Abstract
A production area repeatedly implicated in oyster-related gastroenteritis in France was studied for several months over 2 years. Outbreaks and field samples were analyzed by undertaking triplicate extractions, followed by norovirus (NoV) detection using triplicate wells for genomic amplification. This approach allowed us to demonstrate that some variabilities can be observed for samples with a low level of contamination, but most samples analyzed gave reproducible results. At the first outbreak, implicated oysters were collected at the beginning of the contamination event, which was reflected by the higher NoV levels during the first month of the study. During the second year, NoV concentrations in samples implicated in outbreaks and collected from the production area were similar, confirming the failure of the shellfish depuration process. Contamination was detected mainly during winter-spring months, and a high prevalence of NoV GI contamination was observed. A half-life of 18 days was calculated from NoV concentrations detected in oysters during this study, showing a very slow decrease of the contamination in the production area. Preventing the contamination of coastal waters should be a priority.
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References
Afnor (2011). Methode d’analyse en santé animale, PCR, part 2: Exigences et recommandations pour le développement et la validation de la PCR en santé animale. XPU47-600-2.
Araud, E., DiCaprio, E., Ma, Y., Lou, F., Gao, Y., Kingsley, D., et al. (2016). Thermal inactivation of enteric viruses and bioaccumulation of enteric foodborne viruses in live oysters (Crassostrea virginica). Applied and Environmental Microbiology, 82, 2099–2986.
Atmar, R. L., Neill, F. H., Romalde, J. L., Le Guyader, F., Woodley, C. M., Metcalf, T. G., et al. (1995). Detection of Norwalk virus and hepatitis A virus in shellfish tissues with the PCR. Applied and Environmental Microbiology, 61, 3014–3018.
Atmar, R. L., Opekum, A. R., Gilger, M. A., Estes, M. K., Crawford, S. E., Neill, F. H., et al. (2008). Norwalk virus shedding after experimental human infection. Emerging and Infectious Disease, 14, 1553–1557.
Atmar, R. L., Opekum, A. R., Gilger, M. A., Estes, M. K., Crawford, S. E., Neill, F. H., et al. (2014). Determination of the 50% human infectious dose for Norwalk virus. Journal of Infectious Disease, 209, 1016–1022.
Boxman, I., Verhoef, L., Vennema, H., Ngui, S., Friesema, I. H., Whiteside, C., et al. (2016). International linkage of two food-borne hepatitis A clusters through traceback of mussels, the Netherlands, 2012. Eurosurveillance, 21(3), 30113.
Choi, C., & Kingsley, D. H. (2016). Temperature-dependent persistence of human norovirus within oysters (Crassostrea virginica). Food and Environmental Virology, 8, 141–147.
da Silva, A., Le Saux, J.-C., Parnaudeau, S., Pommepuy, M., Elimelech, M., & Le Guyader, F. S. (2007). Evaluation of removal of noroviruses during wastewater treatment, using real-time reverse transcription-PCR: Different behaviors of genogroups I and II. Applied and Environmental Microbiology, 73, 7891–7897.
Dore, B., Keaveney, S., Flannery, J., & Rajko-Nenow, P. (2010). Managment of health risks associated with oysters harvested from a norovirus contaminated area, Ireland, February–March 2010. Eurosurveillance, 15, 10–13.
Drouaz, N., Schaeffer, J., Farkas, T., Le Pendu, J., & Le Guyader, F. S. (2016). Tulane virus as a potential surrogate to mimic norovirus behavior in oysters. Applied and Environmental Microbiology, 81, 5256–8249.
Grodzki, M., Schaeffer, J., Piquet, J.-C., Le Saux, J.-C., Chevé, J., Ollivier, J., et al. (2014). Bioaccumulation efficiency, tissue distribution, and environmental occurrence of hepatitis E virus in bivalve shellfish from France. Applied and Environmental Microbiology, 80, 4269–4276.
ISO/TS 15216-1 (2013). Microbiology of food and animal feed-horizontal method for determinaion of hepatitis A virus and norovirus in food using real-time RT-PCR, part 1: Method for quantification.
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 of Clinical Microbiology, 41, 154–157.
Le Guyader, F. S., Atmar, R. L., & Le Pendu, J. (2012). Transmission of viruses through shellfish: When specific ligands come into play. Current Opinion in Virology, 2, 103–110.
Le Guyader, F. S., Le Saux, J.-C., Ambert-Balay, K., Krol, J., Serais, O., Parnaudeau, S., et al. (2008). Aichi virus, norovirus, astrovirus, enterovirus and rotavirus involved in clinical cases from a French oyster-related gastroenteritis outbreak. Journal of Clinical Microbiology, 46, 4011–4017.
Le Guyader, F. S., Parnaudeau, S., Schaeffer, J., Bosch, A., Loisy, F., Pommepuy, M., et al. (2009). Detection and quantification of noroviruses in shellfish. Applied and Environmental Microbiology, 74, 618–624.
Loisy, F., Atmar, R. L., Guillon, P., Le Cann, P., Pommepuy, M., & Le Guyader, F. S. (2007). Real-time RT-PCR for norovirus screening in shellfish. Journal of Virological Methods, 123, 1–7.
Maalouf, H., Schaeffer, J., Parnaudeau, S., Le Pendu, J., Atmar, R. L., Crawford, S. E., et al. (2011). Strain-dependent norovirus bioaccumulation in oysters. Applied and Environmental Microbiology, 77, 3189–3196.
Metcalf, T., Melnick, J. L., & Estes, M. K. (1995). Environmental Microbiology: From detection of virus in sewage and water by isolation to identification by molecular biology-a trip of over 50 years. Annual Review in Microbiology, 49, 461–487.
Miura, T., Lhomme, S., Le Saux, J.-C., Le Mehaute, Ph, Guillois, Y., Couturier, E., et al. (2016). Detection of hepatitis E virus in sewage after an outbreak on a French island. Food and Environmental Virology, 8, 194–199.
Polo, D., Alvarez, C., Diez, J., Darriba, S., Longa, A., & Romalde, J. L. (2014). Viral elimination during commercial depuration of shellfish. Food Control, 43, 206–212.
Ramani, S., Atmar, R. L., & Estes, M. K. (2014). Epidemiology of human noroviruses and updates on vaccine development. Current Opinion in Gastroenterology, 30, 25–33.
Robilotti, E., Deresinski, S., & Pinsky, B. A. (2015). Norovirus. Clinical and Microbiology Review, 28, 134–164.
Svraka, S., Duizer, E., Vennema, H., de Bruin, E., van der Veer, B., Dorredteijn, B., et al. (2007). Etiological role of viruses in outbreaks of acute gastroenteritis in the Netherlands from 1994 through 2005. Journal of Clinical Microbiology, 45, 1389–1394.
Teunis, P. F. M., Moe, C. L., Liu, P., Miller, S. E., Lindesmith, L., Baric, R. S., et al. (2008). Norwalk virus: How infectious is it? Journal of Medical Virology, 80, 1468–1476.
Teunis, P. F. M., Sukhrie, F. H. A., Vennema, H., Bogerman, J., Beersma, M. F. C., & Koopmans, M. P. G. (2015). Shedding of norovirus in symptomatic and asymptomatic infections. Epidemiology and Infection, 143, 1710–1717.
Uhrbrand, K., Myrmel, M., Maunula, L., Vainio, K., Trebbien, R., Norrung, B., et al. (2010). Evaluation of a rapid method for recovery of norovirus and hepatitis A virus from oysters and blue mussels. Journal of Virological Methods, 169, 70–78.
Vinje, J. (2015). Advances in laboratory methods for detection and typing of norovirus. Journal of Clinical Microbiology, 53, 373–381.
Woods, J. W., & Burkhardt, W., III. (2010). Occurence of norovirus and hepatitis A virus in US oysters. Food and Environmental Virology, 2, 176–182.
Yu, Y., Cai, H., Hu, L., Lei, R., Pan, Y., Yan, S., et al. (2015). Molecular epidemiology of oyster-related human noroviruses and their global genetic diversity and temporal-geographical distribution from 1983 to 2014. Applied and Environmental Microbiology, 81, 7615–7624.
Acknowledgments
This study was supported by grants 2013 and 2015 from DGAl (Direction Generale de l’Alimentation) and by the EU-H2020 Grant N°643476 COMPARE. We are grateful to Catherine McLeod (Seafood Safety Assessment) for a critical review of the manuscript.
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Cécile Le Mennec and Sylvain Parnaudeau have contributed equally to the work.
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Le Mennec, C., Parnaudeau, S., Rumebe, M. et al. Follow-Up of Norovirus Contamination in an Oyster Production Area Linked to Repeated Outbreaks. Food Environ Virol 9, 54–61 (2017). https://doi.org/10.1007/s12560-016-9260-6
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DOI: https://doi.org/10.1007/s12560-016-9260-6