Involvement of herpesviruses in cases of abortion among water buffaloes in southern Italy.

A six-year study on water buffaloes from the Campania Region (Southern Italy) was conducted to evaluate the presence of bovine/bubaline herpesviruses in cases of abortion. A total of 244 buffalo foetuses were analysed by real-time PCR to detect the presence of: bovine alphaherpesvirus 1(BoHV-1), bubaline alphaherpesvirus 1 (BuHV-1), bovine alphaherpesvirus 2 (BuHV-2), and bovine gammaherpesvirus 4 (BoHV-4). The foetuses of 14 water buffaloes that showed abortions were positive for BuHV-1 (4 animals) and/or BoHV-4 (11 animals), with one of these cases showing co-infection with BuHV-1 and BoHV-4. This study reports the first identification of BoHV-4 in water buffaloes. Cases of abortion were analysed using both molecular and cultural assays for the presence of other pathogens. In nearly all the abortion cases positive for BoHV-4, the virus was identified as a co-infecting agent together with other microorganisms, whereas in two abortion cases, it was the only pathogen found.

Prevalence of Infection with Porcine Circovirus Types 2 and 3 in the Wild Boar Population in the Campania Region (Southern Italy).

In recent years, porcine circovirus (PCV) infection has been documented as an important and emerging health concern for livestock and wildlife worldwide. The purpose of the present study was to assess the molecular prevalence of PCV-2 and PCV-3 and to clarify the epidemiological role of wild boars in the circulation of this virus in Campania, Southern Italy. For this purpose, samples from several organs were collected during the hunting season 2017-2018 from 148 wild boars in the Campania region. Quantitative real-time PCR was used for the detection and quantification of PCV-2 and PCV-3 genomes. The combined prevalence of PCV-2 and PCV-3 was 74.32% in the wild boars tested. The proportions of wild boars positive for PCV-2 or PCV-3, or coinfected, were 47.30%, 49.32%, and 22.30%, respectively. No link was detected between PCV positivity and location, but gender was a risk factor for the disease (female; p < 0.0001; OR 0.29). Furthermore, our study provides a snapshot of PCV-2 and PCV-3 circulation in wild boars in the Campania region: our findings can help us to better understand the role of wildlife in PCV circulation.

Quantitative Real-Time PCR and Digital PCR to Evaluate Residual Quantity of HAV in Experimentally Depurated Mussels.

Kinetics of hepatitis A virus (HAV) accumulation and depuration from mussels (Mytilus galloprovincialis) was studied in an experimental depuration system. Different parameters likely to influence the rate of virus accumulation and elimination were evaluated. Analyses were carried out by both real-time RT-qPCR and digital PCR. Results demonstrated that the animals start to concentrate the virus already after one hour and reach the maximum level of contamination in 6 h of experiment. With respect to depuration, HAV showed a rapid reduction of the concentration (89%) during the first 24-48 h of experiment and a very slow virus decrement in the following days with a 1% residual RNA at the ninth day of depuration. When process parameters likely to increase the depuration rate (presence of ozone, microalgal feeding, presence of lactic bacteria, pre-treatment with digestive enzymes) were tested, no significant differences in the kinetics were observed. Only treatment with pancreatin seemed to positively affect depuration in the first two days of the experiment.

Detection of Hepatitis A Virus and Other Enteric Viruses in Shellfish Collected in the Gulf of Naples, Italy.

To assess the quality of shellfish harvest areas, bivalve mollusk samples from three coastal areas of the Campania region in Southwest Italy were evaluated for viruses over a three-year period (2015-2017). Screening of 289 samples from shellfish farms and other locations by qPCR and RT-qPCR identified hepatitis A virus (HAV; 8.9%), norovirus GI (NoVGI; 10.8%) and GII (NoVGII; 39.7%), rotavirus (RV; 9.0%), astrovirus (AsV; 20.8%), sapovirus (SaV; 18.8%), aichivirus-1 (AiV-1; 5.6%), and adenovirus (AdV, 5.6%). Hepatitis E virus (HEV) was never detected. Sequence analysis identified HAV as genotype IA and AdV as type 41. This study demonstrates the presence of different enteric viruses within bivalve mollusks, highlighting the limitations of the current EU classification system for shellfish growing waters.

Comparison of automated extraction methods for the detection of hepatitis A virus and norovirus in mussels.

Noroviruses and hepatitis A virus are the pathogens most frequently involved in non-bacterial gastroenteritis and hepatitis worldwide. They are mainly transmitted via the faecal-oral route, direct person-to-person contact or through the consumption of contaminated water and foods. In food virology, detection methods of these viruses are currently based on real-time reverse transcription-polymerase chain reaction (RT-PCR). A crucial step in this process is the acid nucleic extraction, since its performance can negatively influence viral detection and thus give false negative results. The aim of this study is to evaluate the efficiency of 2 automated extraction systems, MagMAX Express and EZ1 Advanced XL, in recovering hepatitis A virus and norovirus RNA from mussels. In the present study, we used mengovirus as a process control to assess the efficiency of the extraction process. Samples were tested for mengovirus, hepatitis A, and norovirus by real time one-step RT-PCR assay. Our data indicates the MagMax Express is the better system to extract hepatitis A and norovirus RNA from mussels since its extraction efficiency was higher (p<0.05) than EZ1 Advanced XL.

Prevalence of Foodborne Viruses in Mussels in Southern Italy.

In this study, the prevalence of various enteric viruses in Mytilus galloprovincialis (Mediterranean mussel) belonging to class A and class B mollusc-harvesting areas in the Campania region in southern Italy was evaluated. One hundred and eight mussels were analysed using real-time reverse transcription PCR during a 2-year collection period (2014-2015) to detect the following viruses: human norovirus (genogroups I and II), rotavirus, astrovirus, sapovirus, aichivirus, hepatitis A virus and hepatitis E virus. Overall, 50.93% of mussels were contaminated by at least one of the tested viruses. Of these virus-positive mussels, 63.63% were contaminated by two or more viruses. In 2014, only three of the eight investigated viruses were detected: astrovirus, sapovirus and aichivirus, whereas in 2015, seven of the eight viruses were detected (only hepatitis E virus was not identified). Astrovirus was the most frequently detected virus in both sampling periods. In 2014, sapovirus was detected at the same frequency as astrovirus (16.00%), followed by aichivirus (8%). In 2015, astrovirus (32.53%) was most frequently detected, followed by norovirus GII (26.50%), sapovirus (18.07%), hepatitis A virus (16.87%), rotavirus (16.87%), aichivirus (13.25%) and norovirus GI (12.05%).This study describes, for the first time, the presence of aichivirus and sapovirus in mussels in Italy.