Tracing enteric viruses in the European berry fruit supply chain.

In recent years, numerous foodborne outbreaks due to consumption of berry fruit contaminated by human enteric viruses have been reported. This European multinational study investigated possible contamination routes by monitoring the entire food chain for a panel of human and animal enteric viruses. A total of 785 samples were collected throughout the food production chain of four European countries (Czech Republic, Finland, Poland and Serbia) during two growing seasons. Samples were taken during the production phase, the processing phase, and at point-of-sale. Samples included irrigation water, animal faeces, food handlers' hand swabs, swabs from toilets on farms, from conveyor belts at processing plants, and of raspberries or strawberries at points-of-sale; all were subjected to virus analysis. The samples were analysed by real-time (reverse transcription, RT)-PCR, primarily for human adenoviruses (hAdV) to demonstrate that a route of contamination existed from infected persons to the food supply chain. The analyses also included testing for the presence of selected human (norovirus, NoV GI, NoV GII and hepatitis A virus, HAV), animal (porcine adenovirus, pAdV and bovine polyomavirus, bPyV) and zoonotic (hepatitis E virus, HEV) viruses. At berry production, hAdV was found in 9.5%, 5.8% and 9.1% of samples of irrigation water, food handlers' hands and toilets, respectively. At the processing plants, hAdV was detected in one (2.0%) swab from a food handler's hand. At point-of-sale, the prevalence of hAdV in fresh raspberries, frozen raspberries and fresh strawberries, was 0.7%, 3.2% and 2.0%, respectively. Of the human pathogenic viruses, NoV GII was detected in two (3.6%) water samples at berry production, but no HAV was detected in any of the samples. HEV-contaminated frozen raspberries were found once (2.6%). Animal faecal contamination was evidenced by positive pAdV and bPyV assay results. At berry production, one water sample contained both viruses, and at point-of-sale 5.7% and 1.3% of fresh and frozen berries tested positive for pAdV. At berry production hAdV was found both in irrigation water and on food handler's hands, which indicated that these may be important vehicles by which human pathogenic viruses enter the berry fruit chain. Moreover, both zoonotic and animal enteric viruses could be detected on the end products. This study gives insight into viral sources and transmission routes and emphasizes the necessity for thorough compliance with good agricultural and hygienic practice at the farms to help protect the public from viral infections.

[1]  S R Petterson,et al.  Microbial Risks from Wastewater Irrigation of Salad Crops: A Screening‐Level Risk Assessment , 2001, Water environment research : a research publication of the Water Environment Federation.

[2]  I. Miettinen,et al.  Norovirus Outbreaks from Drinking Water , 2005, Emerging infectious diseases.

[3]  R. Girones,et al.  Development of a quantitative PCR assay for the quantitation of bovine polyomavirus as a microbial source-tracking tool. , 2010, Journal of virological methods.

[4]  S. Rutjes,et al.  Nucleic Acid Amplification-Based Methods for Detection of Enteric Viruses: Definition of Controls and Interpretation of Results , 2011, Food and Environmental Virology.

[5]  R. Girones,et al.  Multicenter Collaborative Trial Evaluation of a Method for Detection of Human Adenoviruses in Berry Fruit , 2012, Food Analytical Methods.

[6]  R. Girones,et al.  Environmental Factors Influencing Human Viral Pathogens and Their Potential Indicator Organisms in the Blue Mussel, Mytilus edulis: the First Scandinavian Report , 2002, Applied and Environmental Microbiology.

[7]  J. Brassard,et al.  Detection of Human Food-Borne and Zoonotic Viruses on Irrigated, Field-Grown Strawberries , 2012, Applied and Environmental Microbiology.

[8]  S F Bloomfield,et al.  Effects of Cleaning and Disinfection in Reducing the Spread of Norovirus Contamination via Environmental Surfaces , 2004, Infection Control & Hospital Epidemiology.

[9]  O. Traoré,et al.  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. , 2006, International journal of food microbiology.

[10]  Sang-Jong Kim,et al.  Enteric Viruses in Raw Vegetables and Groundwater Used for Irrigation in South Korea , 2009, Applied and Environmental Microbiology.

[11]  R. Vilaginés,et al.  Glass Wool for Virus Concentration at Ambient Water pH Level , 1993 .

[12]  M. Bouwknegt,et al.  Hepatitis E virus RNA in commercial porcine livers in The Netherlands. , 2007, Journal of food protection.

[13]  M. Muehlen,et al.  Major Outbreak of Hepatitis A Associated with Orange Juice among Tourists, Egypt, 2004 , 2007, Emerging infectious diseases.

[14]  Larry R. Beuchat,et al.  Pathogenic Microorganisms Associated with Fresh Produce. , 1996, Journal of food protection.

[15]  M H Zwietering,et al.  Thermal stability of structurally different viruses with proven or potential relevance to food safety , 2012, Journal of applied microbiology.

[16]  B. Robertson,et al.  A broadly reactive one-step real-time RT-PCR assay for rapid and sensitive detection of hepatitis E virus. , 2006, Journal of virological methods.

[17]  H. Vennema,et al.  Norovirus on swabs taken from hands illustrate route of transmission: a case study. , 2009, Journal of food protection.

[18]  I. Pavlik,et al.  Detection and Phylogenetic Characterization of Human Hepatitis E Virus Strains, Czech Republic , 2011, Emerging infectious diseases.

[19]  M. Estes,et al.  Norovirus gastroenteritis. , 2009, The New England journal of medicine.

[20]  M. Bouwknegt,et al.  Occurrence of Human Enteric Viruses in Commercial Mussels at Retail Level in Three European Countries , 2012, Food and Environmental Virology.

[21]  M. Elimelech,et al.  Evaluation of Removal of Noroviruses during Wastewater Treatment, Using Real-Time Reverse Transcription-PCR: Different Behaviors of Genogroups I and II , 2007, Applied and Environmental Microbiology.

[22]  F. Stagnitti,et al.  Is the risk of illness through consuming vegetables irrigated with reclaimed wastewater different for different population groups? , 2006, Water science and technology : a journal of the International Association on Water Pollution Research.

[23]  I. Di Bartolo,et al.  Widespread diffusion of genotype 3 hepatitis E virus among farming swine in Northern Italy. , 2008, Veterinary microbiology.

[24]  D. Graham,et al.  Norwalk Virus Shedding after Experimental Human Infection , 2008, Emerging infectious diseases.

[25]  L. Maunula,et al.  An outbreak of calicivirus associated with consumption of frozen raspberries , 1999, Epidemiology and Infection.

[26]  M. Bouwknegt,et al.  Hepatitis E Virus in Pork Production Chain in Czech Republic, Italy, and Spain, 2010 , 2012, Emerging infectious diseases.

[27]  M. Bouwknegt,et al.  Persistence of human norovirus in reconstituted pesticides--pesticide application as a possible source of viruses in fresh produce chains. , 2013, International journal of food microbiology.

[28]  M. Pommepuy,et al.  Real-time RT-PCR for norovirus screening in shellfish. , 2005, Journal of virological methods.

[29]  L. Baert,et al.  Review: norovirus prevalence in Belgian, Canadian and French fresh produce: a threat to human health? , 2011, International journal of food microbiology.

[30]  R. Baric,et al.  Norovirus Infectivity in Humans and Persistence in Water , 2011, Applied and Environmental Microbiology.

[31]  M. Carter Enterically infecting viruses: pathogenicity, transmission and significance for food and waterborne infection , 2005, Journal of applied microbiology.

[32]  S. Sattar,et al.  Norovirus cross-contamination during food handling and interruption of virus transfer by hand antisepsis: experiments with feline calicivirus as a surrogate. , 2004, Journal of food protection.

[33]  Marta Hernández,et al.  Design and Application of Nucleic Acid Standards for Quantitative Detection of Enteric Viruses by Real-Time PCR , 2011, Food and Environmental Virology.

[34]  M. Bouwknegt,et al.  Persistence of human norovirus GII.4 and GI.4, murine norovirus, and human adenovirus on soft berries as compared with PBS at commonly applied storage conditions. , 2012, International journal of food microbiology.

[35]  A. Bosch,et al.  Development, Evaluation, and Standardization of a Real-Time TaqMan Reverse Transcription-PCR Assay for Quantification of Hepatitis A Virus in Clinical and Shellfish Samples , 2006, Applied and Environmental Microbiology.

[36]  George Papageorgiou,et al.  Surveillance of adenoviruses and noroviruses in European recreational waters , 2010, Water Research.

[37]  Martin D'Agostino,et al.  Virus hazards from food, water and other contaminated environments , 2011, FEMS microbiology reviews.

[38]  Joanne Hewitt,et al.  Influence of wastewater treatment process and the population size on human virus profiles in wastewater. , 2011, Water research.

[39]  P. Taylor,et al.  An outbreak of hepatitis A associated with consumption of raw blueberries , 2003, Epidemiology and Infection.

[40]  L. Baert,et al.  The reduction of murine norovirus 1, B. fragilis HSP40 infecting phage B40-8 and E. coli after a mild thermal pasteurization process of raspberry puree. , 2008, Food microbiology.

[41]  N. Cook,et al.  Harmonised Investigation of the Occurrence of Human Enteric Viruses in the Leafy Green Vegetable Supply Chain in Three European Countries , 2012, Food and Environmental Virology.

[42]  L. Baert,et al.  Survival and transfer of murine norovirus 1, a surrogate for human noroviruses, during the production process of deep-frozen onions and spinach. , 2008, Journal of food protection.

[43]  J. Rodriguez-Manzano,et al.  Development of a qPCR assay for the quantification of porcine adenoviruses as an MST tool for swine fecal contamination in the environment. , 2009, Journal of virological methods.

[44]  Marta Hernández,et al.  Construction and Analytical Application of Internal Amplification Controls (IAC) for Detection of Food Supply Chain-Relevant Viruses by Real-Time PCR-Based Assays , 2011 .

[45]  Marta Hernández,et al.  Analytical Application of a Sample Process Control in Detection of Foodborne Viruses , 2011 .

[46]  P. Gallian,et al.  Pig liver sausage as a source of hepatitis E virus transmission to humans. , 2010, The Journal of infectious diseases.

[47]  Barry S. Michaels,et al.  Outbreaks where food workers have been implicated in the spread of foodborne disease. Part 4. Infective doses and pathogen carriage. , 2008, Journal of food protection.

[48]  X. Meng,et al.  From barnyard to food table: the omnipresence of hepatitis E virus and risk for zoonotic infection and food safety. , 2011, Virus research.

[49]  T. Korhonen,et al.  Multiple norovirus outbreaks linked to imported frozen raspberries , 2011, Epidemiology and Infection.

[50]  A. Fiore,et al.  Hepatitis A transmitted by food. , 2004, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[51]  F. Martelli,et al.  Hepatitis E Virus in Pork Food Chain, United Kingdom, 2009–2010 , 2012, Emerging infectious diseases.