M. H., Nainan, O. V., Margolis, H. S., & Gunn, R. A. (1991). A multistate outbreak of hepatitis A caused by the consumption of raw oysters. American Journal of Public Health, 81, 1268–1272.CrossRefPubMedGoogle Scholar
  13. Ellender, R. D., Mapp, J. B., Middlebrooks, B. C., Cooks, D. W., & Cake, E. W. (1980). Natural enterovirus and fecal coliform contamination of Gulf coast oysters. Journal of Food Protection, 43, 105–110.Google Scholar
  14. Flannery, J., Keaveney, S., & Dore, W. (2009). Use of FRNA bacteriophage to indicate the risk of contamination in Irish oysters. Journal of Food Protection, 72, 2358–2362.PubMedGoogle Scholar
  15. Frost, H. W. (1925). Report of committee on the sanitary control of the shellfish industry in the United States. Public Health Reports, 53(Suppl), 1–17.Google Scholar
  16. Gardner, S. N., Kuczmarski, T. A., Vitalis, E. A., & Slezar, T. R. (2003). Limitations of Taqman PCR for detecting divergent viral pathogens illustrated by hepatitis A, B, C, and E viruses and human immunodeficiency virus. Journal of Clinical Microbiology, 41, 2417–2427.CrossRefPubMedGoogle Scholar
  17. Goyal, S. M., Gerba, C. P., & Melnick, J. L. (1979). Human enteroviruses in oysters and their overlying waters. Applied and Environmental Microbiology, 37, 572–581.PubMedGoogle Scholar
  18. Hall, T. (1999). BioEdit, a user friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acid Symposium Series, 41, 95–98.Google Scholar
  19. Hutin, Y. J. F., Pool, V., Cramer, E. H., Nainan, O. V., Weth, J., Williams, I. T., et al. (1999). A multistate, foodborne outbreak of hepatitis A. New England Journal of Medicine, 340, 595–602.CrossRefPubMedGoogle Scholar
  20. Jansen, R., Siegl, G., & Lemon, S. M. (1990). Molecular epidemiology of human hepatitis A virus defined by an antigen-capture polymerase chain reaction method. Proceedings of the National Academy of Sciences United States of America, 87, 2867–2871.CrossRefGoogle Scholar
  21. Joshi, M. S., Walimbe, A. M., & Chitambar, S. D. (2008). Evaluation of genomic regions of hepatitis A virus for phylogenetic anlaysis: Suitability of the 2C region for genotyping. Journal of Virological Methods, 153, 36–42.CrossRefPubMedGoogle Scholar
  22. Kageyama, T., Kojima, S., Shinohara, M., Uchida, K., Fukushi, S., Hoshino, F. B., et al. (2003). Broadly reactive and highly sensitive assay for Norwalk-like viruses based on real-time quantitative reverse transcription-PCR. Journal Clinical Microbiology, 41, 1548–1557.CrossRefGoogle Scholar
  23. Kingsley, D., Mead, G. K., & Richards, G. P. (2002). Detection of both hepatitis A and Norwalk-like virus in imported clams associated with food-borne illnesses. Applied and Environmental Microbiology, 68, 3914–3918.CrossRefPubMedGoogle Scholar
  24. Koopmans, M., & Duizer, E. (2004). Foodborne viruses: An emerging problem. International Journal of Food Microbiology, 90, 23–41.CrossRefPubMedGoogle Scholar
  25. Kroneman, A., Verhoef, L., Harris, J., Vennema, H., Duizer, E., van Duynhoven, Y., et al. (2008). Analysis of integrated virological and epidemiological reports of norovirus outbreaks collected within the foodborne viruses in Europe network from 1 July 2001 to 30 June 2006. Journal of Clinical Microbiology, 46, 2959–2965.CrossRefPubMedGoogle Scholar
  26. Lees, D. (2000). Viruses and bivalve shellfish. International Journal of Food Microbiology, 59, 81–116.CrossRefPubMedGoogle Scholar
  27. Lowther, J. A., Avant, J. M., Gizynski, K., Rangdale, R. E., & Lees, D. N. (2010). Comparison between quantitative real-time reverse transcription PCR results for norovirus in oysters and self-reported gastroenteric illness in restaurant customers. Journal of Food Protection, 73, 305–311.PubMedGoogle Scholar
  28. Lynch, M., Painter, J., Woodruff, R., & Braden, C. (2006). Surveillance for foodborne disease outbreaks—United States—1998–2002. MMWR, 55, 1–34.PubMedGoogle Scholar
  29. Mead, P. S., Slutsker, L., Dietz, V., McCaig, L. F., Bresee, J. S., Shapiro, C., et al. (1999). Food-related illness and death in the United States. Emerging Infectious Disease, 5, 607–625.CrossRefGoogle Scholar
  30. Mullendore, J. L., Sobsey, M. D., & Shieh, Y. C. (2001). Improved method for the recovery of hepatitis A virus from oysters. Journal of Virological Methods, 94, 25–35.CrossRefPubMedGoogle Scholar
  31. Nainan, O. V., Armstrong, G. L., Han, X.-H., Williams, I., Bell, B. P., & Margolis, H. S. (2005). Hepatitis a molecular epidemiology in the United States, 1996–1997: Sources of infection and implications of vaccination policy. Journal of Infectious Disease, 191, 957–963.CrossRefGoogle Scholar
  32. Nainan, O. V., Xia, G., Vaughan, G., & Margolis, H. S. (2006). Diagnosis of a hepatitis A virus infection: A molecular approach. Clinical Microbiology Reviews, 19, 63–79.CrossRefPubMedGoogle Scholar
  33. Pan, C., & Schnurr, D. (2008). Viral gastroenteritis (norovirus) outbreaks: 2007–2008. Resource document. Viral and Rickettsial Disease Laboratory, California Department of Public Health. (norovirus activity 2007-2008).
  34. Patel, M. M., Hall, A. J., Vinje, J., & Parashar, U. D. (2009). Noroviruses: A comprehensive review. Journal of Clinical Virology, 44, 1–8.CrossRefPubMedGoogle Scholar
  35. Richards, G. P., Watson, M. A., Fankhauser, R. L., & Monroe, S. S. (2004). Genogroup I and genogroup II norovirus detection in stool samples by real-time reverse transcription PCR using highly degenerative universal primers. Applied and Environmental Microbiology, 70, 7179–7184.CrossRefPubMedGoogle Scholar
  36. Robertson, B. H., Jansen, R. W., Khanna, B., Totsuka, A., Nainan, O. V., Siegl, G. A., et al. (1992). Genetic relatedness of hepatitis A virus strains recovered from different geographical regions. Journal of General Virology, 73, 1365–1377.CrossRefPubMedGoogle Scholar
  37. Schwab, K. J., Estes, M. K., Neill, F. H., & Atmar, R. L. (1997). Use of heat release and an internal RNA standard control in reverse transcription-PCR detection of Norwalk virus from stool samples. Journal of Clinical Microbiology, 35, 511–514.PubMedGoogle Scholar
  38. Shieh, Y. C., Khudyakov, Y. E., Xia, G., Ganova-Raeva, L. M., Khambaty, F., Woods, J. W., et al. (2007). Molecular confirmation of oysters as the vector for hepatitis A in a 2005 multistate outbreak. Journal Food Protection, 70, 145–150.Google Scholar
  39. Shieh, Y. C., Woods, J. W., & Calci, K. R. (2003). Molecular surveillance of enterovirus and Norwalk-like virus in oysters relocated to a municipal sewage-impacted gulf estuary. Applied and Environmental Microbiology, 69, 7130–7136.CrossRefPubMedGoogle Scholar
  40. Siebenga, J., Kronemen, A., Vennema, H., Duizer, E., & Koopmans, M. (2008). Food-borne viruses in Europe Network Report: The norovirus GII.4 2006b (for U.S. named Minerva-like, for Japan Kobe 034-like, for U.K. V6) variant now dominant in early seasonal surveillance. Eurosurveillance, 13, 1–4.Google Scholar
  41. Tamura, K., Dudley, J., Nei, M., & Kumar, S. (2007). Molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 8, 1596–1599.CrossRefGoogle Scholar
  42. Terio, V., Martella, P., Moschidou, P., Pinto, P. D., Tantillo, G., & Buonavoglia, C. (2010). Norovirus in retail shellfish. Food Microbiology, 27, 29–32.CrossRefPubMedGoogle Scholar
  43. Zheng, D., Ando, T., Fankhauser, R. R., Beard, R. S., Glass, R. I., & Monroe, S. S. (2006). Norovirus classification and proposed strain nomenclature. Virology, 346, 312–323.CrossRefPubMedGoogle Scholar

Copyright information

© U. S. Food and Drug Administration 2010

Authors and Affiliations

  1. 1.FDA Gulf Coast Seafood LaboratoryDauphin IslandUSA

Personalised recommendations