Viral Contamination Research Articles

SARS-CoV-2 transmission via maritime cold chains: A statistical analysis of nucleic acid detection results of cold chain food imported from Fuzhou ports

Numerous epidemic outbreaks related to cold chains have occurred since the coronavirus disease 2019 (COVID-19) outbreak, suggesting the potential danger of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission through cold chain foods (CCFs). By analyzing SARS-CoV-2 RNA contamination of CCFs imported from Fuzhou ports, this study evaluated the contamination and transmission of SARS-CoV-2 RNA via maritime cold chains, with the aim of provide suggestions for CCFs supervision and public health management. The statistical analysis included 131,385 samples. The majority of the CCFs imported into Fuzhou ports was aquatic raw food that originated in Southeast Asia (57.08 %), South America (19.87 %), and South Asia (11.22 %). South Asia had the highest positivity rate of 0.37 %, followed by Southeast Asia (0.21 %) and South America (0.08 %). The positivity rate showed that the outer packaging of CCFs was the most easily contaminated, accounting for 81.33 % of all positive samples. This suggested that CCFs storage and loading processes were the weak links vulnerable to SARS-CoV-2 contamination. The positivity rates in outer packaging, inner packaging, and content of raw food were 0.48 %, 0.08 %, and 0.05 %, respectively, which were obviously higher than those of processed and refined food. This indicated that increasing the mechanization of factories and implementing sensible worker management practices may decrease viral contamination. The monthly positivity rates varied widely from 0 % (March 2021) to 0.40 % (January 2021), with an average of 0.19 %. The positivity rates in outer packaging, inner packaging and content of crustaceans from Southeast Asia were 2.47 %, 0.41 %, and 0.69 %, which were approximately 5–14 times higher than those of fish and cephalopods. Meanwhile, the monthly detection number show that SARS-CoV-2 epidemic prevention strategies affected the trade of imported CCFs.

Potential risk zones and climatic factors influencing the occurrence and persistence of avian influenza viruses in the environment of live bird markets in Bangladesh

Live bird markets (LBMs) are critical for poultry trade in many developing countries that are regarded as hotspots for the prevalence and contamination of avian influenza viruses (AIV). Therefore, we conducted weekly longitudinal environmental surveillance in LBMs to determine annual cyclic patterns of AIV subtypes, environmental risk zones, and the role of climatic factors on the AIV presence and persistence in the environment of LBM in Bangladesh. From January 2018 to March 2020, we collected weekly fecal and offal swab samples from each LBM and tested using rRT-PCR for the M gene and subtyped for H5, H7, and H9. We used Generalized Estimating Equations (GEE) approaches to account for repeated observations over time to correlate the AIV prevalence and potential risk factors and the negative binomial and Poisson model to investigate the role of climatic factors on environmental contamination of AIV at the LBM. Over the study period, 37.8% of samples tested AIV positive, 18.8% for A/H5, and A/H9 was, for 15.4%. We found the circulation of H5, H9, and co-circulation of H5 and H9 in the environmental surfaces year-round. The Generalized Estimating Equations (GEE) model reveals a distinct seasonal pattern in transmitting AIV and H5. Specifically, certain summer months exhibited a substantial reduction of risk up to 70–90% and 93–94% for AIV and H5 contamination, respectively. The slaughtering zone showed a significantly higher risk of contamination with H5, with a three-fold increase in risk compared to bird-holding zones. From the negative binomial model, we found that climatic factors like temperature and relative humidity were also significantly associated with weekly AIV circulation. An increase in temperature and relative humidity decreases the risk of AIV circulation. Our study underscores the significance of longitudinal environmental surveillance for identifying potential risk zones to detect H5 and H9 virus co-circulation and seasonal transmission, as well as the imperative for immediate interventions to reduce AIV at LBMs in Bangladesh. We recommend adopting a One Health approach to integrated AIV surveillance across animal, human, and environmental interfaces in order to prevent the epidemic and pandemic of AIV.

Virus filtration using small pore virus filter in downstream processing of biotherapeutic products: The effect of operating pressure

Virus filtration is a robust and effective method to remove potential virus contaminants. Planova 20 N, a virus filter form Asahi Kasei Bioprocess, has been widely used in the manufacturing process of biotherapeutics. Previous studies have shown that parvovirus removal by Planova 20 N can be impacted by low operation pressure and depressurization. Therefore, it is critical to define an operating pressure range for robust virus removal. In this work, the effect of pressure combined with depressurization on virus removal by Planova 20 N was investigated. Our studies showed that effective virus removal can be achieved in the pressure range from 0.7 bar to 1.6 bar. The data also suggest that re-starting with higher pressure after depressurization is highly desirable for large-scale manufacture to mitigate virus leakage risk. In addition, skipping buffer flush post mainstream filtration minimizes the likelihood of depressurization.

Risk assessment of norovirus and hepatitis A virus in strawberries imported into China.

Norovirus (NoV) and hepatitis A virus (HAV) pose a considerable health risk worldwide. In recent years, many cases of virus infection caused by virus-contaminated strawberries have occurred worldwide. This study applied a critical control point system to analyze the main hazards during the production and marketing of strawberries imported into China and explore the key control points in the whole process. To further evaluate the risks in the supply chain, the established quantitative microbial risk assessment (QMRA) was used to determine the probability that residents would be infected with viruses after consuming imported strawberries. It was found that the risk of virus contamination from imported strawberries was low, and the virus contamination mainly results from water resources and personnel. This research helps the regulatory authorities formulate strategies to ensure the long-term microbial safety of imported strawberries. In addition, the methods may prove useful in evaluating the risks of other agricultural produce.

COVID‑19 Pandemic: Effect of Specific and Non-Specific Prevention Measures on the Risk of SARS‑CoV‑2 Infection in Employees of Healthcare Organizations

From the very onset of SARS‑CoV‑2 spreading, active involvement in the COVID‑19 epidemic process made the healthcare professionals (HCPs) a vulnerable group with higher risks of contracting the disease, severe course and fatal outcome.The aim. We aimed at studying the impact of specific and non-specific preventive measures on the risk of SARS‑CoV‑2 infection among the HCPs in a large industrial region settings during the COVID‑19 pandemic.Materials and methods. We analyzed the data obtained using 1 905 questionnaires for the personnel of infectious hospitals for treatment of COVID‑19 patients and non-core healthcare facilities in a large industrial region during the COVID‑19 pandemic, 100 questionnaires on adherence of the employees to hand hygiene and antiseptics, the results of the tests for specific IgG to the SARS‑CoV‑2 nucleocapsid (2 200 samples), as well as the results of assessment of viral and bacterial contamination of the outer surface of the personal protective equipment for infectious hospital staff (108 specimens).Results and discussion. In the course of the study, an effect of failure in the implementation of some specific and non-specific preventive measures on an increase in the incidence of COVID‑19 in various healthcare professionals has been identified. The COVID‑19 pandemic once again emphasized the need to save lives and health of the HCPs as socially significant category of citizens under the spread of pathogens with high epidemic potential, applying a complex of specific and non-specific measures that are not mutually exclusive.

Severe acute respiratory coronavirus virus 2 (SARS-CoV-2) RNA and viable virus contamination of hospital emergency department surfaces and association with patient coronavirus disease 2019 (COVID-19) status and aerosol-generating procedures.

Emergency departments are high-risk settings for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) surface contamination. Environmental surface samples were obtained in rooms with patients suspected of having COVID-19 who did or did not undergo aerosol-generating procedures (AGPs). SARS-CoV-2 RNA surface contamination was most frequent in rooms occupied by coronavirus disease 2019 (COVID-19) patients who received no AGPs.

Persistence of Coronavirus on Surface Materials and Its Control Measures Using Nonthermal Plasma and Other Agents.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been responsible for the initiation of the global pandemic since 2020. The virus spreads through contaminated air particles, fomite, and surface-contaminated porous (i.e., paper, wood, and masks) and non-porous (i.e., plastic, stainless steel, and glass) materials. The persistence of viruses on materials depends on porosity, adsorption, evaporation, isoelectric point, and environmental conditions, such as temperature, pH, and relative humidity. Disinfection techniques are crucial for preventing viral contamination on animated and inanimate surfaces. Currently, there are few effective methodologies for preventing SARS-CoV-2 and other coronaviruses without any side effects. Before infection can occur, measures must be taken to prevent the persistence of the coronavirus on the surfaces of both porous and non-porous inanimate materials. This review focuses on coronavirus persistence in surface materials (inanimate) and control measures. Viruses are inactivated through chemical and physical methods; the chemical methods particularly include alcohol, chlorine, and peroxide, whereas temperature, pH, humidity, ultraviolet irradiation (UV), gamma radiation, X-rays, ozone, and non-thermal, plasma-generated reactive oxygen and nitrogen species (RONS) are physical methods.

Methods for virus recovery from environmental surfaces to monitor infectious viral contamination

Accurate quantification of infectious contaminants on environmental surfaces, particularly infectious viruses, is essential for contact transmission risk assessment; however, difficulties in recovering viruses from surfaces using swabs complicates this quantification process. Herein, we identified the factors that significantly affected virus recovery rates and developed an ideal swab method that yielded the highest rate of virus recovery. We comprehensively analyzed the effects of swab type (cotton/polyester), swab water content (wet/dry conditions), surface material, and surface area on the rates of viral RNA and infectious virus recovery.The virus recovery rate was significantly lower than the viral RNA recovery rate (P < 0.01), indicating difficulty in the quantification of infectious viruses. The virus recovery rate was significantly higher under wet conditions than that under dry conditions (P < 0.006), and the virus recovery rate obtained using cotton swabs was significantly higher than that using polyester swabs (P < 0.0001). Furthermore, the virus recovery rate had a strong negative correlation (correlation coefficient >0.8) with the target surface area. The maximum surface area where the virus recovery rate was ≥10% (MSA-10%) was identified as the maximum quantifiable area. For influenza virus recovery, MSA-10% on polyvinyl chloride (PVC) sheet, PVC leather, stainless steel, silicone, glass, and polycarbonate surfaces was 66.7, 193, 60.2, 144, 105, and 15.6 cm2, respectively. For feline calicivirus recovery, MSA-10% on PVC sheet, PVC leather, stainless steel, silicone, glass, and polycarbonate surfaces was 210, 111, 2120, 250, 322, and 180 cm2, respectively.The most accurate and ideal method for quantifying infectious viruses on environmental surfaces with the highest recovery rates meets three specifications: “wet conditions,” “the use of cotton swabs,” and “a target surface area of approximately 10 cm2.

Risk factors for H9N2 contamination in live bird markets of the Greater Jakarta, Indonesia

Low Pathogenicity Avian Influenza virus subtype H9N2 causes decreased egg production and two of the four defined H9N2 poultry lineages (G1 and Y280) have been associated with human infections. The high positive test results for the H9N2 virus in the LBMs of the Greater Jakarta area in 2021 can be a source of transmission for the surrounding area. This study aims to determine the prevalence and risk factors that play a role in the H9N2 contamination in the LBMs to obtain a multivariate analysis model.&lt;strong&gt; &lt;/strong&gt;A cross-sectional study was conducted on 87 vendors and 124 broiler chickens from 44 LBMs randomly selected in Tangerang City, North Jakarta, and Bekasi City. Samples were taken as equipment swabs at the stall of poultry carcass vendors, tracheal swabs, and cloacal swabs of broiler chicken. The test used was Real-Time Reverse Transcription Polymerase Chain Reaction. Data analysis was performed using univariate, bivariate, and multivariate analysis with logistic regression.&lt;strong&gt; &lt;/strong&gt;The prevalence of H9N2 virus contamination at the LBM level in Greater Jakarta in 2022 was 77.27% (95% CI, α 7%), while the prevalence at the vendor level was 54.02% (95% CI, α 5%). The risk factors for H9N2 virus contamination with a significant p-value&amp;lt;0.05 include the positive results on equipment swab samples, positive results on both equipment and broiler swabs, and the use of carcass sink. The multivariate analysis model of H9N2 virus contamination at the vendor level was Logit P (H9N2=1 | x) = 0.29924 - 2.42176PPE + 1.58691carcass sink. The risk factor contributing to increase H9N2 contamination is using carcass sinks, while PPE is the one that reduces H9N2 contamination.

Toward better monitoring of human noroviruses and F-specific RNA bacteriophages in aquatic environments using bivalve mollusks and passive samplers: A case study.

Monitoring pathogenic enteric viruses in continental and marine water bodies is essential to control the viral contamination of human populations. Human Noroviruses (NoV) are the main enteric viruses present in surface waters and foodstuff. In a context of global change, it is currently a challenge to improve the management of viral pollutions in aquatic environments and thereby limit the contamination of vulnerable water bodies or foodstuffs. The aim of this study is to evaluate the potential of specific accumulation systems for improving the detection of NoV in water bodies, compared to direct water analyses. Passive samplers (Zetapor filters) and three species of bivalve molluscan shellfish (BMS) (Dreissena polymorpha, Mytilus edulis and Crassostreas gigas) were used as accumulation systems to determine their performance in monitoring continental and marine waters for viruses. F-specific RNA bacteriophages (FRNAPH) were also analyzed since they are described as indicators of NoV hazard in many studies. During a one-year study in a specific area frequently affected by fecal pollution, twelve campaigns of exposure of passive samplers and BMS in continental and coastal waters were conducted. Using suitable methods, NoV (genome) and FRNAPH (infectious and genome) were detected in these accumulation systems and in water at the same time points to determine the frequency of detection but also to gain a better understanding of viral pollution in this area. The reliability of FRNAPH as a NoV indicator was also investigated. Our results clearly showed that BMS were significantly better than passive samplers and direct water analyses for monitoring NoV and FRNAPH contamination in water bodies. A dilution of viral pollution between the continental and the coastal area was observed and can be explained by the distance from the source of the pollution. Viral pollution is clearly greater during the winter period, and stakeholders should take this into consideration in their attempts to limit the contamination of food and water. A significant correlation was once again shown between NoV and FRNAPH genomes in BMS, confirming the reliability of FRNAPH as a NoV indicator. Moreover, a strong correlation was observed between NoV genomes and infectious FRNAPH, suggesting recent viral pollution since infectious particles had not been inactivated at sufficient levels in the environment. More generally, this study shows the value of using BMS as an active method for improving knowledge on the behavior of viral contamination in water bodies, the ranking of the contamination sources, and the vulnerability of downstream water bodies.

Possibilities of using mussels (Mytilus galloprovincialis) to predict rotavirus contamination in Albania.

Rotaviruses are non-enveloped viruses that each consist of 11 double-stranded RNA molecules. These viruses are able to persist in the environment, and therefore play a fundamental role in the epidemiology of gastroenteritis and severe diarrhoea in children worldwide. While mussels have been primarily used as indicators of chemical pollution, they can also be used to monitor viral contamination. The purpose of this study was to demonstrate that the Mytilus galloprovincialis mussel can also be used to detect microbial contamination, owing to its tendency to naturally concentrate viruses and other pathogens. A total of 102 Mytilus galloprovincialis mussel samples from Albania were collected over a three-year period: 37 samples off the Cape of Stillo in 2015, 39 samples from Butrinti Lake in 2019 and 26 samples from Butrinti Lake in 2021. The presence of rotavirus in the Cape of Stillo samples in 2015 was noted in 47% of samples from site 1, 33% from site 2, and 52% from site 3. In Butrinti Lake the percentage of infected individuals in 2019 was 33% from site 1, 41% from site 2, and 33% from site 3, whereas in 2021, it was 50% from site 1, 19% from site 2, and 0% from site 3. In total the percentage of infected individuals off the Cape of Stillo in 2015 was 44%, in Butrinti Lake in 2019 it was 36%, and in Butrinti Lake in 2021 it was 23 %. These results indicate the presence of rotavirus in the shellfish specimens tested, and further analysis is needed to assess the potential health risks associated with consuming these shellfish. This study also indicates that mussels can be used in marine virological surveillance programmes.

Novel Recombinant Rabbit Hemorrhagic Disease Virus 2 (RHDV2) is Circulating in China within 12 Months after Original RHDV2 Arrival

Rabbit hemorrhagic disease (RHD) causes lethal fulminant hepatitis in rabbits. Two different genotypes (GI.1 and GI.2) responsible for RHD are reported. GI.2 was first detected in France in 2010 and subsequently spread to other countries in Europe. In April 2020, GI.2 was detected in China. In this study, we report a novel recombinant strain of fatal rabbit hemorrhagic disease virus 2 (RHDV2 or GI.2) detected from domestic rabbits in three provinces in China in 2020–2021. Full-length genomic analysis has revealed that the recombinant virus contained an RHDV2 capsid gene and nonstructural genes from an unclassified lagovirus genotype. This type of virus emerged and circulated throughout China within a year after the initial detection of the original RHDV2. Compared with the original strain, the new virus showed a longer infected time and lower mortality rate but almost the same viral load at the moribund stage of infection. This might have resulted in high virus contamination in the environment, facilitating virus transmission. As the consequences of the presence of novel recombinant strains are unpredictable, the circulation of the novel variant in the population should be carefully monitored in China.

Progress and prospect of clinical application of anti-rabies virus monoclonal antibody preparation

Rabies is a severe infectious disease caused by the rabies virus, which seriously damages the central nervous system. Once it occurs, the fatality rate is close to 100%. The World Health Organization's position paper on rabies vaccines recognizes that rabies immunoglobulin (RIG) should be used for post-exposure prophylaxis (PEP) in all category Ⅲ exposure for the first time, as well as in category Ⅱ exposure that suffer from severe immune deficiency, long-term massive use of immunosuppressants, and head and face exposure. The anti-rabies virus monoclonal antibody has high purity and specific activity, can be produced on a sustainable scale, and has no risk of blood source virus contamination. Preclinical pharmacodynamic studies and clinical trial results of the anti-rabies virus monoclonal antibody preparation have confirmed that the preparation has a broad-spectrum neutralization effect on the rabies virus. Additionally, its combined application with the vaccine has little impact on the active immunity of the vaccine. Therefore, the anti-rabies virus monoclonal antibody preparation shows great potential for clinical application in PEP.

Evaluation of a novel chlorine dioxide-based packaging technology to reduce human enteric virus contamination on refrigerated tomatoes and blueberries

IntroductionChlorine dioxide (ClO2) is a promising antimicrobial with various food applications, one of those being inclusion in packaging. The purpose of this study was to evaluate a novel ClO2-based antimicrobial packaging system (InvisiShield™) for its efficacy against human norovirus (hNoV) and hepatitis A virus (HAV) in refrigerated fresh produce.MethodsGrape tomatoes or blueberries were placed in polypropylene trays and selectively inoculated with 6.0 log10 hNoV Genome Equivalent Copies (GEC; 20% stool suspension) or 6.2 log10 HAV GEC (cell culture lysate). Trays were heat sealed with a three-phase polymer film consisting of a base, channeling agent, and the ClO2 active (treatment); or control (no active) film and stored at 7°C for 24, 48 h, and 7 days. At each timepoint, the product was collected and processed for virus concentration using the sequential steps of elution and polyethylene glycol precipitation. Viruses in extracts were quantified using RNase-RT-qPCR.Results and discussionLog10 reductions (LR) in hNoV GEC for tomatoes were 2.2 ± 1.3, 2.9 ± 0.7, and 3.6 ± 0.3, after 24, 48 h and 7 days, respectively. For blueberries, hNoV LR were 1.4 ± 0.7, 1.7 ± 0.5, and 2.7 ± 0.2 GEC, respectively. Hepatitis A virus GEC LR were 0.4 ± 0.2, 1.0 ± 0.1, and 2.1 ± 0.7 for tomatoes, and 0.1 ± 0.2, 1.2 ± 0.4, and 3.2 ± 0.2 for blueberries, after 24, 48 h and 7 days, respectively. Position of the fruit in the tray did not affect inactivation (p &amp;gt; 0.05). Sensory analysis on the treated tomato products revealed no statistically significant difference in appearance, flavor and texture attributes compared to the control. This novel ClO2-based antimicrobial packaging system effectively reduced concentrations of hNoV and HAV, as evaluated using reduction in GEC as proxy for infectivity, on grape tomatoes and blueberries after one day, with efficacy improving over 7 days of refrigerated storage. This technology shows promise as an antiviral treatment as applied to refrigerated fresh produce items.

Waterborne viruses in urban groundwater environments

The proportion of global population using urban aquifers as drinking water sources increases every year and indeed the groundwater quality is not monitored adequately. Although norovirus has been identified as the first cause of groundwater-related outbreaks, the surveillance of waterborne viruses has been rather neglected. From ageing or disrupted sewer systems, occasional sewer discharges (e.g. combined sewer overflows, storm runoff), to poorly managed reclaimed water infiltration practices, multiple are the pathways that cause groundwater quality deterioration. This study revises the main viral contamination sources and the factors affecting viral contamination of groundwater bodies in terms of transport, inactivation, and survival of the viral particles. It also summarizes the methods used for those reporting the presence of human viruses in urban groundwaters. A total of 36 articles have been included in the method survey spanning a period of 24 years (1999–2022). There is a need of systematic monitoring considering representative set of waterborne pathogens. The evaluation of the presence of human adenovirus seems a useful tool to predict the presence of other waterborne pathogens in groundwater. Large volume sampling methods, but also new passive sampling methodologies applied to groundwater, coupled to target massive sequencing approaches may elucidate the range of pathogens capable of contaminating urban groundwaters for further evaluation of risk.

Advanced Molecular Docking Approach to Simulate Intermolecular Interaction and Binding Affinity Energy of Four Key Bio-Compounds Against Angiotensin Converting Enzyme 2 (ACE2) Receptor

The scientific community has been mobilized in recent years at the international level to find concrete solution, in order to elaborate vaccines against Covid-19 with less side-effects and thus to overcome this pandemic situation. However, despite the fact that the actual situation is partially under control, the issue of variants remains very problematic until this day. Bioinformatics as an emerging discipline has allowed scientists to understand the pathophysiological mechanism of COVID-19, a disease characterized by its complexity, as well as the comprehension of other serious infectious diseases. In this context, this modest work aimed to explore for the first time the possible binding process of four phyto-compounds against Angiotensin Converting Enzyme 2 (ACE2) receptor using molecular docking approach. The molecular docking analysis suggested that our compounds may possibly interact with ACE2 receptor principally via hydrogen bound and hydrophobic contact, but with an antagonistic way. Data of this study also suggested that compounds 2 and 3 have the best and an exceptional equal binding energy score (−6.8 kcal/mol), among the four tested compounds. This research could be considered a primordial and initial step for future preclinical and clinical experiments investigating SARS-CoV-2 variants, especially to better understand the mechanism of action of ACE2 receptor and the mode of contamination of host cells by this virus.

Biosecurity and Mitigation Strategies to Control Swine Viruses in Feed Ingredients and Complete Feeds.

No system nor standardized analytical procedures at commercial laboratories exist to facilitate and accurately measure potential viable virus contamination in feed ingredients and complete feeds globally. As a result, there is high uncertainty of the extent of swine virus contamination in global feed supply chains. Many knowledge gaps need to be addressed to improve our ability to prevent virus contamination and transmission in swine feed. This review summarizes the current state of knowledge involving: (1) the need for biosecurity protocols to identify production, processing, storage, and transportation conditions that may cause virus contamination of feed ingredients and complete feed; (2) challenges of measuring virus inactivation; (3) virus survival in feed ingredients during transportation and storage; (4) minimum infectious doses; (5) differences between using a food safety objective versus a performance objective as potential approaches for risk assessment in swine feed; (6) swine virus inactivation from thermal and irradiation processes, and chemical mitigants in feed ingredients and complete feed; (7) efficacy of virus decontamination strategies in feed mills; (8) benefits of functional ingredients, nutrients, and commercial feed additives in pig diets during a viral health challenge; and (9) considerations for improved risk assessment models of virus contamination in feed supply chains.

Evaluation of a viral transcriptome Next Generation Sequencing assay as an alternative to animal assays for viral safety testing of cell substrates

The viral safety of biological products is ensured by tests throughout the production chain, and, for certain products, by steps in the manufacturing process enabling the elimination or inactivation of viruses. Current testing programs include sample inoculation in animals and embryonic eggs. Following the 3Rs principles of replacement, reduction, and refinement of animal-use methods, such techniques are intended to be replaced not only for ethical reasons but also because of their inherent technical limitations, their long turnaround times, and their limits in virus detection. Therefore, we have compared the limit and range of sensitivity of in vivo tests used for viral testing of cells with a transcriptomic assay based on Next Generation Sequencing (NGS). Cell cultures were infected with a panel of nine (9) viruses, among them only five (5) were detected, with variable sensitivity, by in vivo tests. The transcriptomic assay was able to detect one (1) infected cell among 103 to 107 non-infected cells for all viruses assessed, including those not detected by the conventional in vivo tests. Here we show that NGS extends the breath of detection of viral contaminants compared to traditional testing. Collectively, these results support the replacement of the conventional in vivo tests by an NGS-based transcriptomic assay for virus safety testing of cell substrates.

The Essential Points Regarding Laboratory Biosafety to Prevent the Spread of SARS-CoV-2

: A current biosafety and biosecurity crisis, SARS-CoV-2/COVID-19, threatens public health, making it crucial to assess and follow optimal laboratory conditions to prevent any contamination and spread of the lethal virus. Much medical staff have been lost to the COVID-19 pandemic. The dimensions of laboratory biosafety were examined in this study to reduce risks associated with COVID-19 treatment. In addition, this research primarily focuses on assessing laboratory biosafety regarding emerging coronaviruses. This research will determine which aspects need to be addressed to address the risks and implement them in the correct order and at the right time to be as educational as possible.

Protection of biomanufacturing processes from virus contamination through upstream virus filtration of cell culture media.

Cell-based manufacturing processes have occasionally been exposed to adventitious viruses, leading to manufacturing interruptions and unstable supply situations. The rapid progress of advanced therapy medicinal products needs innovative approaches to avoid any unwelcome reminder of the universal presence of viruses. Here, we investigated upstream virus filtration as a clearance step for any product too complex for downstream interventions. Culture media virus filtration was investigated with respect to virus clearance capacities under extreme conditions such as high process feed loading (up to ~19,000 L/m²), long duration (up to 34 days), and multiple process interruptions (up to 21 h). The small nonenveloped Minute virus of mice was used as relevant target virus, and as worse-case challenge for the investigated virus filters with a stipulated pore-size of about 20 nm. Certain filters-especially of the newer second generation-were capable of effective virus clearance despite the harsh regimen they were subjected to. The biochemical parameters for un-spiked control runs showed the filters to have no measurable impact on the composition of the culture media. Based on these findings, this technology seems to be quite feasible for large volume premanufacturing process culture media preparations.