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.
•Berry fruit food chain was monitored for enteric viruses in European countries.•Contamination of irrigation water and food handlers' hands by viruses was observed.•Improved hygiene conditions are needed to control viruses at soft fruit farms.
The northern European wild boar population has increased during the last decade. Highest wild boar numbers in Finland have been reported in the southeastern part near the Russian border. Wild boars ...may be infected with several human and animal pathogens. In this study, we investigated the presence of important foodborne pathogens in wild boars hunted in 2016 in Finland using serology, PCR and culturing. Seroprevalence of
Salmonella
(38%) and
Yersinia
(56%) infections was high in wild boars. Antibodies to hepatitis E virus,
Toxoplasma gondii
and
Brucella
were found in 18%, 9% and 9% of the wild boars, respectively.
Trichinella
antibodies were detected in 1% of the animals. We recorded no differences in the seroprevalence between males and females. However,
Yersinia
and
T. gondii
antibodies were detected significantly more often in adults than in young individuals.
Listeria monocytogenes
(48%) and
stx
-positive
Escherichia coli
(33%) determinants were frequently detected in the visceral organs (spleen and kidneys) by PCR.
Yersinia pseudotuberculosis
O:1 and
L. monocytogenes
2a and 4b were identified by culturing from the PCR-positive samples.
Brucella suis
biovar 2 was isolated from visceral organs. No African swine fever, classical swine fever or Aujeszky’s disease were detected in the wild boars. Our study shows that wild boars are important reservoirs of foodborne pathogens.
The detection of fecal viral pathogens in water is hampered by their great variety and complex analysis. As traditional bacterial indicators are poor viral indicators, there is a need for alternative ...methods, such as the use of somatic coliphages, which have been included in water safety regulations in recent years. Some researchers have also recommended the use of reference viral pathogens such as noroviruses or other enteric viruses to improve the prediction of fecal viral pollution of human origin. In this work, phages previously tested in microbial source tracking studies were compared with norovirus and adenovirus for their suitability as indicators of human fecal viruses. The phages, namely those infecting human-associated
strain GA17 (GA17PH) and porcine-associated
strain PG76 (PGPH), and the human-associated crAssphage marker (crAssPH), were evaluated in sewage samples and fecal mixtures obtained from different animals in five European countries, along with norovirus GI + GII (NoV) and human adenovirus (HAdV). GA17PH had an overall sensitivity of ≥83% and the highest specificity (>88%) for human pollution source detection. crAssPH showed the highest sensitivity (100%) and specificity (100%) in northern European countries but a much lower specificity in Spain and Portugal (10 and 30%, respectively), being detected in animal wastewater samples with a high concentration of fecal indicators. The correlations between GA17PH, crAssPH, or the sum of both (BACPH) and HAdV or NoV were higher than between the two human viruses, indicating that bacteriophages are feasible indicators of human viral pathogens of fecal origin and constitute a promising, easy to use and affordable alternative to human viruses for routine water safety monitoring.
Drinking water outbreaks occur worldwide and may be caused by several factors, including raw water contamination, treatment deficiencies, and distribution network failure. This study describes two ...drinking water outbreaks in Finland in 2016 (outbreak I) and 2018 (outbreak II). Both outbreaks caused approximately 450 illness cases and were due to drinking water pipe breakage and subsequent wastewater intrusion into the distribution system. In both outbreaks, the sapovirus was found in patient samples as the main causative agent. In addition, adenoviruses and
(outbreak I), and noroviruses, astroviruses, enterotoxigenic and enterohemorragic
(ETEC and EHEC, respectively) and
(outbreak II) were detected in patient samples. Water samples were analyzed for the selected pathogens largely based on the results of patient samples. In addition, traditional fecal indicator bacteria and host-specific microbial source tracking (MST) markers (GenBac3 and HF183) were analyzed from water. In drinking water, sapovirus and enteropathogenic
(EPEC) were found in outbreak II. The MST markers proved useful in the detection of contamination and to ensure the success of contaminant removal from the water distribution system. As mitigation actions, boil water advisory, alternative drinking water sources and chlorination were organized to restrict the outbreaks and to clean the contaminated distribution network. This study highlights the emerging role of sapoviruses as a waterborne pathogen and warrants the need for testing of multiple viruses during outbreak investigation.
► Methods for detection of NoV from lettuce, ham and raspberries were evaluated. ► NoV detection from ready-to-eat foods was carried out using real-time RT-PCR. ► Ultrasentrifugation showed the best ...NoV recovery efficiencies in lettuce and ham. ► PEG precipitation recovered the highest yield of NoVs from raspberries. ► NoV recoveries of 19, 47 and 28% for lettuce, ham and berries were obtained.
Although noroviruses play a significant role in causing foodborne illness in developed countries, no standardised method for detecting noroviruses in foodstuffs is currently available. This study compared four virus recovery methods based on ultrafiltration, immunomagnetic separation, ultracentrifugation and PEG precipitation techniques using identical real-time RT-PCR protocols for detection of RNA in eluates from lettuce, sliced ham and raspberries inoculated artificially with genogroup II norovirus. Noroviruses in all the food source matrices were successfully detected by all four methods. Ultracentrifugation yielded the highest recovery efficiencies in lettuce and ham, whereas PEG precipitation recovered the highest yield of noroviruses from raspberries. The repeatability of the results and the applicability of the methods to all food matrices were best with PEG precipitation, which had average virus recoveries of 19%, 47% and 28% for lettuce, ham and raspberries (viral RNA in dilution 1:10), respectively. In each case, a tenfold dilution of the extracted RNA clearly reduced the level of PCR inhibitors, which were released from raspberries in particular. The results of this study show that the detection of noroviruses in food is challenging, and more efforts to develop sensitive methods are still needed to detect noroviruses in food containing viruses in low numbers.
The main animal reservoirs of zoonotic hepatitis E virus (HEV) are domestic pigs and wild boars, but HEV also infects cervids. In this study, we estimated the prevalence of HEV in Finnish cervid ...species that are commonly hunted for human consumption. We investigated sera from 342 European moose (
Alces alces
), 70 white-tailed deer (
Odocoileus virginianus
), and 12 European roe deer (
Capreolus capreolus
). The samples had been collected from legally hunted animals from different districts of Finland during 2008–2009. We analysed the samples for total anti-HEV antibodies using a double-sandwich ELISA assay. Seropositive sera were analysed with RT-qPCR for HEV RNA. HEV seroprevalence was 9.1% (31/342) in moose and 1.4% (1/70) in white-tailed deer. None of the European roe deer were HEV seropositive (0/12). No HEV RNA was detected from samples of seropositive animals. HEV seropositive moose were detected in all districts. Statistically, HEV seroprevalence in moose was significantly higher (
p
< 0.05) in the North-East area compared to the South-West area. The highest HEV seroprevalence (20.0%) in district level was more than six times higher than the lowest (3.1%). We demonstrated the presence of total anti-HEV antibodies in European moose and white-tailed deer in Finland. Our results suggest that HEV is circulating among the moose population. Infections may occur also in white-tailed deer. We were the first to report a HEV seropositive white-tailed deer from Europe. Further studies are needed to demonstrate the HEV genotypes in cervids in Finland and to evaluate the importance of the findings in relation to food safety.
Norovirus outbreaks from drinking water Maunula, Leena; Miettinen, Ilkka T; von Bonsdorff, Carl-Henrik
Emerging infectious diseases,
11/2005, Letnik:
11, Številka:
11
Journal Article
Recenzirano
Odprti dostop
As part of an intensified monitoring program for foodborne disease outbreaks in Finland, waterborne outbreaks were investigated for viruses. The diagnostic procedure included analysis of patients' ...stool samples by electron microscopy and reverse transcription-polymerase chain reaction (RT-PCR) for noroviruses and astroviruses. When these test results were positive for a virus, the water sample was analyzed. Virus concentration was based on positively charged filters from 1-L samples. Of the total 41 waterborne outbreaks reported during the observation period (1998-2003), samples from 28 outbreaks were available for analysis. As judged by RT-PCR results from patient samples, noroviruses caused 18 outbreaks. In 10 outbreaks, the water sample also yielded a norovirus. In all but 1 instance, the amplicon sequence was identical to that recovered from the patients. The ubiquity of waterborne norovirus outbreaks calls for measures to monitor water for viruses.
An increased number of suspected outbreaks of gastroenteritis linked to bathing water were reported to the Finnish food- and waterborne outbreak (FWO) registry in July and August 2014. The ...investigation reports were assessed by a national outbreak investigation panel. Eight confirmed outbreaks were identified among the 15 suspected outbreaks linked to bathing water that had been reported to the FWO registry. According to the outbreak investigation reports, 1,453 persons fell ill during these outbreaks. Epidemiological and microbiological data revealed noroviruses as the main causative agents. During the outbreaks, exceptionally warm weather had boosted the use of beaches. Six of eight outbreaks occurred at small lakes; for those, the investigation strongly suggested that the beach users were the source of contamination. In one of those eight outbreaks, an external source of contamination was identified and elevated levels of faecal indicator bacteria (FIB) were noted in water. In the remaining outbreaks, FIB analyses were insufficient to describe the hygienic quality of the water. Restrictions against bathing proved effective in controlling the outbreaks. In spring 2015, the National Institute for Health and Welfare (THL) and the National Supervisory Authority for Welfare and Health (Valvira) published guidelines for outbreak control to prevent bathing water outbreaks.
Seven major food- and waterborne norovirus outbreaks in Western Finland during 2014-2018 were re-analysed. The aim was to assess the effectiveness of outbreak investigation tools and evaluate the ...Kaplan criteria. We summarised epidemiological and microbiological findings from seven outbreaks. To evaluate the Kaplan criteria, a one-stage meta-analysis of data from seven cohort studies was performed. The case was defined as a person attending an implicated function with diarrhoea, vomiting or two other symptoms. Altogether, 22% (386/1794) of persons met the case definition. Overall adjusted, 73% of norovirus patients were vomiting, the mean incubation period was 44 h (4 h to 4 days) and the median duration of illness was 46 h. As vomiting was a more common symptom in children (96%, 143/149) and diarrhoea among the elderly (92%, 24/26), symptom and age presentation should drive hypothesis formulation. The Kaplan criteria were useful in initial outbreak assessments prior to faecal results. Rapid food control inspections enabled evidence-based, public-health-driven risk assessments. This led to probability-based vehicle identification and aided in resolving the outbreak event mechanism rather than implementing potentially ineffective, large-scale public health actions such as the withdrawal of extensive food lots. Asymptomatic food handlers should be ideally withdrawn from high-risk work for five days instead of the current two days. Food and environmental samples often remain negative with norovirus, highlighting the importance of research collaborations. Electronic questionnaire and open-source novel statistical programmes provided time and resource savings. The public health approach proved useful within the environmental health area with shoe leather field epidemiology, combined with statistical analysis and mathematical reasoning.
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