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Enteric viruses have been described as important contaminants in fresh and ready-to-eat foods such as sandwiches, deli meat and dairy products. This is a cross-sectional randomized survey to estimate the prevalence of norovirus and human adenovirus (HAdV) from 100 Brazilian artisanal raw milk cheese samples (Minas and Coalho) obtained from different agroindustries in four producing regions in the states of Minas Gerais and one in Piaui, respectively. From October 2017 to April 2018, norovirus genogroups I and II and HAdV were investigated in these cheese samples by RT-qPCR and qPCR, respectively. Viruses were detected in 43 samples, being 26 norovirus GI strains, 14 HAdV, and 3 both viruses. Norovirus GII strains were not detected. Viral concentrations ranged from 6.17 × 104 to 1.44 × 107 genome copies/L–1 and murine norovirus 1 used as internal process control showed 100% success rate of recovery with efficiency of 10%. There was a trend towards a higher positivity rate for both viruses in the rainy season, and HAdV were more commonly found among samples with higher fecal coliform counts. This study is a first step in assessing the risk that this contamination may pose to the consumer of raw products as well as emphasizing the need for good manufacturing practices, quality control systems in the dairy industry and markets. As a randomized survey, we established baseline figures for viruses’ prevalence in five types of ready-to-eat raw milk artisanal Brazilian cheese, to allow any monitoring trends, setting control targets and future local risk analyses studies.

The viral RNA of SARS-Coronavirus-2 is known to be contaminating municipal wastewater. We aimed to assess if COVID-19 disease is spreading through wastewater. We studied the amount of viral RNA in raw sewage and the efficiency of the sewage treatment to remove the virus. Sewage water was collected before and after the activated sludge process three times during summer 2020 from three different sewage treatment plants. The sewage treatment was efficient in removing SARS-CoV-2 viral RNA. Each sewage treatment plant gathered wastewater from one hospital, of which COVID-19 admissions were used to describe the level of disease occurrence in the area. The presence of SARS-CoV-2 viral RNA-specific target genes (N1, N2, and E) was confirmed using RT-qPCR analysis. However, hospital admission did not correlate significantly with viral RNA. Moreover, viral RNA loads were relatively low, suggesting that sewage might preserve viral RNA in a hot climate only for a short time.

Herein, we report that nosocomial infection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) may be mitigated by using surgical masks and closed looped ventilation for both non-critical and critical patients. These preventive measures resulted in no viral contamination of surfaces in negative pressure environments.

Goal. To carry out diagnostics and to establish ways of spreading cucumber mosaic virus on the territory of Ukraine. Methods. A range of methods was used, which included visual diagnostics, enzyme immunoassay, method in various modifications, electron microscopy method, and statistical data processing method. Results. Monitoring of crops in different climatic regions of Ukraine for visual symptoms was carried out. Enzyme immunoassay revealed that the symptoms on the plants were caused by cucumber mosaic virus. Using electron microscopy, the morphological properties of the virus were studied, and spherical viral particles with a size of 29 nm were detected. Commercial seeds of different varieties of cucumbers, zucchini, tomatoes, and vegetable peppers were tested for the possibility of their viral contamination. Commercial seeds were found to be contaminated with cucumber mosaic virus antigens. Conclusions. Plants of the Cucurbitaceae and Solanaceae families were selected according to their characteristic symptoms. The presence of cucumber mosaic virus in Ukraine has been proved using electron microscopy and enzyme immunoassay. Virus-infected plants were diagnosed in agrocnosis of Vinnytsia, Odesa, and Kherson regions. Various commercial seed varieties of Cucurbitaceae and Solanaceae were tested for viral contamination. It was found that 37% of them were infected with cucumber mosaic virus and therefore became a potentially dangerous reservoir for viral infection and, as a consequence, a serious cause of outdoor vegetable disease.

Aims: This study aims to investigate the presence and spatial-seasonal variability of human and fish viruses in coastal marine systems using Ravenna’s harbour area (Adriatic Sea, Italy) as a model. Methods and Results: Human viruses (noroviruses and hepatitis A virus) and one of the most threatening finfish pathogens, the nervous necrosis virus (NNV), were investigated in mussels living inside and offshore Ravenna’s harbour. Thirty-three and 36·7% of tested mussel samples resulted contaminated by human and fish viruses respectively. A different spatial-seasonal distribution was observed. Human viruses were detected mainly in inner port sites during colder months, while NNV was detected in both inside and offshore of Ravenna’s harbour, mainly during warmer months. Conclusions: The presence of human viruses in the inner port close to the city centre could be attributed to wastewaters carrying pathogens in the port environment and this arises public health concerns, however, the presence of these viruses limited to the canal port during the winter can greatly reduce the risk to human health. Regarding NNV, the accumulation and release of viable virus by mussels, could represent a viral source for susceptible finfish. These findings reflect the different epidemiological features of these infections and indicate the importance to choose the correct indicator to monitor viral contaminations. Significance and Impact of the Study: The high frequency of viral contamination pointed out in the study stresses the imperative to monitor the viral presence in all coastal habitats where the high natural value meets several recreational and commercial activities such as the Ravenna’s harbour area. Particularly, this study could represent a novel starting point for the development of a more structured bio-monitoring program, in order to ensure improved environmental management and safety of coastal areas.

Objective:To characterize the presence and magnitude of viruses in the air and on surfaces in the rooms of hospitalized patients with respiratory viral infections, and to explore the association between care activities and viral contamination.Design:Prospective observational study.Setting:Acute-care academic hospital.Participants:In total, 52 adult patients with a positive respiratory viral infection test within 3 days of observation participated. Healthcare workers (HCWs) were recruited in staff meetings and at the time of patient care, and 23 wore personal air-sampling devices.Methods:Viruses were measured in the air at a fixed location and in the personal breathing zone of HCWs. Predetermined environmental surfaces were sampled using premoistened Copan swabs at the beginning and at the end of the 3-hour observation period. Preamplification and quantitative real-time PCR methods were used to quantify viral pathogens.Results:Overall, 43% of stationary and 22% of personal air samples were positive for virus. Positive stationary air samples were associated with ≥5 HCW encounters during the observation period (odds ratio [OR], 5.3; 95% confidence interval [CI], 1.2–37.8). Viruses were frequently detected on all of the surfaces sampled. Virus concentrations on the IV pole hanger and telephone were positively correlated with the number of contacts made by HCWs on those surfaces. The distributions of influenza, rhinoviruses, and other viruses in the environment were similar.Conclusions:Healthcare workers are at risk of contracting respiratory virus infections when delivering routine care for patients infected with the viruses, and they are at risk of disseminating virus because they touch virus-contaminated fomites.

This survey was conducted to estimate the incidence and level of potential viral contamination in commercially collected porcine plasma. Samples of spray dried porcine plasma (SDPP) were collected over a 12- month period from eight spray drying facilities in Spain, England, Northern Ireland, Brazil, Canada, and the United States. In this survey, viral load for several porcine pathogens including SVA, TGEV, PRRSV (EU and US strains), PEDV, PCV-2, SIV, SDCoV and PPV were determined by qPCR. Regression of Ct on TCID50 of serial diluted stock solution of each virus allowed the estimate of potential viral level in SDPP and unprocessed liquid plasma (using typical solids content of commercially collected porcine plasma). In this survey SVA, TGEV or SDCoV were not detected in any of the SDPP samples. Brazil SDPP samples were free of PRRSV and PEDV. Samples of SDPP from North America primarily contained the PRRSV-US strain while the European samples contained the PRRSV-EU strain (except for one sample from each region containing a relatively low estimated level of the alternative PRRSV strain). Estimated viral level tended to be in the range from 10 TCID50 to 10 TCID50. Estimated level of SIV was the exception with a very low incidence rate but higher estimated viral load 10 TCID50. In summary, the incidence of potential viral contamination in commercially collected porcine plasma was variable and estimated virus level in samples containing viral DNA/RNA was relatively low compared with that occurring at the peak viremia during an infection for all viruses or when considering the minimal infectious dose for each of them.

A novel coronavirus (SARS-CoV-2) has caused more than 150 million confirmed infections worldwide, while it is not clear whether it affects the coastal waters. This paper proposed a biophysical model based on 16 scenarios with different virus half-life parameters to assess potential viral contamination from 25 municipal sewage outfalls into the Bohai Sea. Viral concentration maps showing spatial and temporal changes are provided based on a biophysical model under multiple scenarios. Results demonstrate that adjacent sea areas can become exposed to SARS-CoV-2 via water-borne transport from outfalls, with a higher risk in winter, because SARS-CoV-2 can be highly stable at low temperature. As coastal waters are the ultimate sink for wastewater and the epidemic will last for long time, this work is of great importance to raise awareness, identify vulnerable areas for marine mammals, and avoid the risk of exposure of tourists at bathing beach.

Mussels can be affected by environmental contaminants, as non-dioxin-like polychlorinated biphenyls (NDL-PCBs), polycyclic aromatic hydrocarbons (PAHs) and cadmium (Cd). Moreover, mussels may concentrate human enteric viruses, like noroviruses (NoVGI/GII), astrovirus (AsV) and rotavirus (RV). Herein, to establish a relationship between environmental and viral contamination, with the aim to ensure human food safety, both chemical and microbiological analysis were carried out in mussels Mytilus galloprovincialis, farmed in Campania region (Italy). Chemical analysis revealed ranges below the European maximum limits, and were: ∑6 NDL-PCBs (28, 52, 101, 138, 153, 180) 0.579–16.857 ng g−1 wet weight (ww); BaP LOQ (

For many years in vivo assays have been a corner stone in safety testing of vaccines for human use. However, there is now an increasing regulatory focus on replacement, reduction and refinement of methods involving animal use. Accordingly, European Pharmacopoeia (Ph.Eur.) monographs and chapters are currently being revised to reduce or discontinue the use of animals in safety and other testing, when such in vivo tests are not absolutely necessary to facilitate risk mitigation. In the current study, a risk assessment of extraneous agents in viral vaccine production has been carried out and it is concluded that only the handling procedures carried out by the technical personnel pose a risk for extraneous viral contamination. A list of named, potentially virulent contaminating viruses, which may have been introduced by these procedures, has been generated. Each of the viruses on this list has been evaluated for possible persistence during the production processes, and it has for all of these been concluded that, if at all present, they only present a negligible risk of introducing extraneous agents in the final product. The overall conclusion of the risk assessment of our vaccine production process is that it justifies the discontinuation of the current in vivo testing, and furthermore demonstrates that there is no need to substitute these in vivo assays with novel in vitro methods.

Simple Summary The African swine fever (ASF) virus is one of the deadliest viruses plaguing the global swine industry. There are no effective treatments for pigs becoming infected with the ASF virus. Eliminating infected pigs is the only effective method to tackle an outbreak. Whole farm biosecurity is therefore critical to prevent the virus from entering in the first place. In recent years, feed and feed ingredients have been identified as potential vectors of viral diseases including ASF. The objective of this research was to simulate contamination of feed by ASF virus in a laboratory setting and use this simulation to test the efficacy of some select feed additives for reducing the amount and viability of this virus in feed. The main result shows that the inoculated virus in feed disappeared in a shorter time when the feed had been treated with the compounds. We conclude that these feed additives could be adopted as part of a comprehensive biosecurity system for pig holdings. Pigs might be protected from feed possibly contaminated by this virus, and the pork supply will thus be less likely to be impacted by this viral disease. Abstract African swine fever virus is one of the most highly contagious and lethal viruses for the global swine industry. Strengthening biosecurity is the only effective measure for preventing the spread of this viral disease. The virus can be transmitted through contaminated feedstuffs and, therefore, research has been conducted to explore corresponding mitigating measures. The purpose of the current study was to test a combination of pure benzoic acid and a blend of nature identical flavorings for their ability to reduce African swine fever viral survival in feed. This virus was inoculated to feed with or without the supplementation of the test compounds, and the viral presence and load were measured by a hemadsorption test and quantitative real time polymerase chain reaction. The main finding was that the combination of pure benzoic acid and nature identical flavorings could expedite the reduction in both viral load and survival in a swine feed. Therefore, this solution could be adopted as a preventive measure for mitigating the risk of contaminated feed by African swine fever virus.

The production of recombinant therapeutic proteins from animal or human cell lines entails the risk of endogenous viral contamination from cell substrates and adventitious agents from raw materials and environment. One of the approaches to control such potential viral contamination is to ensure the manufacturing process can adequately clear the potential viral contaminants. Viral clearance for production of human monoclonal antibodies is achieved by dedicated unit operations, such as low pH inactivation, viral filtration, and chromatographic separation. The process development of each viral clearance step for a new antibody production requires significant effort and resources invested in wet laboratory experiments for process characterization studies. Machine learning methods have the potential to help streamline the development and optimization of viral clearance unit operations for new therapeutic antibodies. The current work focuses on evaluating the usefulness of machine learning methods for process understanding and predictive modeling for viral clearance via a case study on low pH viral inactivation.

Events of viral contaminations occurring during the production of biopharmaceuticals have been publicly reported by the biopharmaceutical industry. Upstream raw materials were often identified as the potential source of contamination. Viral contamination risk can be mitigated by inactivating or eliminating potential viruses of cell culture media and feed solutions. Different methods can be used alone or in combination on raw materials, cell culture media, or feed solutions such as viral inactivation technologies consisting mainly of high temperature short time, ultraviolet irradiation, and gamma radiation technologies or such as viral removal technology for instance nanofiltration. The aim of this review is to present the principle, the advantages, and the challenges of high temperature short time (HTST) technology. Here, we reviewed effectiveness of HTST treatment and its impact on media (filterability of media, degradation of components), on process performance (cell growth, cell metabolism, productivity), and product quality based on knowledge shared in the literature.

Avian influenzas, Ebola, Nipah, Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is an RNA virus covered by a lipid bilayer, are directly affecting people worldwide. On the other hand, in addition to the main spread source (human contact) of SARS-CoV-2, consumers have started to think about whether foods are dangerous in terms of SARS-CoV-2 spread. The consumption of wild animals as well as the possible contamination of SARS-CoV-2 in fresh and frozen foods have caused concern and increased awareness among consumers. A heating process >70°C is being suggested to eliminate viral contamination risk. Cutting tools, slicing machines, and food-contact surfaces including stainless steel, aluminum, or glass must be regularly sanitized. The sous vide cooking method, which is based on cooking under vacuum and with pH treatments in the range of 3 and 10, could be advised in this risky period for decreasing contamination risk in food. Also, recent studies have shown that nanotechnology applications such as nanoparticles could be used to combat the SARS-CoV-2 spread, which is 50–200 nm in size. Another suggested technique is cold plasma technology that could damage the protein structure of the virus. Besides these techniques, it is important to boost the immune system. In this regard, recent researches have revealed the importance of honey consumption (1 g/kg per person/day), intake of vitamins, minerals like selenium, and ω-3 fatty acids.

The world faces a severe and acute public health emergency due to the ongoing coronavirus disease 2019 (COVID-19) global pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Healthcare workers are in the front line of the COVID-19 outbreak response and are exposed to the risk of SARS-CoV-2 infection daily. Personal protective equipment (PPE) is their main defense against viral contamination; gloves, visors, face masks, and gown materials are designed to eliminate viral transfer from infected patients. Here, we review research investigating the stability of SARS-CoV-2 and similar viruses on surfaces and highlight opportunities for materials that can actively reduce SARS-CoV-2 surface contamination and associated transmission and improve PPE., Graphical Abstract, Progress and Potential It remains to be seen if any of the existing materials commonly used to fabricate PPE may offer improvements in viral binding and inactivation rate at their surface; for example, comparative viral lifetime data cannot be found for the three types of polymer gloves commonly used clinically. The theoretical basis for describing viral particle interactions at synthetic surfaces is not well developed. One means of identifying materials that does not require such a theoretical framework, and could help to build it, is to use high-throughput polymer microarray screening. There also appears to be an opportunity to develop new polymers for use, i.e., those that bind virus particles strongly and speed the inactivation of adsorbed virus under conditions modeling ambient transmission. In the medium term, there are a number of lines of enquiry for the development of anti-SARS-CoV-2 PPE materials that could reduce contact transfer of infectious virus in clinical and public settings., The world faces a severe and acute public health emergency caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Healthcare workers are on the front line and exposed to the risk of infection daily. Personal protective equipment (PPE) is their main defense against viral contamination. We review research investigating the binding and stability of SARS-CoV-2 and other viruses on surfaces, and highlight opportunities for materials that can actively reduce infectious SARS-CoV-2 surface contamination and associated transmission to improve PPE.

The pandemic due to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has emerged as a serious global public health issue. Since the start of the outbreak, the importance of hand-hygiene and respiratory protection to prevent the spread of the virus has been the prime focus for infection control. Health regulatory organisations have produced guidelines for the formulation of hand sanitisers to the manufacturing industries. This review summarises the studies on alcohol-based hand sanitisers and their disinfectant activity against SARS-CoV-2 and related viruses. The literature shows that the type and concentration of alcohol, formulation and nature of product, presence of excipients, applied volume, contact time and viral contamination load are critical factors that determine the effectiveness of hand sanitisers.

Viral contamination is a major concern for biological products. Therefore, virus testing of raw materials and cells is essential for the safety of the final product. We used high-throughput sequencing to detect viral-like sequences in selected CHO cell lines. Our aim was to test various approaches of sample preparation, to establish a pipeline for metagenomic analysis and to characterize standard viral metagenome of production and parental CHO cell lines. The comparison of the metagenomics composition of the differently prepared samples showed that among four tested approaches sequencing of ribosomal RNA depleted total RNA is the most promising approach. The metagenomics investigation of one production and three parental CHO cell lines of diverse origin did not indicate the presence of adventitious viral agents in the investigated samples. The study revealed an expected background of virus-like nucleic acids in the samples, which originate from remains of expression vectors, endogenized viral elements and residuals of bacteriophages.

Depuration of oysters can effectively reduce levels of E. coli, however, may not be effective in safeguarding against viral contamination (EFSA, 2012). These trials assess the removal of Norovirus Genogroups I and II (NoV GI and GII) and F+RNA bacteriophage genogroup II (FRNAP-II) from oysters under depuration using molecular and viability assay methods. Our results show consistently better removal of NoV GII compared with Nov GI. We found approximately 46% removal of NoV GII at 18°C after 2 days and 60% after 5 days compared with a maximum of 16% NoV GI removal. Twice the rate of NoV GII removal was achieved at 18°C compared with 8°C after 5 days. Results suggest better NoV removal when depuration water salinity is close to that prevailing in the harvesting area. Trials investigating algal feeding, light/dark and disturbance from pump vibration did not show any significant effect. We found that FRNAP-II was more readily removed than NoV. No significant difference was found between the rate of removal (as measured by RT-qPCR) and inactivation (as measured by bioassay) of FRNAP-II. This indicates that reduction in FRNAP-II may be primarily due to physical removal (or destruction) rather than in situ inactivation of the virus.

The problem of viral pollution of water bodies of the environment (OOS) is acute in the world. Until recently, sanitary-virological control of water sources and drinking water is carried out directly by direct determination of viruses, as well as using indirect indicators, the most optimal of which are coliphages [1-3]. However, in recent years, researchers have increasingly used viral markers - RNA and DNA of viruses, which are determined in the RT-PCR reaction, both for monitoring viral contamination of water bodies and for deciphering outbreaks of diseases associated with the use of poorquality water or food products that are unfavorable in the epidemic relation .

A mandatory requirement for cell cultures used in scientific researches and biomanufacturing is the absence of their contamination by viruses. We have described the case of adventitious isolation of mammalian orthoreovirus in the rhesus macaque embryo kidney cells MA-104. PCR analysis for the presence of reovirus RNA of all probable sources of reovirus (trypsin, fetal bovine serum, clinical samples, cell culture) revealed no viral RNA in any of the samples. An important condition for the activation of the reovirus reproduction in the MA-104 cells was the presence of trypsin in the culture medium. The obtained results underscore the urgency of control for the reovirus contamination of chemicals of animal origin and cell cultures. Since reoviruses are associated with human diseases, such control in pharmaceutical production is mandatory.

Viral contamination of the water bodies is neglected area of research in Saudi Arabia. The study, first of its kind, using RT- PCR technique, reports Hepatitis A virus (HAV) and Rotavirus (RV) presence in almost 13% of the samples collected. This report may trigger the detection of accurate viral contamination status in Saudi Arabia’s environmental waters.

Insect expression systems based on baculovirus are widely used for generating recombinant proteins. Here, the infectivity of baculoviruses under the physiological stresses of 'freeze-thaw' and sonication and the baculoviral contamination of recombinant proteins after protein purification were evaluated. Our findings suggest that Nonidet P-40 (NP-40) treatment of baculoviruses completely abolishes their infectivity and that recombinant proteins purified with affinity beads do not include infectious baculoviruses. We therefore suggest that baculovirus is completely inactivated by NP-40 treatment and that recombinant proteins are unlikely to be contaminated with infectious baculoviruses after their affinity purification.

Viral contamination is an inherent risk during the manufacture of biopharmaceuticals. As such, biopharmaceutical companies must demonstrate the viral clearance efficacy of their downstream process steps prior to clinical trials and commercial approval. This is accomplished through expensive and logistically challenging spiking studies, which utilize live mammalian viruses. These hurdles deter companies from analyzing viral clearance during process development and characterization. We utilized a noninfectious minute virus of mice-mock virus particle (MVM-MVP) as a surrogate spiking agent during small scale viral filtration (VF) and anion exchange chromatography (AEX) studies. For VF experiments, in-process mAb material was spiked and processed through Asahi Kasei P15, P20, P35, and BioEX nanofilters. Across each filter type, flux decay profiles and log reduction values (LRVs) were nearly identical for either particle. For AEX experiments, loads were conditioned with various amounts of sodium chloride (9, 20, 23, and 41 mS/cm), spiked with either particle and processed through a Q-SFF packed column. LRV results met our expectations of predicting MVM removal.

Gastroenteritis viruses in wastewater reclamation systems can pose a major threat to public health. In this study, multiple gastroenteritis viruses were detected from wastewater to estimate the viral contamination sources in a wastewater treatment and reclamation system installed in a suburb of Xi&rsquo, an city, China. Reverse transcription plus nested or semi-nested PCR, followed by sequencing and phylogenetic analysis, were used for detection and genotyping of noroviruses and rotaviruses. As a result, 91.7% (22/24) of raw sewage samples, 70.8% (17/24) of the wastewater samples treated by anaerobic/anoxic/oxic (A2O) process and 62.5% (15/24) of lake water samples were positive for at least one of target gastroenteritis viruses while all samples collected from membrane bioreactor effluent after free chlorine disinfection were negative. Sequence analyses of the PCR products revealed that epidemiologically minor strains of norovirus GI (GI/14) and GII (GII/13) were frequently detected in the system. Considering virus concentration in the disinfected MBR effluent which is used as the source of lake water is below the detection limit, these results indicate that artificial lake may be contaminated from sources other than the wastewater reclamation system, which may include aerosols, and there is a possible norovirus infection risk by exposure through reclaimed water usage and by onshore winds transporting aerosols containing norovirus.

The variability of environmental water samples impacts the allowance of one method to be universally ideal for all water types and volumes. Surface and reclaimed waters can be used for crop irrigation and may be referred to as non-traditional irrigation waters as these water types may be associated with a higher risk of microbial contamination compared to groundwater. These waters are typically more microbially and chemically complex than groundwater and have a higher risk of viral contamination. To detect viruses in these water types, an infinite number of variations can be made to traditional recovery methods. This protocol was developed based on a commonly used virus adsorption and elution (VIRADEL) method. Additional steps were included to simplify and efficiently reduce particulates in the viral concentrate and remove DNA from eluted nucleic acids prior to detection. Method alterations allow for volumes up to 40 liters to be processed with consistent recovery of enteric viruses including Aichi virus, hepatitis A virus, and noroviruses belonging to genogroups GI and GII. No inhibition was observed among either surface or reclaimed water samples. This protocol could be utilized in the monitoring of a wide array of irrigation water sources throughout irrigation processes.

Human enteric viruses have properties that are unique from those of bacterial foodborne pathogens. From an epidemiologic perspective, the noroviruses (NoVs) and hepatitis A virus (HAV) are the two most important enteric virus groups transmitted by foodborne routes. Human isolates of HAV comprise a single serotype, and monoclonal antibodies raised to different isolates fail to distinguish the isolates from one another. The rotaviruses usually are transmitted by either waterborne or person-to-person routes, but they can be food borne. This virus group is the leading cause of infantile diarrhea worldwide and is responsible for up to 130 million illnesses and 600,000 to 870,000 deaths per year, with the vast majority of rotavirus associated deaths occurring in developing countries. The contamination of produce items usually occurs before the product reaches food service establishments. The most commonly implicated bivalves are oysters, followed by clams, and it is now estimated that human enteric viruses are the most common disease agents transmitted by molluscan shellfish. Adherence to strict hygienic practices when handling and preparing foods is critical to control viral contamination of ready-to-eat (RTE) food products. A potential control strategy, albeit in developmental stages, would be the detection of viral contamination in foods. Improved and more widespread reporting and investigation of foodborne viral disease outbreaks, and targeted epidemiologic studies to identify the risk factors for viral gastroenteritis, would improve the understanding of attribution.

In mammalian cell culture technology, viral contamination is one of the main challenges; and, so far, various strategies have been taken to remove or inactivate viruses in the cell-line production process. The suitability and feasibility of each method are determined by different factors including effectiveness in target virus inactivation, maintaining recombinant protein stability, easiness-in terms of the process condition, cost-effectiveness, and eco-friendliness. In this research, Taguchi design-of-experiments (DOE) methodology was used to optimize a non-detergent viral inactivation method via considering four factors of temperature, time, pH, and alcohol concentration in an unbiased (orthogonal) fashion with low influence of nuisance factors. Herpes Simplex Virus-1 (HSV1) and Vero cell-line were used as models for enveloped viruses and cell-line, respectively. Examining the cytopathic effects (CPE) in different dilutions showed that pH (4), alcohol (15%), time (120 min), and temperature (25 °C) were the optimal points for viral inactivation. Evaluating the significance of each parameter in the HSV-1 inactivation using Taguchi and ANOVA analyses, the contributions of pH, alcohol, temperature and time were 56.5%, 19.2%, 12%, and 12%, respectively. Examining the impact of the optimal viral treatment condition on the stability of model recombinant protein-recombinant human erythropoietin, no destabilization was detected.

International audience; Viral aerosols can lead to respiratory viral infections with high infectivity. About 90% of people’s time is spent in closed environments. A few studies have pointed out that the ventilation systems in air handling units (AHUs) that treat and transmit a new synthetic clean and conditioned environment can also spread and transport viral particles in buildings. The aim of this work is to study the characterization of adenovirus, a DNA non-enveloped respiratory virus, on the F7 fiberglass filter used in AHUs. In this study, an experimental setup simulating an AHU was used. The SYBR® QPCR, Electrical Low-Pressure Impactor (ELPI™) and Scanning Mobility Particle Sizer (SMPS™) were used to detect, measure and characterize the aerosolized adenovirus solution. The characterization results showed that the nebulized adenovirus could be aerosolized in different forms associated or not with cell debris and proteins. The quantification and level of infectivity of adenovirus demonstrated that viruses passed through filters and remained infectious up- and downstream of the system during the 25 min of aerosolization. This study showed that AHUs should be considered an indoor source of viral contamination. © 2019, Springer Nature B.V.

Pseudorabies virus (PRV) is a zoonotic agent with a wide host range, causing significant economic losses in animal husbandry and potential public health risk globally. The causative agent has recently gained attention due to the inter-species transmission among different species of animals, even human beings. Although PRV's prevalence is found in many species of animals, regardless of whether the strain involved is a vaccine, classical or variant, few lines of evidence for the viral transmission route are available. Here, we reported that viral contamination is associated with the inter-species transmission of PRV. We found that PRV contamination was widely distributed in the environment of pig farms, that viral distribution in the environment is associated with the implementation of biosecurity measures, and that PRV could transmit from pigs to dogs through virally contaminated fomites. Collectively, our findings provide a basis for understanding the ecology and transmission route of PRV and underscore the importance of implementing biosecurity measures to control this disease.

Human enteric virus occurrence in bathing beaches poses a potential health risk to swimmers. They may come from several sources, but the understanding of the seasonal contribution of contamination sources to virus occurrence is still lacking. Here, the surveillance of human enteric viruses at the First Bathing Beach in Qingdao was performed January-December 2018. The occurrence of Enteric viruses, assayed with quantitative polymerase chain reaction (qPCR), was analyzed at temporal and spatial levels to determine the viral contamination sources. The results showed that only Astroviruses (AstVs) and Adenoviruses (HAdVs) were found in the swimming area. Their occurrence correlated significantly with the sewage-polluted area, but HAdVs were only found in autumn and AstVs in spring. Meanwhile, enteric viruses in the swimming area showed significantly higher levels than the surrounding area, particularly AstVs in summer with the swimmer crowd. All these data imply that sewage discharge and swimmers co-contribute to the viral occurrence in a seasonal pattern, with the former being more focused in warm seasons (spring and autumn) and the latter in hot seasons (summer). These results indicate that sewage discharge and crowd swimmers, as unsafe swimming conditions, should be avoided to improve public health at the bathing beaches.

Additive manufacturing has been extensively used during the COVID-19 pandemic to design and produce protection equipment. During clinical examinations using slit lamps, ophthalmologists are at risk of being contaminated by the SARS-CoV-2 virus, and the device itself is exposed to viral contamination. Several solutions have already been proposed for fixing transparent shields on the physician side. Here we propose a 3D-printed device fixed on the chin rest on the patient side, aiming at limiting viral spread both on the lamp itself and towards the physician.

Rotavirus is an enteric pathogen that causes morbidity and mortality in young mammals, including pigs. Outbreaks of rotavirus on commercial farms have a significant economic impact in terms of losses in production. Effective cleaning and disinfection along with good farm management can reduce rotavirus contamination in the environment, and decrease the chance of outbreaks of disease. This study investigated the efficacy of six commercial disinfectants against MS2 bacteriophage and Group A porcine rotavirus, in the presence of high and low levels of organic matter to simulate the farm environment. A phenolic-based disinfectant (Bi-OO-cyst) was effective at all levels of organic matter concentrations. Iodophore-based disinfectants did not have a significant virucidal effect against rotavirus under any conditions. For peroxygen compound-based disinfectants and glutaraldehyde-based disinfectants, organic matter load made a significant difference in reducing efficacy. This highlights the importance of thorough cleaning with detergent before disinfection to reduce viral contamination on the farm and decrease rotavirus disease incidence in pigs.Infection of rotavirus has a negative impact on the health and growth of pigs in production. Given that the virus is transmitted faecal-orally, use of an effective disinfectant on farm, which works even in high organic matter, has the potential to save costs in terms of outbreaks of disease and viral contamination. Here, we test a number of commercial disinfectants of which one a phenolic compound, Bio-OO-cyst, shows effectivity even in high organic matter, implying its use could have a huge impact in reducing viral contamination and preventing losses in production.

In a cluster of hepatitis A infections that occurred in Nagano Prefecture in 2017, hepatitis A virus (HAV) was detected in asari clams (reference food) and the patients' fecal samples. Initially, the asari clams were suspected to be the infection source. However, the exact infection route remained unknown because a patient who had not consumed an asari clam dish also developed the disease. Suspecting a secondary infection originating from the asari clams, we investigated the presence of HAV genomes in water used for washing and soaking the frozen asari clams and detected HAV in the soaking water. These results suggest that soaking water is a risk factor for secondary contamination because of the leakage of HAV accumulated in midgut gland of the asari clam. During the asari clam sand removal process, the water used to clean asari clams spread across a wide area in a concentric fashion, raising concerns that this process may aggravate contamination. In addition to HAV, diarrhea viruses, such as norovirus, have often been detected in bivalves, including asari clams. Thus, handling these foodstuffs requires adequate care.

Various mammalian cell lines are used as substrates for drug production without safety issues concerning viral contamination. However, viral contamination events in good manufacturing practice (GMP) cell culture processes, while rare, do sometimes occur. When contamination happens, it can result in serious consequences, including supply risk of life-saving drugs and substantial financial loss. To mitigate the potential risk of viral contamination, one approach taken by the industry is to implement preventative measures upstream. High-temperature short-time (HTST) treatment of culture media, at the point of use, was implemented as a virus barrier following murine minute virus (MMV) contamination. In recent years, nanofiltration, commonly used in downstream purification processes, has been evaluated for potential use as a virus barrier alternative to HTST. Several companies shared their data and experience in evaluating nanofiltration for viral barrier purpose upstream in Session 1, Part 2: Virus Barrier. These presentations are summarized below. LAY ABSTRACT: Viral contamination events in GMP cell culture processes, while rare, do sometimes occur. When contamination happens, it can result in serious consequences, including supply risk of life-saving drugs and substantial financial loss. To mitigate the potential risk of viral contamination, one approach taken by the industry is to implement preventative measures upstream. Several companies shared their data and experience in evaluating virus-retentive filtration for viral barrier purpose upstream.

This study evaluated the impact of sewage discharge in recreational coastal marine environments of Niteroi, Rio de Janeiro, Brazil, over a six-month period by the detection of waterborne enteric viruses. Ten-liter water samples were collected in four beaches from January to July 2017. Viruses were concentrated by an organic flocculation and human adenoviruses (HAdV), polyomavirus (JCPyV), and Hepatitis A virus (HAV) detected by qPCR. Forty-eight water samples were collected, being 43% positive for HAdV and 23% for JCPyV; only one sample was positive for HAV. Viruses were detected in all sampling sites, including in areas suitable for bathing according to the current bacterial standards. The results herein provide an overview of the viral contamination of beaches used for recreational purposes. The viral presence in the sampled areas indicates the need for more rigid effluent discharge controls in these areas, as sewage represents a possible transmission risk for waterborne viral diseases.

Recombinant protein therapeutics, vaccines, and plasma products have a long record of safety. However, the use of cell culture to produce recombinant proteins is still susceptible to contamination with viruses. These contaminations cost millions of dollars to recover from, can lead to patients not receiving therapies, and are very rare, which makes learning from past events difficult. A consortium of biotech companies, together with the Massachusetts Institute of Technology, has convened to collect data on these events. This industry-wide study provides insights into the most common viral contaminants, the source of those contaminants, the cell lines affected, corrective actions, as well as the impact of such events. These results have implications for the safe and effective production of not just current products, but also emerging cell and gene therapies which have shown much therapeutic promise.

Controlling viral contamination is an important issue in the process development of monoclonal antibodies (MAbs) produced from mammalian cell lines. Virus filtration (VF) has been demonstrated to be a robust and effective clearance step which can provide ≥4 logs of reduction via size exclusion. The minimization of VF area by increasing flux and filter loading is critical to achieving cost targets as VFs are single use and often represent up to 10% of total purification costs. The research presented in this publication describes a development strategy focused on biophysical attributes of product streams that are directly applicable to VF process performance. This article summarizes a case study where biophysical tools (high-pressure size exclusion chromatography, dynamic light scattering, and absolute size exclusion chromatography) were applied to a specific MAb program to illustrate how changes in feed composition (pH, sodium chloride concentration, and buffer salt type) can change biophysical properties which correlate with VF performance. The approach was subsequently refined and expanded over the course of development of three MAbs where performance metrics (i.e., loading and flux) were evaluated for two specific virus filters (Viresolve Pro and Planova 20N) during both unspiked control runs and virus clearance experiments. The analyses of feed attributes can be applied to a decision tree to guide the recommendation of a VF filter and operating conditions for use in future MAb program development. The understanding of the biophysical properties of the feed can be correlated to virus filter performance to significantly reduce the mass of product, time, and costs associated with virus filter step development.

Norovirus (NoV) gastroenteritis outbreaks appear frequently in food service operations (FSOs), such as in restaurants and canteens. In this study the presence of NoV and adenovirus (AdV) genomes was investigated on the surfaces of premises, especially in kitchens, of 30 FSOs where foodborne gastroenteritis outbreaks were suspected. The objective was to establish a possible association between the presence of virus genomes on surfaces and a visual hygienic status of the FSOs. NoV genome was found in 11 and AdV genome in 8 out of 30 FSOs. In total, 291 swabs were taken, of which 8.9% contained NoV and 5.8% AdV genome. The presence of NoV genomes on the surfaces was not found to associate with lower hygiene level of the premises when based on visual inspection; most (7/9) of the FSOs with NoV contamination on surfaces and a completed evaluation form had a good hygiene level (the best category). Restaurants had a significantly lower proportion of NoV-positive swabs compared to other FSOs (canteens, cafeteria, schools etc.) taken together (p = 0.00014). The presence of a designated break room for the workers was found to be significantly more common in AdV-negative kitchens (p = 0.046). Our findings suggest that swabbing is necessary for revealing viral contamination of surfaces and emphasis of hygiene inspections should be on the food handling procedures, and the education of food workers on virus transmission. Electronic supplementary material The online version of this article (doi:10.1007/s12560-017-9291-7) contains supplementary material, which is available to authorized users.