Small, non-enveloped RNA viruses belonging to the genera Sapovirus, Kobuvirus, and Mamastrovirus are usually associated with gastroenteritis in humans and animals. These enteric pathogens are considered potential zoonotic agents. In this study, the prevalence and genetic diversity of sapoviruses (SaVs), kobuviruses (KoVs), and astroviruses (AstVs) in asymptomatic pigs were investigated using a PCR approach. KoV was found to be the most prevalent virus (87.3 %), followed by AstV (34.2 %) and SaV (10.2 %). Interestingly, the intra- and inter-cluster distances between porcine SaV capsid sequences revealed one strain (P38/11/CZ) that formed a new genotype within genogroup III of porcine SaVs, and it is tentatively called "P38/11-like" genotype. Moreover, this is the first report of porcine kobuvirus detection on Czech pig farms. The high prevalence rate of gastroenteritis-producing viruses in clinically healthy pigs represents a continuous source of infection of pigs, and possibly to humans.
The global SARS-CoV-2 pandemic dictates that anti-contagion strategies should become matters of essential routine in everyday life. Fomite transference is one of the routes of transmission that has been considered for this virus. However, the risks associated with contaminated surfaces of food packaging kept in refrigerators have not yet been adequately assessed. In this study, a surrogate virus, Alphacoronavirus 1, was used to investigate the persistence of coronavirus dried on a plastic carrier at 4 °C. Techniques of wet wiping, with or without disinfectant saturation, were employed to evaluate their effectiveness in the elimination of the virus. If not wiped, the loss of infectivity of the virus on plastic surfaces was, on average, 0.93 log 10 (i.e. 83%) per day of storage at 4 °C. Wiping with water-saturated material reduced the initial virus titre on the plastic carrier by 2.4 log 10 (99.6%); the same results were achieved through wiping with bactericidal wipes containing ethanol. Wipes saturated with a combination of disinfectant agents (didecyl-dimethyl-ammonium chloride, hydrogen peroxide) decreased the virus titre still more efficiently, by 3.8 log 10 (99.98%) and also significantly prevented further transfer of the virus to a secondary surface through wiping. Thus SARS-CoV-2 transmission potential via contaminated plastic packaging and food may be efficiently eliminated by wet-wiping, especially when wipes saturated with specific disinfectants are used.
This study contributes to the establishment of general guidelines for animal virus lyophilization, with particular respect to differences in virus structure.
The detection and quantification of enteric RNA viruses is based on isolation of viral RNA from the sample followed by quantitative reverse transcription polymerase chain reaction (RT-qPCR). To control the whole process of analysis and in order to guarantee the validity and reliability of results, process control viruses (PCV) are used. The present article describes the process of preparation and use of such PCV– MS2 phage-like particles (MS2 PLP) – in RT-qPCR detection and quantification of enteric RNA viruses. The MS2 PLP were derived from bacteriophage MS2 carrying a unique and specific de novo-constructed RNA target sequence originating from the DNA of two extinct species. The amount of prepared MS2 particles was quantified using four independent methods – UV spectrophotometry, fluorimetry, transmission electron microscopy and a specifically developed duplex RT-qPCR. To evaluate the usefulness of MS2 PLP in routine diagnostics different matrices known to harbor enteric RNA viruses (swab samples, liver tissue, serum, feces, and vegetables) were artificially contaminated with specific amounts of MS2 PLP. The extraction efficiencies were calculated for each individual matrix. The prepared particles fulfill all requirements for PCV – they are very stable, non-infectious, and are genetically distinct from the target RNA viruses. Due to these properties they represent a good morphological and physiochemical model. The use of MS2 PLP as a PCV in detection and quantification of enteric RNA viruses was evaluated in different types of matrices.
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