Quantitative levels of norovirus and hepatitis A virus in bivalve molluscs collected along the food chain in the Netherlands, 2013-2017.

Contamination of bivalve molluscs with viruses is well recognized as a food safety risk. A microbiological criterion for norovirus (NoV) and hepatitis A virus (HAV) in shellfish, however, does not exist in the European Union currently. The aim of this study was to evaluate the contamination levels of these viruses for fluctuation over a long period (2013-2017) in oyster (n = 266) and mussel samples (n = 490) using a method based on ISO/TS 15216-1: 2013. Samples were taken at different points in the food chain, either directly post-harvest, at Dutch dispatch centers or in retail stores, from September until March of each year. Altogether, 53.1% of the mussel and 31.6% of the oyster samples tested positive for NoV RNA. Simultaneous presence of NoV GI and GII RNA was observed in 31.6% of mussel and 10.2% of oyster samples. Contamination levels in NoV positive mussel samples collected post-harvest from B-areas were significantly higher than in those collected post-harvest from A-areas, or at dispatch centers or retail stores. Levels in oysters from dispatch were significantly lower than those collected in retail stores. Ready for sale mussels and oysters contained 2.04 and 1.76 mean log10 transformed NoV genome copies/gram (gc/g), respectively. GII levels were at a constant level in ready for sale mussels throughout all sampling periods in the study. This seemed to be true for oysters as well. HAV RNA was detected in only one of the tested mussel samples (n = 392) (typed HAV 1A) and in none of the tested oyster samples (n = 228). Critical evaluation of NoV and HAV levels in shellfish can be of help for risk assessment and risk management actions.

Detection and quantification of hepatitis E virus RNA in ready to eat raw pork sausages in the Netherlands.

The aim of the present study was to assess raw pork sausages collected on the Dutch market for the presence of hepatitis E virus (HEV) RNA. 46 of 316 (14.6%) products sampled from Dutch retail stores in 2017-2019 were positive for HEV RNA. HEV RNA was detected in 10.8% of "cervelaat" (n = 74), 18.5% of salami (n = 92), 26.1% of "metworst" (n = 46), 16.3% of "snijworst" (n = 43) samples. This was significantly more often than in other raw pork sausages like dried sausages, fuet or chorizo (3.3%, n = 61). The percentage of HEV RNA positive products was not significantly different for products sold as either sliced or unsliced deli meat. The average viral load in positive tested products was 2.76 log10 genome copies per 5 g, incidentally reaching up to 4.5 log10 genome copies per 5 g. The average HEV RNA level was significantly higher in samples collected in 2017 than those in samples collected in 2018, and most of the samples in 2019. Typing by sequence analysis was successful for 33 samples, all revealing genotype 3c. The results support recent epidemiological studies that identified specific raw pork sausages as risk factor for hepatitis E virus infection in the Netherlands. Persons at risk, including Dutch transplant recipients, have been advised to avoid the consumption of raw pork sausages. The study warrants a continuation of monitoring to follow the HEV RNA levels in pork products for use in risk assessments and risk management.