EXPERIMENTAL CONTAMINATION OF CHAMELEA GALLINA WITH MURINE NOROVIRUS AND EFFECTIVENESS OF DEPURATION

Main Article Content

M. BERTI
L. TEODORI
O. PORTANTI
A. LEONE
I. CARMINE
N. FERRI
P. VISCIANO
M. SCHIRONE
G. SAVINI

Keywords

Norovirus, clams, depuration, tissue culture, RT-PCR

Abstract

Human Norovirus has been reported as the major non-bacterial cause of human gastroenteritis due to the consumption of contaminated bivalve mollusks. The European legislation established microbiological criteria only for bacteria (Salmonella spp.  and Escherichia coli),  while no viruses have still been considered. In this study,  samples of Chamelea gallina were harvested along the Central  Adriatic coasts (Italy) and artificially contaminated with  Murine  norovirus-1 (MNV-1) up to a  final concentration of 103TCID50/ml  in water.  They were subject to a  depuration process in a  closed-circuit system using both ozone and ultraviolet light. Four experimental trials (100 specimens/trial) were performed and, at the end of depuration, the digestive glands of mollusks were examined by means of two methods –namely, RT-PCR and tissue culture. The results of RT-PCR ranged from 103.17 to 104.60 TCID50/ml, and the constant presence of MNV-1 was confirmed by the tissue culture as well. In conclusion,  no significant viral reduction was obtained, but the contaminated bivalve mollusks remained infectious until the end of the depuration treatment. The proper cooking of live bivalve mollusks could be considered the most important preventive measure against this sanitary risk.

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References

Baert L., Wobus C. E., Van Coillie E., Thackray L. B., Debevere J. and Uyttrndaele M. 2008. Detection of murine norovirus 1 by using plaque assay, transfection assay, and Real-Time Reverse Transcription-PCR before and after heat exposure. Appl. Environ. Microbiol. 74(2):543-546.

Campos C.J.A., Goblick G., Lee R., Wittamore K. and Lees D.N. 2017. Determining the zone of impact of norovirus contamination in shellfish production areas through microbiological monitoring and hydrographic analysis. Water Res. 124:556-565.

Chan M.C.W., Hu Y., Chen H., Podkolzin A.T., Zaytseva E.V., Komano J. et al. 2017. Global spread of Norovirus GII.17 Kawasaki 308, 2014-2016. Emerg. Infect. Dis. 23(8), 1350-1354.

Cheng H.Y., Hung M.N., Chen W.C., Lo Y.C., Su Y.S., Wei H.Y. et al. 2017. Ice-associated norovirus outbreak predominantly caused by GII.17 in Taiwan, 2015. BMC Public Health 17(870):1-8.

Colangeli P., Iannetti S., Ruocco L., Forlizzi L., Cioci D. and Calistri P. 2013. The Italian information system on zoonoses data collection. Zoonoses Public Health. 60(2):182-188.

EC 2004a. Commission Regulation (EC) No 854/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for food of animal origin. Off. J. Eur. Union L226:83-127.

EC 2004b. Commission Regulation (EC) No 853/2004 of the European Parliament and of the Council of 29 April 2004 laying down specific hygiene rules for on the hygiene of foodstuffs. Off. J. Eur. Union L226:22-82.

EC 2005. Commission Regulation (EC) No 2073/2005 of 15 November 2005 on microbiological criteria for foodstuffs. Off. J. Eur. Union L338:1-26.

EFSA (European Food Safety Authority) and ECDC (European Centre for Disease, Prevention and Control) 2017. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2016. EFSA J., 15(12), 5077:1-228.

Fusco G., Di Bartolo I., Cioffi B., Ianiro G., Palermo P., Monini M. and Amoroso M.G. 2017. Prevalence of foodborne viruses in mussels in Southern Italy. Food Environ. Virol. 9(2):187-194.

Hassard F., Sharp J. H., Taft H., LeVay L., Harris J.P., McDonald J.E. et al. 2017. Critical review on the public health impact of Norovirus contamination in shellfish and the environment: a UK perspective. Food Environ. Virol. 9:123-141.

Ilic N., Velebit B., Teodorovic V., Djordjevic V., Karabasil N., Vasilev D., Djuric S., Adzic B. and Dimitrijevic M. 2017. Influence of environmental conditions on Norovirus presence in mussels harvested in Montenegro. Food Environ. Virol. 9(4):406-414.

Imamura S., Haruna M., Goshima T., Kanezashi H., Okada T. and Akimoto K. 2016. Application of next-generation sequencing to evaluate the profile of Noroviruses in pre-and post-depurated oysters. Foodborne Pathog. Dis. 13(10):559-565.

Jeon S.B., Seo D.J., Oh H., Kingsley D.H. and Choi C. 2017. Development of one-step reverse transcription loop-mediated isothermal amplification for norovirus detection in oysters. Food Control 73:1002-1009.

Kim S.H., Shahbaz H.M., Park D., Chun S., Lee W., Oh J.W., Lee D.U. and Park J. 2017. A combined treatment of UV-assisted TiO2 photocatalysis and high hydrostatic pressure to inactivate internalized murine norovirus. Innov. Food Sci. Emerg. 39:188-196.

Le Mennec C., Pamaudeaus S., Rumebe M., Le Saux J.C., Piquet J.C. and Le Guyader S.F. 2017. Follow-up of Norovirus contamination in an oyster production area linked to repeated outbreaks. Food Environ. Virol. 9(1):54-61.

Leal Diego A.G., Dores Ramos A.P., Marques Souza D.S., Durigan M., Greinert-Goulart J.A., Moresco V. et al. 2013. Sanitary quality of edible bivalve mollusks in Southeastern Brazil using an UV based depuration system. Ocean Coast. Manage. 72:93-100.

Leroux-Roels G., Cramer J.P., Mendelman P.M., Sherwood J., Clemens R., Aerssens A. et al. 2018. Safety and immunogenicity of different formulations of norovirus vaccine candidate in healthy adults: a randomized, controlled, double-blind clinical trial. J. Infect. Dis. 217(4):597-607.

McLeod C., Polo D., Le Saux J.C. and Le Guyader F.S. 2017. Depuration and relaying: a review on potential removal of Norovirus from oysters. Compr. Rev. Food Sci. F. 16:692-706.

Papapanagiotou E.P. 2017. Foodborne Norovirus State of Affairs in the EU Rapid Alert System for Food and Feed. Vet. Sci. 4:61.

PPRIC 2015-2018. Piano Pluriennale Regionale Integrato dei Controlli della Sanità Pubblica Veterinaria e Sicurezza Alimentare della Regione Abruzzo 2015–2018. 3rd Edn. 1-892.

Polo D., Álvarez C., Díez J., Darriba S., Longa Á. and Romalde J.L. 2014a. Viral elimination during commercial depuration of shellfish. Food Control 43:206-212.

Polo D., Feal X., Varela M.F., Monteagudo A. and Romalde J.L. 2014b. Depuration kinetics of murine norovirus in shellfish. Food Res. Int. 64:182-187.

Predmore A., Sanglay G., Li J. and Lee K. 2015. Control of human norovirus surrogates in fresh foods by gaseous ozone and a proposed mechanism of inactivation. Food Microbiol. 50:118-125.

Reed L.J. and Müench H. 1938. A simple method of estimating fifty percent endpoints. Am. J. Epidemiol. 27:493-497.

Savini G., Casaccia C., Barile N.B., Paoletti M. and Pinoni C. 2009. Norovirus in bivalve molluscs: a study of the efficacy of the depuration system. Veterinaria Italiana 45(4):535-539.

Souza D.S.M., Piazza R.S., Pilotto M.R., Nascimento M.A., Moresco V., Taniguchi S. et al. 2013. Virus, protozoa and organic compounds decay in depurated oysters. Int. J. Food Microbiol. 167(3):337-345.

Suffredini E., Iaconelli M., Equestre M., Valdazo-González B., Ciccaglione A.R., Marcantonio C., Della Libera S., Bignami F. and La Rosa G. 2018. Genetic diversity among genogroup II noroviruses and progressive emergence of GII.17 in wastewaters in Italy (2011-2016) revealed by next-generation and sanger sequencing. Food Environ. Virol. 10(2):141-150.

Trivedi T.K., Desai R., Hall A.J., Patel M., Parashar U.D. and Lopman B.A. 2013. Clinical characteristics of norovirus-associated deaths: a systematic literature review. Am. J. Infect. Control 41:654-657.

Varela M.F., Polo D. and Romalde J.L. 2016. Prevalence and genetic diversity of human sapoviruses in shellfish from commercial production areas in Galicia, Spain. Appl. Environ. Microbiol. 82(4):1167-1172.