Screening of Ready-to-Eat Meat Products for Hepatitis E Virus in Switzerland.

Seroprevalence data for pig herds suggested that there must be a relevant reservoir for hepatitis E virus (HEV) in Switzerland. To know more about the viral presence in ready-to-eat meat products, we screened pork liver sausages and raw meat sausages from the Swiss retail market for the presence of HEV. Testing was performed with a detection method where the virus extraction step was optimized. As for the performance of the improved method, the mean recovery rate for the mengovirus process control was 24.4%, whereas for HEV-inoculated sample matrices between 10.4 and 100% were achieved. The limit of detection was about 1.56 × 103 and 1.56 × 102 genome copies per gram for liver sausages and raw meat sausages, respectively. In the screening programme, HEV-RNA was detected in 10 of total 90 (11.1%) meat products, 7 of 37 (18.9%) liver sausages, and 3 of 53 (5.7%) raw meat sausages. Virus loads of up to 5.54 log10 HEV genome copies per gram were measured. All sequences retrieved from positive samples belonged to HEV genotype 3. The significance of the presented work was a current overview of the HEV prevalence in ready-to-eat meat products on the Swiss retail marked and an improvement of the extraction efficiency of the HEV detection method.

Improved detection of Rhodococcus coprophilus with a new quantitative PCR assay.

Agricultural practices, such as spreading liquid manure or the utilisation of land as animal pastures, can result in faecal contamination of water resources. Rhodococcus coprophilus is used in microbial source tracking to indicate animal faecal contamination in water. Methods previously described for detecting of R. coprophilus in water were neither sensitive nor specific. Therefore, the aim of this study was to design and validate a new quantitative polymerase chain reaction (qPCR) to improve the detection of R. coprophilus in water. The new PCR assay was based on the R. coprophilus 16S rRNA gene. The validation showed that the new approach was specific and sensitive for deoxyribunucleic acid from target host species. Compared with other PCR assays tested in this study, the detection limit of the new qPCR was between 1 and 3 log lower. The method, including a filtration step, was further validated and successfully used in a field investigation in Switzerland. Our work demonstrated that the new detection method is sensitive and robust to detect R. coprophilus in surface and spring water. Compared with PCR assays that are available in the literature or to the culture-dependent method, the new molecular approach improves the detection of R. coprophilus.

Detection and frequency of Cronobacter spp. (Enterobacter sakazakii) in different categories of ready-to-eat foods other than infant formula.

Two hundred sixty eight samples of ready-to-eat foods from retail shops were screened for the presence of Cronobacter by selective enrichment followed by plating on three chromogenic agars (ESIA, ESPM and DFI). Cronobacter was isolated from 14/23 samples of sprouts and fresh herbs/salads (60.9%), 7/26 samples of spices and dried herbs (26.9%) and 3/42 confectionery samples (7.1%). In cases where repeat samples were available, foods positive for Cronobacter were retested twice. In total, 54 Cronobacter isolates from 24 foods were recovered and genetic fingerprint patterns generated using PFGE. Identical PFGE-profiles were generated for Cronobacter isolates from five samples of two confectionery products obtained from a particular bakery shop over a period of 11 months. This may indicate a persistent contamination of the production site. For all other isolates, no clustering by phylogenetic analysis of PFGE-profiles was observed, indicating the sporadic nature of Cronobacter in ready-to-eat foods. Enterobacterial counts varied from a maximum value of 2.9 x 10(7) CFU/g (in dill) to a minimum value of <10 CFU/g (in confectionery and dried herbs/spices). There was no correlation between Enterobacterial count and the presence of Cronobacter. Cronobacter may be regularly imported into private households via ready-to-eat foods.