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Background: The risk of SARS-CoV-2 transmission to health care workers in intensive care units (ICU) and the contribution of airborne and fomites to SARS-CoV-2 transmission remain unclear. To assess the rate of air and surface contamination and identify risk factors associated with this contamination in patients admitted to the ICU for acute respiratory failure due to SARS-CoV-2 pneumonia. Methods: Prospective multicentric non-interventional study conducted from June 2020 to November 2020 in 3 French ICUs. For each enrolled patient, 3 predefined surfaces were swabbed, 2 air samples at 1 m and 3 m from the patient’s mouth and face masks of 3 health care workers (HCW) were collected within the first 48 h of SARS-CoV-2 positive PCR in a respiratory sample. Droplet digital PCR and quantitative PCR were performed on different samples, respectively. Results: Among 150 included patients, 5 (3.6%, 95%CI: 1.2% to 8.2%) had positive ddPCR on air samples at 1 m or 3 m. Seventy-one patients (53.3%, CI95%: 44.5% to 62.0%) had at least one surface positive. Face masks worn by HCW were positive in 6 patients (4.4%, CI: 1.6% to 9.4%). The threshold of RT-qPCR of the respiratory sample performed at inclusion (odds ratio, OR= 0.88, 95%CI: 0.83 to 0.93, p

HAdV and HBoV are frequently found in different water sources throughout the year. Due to their health impact on humans, continuous monitoring of these viruses is considered an important tool to evaluate their circulation within society. This study investigates the prevalence of HAdV and HBoV in different environmental samples collected from the western Nile Delta of Egypt. In total, 264 samples were collected from treated and untreated sewage (48), sewage sludge (24), drainage water (96), and drainage sediment (96) from December 2019 to November 2021. Molecular detection was performed to detect both viruses in the collected samples. The results indicated that HAdV was more prevalent than HBoV in all types of sewage samples, with the highest detection rate in untreated sewage. Both viruses were relatively lower in sludge sewage samples, while in drainage water and sediment samples were detected with similar frequencies, although the detection rates were generally lower than those observed in sewage samples. Moreover, no significant seasonal distribution of both viruses was observed throughout the year. The results of this study emphasize the importance of monitoring the presence of these viruses in environmental samples with proper treatment and disposal of sewage to ensure public health and safety. HIGHLIGHTS The first study that shows the prevalence of enteric viruses in the aquatic environment in the western delta region of Egypt.; HAdV and HBoV were detected in treated and untreated wastewater samples.; Both viruses did not show any significant seasonal variation.;

Objectives: Interpreting real-time reverse transcription-polymerase chain reaction (rRT-PCR) results for human avian influenza A virus (AIV) detection in contaminated settings like live bird markets (LBMs) without serology or viral culture poses a challenge. Methods: During February-March 2012 and November 2012-February 2013, we screened workers at nine LBMs in Dhaka, Bangladesh, to confirm molecular detections of AIV RNA in respiratory specimens with serology. We tested nasopharyngeal (NP) and throat swabs from workers with influenza-like illness (ILI) and NP, throat, and arm swabs from asymptomatic workers for influenza virus by rRT-PCR and sera for seroconversion and antibodies against HPAI A(H5N1) and A(H9N2) viruses. Results: Among 1273 ILI cases, 34 (2.6%) had A(H5), 56 (4%) had A(H9), and six (0.4%) had both A(H5) and A(H9) detected by rRT-PCR. Of 192 asymptomatic workers, A(H5) was detected in eight (4%) NP and 38 (20%) arm swabs. Of 28 ILI cases with A(H5) or A(H9) detected, none had evidence of seroconversion, but one (3.5%) and 12 (43%) were seropositive for A(H5) and A(H9), respectively. Conclusion: Detection of AIV RNA in respiratory specimens from symptomatic and asymptomatic LBM workers without evidence of seroconversion or virus isolation suggests environmental contamination, emphasizing caution in interpreting rRT-PCR results in high viral load settings.

• We investigated molecular detections of avian influenza A virus (AIV) RNA with serology. • AIV RNA detections in contaminated settings may suggest environmental contamination. • Influenza-like-illness symptom frequency was similar between live bird market workers with and without AIV RNA. • Symptomatic and asymptomatic workers exhibited comparable AIV RNA detection patterns. • Relying on real-time reverse transcription-polymerase chain reaction detections can overestimate AIV infections. Interpreting real-time reverse transcription-polymerase chain reaction (rRT-PCR) results for human avian influenza A virus (AIV) detection in contaminated settings like live bird markets (LBMs) without serology or viral culture poses a challenge. During February-March 2012 and November 2012-February 2013, we screened workers at nine LBMs in Dhaka, Bangladesh, to confirm molecular detections of AIV RNA in respiratory specimens with serology. We tested nasopharyngeal (NP) and throat swabs from workers with influenza-like illness (ILI) and NP, throat, and arm swabs from asymptomatic workers for influenza virus by rRT-PCR and sera for seroconversion and antibodies against HPAI A(H5N1) and A(H9N2) viruses. Among 1273 ILI cases, 34 (2.6%) had A(H5), 56 (4%) had A(H9), and six (0.4%) had both A(H5) and A(H9) detected by rRT-PCR. Of 192 asymptomatic workers, A(H5) was detected in eight (4%) NP and 38 (20%) arm swabs. Of 28 ILI cases with A(H5) or A(H9) detected, none had evidence of seroconversion, but one (3.5%) and 12 (43%) were seropositive for A(H5) and A(H9), respectively. Detection of AIV RNA in respiratory specimens from symptomatic and asymptomatic LBM workers without evidence of seroconversion or virus isolation suggests environmental contamination, emphasizing caution in interpreting rRT-PCR results in high viral load settings. [ABSTRACT FROM AUTHOR]

To ensure the safety and to secure the approval of injectable medicinal products based on antigen-specific immunoglobulins of animal origin, it is necessary to exclude their contamination with adventitious human pathogens. Ensuring the viral safety of heterologous immunoglobulins presents a major challenge, because the State Pharmacopoeia of the Russian Federation, 14 edition, lacks production stage-specific viral safety requirements for such medicinal products. The aim of the study was to analyse the requirements set forth in general and individual monographs of the State Pharmacopoeia of the Russian Federation, the European Pharmacopoeia, (10th edition), the British Pharmacopoeia (2019), the United States Pharmacopoeia (USP 43–NF 38), the Japanese Pharmacopoeia (17th edition), as well as the recommendations of the European Medicines Agency and the World Health Organisation concerning the viral safety of medicinal products for human use based on heterologous antigen-specific immunoglobulins. The authors analysed regulatory requirements for the following: serum/plasma-producing animals; immunisation antigens for the animals; quarantine of the animals; viral contamination tests for immune animal serum/plasma pools; model viruses to validate viral inactivation/removal processes at different stages of vaccine production; viral load reduction at each inactivation/ removal step; testing of materials obtained at critical production stages. The authors drafted sections for quality standards on production stage-specific measures to minimise the viral contamination risk of medicinal products for human use based on heterologous immunoglobulins, which they proposed for inclusion to the State Pharmacopoeia of the Russian Federation.

Mussels filter large amounts of water to extract nutrients; therefore, they can concentrate and accumulate in their tissues infectious agents, and vectors of enteric diseases. The aim of this study was to assess mussel contamination by Norovirus genogroups I and II in the Cherrat estuary to determine the public health risk linked to their consumption. Mussels (Mytilus galloprovincialis) were collected (n=52 samples; 12 mussels/sample) at four sites (S1 and S2 located on the right rocky bank / S3 and S4, located on the rocky left bank) in the Cherrat estuary (Casa-Settat region), Morocco, during 13 months, from March 2019 to March 2020. Norovirus was detected and quantified by real-time Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Norovirus genogroups I and II were detected in 17.30% and 94.23% of mussel samples, respectively. Contamination by Norovirus (genogroups I and II) was not correlated with seasonal factors (month and rainfall), and Norovirus prevalence was comparable among the four sampling sites. Consumption of raw or undercooked mussels contaminated with Norovirus can cause gastroenteritis, which represents a potential risk to human health. The present study would be helpful to control and manage the potential risk to the public health of the Moroccan population due to the degradation of water quality continuously impacted by runoff, the urban wastewater treatment system malfunctions, and overflows from nearby sewage systems. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

A significant decrease in norovirus prevalence and concentration was observed in oyster production areas in Ireland during winter 2020 to 2021. Oyster production areas impacted by human wastewater discharges that had been undergoing norovirus surveillance since 2018 were investigated. Samples collected in the winter seasons of 2018 to 2019 and 2019 to 2020, prior to when the COVID-19 pandemic interventions were applied, showed a prevalence of 94.3 and 96.6%, respectively, and geometric mean concentrations of 533 and 323 genome copies per g, respectively. These values decreased significantly during the winter of 2020 to 2021 (prevalence of 63.2% and geometric concentration of below the limit of quantification), coinciding with the control measures to mitigate the transmission of severe acute respiratory syndrome coronavirus 2 of the genus Betacoronavirus. Divergence between norovirus GI and GII prevalence and concentrations was observed over the 3-year monitoring period. Norovirus GII was the dominant genogroup detected in winter 2020 to 2021, with over half of samples positive, although concentrations detected were significantly lower than prepandemic winters, with a geometric mean concentration of below the limit of quantification. Oyster surveillance revealed the norovirus rate during the COVID-19 pandemic. SARS-CoV-2 mitigation measures had an indirect impact on norovirus transmission. A decrease in norovirus levels was seen in 2020 to 2021 compared with prepandemic winters. Norovirus concentrations were significantly lower than in previous winter periods. [ABSTRACT FROM AUTHOR]

The risk of SARS-CoV-2 transmission to health care workers in intensive care units (ICU) and the contribution of airborne and fomites to SARS-CoV-2 transmission remain unclear. To assess the rate of air and surface contamination and identify risk factors associated with this contamination in patients admitted to the ICU for acute respiratory failure due to SARS-CoV-2 pneumonia. Prospective multicentric non-interventional study conducted from June 2020 to November 2020 in 3 French ICUs. For each enrolled patient, 3 predefined surfaces were swabbed, 2 air samples at 1 m and 3 m from the patient's mouth and face masks of 3 health care workers (HCW) were collected within the first 48 h of SARS-CoV-2 positive PCR in a respiratory sample. Droplet digital PCR and quantitative PCR were performed on different samples, respectively. Among 150 included patients, 5 (3.6%, 95%CI: 1.2% to 8.2%) had positive ddPCR on air samples at 1 m or 3 m. Seventy-one patients (53.3%, CI95%: 44.5% to 62.0%) had at least one surface positive. Face masks worn by HCW were positive in 6 patients (4.4%, CI: 1.6% to 9.4%). The threshold of RT-qPCR of the respiratory sample performed at inclusion (odds ratio, OR= 0.88, 95%CI: 0.83 to 0.93, p < 0.0001) and the presence of diarrhea (OR= 3.28, 95%CI: 1.09 to 9.88, p = 0.037) were significantly associated with the number of contaminated surfaces. In this study, including patients admitted to the ICU for acute respiratory failure " contact route " of transmission, i.e. through fomites, seems dominant. While presence of SARS-CoV-2 in the air is rare in this specific population, the presence of diarrhea is associated to surface contamination around Covid patients. [ABSTRACT FROM AUTHOR]

BackgroundHand drying is an essential step of hand hygiene, helping remove microbes remaining on hands following handwashing. However, it is unclear whether particles dispersed or aerosolized during hand drying can also have an impact on microbe dissemination and so pose an infection risk.MethodsWe used a PR772 bacteriophage to investigate whether microorganisms remaining on hands can disperse in the washroom environment and contaminate facemasks of others sharing the same space, as a surrogate for virus inhalation risk. Hand drying using either a jet air dryer or paper towels were performed, and mask contamination by splattering and droplet deposition was investigated, up to 15 min following each procedure.ResultsFacemask contamination by splattering was 10-fold higher when a jet air dryer was used, compared with hand drying by paper towels, for both the person performing the hand drying and for standby users stationed at 1 and 2 m distance. Facemask contamination by droplet/aerosols deposition was higher in the first 5 min following hand drying, for both methods; however, virus load was significantly higher when a jet air dryer was used. In the jet air dryer assays, facemask contamination increased at 15 min post-hand drying, suggesting aerosolization of small particles that remain airborne for longer.ConclusionWhen using a jet air dryer, virus contamination dispersed further and for a longer period of time (up to 15 min post hand-drying). The method chosen for hand drying can potentially impact the airborne dissemination of microbial pathogens, including respiratory virus, and so potentially increase the risk of exposure and infection for other washroom users.

The selection of suitable cell cultures for use in the biological industry as well as research and diagnostic studies is critical. One of the factors affecting cell culture that can affect the results of studies is the contamination with viral agents. Therefore, efforts to preserve the health of cell cultures from contamination sound logical, and the use of virus-free cells is vital in research and diagnostic studies as well as in the manufacturing industries. For this purpose, it is crucial to use fast and correct diagnostic methods to detect the presence of critical viral contaminants in cells. Moreover, the use of susceptible diagnostic methods is also doubly important, especially in the case of contaminants that may remain hidden. Therefore, in this study, the BHK-21C5 cell line, one of the most widely used cells in the production and quality control of biological products and virological studies, was examined in terms of contamination with the most important viruses such as BVD and BLV. Detection of possible contaminants by using two-step RT-PCR to detect the 5' UTR portion of the BVD virus. Moreover, Nested PCR was carried out to detect the BLV virus using the gp51 env gene region. Also, an Flk cell line and Hela cell line that were consistently contaminated with the BLV virus were used as positive controls in Nested PCR. Due to the absence of bands in the BHK-21C5 cell column and the bandwidth observed in the positive control column (BVDV) in the range of 283 bp, non-contaminating of the cell clone with the BVD virus was proved. Also, no band was observed in well related to BHK-21C5 cell, and no cell clone contamination with BLV was confirmed, and in wells related to the positive controls (Flk and Hela cells), the bands have seen in the range 444bp. So, the results showed that no obvious or covert viral contamination effect was observed in the cell clone studied. Hence, the use of this cell line seems unobstructed in the quality control and production of biological products and research and diagnostic studies. [ABSTRACT FROM AUTHOR]

Early physical therapy rehabilitation protocols have shown improved functional outcomes and a reduction in mortality rates. Virtual rehabilitation (VR) has allowed practitioners to provide safe treatment without any risk of viral contamination. This review aimed to evaluate the importance of physical therapy and rehabilitation during and after the infection of coronavirus (COVID-19). To do so, four electronic databases such as PubMed, ResearchGate, Science Direct, and Google Scholars were searched to retrieve articles published after the outbreaks of the COVID-19 pandemic. Six studies were included in the final review. It is a compilation and examination of physical therapy treatments provided by various hospitals and organizations during and after COVID- 19. The present review throws light on physical therapy, going hand in hand with VR, to play a promising role in patients with mild to moderate symptoms, thus improving their quality of life. The necessary references, as well as the statistics gathered from the published articles, are available. A total of 12 046 patients were included. Of the total patients, 3085 (25.6%) had fatigue symptoms and 1866 (15.5%) had arthralgia/myalgia. Within an hour of being in the prone position, patients were found to have an increase in the oxygen saturation (SpO2) level from 94% to 98%. The pace of breathing also improved, dropping from 31 to 22 per minute. Physical therapy and VR have improved daily activities while assisting in the recovery of health, lifestyle, and lowering post-illness physical and mental deficiencies. [ABSTRACT FROM AUTHOR]

During the Covid-19 pandemic, location of the SARS-CoV-2 infected patients inside the hospital is a major issue to prevent viral cross-transmission. The objective of this study was to evaluate the risk of contamination through aerosol by using a global approach of the multiple environmental parameters to simulate, including seasonal context. A computational fluid dynamic (CFD) simulation based on the Lattice Boltzmann Method approach was used to predict airflow on the entire floor of a private hospital in Paris. The risk of contamination outside the rooms was evaluated by using a water vapor mass fraction tracker. Finally, the air contamination was estimated by a “cough model” producing several punctual emissions of contaminated air from potentially infected patients. In a winter configuration, the simulation showed a well-balanced ventilation on the floor and especially inside the rooms. After cough emissions from COVID-positive rooms, no significant contamination was observed in the circulation area, public waiting space and nurse office. On the contrary, in a summer configuration, the temperature difference due to the impact of the sun radiation between both sides of the building created additional air transport increasing the contamination risk in neighboring rooms and public spaces. Airborne spread was limited to rooms during winter conditions. On the contrary, during summer conditions, market airflow with potentially contaminated air coming from rooms located on the side of the building exposed to solar radiation was evidenced. These observations have implications to locate infected patients inside the building and for the conception of future health care structures.

Background: Closed system drug-transfer devices (CSTD) allow the reconstitution of hazardous drugs into infusion bags, while preserving the sterility of the product and preventing the escape of liquids and aerosols into the environment. Air-cleaning technology CSTD is based on an activated carbon filter and a membrane which enable maintaining the drug sterile by filtration of air entering the vial during pressure equalization. Objective: The study aimed to investigate if an air-cleaning CSTD can prevent liquid viral contamination by human coronavirus OC43 (HCoV-OC43). Methods: Chemfort™ CSTD with (intact) or without (control) a Toxi-Guard system was used to transfer liquids between an IV bag and an empty vial (a total of 5 liquid transfers) inside a sealed glove box contaminated by HCoV-OC43 aerosols. In addition, the vial adaptor was challenged by direct spray of HCoV-OC43 solution on the septum and filter areas. HCoV-OC43 RNA was extracted from samples of the transferred liquid and compared between the devices with or without a Toxi-Guard system. Results: Use of a CSTD with the Toxi-Guard system resulted in non-detectable cycle threshold (CT) values, indicative of no detectable HCoV-OC43RNA in the transferred liquid, even when the septa and filter areas were directly sprayed with HCoV-OC43 stock solution. In contrast, use of the CSTD with no Toxi-Guard system resulted in a detectable CT value of the transferred liquid. Conclusions: Using Chemfort CSTD with integral Toxi-Guard technology can prevent the introduction of microbial and airborne contaminants into the fluid path, thus potentially protecting patients from infection. [ABSTRACT FROM AUTHOR]

In the late 1930s, the 17D vaccine against yellow fever was produced in record time. 17D was and is an excellent vaccine. Its rapid diffusion led, however, to several problems, the most important among them being the 1942 massive contamination of the vaccine distributed to the US Army by the hepatitis B virus. The US part of this story is relatively well-known, but its Brazilian part much less so. In 1940, scientists who were producing the 17D vaccine in Rio de Janeiro found that it was contaminated by an "icterus virus" that originated in normal human serum. They solved this problem through the exclusion of human serum from vaccine production, but failed to persuade their US colleagues to do the same. The Rio experts, aware of the potential pitfalls of a new technology, carefully supervised the consequences of their vaccination campaigns. They were thus able to rapidly spot problems and eliminate them. By contrast, US scientists, persuaded of their technical superiority and distrustful of warnings that originated from a "less developed" country, neglected to implement basic public health rules. A major disaster followed. (Am J Public Health. 2021;111(9): 1654–1660. https://doi.org/10.2105/AJPH.2021.306313) [ABSTRACT FROM AUTHOR]

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.

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. [ABSTRACT FROM AUTHOR]

In this report, we describe the detection of bovine viral diarrhea virus (BVDV) contamination in commercial animal-derived sera and vaccines against animal viral pathogens on the market in Poland. Antibodies against BVDV were detected in 4/45 sera samples (8.9%) using an ELISA test. The presence of BVDV antigen was found using ELISA in 3/45 serum samples (6.6%) and 18/172 vaccine samples (10.5%). An RT -PCR was conducted using primers targeting two genome regions, the five prime untranslated region (5'UTR) and N-terminal protease (Npro). BVDV RNA was detected in 33/45 (73.3%) of sera, and 11/172 samples (6.4%) of collected vaccines, of which one vaccine did not declare BVDV strain in its composition. A single serum showed the presence of an infectious virus and only one was contaminated with all 3 species of BVDV. The most frequent species in sera was BVDV-3 (75.5%), whereas in vaccines only BVDV-1 was identified. Sequence analysis showed that the tested commercial sera and one vaccine were contaminated by six genotypes of BVDV: -1a, -1b, -1c, -1d, -2a, and -3. Identification of BVDV and its genetic material in animal-derived products is important due to the possibility of pestivirus transmission as well as the chance of falsifying the results of a diagnostic test. It also demonstrates the necessity of rigorous monitoring of the bioproducts used at the laboratory and industry level. [ABSTRACT FROM AUTHOR]

The Covid-19 pandemic has markedly changed our practices. This article analyses the risks of contamination among healthcare professionals (HCPs) during laparoscopic surgery on patients with Covid-19. Harmful effects of aerosols from a pneumoperitoneum, with the virus present, have not yet been quantified. Measures for the protection of HCPs are an extrapolation of those taken during other epidemics. They must still be mandatory to minimise the risk of viral contamination. Protection measures include personal protection equipment for HCPs, adaptation of surgical technique (method for obtaining pneumoperitoneum, filters, preferred intracorporeal anastomosis, precautions during the exsufflation of the pneumoperitoneum), and organisation of the operating room. [ABSTRACT FROM AUTHOR]

Enteroviruses are ubiquitous pathogens that are present worldwide and they are able to survive for long periods in a marine environment. They are transmitted through water and food contaminated with stools. The purpose of this study was to evaluate the prevalence of enteroviruses in oysters collected from Oualidia lagoon, using real-time RT-PCR. To achieve this objective, 624 oysters representing 52 batches were collected and tested for viral contamination using real-time polymerase chain reaction. The results show that 32.69% of the samples tested were positive for enteroviruses. The spatial distribution of the enteroviruses was statistically significant. Moreover, a prominent seasonal variation of enterovirus contamination was observed in this study. The consumption of oysters contaminated with this type of virus presents a major risk to human health, By causing serious illnesses such as gastroenteritis, hepatitis, and poliomyelitis, the presence of enteroviruses in oyster production areas represents a potential health risk. [ABSTRACT FROM AUTHOR]

Abstract Assessment of viral contamination is essential for monitoring the microbial quality of water and protection of public health, as human virus presence is not accurately determined using bacterial indicators. Currently, the time required for conventional viral testing means that water contaminated with human pathogens may be used (e.g. for drinking, recreation or irrigation) days before results are available. Here we report a new rapid method for coliphage enumeration, the QuantiPhage (QP) assay. The novelty of the assay is the use of cellulose absorbent pad materials to support coliphage growth and colorimetric detection, in place of agar that is used in the plaque assay. In addition to saving time associated with agar preparation and tempering, the QP assay enabled enumeration of somatic coliphages in 1.5–2 h and F+ coliphages in 2.5–3 h. The assays were highly sensitive, with a lower detection limit of 1 plaque forming unit (PFU) per mL where 1 mL sample volumes were analysed, and 1 PFU per 10 mL where 10 mL sample volumes were analysed. This is the first rapid culture assay to enable low numbers of coliphages to be reliably detected and to produce directly equivalent results to agar-based plaque assays. A novel gelatin-immobilisation method is also reported, that reduces time to prepare bacterial cells from ∼20 h to 40–60 min (depending on the assay format), and provides a ready to use form of cells, that is compatible with rapid detection and kit formats. When applied to analysis of somatic coliphages in wastewater samples and surface water samples, mean differences in results of the QP assay and the conventional plaque assay were not statistically significant (mean difference ≤ 0.15 log 10 PFU/L and 0.5 PFU/10 mL respectively, P > 0.05). The QP is a valuable tool for assessing microbial water quality, which may assist in improving the management of water resources. Graphical abstract Image 1 Highlights • Rapid, sensitive (1 PFU/10 mL) and simple culture method for enumerating coliphages. • Somatic coliphages can be detected in 1.5 h (>100 PFU) to 2 h (1–100 PFU). • F+ coliphages can be detected in 2.5 h (>100 PFU) to 3 h (1–100 PFU). • Use with freshly cultured or immobilised (ready to use) host bacteria. • Equivalent results to the conventional plaque assay, but in a much faster time. [ABSTRACT FROM AUTHOR]

Abstract Increased levels of norovirus contamination in oysters were reportedly associated with a gastroenteritis epidemic occurring upstream of an oyster farming area. In this study, we monitored the norovirus concentration in oysters weekly between November 2014 and March 2015 and investigated the statistical relationship between norovirus genogroup II (GII) concentrations in oyster and sewage samples and the number of gastroenteritis cases in the area using cross-correlation analysis. A peak correlation coefficient (R = 0.76) at a time lag of +1 week was observed between the number of gastroenteritis cases and norovirus GII concentrations in oysters, indicating that oyster contamination is correlated with the number of gastroenteritis cases with a 1-week delay. Moreover, weekly variations in norovirus GII genotypes in oysters were evaluated using pyrosequencing. Only GII.3 was detected in November and December 2014, whereas GII.17 and GII.4 were present from January to March 2015. GII.17 Kawasaki 2014 strains were detected more frequently than GII.4 Sydney 2012 strains in oyster samples, as previously observed in stool and sewage samples collected during the same study period in Miyagi, Japan. Our observations indicate that there is a time lag between the circulation of norovirus genotypes in the human population and the detection of those genotypes in oysters. Highlights • Oyster norovirus GII are associated with gastroenteritis cases with a 1-week delay. • Weekly variation in norovirus GII genotypes was observed in oysters. • Prevalent strains in oysters belonged to GII.3 or GII.17 Kawasaki 2014 lineage. • Strains prevalent in oysters were found in both gastroenteritis cases and sewage. [ABSTRACT FROM AUTHOR]

Abstract: Since 2012, lumpy skin disease virus (LSDV) has been spreading from the Middle East to south‐east Europe and Russia. Although vaccination campaigns have managed to contain LSDV outbreaks, the risk of further spread is still high. The most likely route of LSDV transmission in short distance spread is vector‐borne. Several arthropod species have been suggested as potential vectors, but no proven vector has yet been identified. To check whether promiscuous‐landing synanthropic flies such as the common housefly (Musca domestica) could be involved, we carried out entomological trapping at the site of a recent LSDV outbreak caused by a vaccine‐like LSDV strain. The presence of vaccine‐like LSDV DNA was confirmed by the assay developed herein, the assay by Agianniotaki et al. (2017) and RPO30 gene sequencing. No evidence of field LSDV strain circulation was revealed. In this study, we discovered that M. domestica flies carried vaccine‐like LSDV DNA (Ct > 25.5), whereas trapped stable flies from the same collection were negative for both field and vaccine LSDV. To check whether flies were contaminated internally and externally, 50 randomly selected flies from the same collection were washed four times and tested. Viral DNA was mainly detected in the 1st wash fluid, suggesting genome or even viral contamination on the insect cadaver. In this study, internal contamination in the insect bodies without differentiation between the body locations was also revealed; however, the clinical relevance for mechanical transmission is unknown. Further work is needed to clarify a role of M. domestica in the transmission of LSDV. To our knowledge, this is the first report demonstrating that an attenuated LSD vaccine strain has been identified in Russian cattle given the ban on the use of live attenuated vaccines against LSDV. [ABSTRACT FROM AUTHOR]

Abstract Blood transfusion safety has been increasingly improving during the past two decades. However, threats from both known and emerging pathogens require continual improvement and re-assessment of blood safety measures. In this respect, we are currently witnessing the broader implementation of Pathogen reduction technology (PRT) for blood complements. These methods, combined with existing safety measures, have helped to reduce the pathogen risks of transfusion-transmitted infections. Currently multiple reviews have compared levels of inactivation between different commercialized PRTs. However, to analyze levels of pathogen inactivation, it is necessary to understand the dynamics of infectivity as well as the modes of disease transmission by blood transfusion for various pathogens. It is well known that contributing variables include donor characteristics through the processing of blood components to ultimately the recipient characteristics, which create enormous variability in overall outcomes relative to disease transmission. The aim of this paper is to discuss bacterial and viral contamination of blood components in order to determine adequate levels of efficacy and subsequent disease transmission safety of current pathogen inactivation protocols that are designed to reduce the risk of transfusion-transmitted infections. In such a conceptual analysis, however, it is important to understand several contributing factors including the measurement of pathogen load in blood products and the dynamics, infectivity and disease transmission of various pathogens via transfusion of blood components and products. In many cases, the log reduction values observed do not truly reflect the extent of reduction in the levels of infectivity that are observed clinically. Results from clinical trials and hemovigilance programs upon routine implementation of PRT methods provide a more direct insight into effectiveness with regard to clinical relevance of in vitro spiking studies. These issues are briefly addressed in this manuscript. [ABSTRACT FROM AUTHOR]

Personal protective equipment (PPE) is an element of biosecurity intended to prevent the access or spread of diseases in farms. Nevertheless, to date no extensive reports exist about the effectiveness of different available PPE on farms. Thus, our aim was to estimate the degree of protection of PPE from viral contamination during farm visits. Two farms, infected with Aleutian mink disease virus and porcine circovirus–2 respectively, were visited by six visitors wearing different combinations of PPE: coveralls with hood and bootcovers, both with a certified barrier to infective agents (certified PPE group) and non-certified bootcover and coverall without hood (non-certified PPE group). Seventy-two swab samples from PPE and both hair and street clothes under PPE were taken after the visit and analysed by qPCR. Our results reveal viral exposure during visits, and the external protections of body and shoes were contaminated in all cases (24/24). In addition, protection from viral contamination varied noticeably according to the biosecurity elements used. A higher number of positives were detected in the non-certified PPE group than in the certified PPE group, both in elements under external protections (14/18 vs 3/18) and also in hair (4/6 vs 0/6). In fact, non-certified bootcovers broke during visits, resulting in viral contamination of the internal elements under them; these are consequently not suitable for using with wrinkled surfaces usually found in farm facilities. Thus, certified coveralls should be used in order to prevent contaminations, and workers and personnel of farms should be trained in their proper use. qPCR is a useful tool in the risk management of biosecurity programmes, and our results may serve as a model to evaluate different biosecurity measures. [ABSTRACT FROM AUTHOR]

ABSTRACT: 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. [ABSTRACT FROM AUTHOR]

Coxsackie B4 (CV-B4), is a major cause of viral myocarditis, dilated cardiomyopathy, and pancreatitis. Like other human enteroviruses, CV-B4 is ubiquitous, excreted in the stool, transmitted by fecal-oral route, and persists in the environment. In the context of studies on CV-B4 infection, it is important to investigate how this virus can be eliminated and to show the possibility of contamination risk with a CV-B4 E2 infected Swiss albino mice. Swiss albino female mice were inoculated with CV-B4 E2 strain and divided in two groups: the first was intraperitoneally (I.P.) infected; the second was orally infected. In order to study the CV-B4 E2 infection in mice, total RNA was extracted from thymus, spleen, pancreas, and intestine, and viral genome was detected using semi-nested (RT-PCR). To further demonstrate infection or immunization of mice, Sera obtained from infected mice were assayed in vitro for their neutralizing antibody. To detect virus in stool of infected mice, stool samples were collected at different post-infection (p.i.) times. Neutralizing antibodies were detectable all along the follow-up period (Day 0, 1, 3, 7, 9, 17, 22, 30, 45, 60 p.i.) in I.P and oral infected mice. Our results showed that when mice were inoculated successively at day 0 and day 8, neutralizing activity was increased in I.P route more than in the oral route. Viral isolation in HEp-2 cells showed negative results. Stool viral analyses reveal a low detection of the CV-B4 E2 genome for all infected mice. In conclusion, our experiments demonstrated that there are no risks linked with the stool of CV-B4 E2 of Swiss albino mice. It would be interesting to characterize the inhibitors of the virus infectivity in these biological samples (stool) and investigate their targets and mechanisms of action. [ABSTRACT FROM AUTHOR]

A variety of biological and chemical contaminants can adversely impact cells in culture, ranging from outright destruction of the culture, mutation, phenotypic changes to relatively minor changes in morphology, or growth rate. There are various approaches to detecting and mitigating the risk of biological or microbial contaminants in cell cultures, and these are discussed in this article. Chemical contaminants typically arise from improper handling or sourcing of cell culture reagents, glassware, or other types of consumables. These and other sources of chemical contaminants of cell cultures are discussed. The occurrence of chemical contamination is mitigated through adherence to best practices in sourcing and handling of such materials and by avoiding the use of volatile solvents within incubators that are used for maintaining cell cultures. [ABSTRACT FROM AUTHOR]

New regulatory and consumer demands highlight the importance of animal feed as a part of our national food safety system. Porcine epidemic diarrhea virus (PEDV) is the first viral pathogen confirmed to be widely transmissible in animal food. Because the potential for viral contamination in animal food is not well characterized, the objectives of this study were to 1) observe the magnitude of virus contamination in an animal food manufacturing facility, and 2) investigate a proposed method, feed sequencing, to decrease virus decontamination on animal food-contact surfaces. A U.S. virulent PEDV isolate was used to inoculate 50 kg swine feed, which was mixed, conveyed, and discharged into bags using pilot-scale feed manufacturing equipment. Surfaces were swabbed and analyzed for the presence of PEDV RNA by quantitative real-time polymerase chain reaction (qPCR). Environmental swabs indicated complete contamination of animal food-contact surfaces (0/40 vs. 48/48, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05) and near complete contamination of non-animal food-contact surfaces (0/24 vs. 16/18, positive baseline samples/total baseline samples, positive subsequent samples/total subsequent samples, respectively; P < 0.05). Flushing animal food-contact surfaces with low-risk feed is commonly used to reduce cross-contamination in animal feed manufacturing. Thus, four subsequent 50 kg batches of virus-free swine feed were manufactured using the same system to test its impact on decontaminating animal food-contact surfaces. Even after 4 subsequent sequences, animal food-contact surfaces retained viral RNA (28/33 positive samples/total samples), with conveying system being more contaminated than the mixer. A bioassay to test infectivity of dust from animal food-contact surfaces failed to produce infectivity. This study demonstrates the potential widespread viral contamination of surfaces in an animal food manufacturing facility and the difficulty of removing contamination using conventional feed sequencing, which underscores the importance for preventing viruses from entering and contaminating such facilities. [ABSTRACT FROM AUTHOR]

Ultraviolet (UV) irradiation is being considered for protection against viral contamination in cell culture media. Hydrogen peroxide (H 2 O 2 ) produced by UV irradiation has been suggested as the cause for poor cell growth in irradiated media, but this hypothesis has not been carefully evaluated. The impact of H 2 O 2 on Chinese Hamster Ovary (CHO) cell culture medium was compared with the impact of UV irradiation. Media composition was analyzed via nuclear magnetic resonance (NMR) spectroscopy and liquid chromatography mass spectrometry (LCMS), and cell growth in treated media was also evaluated. Although addition of H 2 O 2 to medium caused significant changes in pyruvate, formate, acetate, and sarcosine concentrations, there was less effect on CHO cell growth compared with irradiation. UV irradiation caused other changes in composition that did not occur as a result of H 2 O 2 addition. Catalase inhibited the effects of adding H 2 O 2 to the media, but catalase added before irradiation did not affect most irradiation-induced changes, even though catalase retained activity. In conclusion we found that while H 2 O 2 , which can be generated as a result of UV-irradiation, may be the cause of some changes in medium composition, it does not directly account for impaired CHO cell growth after high UV doses. [ABSTRACT FROM AUTHOR]

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’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.

Microbial food-borne diseases are still frequently reported despite the implementation of microbial quality legislation to improve food safety. Among all the microbial agents, viruses are the most important causative agents of food-borne outbreaks. The development and application of a new generation of sequencing techniques to test for viral contaminants in fresh produce is an unexplored field that allows for the study of the viral populations that might be transmitted by the fecal-oral route through the consumption of contaminated food. To advance this promising field, parsley was planted and grown under controlled conditions and irrigated using contaminated river water. Viruses polluting the irrigation water and the parsley leaves were studied by using metagenomics. To address possible contamination due to sample manipulation, library preparation, and other sources, parsley plants irrigated with nutritive solution were used as a negative control. In parallel, viruses present in the river water used for plant irrigation were analyzed using the same methodology. It was possible to assign viral taxons from 2.4 to 74.88% of the total reads sequenced depending on the sample. Most of the viral reads detected in the river water were related to the plant viral families Tymoviridae (66.13%) and Virgaviridae (14.45%) and the phage viral families Myoviridae (5.70%), Siphoviridae (5.06%), and Microviridae (2.89%). Less than 1% of the viral reads were related to viral families that infect humans, including members of the Adenoviridae , Reoviridae , Picornaviridae and Astroviridae families. On the surface of the parsley plants, most of the viral reads that were detected were assigned to the Dicistroviridae family (41.52%). Sequences related to important viral pathogens, such as the hepatitis E virus, several picornaviruses from species A and B as well as human sapoviruses and GIV noroviruses were detected. The high diversity of viral sequences found in the parsley plants suggests that irrigation on fecally-tainted food may have a role in the transmission of a wide diversity of viral families. This finding reinforces the idea that the best way to avoid food-borne viral diseases is to introduce good field irrigation and production practices. New strains have been identified that are related to the Picornaviridae and distantly related to the Hepeviridae family. However, the detection of a viral genome alone does not necessarily indicate there is a risk of infection or disease development. Thus, further investigation is crucial for correlating the detection of viral metagenomes in samples with the risk of infection. There is also an urgent need to develop new methods to improve the sensitivity of current Next Generation Sequencing (NGS) techniques in the food safety area. [ABSTRACT FROM AUTHOR]