Wastewater represents the main reusable water source after being adequately sanitized by wastewater treatment plants (WWTP). In this sense, only bacterial quality indicators are usually checked to this end, and human pathogenic viruses usually escape from both sanitization procedures and controls, posing a health risk on the use of effluent waters.In this study, we evaluated a protocol based on aluminium adsorption-precipitation to concentrate several human enteric viruses, including norovirus genogroup I (NoV GI), NoV GII, hepatitis A virus (HAV), astrovirus (HAstV), and rotavirus (RV), with limits of detection of 4.08, 4.64, 5.46 log genomic copies/L, 3.31, and 5.41 log PCR units (PCRU)/L, respectively. Furthermore, the method was applied in two independent laboratories to monitor the presence of NoV GI, NoV GII, and HAV in effluent and influent waters collected from five WWTPs at two different sampling dates.Concomitantly, a viability PMAxx-RT-qPCR was applied to all the samples to get information on the potential infectivity of both influent and effluent waters. The range of the titers in influent waters for NoV GI, NoV GII, RV and HAstV was 4.80-7.56, 5.19-7.31 log genomic copies/L, 5.41-6.52, and 4.59-7.33 log PCRU/L, respectively. In effluent waters, the titers ranged between 4. 08-6.27, 4.64-6.08 log genomic copies/L, <5.51, and 3.31-5.58 log PCRU/L. Moreover, the viral titers detected by viability RT-qPCR showed statistical differences with RT-qPCR alone, suggesting the potential viral infectivity of the samples despite some observed reductions. The proposed method could be applied in ill-equipped laboratories, due to the lack of a requirement for a specific apparatus (i.e., ultracentrifuge).

Aim To improve the efficacy of intercalating dyes to distinguishing between infectious and inactivated hepatitis A virus (HAV) in food. Methods and Results Different intercalating dyes were evaluated for the discrimination between infectious and thermally inactivated HAV suspensions combining with the RT‐qPCR proposed in the ISO 15216. Among them, PMAxx was the best dye in removing the RT‐qPCR signal from inactivated HAV. Applied to lettuce and spinach, PMAxx–Triton pretreatment resulted in complete removal of the RT‐qPCR signal from inactivated HAV. Likewise, this study demonstrates that this pretreatment is suitable for the discrimination of inactivated HAV in shellfish without further sample dilution. In mussels and oysters, the developed viability RT‐qPCR method reduced the signal of inactivated HAV between 1·7 and 2·2 logs at high inoculation level, and signal was completely removed at low inoculation level. Conclusions This study showed that the use of PMAxx is an important improvement to assess HAV infectivity by RT‐qPCR. It was shown that PMAxx–Triton pretreatment is suitable for the analysis of infectious HAV in complex food samples such as vegetables and shellfish. Significance and Impact of the Study The PMAxx–Triton pretreatment can be easily incorporated to the ISO norm for infectious virus detection.

Monascus fermentation products can be consumed as food or feed supplements or used as food colouring or flavouring agents. In this study, qPCR and Q-TOF LC/MS methods were developed to monitor Monascus ruber and Monascus purpureus growth and extrolite (lovastatin, mevastatin, as well as the regulated mycotoxin, citrinin (CIT)) production, respectively. Wheat kernels were inoculated with one strain of each species during a solid-state fermentation followed over 63 days. Different growth and extrolite production patterns were clearly observed for the 2 tested strains. After 63 days, high lovastatin levels (up to 0.5 mg/g) were reached for M. ruber wheat fermented kernels while M. purpureus only yielded 0.1 mg/g of lovastatin at best, suggesting that M. ruber may be a better candidate for lovastatin production in a wheat-based model. Mevastatin levels were low and stable for both species. However, M. ruber fermented wheat kernels also contained the highest CIT content, up to 4.2 μg/g, i.e. at levels above the regulation threshold set by the European Union. CIT toxicity was then determined in vitro on bovine kidney cells, showing an IC10 of 6.10-4 M. At maximum concentrations encountered during solid-state fermentation, CIT toxicity was very low under chronic exposure.