Viruses in food products

Data on viral food contaminants that are actually or potentially capable of realizing the food route of infection are presented. The main sources of infection of food with viruses are named: human waste / faeces, contaminated food processing facilities, animals-carriers of zooanthroponotic infections. The groups of viruses transmitted through food are characterized: 1) gastroenteritis pathogens – Sapporo and Norwalk viruses from the family Caliciviridae; Rotavirus A from the family Reoviridae; Mammastroviruses 1, 6, 8 and 9 from the family Astroviridae; Human mastadenovirus F from the family Adenoviridae; Aichivirus A from the family Picornaviridae; 2) Hepatovirus A from the family Picornaviridae and Orthohepevirus A from the family Hepeviridae (with replication in the liver); 3) viruses with replication in the human intestine, which after generalization of the infection affect the CNS – Еnteroviruses B and C from the family Picornaviridae. The stability and survival time of viruses in the environment and food are shown. The main ways of transmission of viruses that are able to enter the human body through infected foods are considered. Influenza A (H1N1) virus has been identified as a possible contaminant in pork and chicken, which without heat treatment can pose a potential risk of human infection. The ability of classical and African swine fever pathogens to remain viable after industrial processing of meat or raw meat has been shown. Families of viruses whose zoopathogenic representatives can contaminate meat products (beef, pork, chicken) are named: Parvoviridae, Anelloviridae, Circoviridae, Polyomaviridae, Smacoviridae. To determine the possible latent infection of people with these viruses, it is necessary to test sera for the presence of specific antibodies. The detection of gyroviruses of the family Anelloviridae and huchismacoviruses of the family Smacoviridae in human faeces may be due to the consumption of infected chicken meat. Data on extraction and concentration methods and methods of virus detection in contaminated food products: PCR (reverse transcription and real-time), ELISA, IСA, electron microscopy, virus isolation in transplanted cell cultures with subsequent identification in serological reactions, NR, IFА, ELISA) or PCR.

Molecular Detection of Rotavirus in Mollusks from the Oued El Maleh Estuary of Mohammedia, Morocco

Viral outbreaks can result from the consumption of contaminated bivalve mollusks. However, despite the regulation related to enteric bacteria in food products, the consumption of raw and undercooked mollusks remains linked to viral epidemics in human populations. Real-time RT-PCR is a highly sensitive approach for detecting and quantifying enteric viruses, and after eliminating enzymatic amplification inhibitors from samples of interest, sensitive and specific tests, like real-time RT-PCR, can facilitate the detection and quantification of a wide range of viruses that are concentrated in mollusk digestive tissues. The aim of the present study was to evaluate the prevalence of Group-A rotaviruses in mussel (Mytilus edulis Linnaeus, 1758) specimens (n=576) collected downstream of the Oued El Maleh Estuary, which is along the coast of Mohammedia City in Morocco, using real-time RT-PCR. Rotavirus A RNA was detected in 37.5% (n=18) of the 48 sample batches, and viral loads ranged from 0.42×101 to 1.8603×104 genomic copies per g digestive tissue. Most (72.22%) of the positive samples were collected during the wet season (September-April), and the probability of detecting rotaviruses was significantly greater during the wet season than during the dry season (P<0.001). Monitoring Rotavirus A and similar viruses in shellfish may help prevent viral contamination and preserve public health.

Post-weaning diarrhea in pigs weaned without medicinal zinc: risk factors, pathogen dynamics, and association to growth rate

Background Porcine post-weaning diarrhea (PWD) has reemerged as an important topic in pig production, as common control strategies based on prophylactic use of antimicrobials and zinc oxide have been deemed unsustainable. The objectives of this study were to estimate the cumulative incidence of porcine post-weaning diarrhea with different etiologies in production systems weaning without zinc oxide and prophylactic antimicrobials, to assess risk factors for post-weaning diarrhea, and to estimate the impact of post-weaning diarrhea on growth rate. A cohort study was conducted at two commercial indoor producers weaning without medicinal zinc oxide and prophylactic antimicrobials. Results Piglets were included at birth (n = 300) and 272 survived until weaning. After insertion to the nursery units, the piglets were clinically examined every day for 14 days, and rectal swabs were collected and analyzed for enterotoxigenic Escherichia coli (ETEC) and rotavirus A. The cumulative incidences of PWD the first 14 days after insertion to the nursery units were 41.8% (CI 33.6, 50.4) and 51.1% (CI 42.3, 60.0) at the two producers, respectively. We found a low incidence of cases associated to ETEC, and detected a substantial proportion of cases associated to rotavirus. We observed a biphasic pattern in the assumed etiology with rotavirus occurring first, and then a shift towards cases associated to ETEC/non-ETEC hemolytic E. coli. Being offspring of older sows was a protective factor for the development of PWD (Hazard ratio = 0.88 [CI 0.78, 0.99] per unit increase in parity of the dam). Low birth weight reduced the post-weaning growth rate (− 5.2 g/day [CI − 7.5, − 2.9] per 100 g decrease in birthweight) and increased the hazard of developing PWD (Hazard ratio for birthweight below 1100 g: 2.30 [CI 1.41–3.74]). The combined effect of having diarrhea for 2 days or more and receiving antimicrobial treatment was associated with an increased average daily weight gain. Conclusions This study suggests novel insights regarding pathogen dynamics and risk factors for PWD in productions not using prophylactic antimicrobials and medicinal zinc. The findings may have important implications for both antimicrobial usage and prevention strategies.

Study of immunogenicity and safety of a candidate rotavirus vaccine based on a recombinant hybrid protein

BACKGROUND: Rotaviruses are the main cause of acute gastroenteritis in children in both developed and developing countries. Vaccination is the only way to prevent severe and fatal course of this disease. Live attenuated viruses-based vaccines currently available can have a number of side effects. A candidate rotavirus vaccine reported is based on a hybrid recombinant protein FliCVP6VP8, which includes a VP6 protein fragment, a rotavirus A VP8 protein fragment, and S. typhimurium FliC flagellin components. AIM: The aim was to evaluate the immunogenicity and safety of а preparation Rotavirus vaccine, recombinant in preclinical studies. MATERIALS AND METHODS: The immunogenicity of vaccine (blood antibody titers, antigen-specific proliferative response of spleen cells) was evaluated in BALB/c mice. The acute and subchronic toxicity, the possible irritating effect, pyrogenicity and the anaphylactic effect and delayed type hypersensitivity were evaluated in laboratory mice, rats, Guinea pigs, and rabbits. RESULTS: Double immunization of mice with the candidate vaccine demonstrated a significant increase in antibody titers in mouse sera compared to that in control mice. Evaluation of antigen-specific proliferative response after double immunization with a candidate vaccine demonstrated a significant increase in the values of stimulated proliferation. Evaluation of safety through acute and chronic toxicity studies demonstrated no toxicity. The immunostimulatory effect of vaccine was demonstrated when evaluating the number of antibody-producing cells with sheep red blood cells as antigens. The number of white blood cells was demonstrated to increase after the prolonged vaccine administration. CONCLUSIONS: The preclinical studies have demonstrated safety of the candidate rotavirus vaccine and its capability to produce the immune response.

Ocurrence of rotavirus and picobirnavirus in wild and exotic avian from amazon forest

The present study reports the occurrence of rotavirus A (RVA), rotavirus D (RVD), rotavirus F (RVF), rotavirus G (RVG), and picobirnavirus (PBV) in fecal specimens of wild (n = 22), and exotic birds (n = 1) from different cities of Pará state. These animals were hospitalized at Veterinary Hospital of the Federal University of Pará, Brazil, in a period from January 2018 to June 2019. The animals exhibited different clinical signs, such as diarrhea, malnutrition, dehydration, and fractures. The results showed 39.1% (9/23) of positivity for RVA by RT-qPCR. Among these, one sample (1/9) for the NSP3 gene of T2 genotype was characterized. About 88.9% (8/9) for the VP7 gene belonging to G1, G3 equine like and G6 genotypes, and 55.5% (5/9) for the VP4 gene of P[2] genotype were obtained. In the current study, approximately 4.5% of the samples (1/23) revealed coinfection for the RVA, RVD and RVF groups. Furthermore, picobirnavirus (PBV) was detected in one of the 23 samples tested, and was classified in the Genogroup I. The findings represent the first report of RVA, RVD, RVF, RVG, and PBV genotypes in wild birds in Brazil, and due to wide distribution it can implies potential impacts of RVs, and PBVs on avian health, and other animals contributing to construction of new knowledge, and care perspectives.

Correlation between prevalence of selected enteropathogens and diarrhea in children: a case-control study in China

Background The application of nucleic acid detection methods improves the ability of laboratories to detect diarrheal pathogens, but it also poses new challenges for the interpretation of the results. It is often difficult to attribute a diarrhea episode to the detected pathogens. Here we investigated the prevalence of 19 enteropathogens among diarrheal and non-diarrheal children and provided support for understanding the clinical significance of the pathogens. Methods A total of 710 fecal samples were collected from children under 5 years old in two different regions of China from May 2017 to March 2018, comprising 383 mild to moderate diarrheal cases and 327 non-diarrheal controls. The enteropathogens were detected using real-time polymerase chain reaction (PCR) or real-time reverse transcription PCR (RT-PCR). Results Enteropathogens were detected in 68.9% of the cases and 41.3% of the controls. Rotavirus A (adjusted OR [aOR], 9.91; 95% CI, 4.99–19.67), norovirus GI and GII (aOR, 3.82; 95% CI, 2.12–6.89), and Campylobacter jejuni (aOR, 20.12; 95% CI, 2.57–157.38) were significantly associated with diarrhea (p &lt; 0.05). Adenovirus, norovirus GII, rotavirus A, and enteroaggregative Escherichia coli (pCVD432) gave lower cycle threshold (Ct) values in the cases than in the controls (p &lt; 0.05). Rotavirus A and norovirus GII were associated with diarrhea when the Ct value were ≤ 30 and ≤ 25, respectively. Conclusions The types and loads of enteropathogens are likely to influence the interpretation of the clinical significance of positive results.

Coinfection of the intestinal tract with Aeromonas hydrophila, Clostridium difficile and Rotavirus - a case report

Most cases of acute diarrhea in adults are of infectious etiology, likely viral and self-limited. Among those with severe diarrhea, however, bacterial causes are responsible for most cases. Apart from the standard stool cultures, to increase the positive yield a novel multiplex molecular test can be performed simultaneously. The authors present a patient with recurrent diarrhea and detection of Aeromonas hydrophila by culturing and Rotavirus and Clostridioides difficile by multiplex molecular test. They discuss and justify which is the most likely actionable pathogen. Good communication between the physicians and interpretation on the multiple positive results in the context of clinical picture and the test employed were important for a better management and favourable outcome of the patient.

Detection of Enteric Viruses and Core Microbiome Analysis in Artisanal Colonial Salami-Type Dry-Fermented Sausages from Santa Catarina, Brazil

Microbial fermentation plays an important role in the manufacturing of artisanal sausages and can have major effects on product quality and safety. We used metagenomics and culture-dependent methods to study the presence of Hepatitis E virus (HEV) and Rotavirus A (RV-A), and fungal and bacterial communities, in artisanal Colonial salami-type dry-fermented sausages in Santa Catarina state, Brazil. Lactic acid bacteria (LAB) and yeast dominated the microbiome. Latilactobacillus sakei and Debaryomyces hansenii were ubiquitous and the most abundant species. The DNA of some foodborne pathogens was found in very low concentrations although viable cells of most of these species were undetectable by cultivation methods. The characteristics of the raw material and hygiene of the artisanal sausage manufacturing process resulted in high loads of beneficial microorganisms and the absence of HEV and RV-A viruses as determined by RT-qPCR assays. In conclusion, high LAB load in sausages was more relevant to preventing pathogen growth than the ripening time and/or physicochemical characteristics. However, the presence of Clostridium spp. and other pathogens in some samples must be taken into account for the development of future preservation methods; appropriate LAB starter cultures and health surveillance are required in the production process to prevent foodborne outbreaks.

Rotavirus A infection in pre- and post-vaccine period: Risk factors, genotypes distribution by vaccination status and age of children in Nampula Province, Northern Mozambique (2015-2019)

Mozambique introduced the monovalent rotavirus vaccine (Rotarix®, GSK Biologicals, Rixensart, Belgium) in September 2015. Previous analysis, showed that Nampula province continues reporting a high frequency of Rotavirus A (RVA) infection and the emergence of G9P[6], G9P[4] and G3P[4] genotypes. This analysis aimed to determine the RVA frequency; risk factors; genotype distribution by vaccination status and age between pre- and post-vaccine periods in children under-five years old with diarrhea in Nampula. A cross-sectional, hospital-based surveillance study was conducted in the Hospital Central de Nampula in Mozambique. Socio-demographic and clinical data were collected to assess factors related to RVA infection in both periods. Stool specimens were screened to detect RVA by ELISA, and positive samples were genotyped. Between 2015 (pre-vaccine period) and 2016–2019 (post-vaccine period), 614 stool specimens were collected and tested for RVA in which 34.9% (67/192) were positive in pre-vaccine period and 21.8% (92/422) in post-vaccine (p = 0.001). In the post-vaccine period, age, year, and contact with different animal species (chicken, duck, or multiple animals) were associated with RVA infection. RVA infection was higher in children partially vaccinated (40.7%, 11/27) followed by the fully vaccinated (29.3%, 56/191) and the unvaccinated (15.3%, 21/137) (p = 0.002). G1P[8] and G9P[4] were common in vaccinated children less than 12 months. The present analysis showed that RVA infection reduced slightly in the post-vaccine period, with a high proportion of infection and genotype diversity in children, under 12 months of age, vaccinated. Further research on factors associated with RVA infection on vaccinated compared to unvaccinated children and vaccination optimization should be done.