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Harmonised Investigation of the Occurrence of Human Enteric Viruses in the Leafy Green Vegetable Supply Chain in Three European Countries

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Abstract

Numerous outbreaks have been attributed to the consumption of raw or minimally processed leafy green vegetables contaminated with enteric viral pathogens. The aim of the present study was an integrated virological monitoring of the salad vegetables supply chain in Europe, from production, processing and point-of-sale. Samples were collected and analysed in Greece, Serbia and Poland, from ‘general’ and ‘ad hoc’ sampling points, which were perceived as critical points for virus contamination. General sampling points were identified through the analysis of background information questionnaires based on HACCP audit principles, and they were sampled during each sampling occasion where as-ad hoc sampling points were identified during food safety fact-finding visits and samples were only collected during the fact-finding visits. Human (hAdV) and porcine (pAdV) adenovirus, hepatitis A (HAV) and E (HEV) virus, norovirus GI and GII (NoV) and bovine polyomavirus (bPyV) were detected by means of real-time (RT-) PCR-based protocols. General samples were positive for hAdV, pAdV, HAV, HEV, NoV GI, NoV GII and bPyV at 20.09 % (134/667), 5.53 % (13/235), 1.32 % (4/304), 3.42 % (5/146), 2 % (6/299), 2.95 % (8/271) and 0.82 % (2/245), respectively. Ad hoc samples were positive for hAdV, pAdV, bPyV and NoV GI at 9 % (3/33), 9 % (2/22), 4.54 % (1/22) and 7.14 % (1/14), respectively. These results demonstrate the existence of viral contamination routes from human and animal sources to the salad vegetable supply chain and more specifically indicate the potential for public health risks due to the virus contamination of leafy green vegetables at primary production.

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References

  • Albinana-Gimenez, N., Miagostovich, M. P., Calgua, B., Huguet, J. M., Matia, L., & Girones, R. (2009). Analysis of adenoviruses and polyomaviruses quantified by qPCR as indicators of water quality in source and drinking-water treatment plants. Water Research, 43(7), 2011–2019.

    Article  PubMed  CAS  Google Scholar 

  • Ambrožič, M., Božič, T., Jevšnik, M., Cook, N., & Raspor, P. (2011). Compliance of proposed Codex Alimentarius guidelines for virus management with principles of good practice. Acta Alimentaria, 40, 364–375.

    Article  Google Scholar 

  • Berger, C. N., Sodha, S. V., Shaw, R. K., Griffin, P. M., Pink, D., Hand, P., et al. (2010). Fresh fruit and vegetables as vehicles for the transmission of human pathogens. Environmental Microbiology, 12(9), 2385–2397. doi:10.1111/j.1462-2920.2010.02297.x.

    Google Scholar 

  • Berto, A., Martelli, F., Grierson, S., & Banks, M. (2012). Hepatitis E virus in pork food chain, United Kingdom. Emerging Infectious Diseases, 18(8), 1358–1360. doi:10.3201/eid1808.111647.

  • Bidawid, S., Farber, J. M., & Sattar, S. A. (2000). Contamination of foods by food handlers: Experiments on hepatitis A virus transfer to food and its interruption. Applied and Environmental Microbiology, 6(7), 2759–2763.

    Article  Google Scholar 

  • Bofill-Mas, S., Albiñana-Gimenez, N., Clemente-Casares, P., Hundesa, A., Rodriguez-Manzano, J., Allard, A., et al. (2006). Quantification and stability of human adenoviruses and polyomavirus JCPyV in wastewater matrices. Applied and Environmental Microbiology, 72, 7894–7896.

    Article  PubMed  CAS  Google Scholar 

  • Bofill-Mas, S., Calgua, B., Clemente-Casares, P., La Rosa, G., Iaconelli, M., Muscillo, M., et al. (2010). Quantification of human adenoviruses in European recreational waters. Food and Environmental Virology, 2, 101–109.

    Article  CAS  Google Scholar 

  • Cheong, S., Lee, C., Song, S. W., Choi, W. C., Lee, C. H., & Kim, S. J. (2009). Enteric viruses in raw vegetables and groundwater used for irrigation in South Korea. Applied and Environmental Microbiology, 75(24), 7745–7751.

    Article  PubMed  CAS  Google Scholar 

  • Costafreda, M. I., Bosch, A., & Pintó, R. M. (2006). Development, evaluation, and standardization of a real-time TaqMan reverse transcription-PCR assay for quantification of hepatitis A virus in clinical and shellfish samples. Applied and Environment Microbiology, 72(6), 3846–3855.

    Article  CAS  Google Scholar 

  • Croci, L., De Medici, D., Scalfaro, C., Fiore, A., & Toti, L. (2002). The survival of hepatitis A virus in fresh produce. International Journal of Food Microbiology, 73(1), 29–34.

    Article  PubMed  CAS  Google Scholar 

  • Croci, L., Dubois, E., Cook, N., de Medici, D., Schultz, A. C., China, B., et al. (2008). Current methods for extraction and concentration of enteric viruses from fresh fruit and vegetables: Towards international standards. Food Analytical Methods, 1, 73–84.

    Article  Google Scholar 

  • D’Agostino, M., Cook, N., Di Bartolo, I., Ruggeri, F. M., Berto, A., Martelli, F., et al. (2012). Multicenter collaborative trial evaluation of a method for detection of human adenoviruses in berry fruit. Food Analytical Methods, 5, 1–7.

    Article  Google Scholar 

  • D’Agostino, M., Cook, N., Rodriguez-Lazaro, D., & Rutjes, S. (2011). Nucleic acid amplification-based methods for detection of enteric viruses: Definition of controls and interpretation of results. Food and Environmental Virology, 2, 55–60.

    Article  Google Scholar 

  • Da Silva, A. K., 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(24), 7891–7897.

    Article  PubMed  Google Scholar 

  • Dentinger, C. M., Bower, W. A., Nainan, O. V., Cotter, S. M., Myers, G., Dubusky, L. M., et al. (2001). An outbreak of hepatitis A associated with green onions. Journal of Infectious Diseases, 183, 1273–276.

    Google Scholar 

  • Diez-Valcarce, M., Cook, N., Hernández, M., & Rodríguez-Lázaro, D. (2011a). Analytical application of a sample process control in detection of foodborne viruses. Food Analytical Methods, 4, 614–618. doi:10.1007/s12161-011-9262-9.

  • Diez-Valcarce, M., Kovač, K., Cook, N., Hernández, M., & Rodríguez-Lázaro, D. (2011b). Construction and analytical application of internal amplification controls (IAC) for detection of food supply chain relevant viruses by real-time PCR-based assays. Food Analytical Methods, 4, 437–445.

    Article  Google Scholar 

  • Dubois, E., Hennechart, C., Deboosère, N., Merle, G., Legeay, O., Burger, C., et al. (2006). Intra-laboratory validation of a concentration method adapted for the enumeration of infectious F-specific RNA coliphage, enterovirus, and hepatitis A virus from inoculated leaves of salad vegetables. International Journal of Food Microbiology, 108(2), 164–171.

    Article  PubMed  CAS  Google Scholar 

  • FAO/WHO [Food and Agriculture Organization of the United Nations/World Health Organization]. (2008). Microbiological hazards in fresh leafy vegetables and herbs: Meeting Report. Microbiological Risk Assessment Series No. 14. Rome. 151 pp.

  • Fino, V. R., & Kniel, K. E. (2008). Comparative recovery of foodborne viruses from fresh produce. Foodborne Pathogens and Disease, 5(6), 819–825.

    Article  PubMed  Google Scholar 

  • Formiga-Cruz, M., Tofiño-Quesada, G., Bofill-Mas, S., Lees, D. N., Henshilwood, K., Allard, A. K., et al. (2002). Distribution of human viral contamination in shellfish from different growing areas in Greece, Spain, Sweden and UK. Applied and Environmental Microbiology, 68(12), 5990–5998.

    Article  PubMed  CAS  Google Scholar 

  • Gandhi, K. M., Mandrell, R. E., & Tian, P. (2010). Binding of virus-like particles of Norwalk virus to romaine lettuce veins. Applied and Environmental Microbiology, 76(24), 7997–8003.

    Article  PubMed  CAS  Google Scholar 

  • Heaton, J. C., & Jones, K. (2008). Microbial contamination of fruit and vegetables and the behaviour of enteropathogens in the phyllosphere: A review. Journal of Applied Microbiology, 104(3), 613–626.

    Article  PubMed  CAS  Google Scholar 

  • Hernandez, F., Monge, R., Jimenez, C., & Taylor, L. (1997). Rotavirus and hepatitis A virus in market lettuce (Latuca sativa) in Costa Rica. International Journal of Food Microbiology, 37, 221–223.

    Article  PubMed  CAS  Google Scholar 

  • Hernroth, B. E., Conden-Hansson, A. C., Rehnstam-Holm, A. S., Girones, R., & Allard, A. K. (2002). Environmental factors influencing human viral pathogens and their potential indicator organisms in the blue mussel, Mytilus edulis: The first Scandinavian report. Applied and Environmental Microbiology, 68(9), 4523–4533.

    Article  PubMed  CAS  Google Scholar 

  • Holtby, I., Tebbutt, G. M., Green, J., Hedgeley, J., Weeks, G., & Ashton, V. (2001). Outbreak of Norwalk-like virus infection associated with salad provided in restaurant. Communicable Disease and Public Health, 4, 305–310.

    PubMed  CAS  Google Scholar 

  • Hundesa, A., Bofill-Mas, S., Maluquer de Motes, C., Rodriguez-Manzano, J., Bach, A., Casas, M., et al. (2010). Development of a quantitative PCR assay for the quantitation of bovine polyomavirus as a microbial source-tracking tool. Journal of Virological Methods, 163(2), 385–389.

    Article  PubMed  CAS  Google Scholar 

  • Hundesa, A., Maluquer de Motes, C., Albinana-Gimenez, N., Rodriguez-Manzano, J., Bofill-Mas, S., Suñen, E., et al. (2009). Development of a qPCR assay for the quantification of porcine adenoviruses as an MST tool for swine fecal contamination in the environment. Journal of Virological Methods, 158(1–2), 130–135.

    Article  PubMed  CAS  Google Scholar 

  • Jothikumar, N., Cromeans, T. L., Robertson, B. H., Meng, X. J., & Hill, V. R. (2006). A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. Journal of Virological Methods, 131(1), 65–71.

    Article  PubMed  CAS  Google Scholar 

  • Kilgore, P. E., Belay, E. D., Hamlin, D. M., Noel, J. S., Humphrey, C. D., Gary, H. E, Jr., et al. (1996). A university outbreak of gastroenteritis due to a small round-structured virus. Application of molecular diagnostics to identify the etiologic agent and patterns of transmission. Journal of Infectious Diseases, 173(4), 787–793.

    Article  PubMed  CAS  Google Scholar 

  • Koopmans, M., von Bonsdorff, C. H., Vinge, J., De Medici, D., & Monroe, S. (2002). Foodborne viruses. FEMS Microbiology Reviews, 26, 187–205.

    PubMed  CAS  Google Scholar 

  • Lees, D. (2000). Viruses and bivalve shellfish. International Journal of Food Microbiology, 59, 81–116.

    Article  PubMed  CAS  Google Scholar 

  • Long, S. M., Adak, G. K., Brien, S. J., & Gillespie, I. A. (2002). General outbreaks of infectious intestinal disease linked with salad vegetables and fruit, England and Wales, 1992–2000. Communicable Disease and Public Health, 5, 101–105.

    PubMed  CAS  Google Scholar 

  • Lowry, P. W., Levine, R., Stroup, D. F., Gunn, R. A., Wilder, M. H., & Konigsberg, C, Jr. (1989). Hepatitis A outbreak on a floating restaurant in Florida, 1986. American Journal of Epidemiology, 129, 155–164.

    PubMed  CAS  Google Scholar 

  • Lynch, M. F., Tauxe, R. V., & Hedberg, C. W. (2009). The growing burden of foodborne outbreaks due to contaminated fresh produce: Risks and opportunities. Epidemiology and Infection, 137(3), 307–315.

    Article  PubMed  CAS  Google Scholar 

  • Makary, P., Maunula, L., Niskanen, T., Kuusi, M., Virtanen, M., Pajunen, S., et al. (2009). Multiple norovirus outbreaks among workplace canteen users in Finland, July 2006. Epidemiology and Infection, 137, 402–407.

    Article  PubMed  CAS  Google Scholar 

  • Maluquer de Motes, C., Clemente-Casares, P., Hundesa, A., Martín, M., & Girones, R. (2004). Detection of bovine and porcine adenoviruses for tracing the source of fecal contamination. Applied and Environmental Microbiology, 70(3), 1448–1454.

    Google Scholar 

  • Martínez-Martínez, M., Diez-Valcarce, M., Hernández, M., & Rodríguez-Lázaro, D. (2011). Design and application of nucleic acid standards for quantitative detection of enteric viruses by real-time PCR. Food and Environmental Virology, 3(2), 92–98.

    Article  PubMed  Google Scholar 

  • Muniain-Mujika, I., Girones, R., & Lucena, F. (2000). Viral contamination of shellfish: Evaluation of methods and analysis of bacteriophages and human viruses. Journal of Virological Methods, 89, 109–118.

    Article  PubMed  CAS  Google Scholar 

  • Newell, D. G., Koopmans, M., Verhoef, L., Duizer, E., Aidara-Kane, A., Sprong, H., et al. (2010). Food-borne diseases - the challenges of 20 years ago still persist while new ones continue to emerge. International Journal of Food Microbiology, 30(139 Suppl 1), S3–S15.

    Article  Google Scholar 

  • Pebody, R. G., Leino, T., Ruutu, P., Kinnunen, L., Davidkin, I., Nohynek, H., et al. (1998). Foodborne outbreaks of hepatitis A in a low endemic country: An emerging problem? Epidemiology and Infection, 120(1), 55–59.

    Article  PubMed  CAS  Google Scholar 

  • Pina, S., Puig, M., Lucena, F., Jofre, J., & Girones, R. (1998). Viral pollution in the environment and shellfish: Human adenovirus detection by PCR as an index of human viruses. Applied and Environmental Microbiology, 64(9), 3376–3382.

    PubMed  CAS  Google Scholar 

  • Renou, C., Moreau, X., Pariente, A., Cadranel, J. F., Maringe, E., Morin, T., et al. (2008). A national survey of acute hepatitis E in France. Alimentary Pharmacology and Therapeutics, 27(11), 1086–1093.

    Google Scholar 

  • Richards, G. P. (2001). Food-borne pathogens: Enteric virus contamination of foods through industrial practices: A primer on intervention strategies. Journal of Industrial Microbiology and Biotechnology, 27(2), 117–125.

    Article  PubMed  CAS  Google Scholar 

  • Rosenblum, L. S., Mirkin, I. R., Allen, D. T., Safford, S., & Hadler, S. C. (1990). A multifocal outbreak of hepatitis A traced to commercially distributed lettuce. American Journal of Public Health, 80, 1075–1079.

    Article  PubMed  CAS  Google Scholar 

  • Scallan, E., Hoekstra, R. M., Angulo, F. J., Tauxe, R. V., Widdowson, M. A., & Roy, S. L. (2011). Foodborne illness acquired in the United States - major pathogens. Emerging Infectious Diseases, 17(7), 7–15.

    Article  PubMed  Google Scholar 

  • Seymour, I. J., & Appleton, H. (2001). Foodborne viruses and fresh produce. Journal of Applied Microbiology, 91(5), 759–773.

    Article  PubMed  CAS  Google Scholar 

  • Steele, M., & Odumeru, J. (2004). Irrigation water as source of foodborne pathogens on fruit and vegetables. Journal of Food Protection, 67(12), 2839–2849.

    PubMed  Google Scholar 

  • Svraka, S., Duizer, E., Vennema, H., de Bruin, E., van der Veer, B., Dorresteijn, 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(5), 1389–1394.

    Article  PubMed  Google Scholar 

  • Vega, E., Garland, J., & Pillai, S. D. (2008). Electrostatic forces control nonspecific virus attachment to lettuce. Journal of Food Protection, 71(3), 522–529.

    PubMed  Google Scholar 

  • Vilagines, P., Sarrette, B., Husson, G., & Vilagines, R. (1993). Glass wool for virus concentration at ambient water pH level. Water Science. Technology, 27(3–4), 299–306.

    Google Scholar 

  • Wei, J., Jin, Y., Sims, T., & Kniel, K. E. (2010). Internalization of murine norovirus 1 by Lactuca sativa during irrigation. Applied and Environmental Microbiology, 77(7), 2508–2512.

    Article  Google Scholar 

  • Wheeler, C., Vogt, T. M., Armstrong, G. L., Vaughan, G., Weltman, A., Nainan, O. V., et al. (2005). An outbreak of hepatitis A associated with green onions. New England Journal of Medicine, 353, 890–897.

    Article  PubMed  CAS  Google Scholar 

  • Widdowson, M. A., Sulka, A., Bulens, S. N., Beard, R. S., Chaves, S. S., Hammond, R., et al. (2005). Norovirus and foodborne disease, United States, 1991–2000. Emerging Infectious Diseases, 11, 95–102.

    Article  PubMed  Google Scholar 

  • Wyn-Jones, A. P., Carducci, A., Cook, N., D’Agostino, M., Divizia, M., Fleischer, J., et al. (2011). Surveillance of adenoviruses and noroviruses in European recreational waters. Water Research, 45(3), 1025–1038.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by a European Community’s Seventh Framework Programme (FP7/2007-2013) under Grant agreement no. KBBE-213178 (Integrated monitoring and control of food-borne viruses in European food supply chains, VITAL) led by the coordination team of Nigel Cook (FERA, UK), Martin D’Agostino (FERA, UK) and Franco M Ruggeri (ISS, Italy). The authors are grateful to the owners of the companies who took part in this study for the kind and skilful collaboration and assistance and to all VITAL members.

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Authors and Affiliations

  1. Environmental Microbiology Unit, Department of Public Health, Medical School, University of Patras, 26500, Patras, Greece

    P. Kokkinos, E. Legaki & A. Vantarakis

  2. National Veterinary Research Institute, Al. Partyzantów 57, 24-100, Puławy, Poland

    I. Kozyra, A. Kaupke & A. Rzeżutka

  3. Scientific Veterinary Institute “Novi Sad”, Rumenacki put 20, 21000, Novi Sad, Serbia

    S. Lazic & T. Petrovic

  4. National Institute for Public Health and the Environment, RIVM, Bilthoven, Netherlands

    M. Bouwknegt, S. Rutjes & A. M. de Roda Husman

  5. Laboratory for Process Microbial Ecology and Bioinspirational Management, Consortium for Industrial Microbiology and Biotechnology, Department of Microbial and Molecular Systems, KU Leuven, Leuven, Belgium

    K. Willems

  6. Scientia Terrae Research Institute, Sint-Katelijne-Waver, Belgium

    K. Willems

  7. Environmental Health Service, Health Service Executive, Sandfield Centre, Ennis, Co., Clare, Ireland

    R. Moloney

  8. Food and Environment Research Agency (FERA), Sand Hutton, York, UK

    M. D’Agostino & N. Cook

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  1. P. Kokkinos
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  2. I. Kozyra
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  3. S. Lazic
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  4. M. Bouwknegt
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  6. K. Willems
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  7. R. Moloney
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  8. A. M. de Roda Husman
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  9. A. Kaupke
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  10. E. Legaki
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  11. M. D’Agostino
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  12. N. Cook
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  13. A. Rzeżutka
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  14. T. Petrovic
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  15. A. Vantarakis
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Correspondence to P. Kokkinos, A. Rzeżutka, T. Petrovic or A. Vantarakis.

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P. Kokkinos, I. Kozyra, and S. Lazic contributed equally to the work.

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Kokkinos, P., Kozyra, I., Lazic, S. et al. Harmonised Investigation of the Occurrence of Human Enteric Viruses in the Leafy Green Vegetable Supply Chain in Three European Countries. Food Environ Virol 4, 179–191 (2012). https://doi.org/10.1007/s12560-012-9087-8

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