Viral Contamination Research Articles

Emerging Begomovirus ToLCNDV Molecular Detection & Chip Grafting Inoculation Transmission Assay under Natural Conditions in Tomato Plants from District Poonch Azad Kashmir & Pakistan

Tomato leaf curl New Delhi virus (ToLCNDV) is a bipartite, emerging Begomovirus species, usually coupled with satellite molecules and affects tomato cultivation in entire Pakistan. Present study was conducted to evaluate and analyze tomato germplasm against ToLCNDV using Chip graft inoculation assay (CGIA) as most accurate and reliable technique. Molecular studies were conducted for confirmation. According to PCR results, viral contamination was confirmed as ToLCNDV was recorded from all tested seedling samples. Further mechanical transmission confirmed that seedlings from ToLCNDV-infected seeds act as inoculum. Based on disease symptoms developed, plants were classified as tolerant, moderately tolerant, moderately susceptible, and susceptible. It is pertinent to mention that the magnitude of the virus impact particularly on hosts different from Solanaceae and Cucurbitaceae, and seed transmission and ToLCNDV qualify as a potential candidate of Quarantine Pest. Present work is novel study demonstrating ToLCNDV as a seed-transmissible virus in tomato plants from Rawalakot Azad Kashmir & this research further provides a fictitious starting point for plant pathologists suggesting practical implications regarding management approaches.

Pork as a Source of Diverse Viral Foodborne Infections: An Escalating Issue

This review synthesizes current knowledge on the risks posed by viral foodborne infections associated with pork, emphasizing their global prevalence and the complexity of managing such pathogens. It covers a range of significant viruses, including hepatitis A and E, norovirus, rotavirus, sapovirus, enterovirus, astrovirus, and enteric adenovirus. The role of pigs as reservoirs for diverse pathogens with zoonotic potential further complicates safety challenges, extending risks to individuals involved in pork production and processing. Various factors influencing viral contamination throughout the meat production chain are explored, from farm-level practices to processing and handling procedures. Emphasis is placed on the critical importance of implementing effective control measures at each stage, including enhanced biosecurity, rigorous hygiene practices, and appropriate thermal processing techniques. Additionally, the need for improved surveillance and detection methods to effectively identify and monitor viral presence in meat products is highlighted. In conclusion, the necessity of adopting a One Health approach that integrates efforts in animal health, food safety, and public health to mitigate the risks of viral foodborne infections associated with meat consumption is underscored. This holistic strategy is essential for safeguarding consumer health and ensuring the safety of the global food supply.

Research progress on environmental stability of SARS-CoV-2 and influenza viruses.

We reviewed research on SARS-CoV-2 and influenza virus detection on surfaces, their persistence under various conditions, and response to disinfectants. Viral contamination in community and healthcare settings was analyzed, emphasizing survival on surfaces influenced by temperature, pH, and material. Findings showed higher concentrations enhance survivability at room temperature, whereas stability increases at 4°C. Both viruses decline in low pH and high heat, with influenza affected by salinity. On various material surfaces, SARS-CoV-2 and influenza viruses demonstrate considerable variations in survival durations, and SARS-CoV-2 is more stable than influenza virus. On the skin, both virus types can persist for ≥2 h. Next, we delineated the virucidal efficacy of disinfectants against SARS-CoV-2 and influenza viruses. In daily life, exposure to ethanol (70%), isopropanol (70%), bleach (10%), or hydrogen peroxide (1-3%) for 15-30 min can effectively inactive various SARS-CoV-2 variants. Povidone-iodine (1 mg/mL, 1 min) or cetylpyridinium chloride (0.1 mg/mL, 2 min) may be used to inactive different SARS-CoV-2 variants in the mouth. Chlorine disinfectants (500 mg/L) or ultraviolet light (222 nm) can effectively inhibit different SARS-CoV-2 variants in public spaces. In conclusion, our study provides a scientific basis and practical guidance for reduction of viral persistence (retention of infectivity) on surfaces and environmental cleanliness.

The Antiviral and Antimalarial Prodrug Artemisinin from the Artemisia Species: A Review

Artemisinin is a truly fascinating drug in many ways. Since the unrestrained procedure of its detection, as an antimalarial drug, artemisinin has received a great deal of consideration. Recently, application of artemisinin-based combination therapy has been broadly applied for treating numerous ailments. Moreover, as an antimalarial compound, artemisinin and its associated compounds have abundant healing efficacy and can be repurposed for additional symptoms, like autoimmune infections, cancer, and viral contaminations. Recently a number of studies have highlighted the significance of the artemisinin-related compounds in SARS-CoV-2 treatment. The current review purposes to present a concise account of the history of the antiviral and antimalarial prodrugs—Artemisinin, from the Artemisia species. It is followed by its antiviral, antimalarial prospective, chemical nature and extraction procedure, photochemistry, mechanism of action, and its clinical trials and patents, and accentuates the significance of the mechanistic studies concerned for therapeutic results, both in viral and malarial circumstances.

Importance of RNase monitoring during large-scale manufacturing and analysis of mRNA-LNP based vaccines

Ribonucleases (RNases) are ubiquitous in nature, being able to cleave a wide range of polyribonucleotides. While the presence of microbial and viral contamination in sterile manufacturing is highly studied and controlled, there are no standardized practices for evaluating RNase in the production facility. Since the COVID-19 pandemic, mRNA-LNP based vaccines have become part of routine large-scale manufacturing. The unstable nature of mRNA poses new challenges to safeguard the working efficacy of mRNA – Lipid nanoparticle (LNP) based vaccines or therapeutics, where the presence of RNase in the formulation process could have a profound impact on the mRNA integrity. In this article, lessons learned are presented with respect to the evaluation of RNase contamination during LNP drug product formulation and analysis. Using sensitive detection methods, the potential presence of RNase in the manufacturing of mRNA-LNPs was investigated. Additionally, capillary gel electrophoresis (CGE) data, used to measure mRNA integrity, demonstrate the quality of the active mRNA substance and importance of suitable RNase control strategies. The results and cases presented in this paper should pave the way forward for evaluation and control strategies dedicated to mRNA-LNP based vaccines and therapeutics.

Removal of virus from hands: a study on the role of washing and drying

Proper hand hygiene is highly important to control pathogens' transmission. Although the effect of washing and drying on hands' bacterial contamination has been reported, studies on viruses are scarce. To assess the viral load of artificially contaminated hands after washing and after drying. 30 volunteers filled a questionnaire about hand hygiene and participated in microbial assays, testing five different drying approaches, using the whole hand methodology to quantify viruses on hands. Bacterial assays were also performed for comparison purposes. For both virus and bacteria contaminations, the washing step promoted a significant reduction of the microbial load, while the drying step only promoted slight reductions regardless of the drying method used. Moreover, hand dryers and paper towels didn't induce the re-contamination of washed hands. Hand washing promoted a reduction of the microbial load in hands, while none of the different drying methods significantly reduced hands microbial load.

Groundwater in Katowice - center of a large urban and deep coal mining area in Poland

The paper presents the groundwater resources of Katowice and the results of studies conducted to identify prospective areas in aquifers with sufficient water resources for the location of public water intakes. The city of Katowice is the capital of Upper Silesia, the largest urban-industrial area in Poland, the region with a highly developed hard coal mining and many other different industries covering almost the entire scope of the economy. Due to long term drainage of mines causing the depletion and deterioration of groundwater and surface water resources, the main municipal water intakes have been located outside Katowice. Most of the pumped mine water is discharged into surface waters, causing significant salinity of water and disallow its use for municipal or industrial purposes in Katowice. The water collection and supply system is very extensive and covers a significant part of the Silesian agglomeration which is why Katowice is supplied with water from surface water intakes located outside the city. According to older published materials, such as the Hydrogeological map of Poland, there were no aquifers in the city that could have been used to supply the city with drinking water, however recent studies have shown that groundwater in the Quaternary aquifers in the western part of the city can be used to supply the city with public water. Less productive, Quaternary aquifers in north-eastern parts of the city may constitute the basis for drinking water supply in emergency situations, when surface intakes cannot be used, such as toxic, radiological or viral contamination or damage to the water supply network eg. as a result of an earthquake or warfare.

Graphene-Based Virus Enrichment Protocol Increases the Detection Sensitivity of Human Norovirus in Strawberry and Oyster Samples.

Human noroviruses (HuNoVs), the most prevalent viral contaminant in food, account for a substantial proportion of nonbacterial gastroenteritis cases. Extensive work has been focused on the diagnosis of HuNoVs in clinical samples, whereas the availability of sensitive detection methods for their detection in food is lacking. Here, we developed a virus enrichment approach utilizing graphene-based nanocomposites (CTAB-rGO-Fe3O4) that does not rely on large instruments and is suitable for on-site food pretreatment. The recovery efficiency of the developed virus enrichment procedure for serially diluted GII.4 norovirus ranged from 10.06 to 72.67% in strawberries and from 2.66 to 79.65% in oysters. Furthermore, we developed a real-time recombinase polymerase amplification (real-time RPA) assay, which can detect as low as 1.22 genome copies µL-1 of recombinant plasmid standard and has no cross-reactivity with genomes of astrovirus, rotavirus, adenovirus, and MS2 bacteriophage. Notably, the combined virus enrichment and real-time RPA detection assay enhanced the detection limits to 2.84 and 37.5 genome copies g-1 in strawberries and oysters, respectively, compared to those of qPCR. Our strategy, the graphene-based virus enrichment method combined with real-time RPA, presents a promising tool for sensitively detecting HuNoVs in food samples.

Monitoring airborne pathogen transmission for enhanced safety at food processing facilities

AbstractFood manufacturing and processing are part of the nation's critical infrastructure. Due to the recent global spread of the SARS-CoV-2 virus, the potential contamination of the food chain and the resulting public health implications are of high consequence to society. The current primary food manufacturing and processing facilities already have various mechanisms such as hazard analysis and critical control point (HACCP) system in place. However, the widespread microbial infections in these facilities raise concerns that they will not only threaten the welfare of food processing workers, but also have a potentially greater consequence on the public if the food is contaminated with an infectious agent.Despite the increasingly recognised role of the environment in the spread of microbes, the effect of air properties remains poorly understood. Heating, ventilation, and air conditioning (HVAC) systems in meat processing facilities not only provide a means of transport for viruses and bacteria but may also deposit them on surfaces where they can survive for days. To maintain a stable and safe food chain supply during the pandemic, the challenges to ensure safe food supply and protect the workers' health must be quickly addressed through sustainable, safe and economic approaches. With these two imminent challenges in mind, the overall goal of this review article is to provide a comprehensive overview of the role of the environment in the impaction and resuspension of bioaerosols, focusing on airborne bacteria and viruses. The review includes the latest results of modeling the spread of microbial aerosols in the airflow and the development of preventive measures to mitigate virus contamination in the unique environment of meat processing operations. By understanding how the environmental factors and seasonality affect the infectivity and spread of airborne pathogens, mitigation measures can be designed to minimise future infections within and beyond these facilities.

Virus inactivation using an electrically conducting virus filter in biopharmaceutical manufacturing process

Biopharmaceutical manufacturing processes using mammalian cells or plasma carry the risk of viral contamination. To mitigate these risks, it is essential to ensure viral clearance during the downstream process. Virus-retentive filters are used for size-based virus filtration, offering robust viral removal of more than 99.99%. However, virus breakthroughs have also been reported during virus filtration under certain conditions. In addition, these virus-retentive filters are disposable to ensure the safety of bioproducts, leading to significant costs and environmental concerns. In this study, innovative electrically conducting virus filters were fabricated using free-standing carbon veils (CV) and used to achieve additional virus inactivation after filtration. The viruses were captured in a CV-assisted virus filter, which was electrically heated using direct current to inactivate the viruses. This electrically conducting virus filter can inactivate viruses and can be reused up to five times. These results demonstrate that electrical conduction through electrical conducting damaged the phage capsid and eliminated the RNA genome, leading to bacteriophage inactivation. Moreover, it was confirmed that the electrically conducting virus filter could be reused up to five times without any changes to its physical or chemical structure. This study contributes to the reduction of process costs and environmental impacts by enabling the reuse of virus filters and enhancing the safety of the virus filtration process by preventing undesired virus breakthroughs.

Minimal influenza virus transmission from touching contaminated face masks: a laboratory study

The risk of virus transmission via the touching of contaminated masks has long been assumed by infection control teams. Yet, robust evidence to support this belief has been lacking. This risk was investigated in a laboratory setting by measuring the amount of viable influenza virus successfully transferred from artificially contaminated medical (surgical) mask surfaces to a human finger used to swipe their outer surface under various experimental conditions. Despite being exposed to high levels of virus contamination on the masks, very little or no viable virus was successfully transferred from the mask to the finger in these experiments.

Assessment of the viral contamination of fecal origin over a wide geographical area using an active approach with Dreissena polymorpha

Biomonitoring appears to be a key approach to assess chemical or microbiological contaminations. The freshwater mussel, Dreissena polymorpha (D. polymorpha), is a suitable tool already used to monitor chemical and, more recently, microbiological pollution. In the present study, we used this sentinel species to monitor viral contamination of fecal origin over a wide geographical distribution. An active approach was implemented based on caging of calibrated and pathogen-free organisms with the same exposure conditions, allowing spatio-temporal comparisons between different water bodies. In addition, different types of sites were selected to investigate the range of environmental concentrations that D. polymorpha are able to translate. Different viral genome targets were measured: norovirus genogroup I and II (NoV GI and GII) and F-specific RNA bacteriophages belonging to the genogroup –I and –II (FRNAPH-I and -II). Total infectious FRNAPH were also monitored.D. polymorpha was able to translate a wide range of concentrations for all the viral targets studied, meaning that this sentinel species can be used for both low and highly anthropised sites. Moreover, D. polymorpha caging proved effective in achieving gradients of viral contamination of fecal origin pressure and to highlight the contribution of tributaries to the main rivers. D. polymorpha provided spatial and temporal variations of the viral contamination. It allowed to highlight the prevalence of the FRNAPH-I and -II genogroups according to the caging site. FRNAPH-II was found to be dominant in urban areas and FRNAPH-I in rural areas. This strategy uses the caging of the sentinel species D. polymorpha on selected sites with standardised analysis methods has proven to be a promising tool for characterizing viral contamination at both large and very fine scales.

Longitudinal study on the multifactorial public health risks associated with sewage reclamation

This year-long research analyzed emerging risks in influent, effluent wastewaters and biosolids from six wastewater treatment plants in Spain’s Valencian Region. Specifically, it focused on human enteric and respiratory viruses, bacterial and viral faecal contamination indicators, extended-spectrum beta-lactamases-producing Escherichia coli, and antibiotic-resistance genes. Additionally, particles and microplastics in biosolid and wastewater samples were assessed. Human enteric viruses were prevalent in influent wastewater, with limited post-treatment reduction. Wastewater treatment effectively eliminated respiratory viruses, except for low levels of SARS-CoV-2 in effluent and biosolid samples, suggesting minimal public health risk. Antibiotic resistance genes and microplastics were persistently found in effluent and biosolids, thus indicating treatment inefficiencies and potential environmental dissemination. This multifaced research sheds light on diverse contaminants present after water reclamation, emphasizing the interconnectedness of human, animal, and environmental health in wastewater management. It underscores the need for a One Health approach to address the United Nations Sustainable Development Goals.

QUESTÕES DE BIOÉTICA NA PANDEMIA DA COVID-19

Bioethics area at the Department of Legal Medicine, Bioethics, Labor Medice and Phisycal and Reabilitation Medicine at São Paulo University Medical School, has been developing ethical researches concerning the conflicts of modern world. The new coronavirus pandemic (COVID-19) has brough several conflicts which might be the most important ones in the last decades. This paper aims at relating some aspects of daily life of those affected by the vírus, besides morbidity and mortality, along with bioethical concepts. The following issues were discussed: autonomy and social inequality, social isolation and intrafamily violence and death and mourning. It is observed that daily conflicts were far beyond the virus contamination. There was an overflow of pandemic side effects. Autonomy, bioethical principle most questioned during pandemic, showed to be thought about only within a relationship bioethics. One can only be able to practice one’s autonomy in an ethical way through an agreement between one´s autonomy and one´s own social responsibility. Social inequalities were widely open during pandemic time thus leading to inequality of treatment resources. Social isolation, which was necessary to stop the virus caused a condition presente in all socieconomic classes: sexual violence against children and adolescents.

Upcycling of medical protective clothing for H2 production through catalytic steam reforming over Ni-impregnated catalysts

Disposing medical protective clothing (MPC) presents a significant challenge due to the potential risk of virus contamination. Catalytic steam reforming using Ni-impregnated catalysts was proposed to upcycle MPC for H2-rich gas production. Parametric studies validated that an optimal steam rate of 42 mL/h and a catalytic reforming temperature of 800 ℃ were ideal for H2-rich gas production. 10 wt% Ni impregnated on diverse supports (HY, ZSM-5, MCM-41, γ-Al2O3, MgO, CaO, and activated carbon) were prepared for steam reforming of MPC to assess the impact of support type on H2 production. Of these Ni-impregnated catalysts, Ni/CaO was found to present the highest gas yield (141.4 mmol/g) with a high H2 proportion of 65.2 vol%, due to its superior catalytic cracking and reforming ability of Ni/CaO, favoring the conversion of MPC intermediates into H2-rich gas. To evaluate the recyclability and reforming activity, the optimized Ni/CaO with and without catalyst regeneration was evaluated throughout the successive tests. It was found that Ni/CaO regenerated throughout the successive tests could more effectively mitigate the decreasing tendency of gas yields, favor H2 production, and reduce tar formation than the catalyst without regeneration. That was due to the removal of carbon deposition by catalyst regeneration, thereby mitigating the catalyst deactivation. In contrast, Ni/CaO without the catalyst regeneration was more responsible for the CO2 reduction, accompanying with a more significant decrease of ƞ (4.8 – 3.5 g CO2 /g H2). Overall, this study demonstrated the feasibility of valorizing medical plastic waste into H2-rich gas using effective Ni-based catalysts.

Fractional‐order modeling of Chikungunya virus transmission dynamics

This article presents two innovative mathematical models for the dynamics of Chikungunya virus contamination by using Caputo fractional derivative. By applying the recently developed numerical technique to find the approximate solutions for the Chikungunya virus system which allowing us for the valuable insights. Through a rigorous analysis of the obtained numerical and graphical solutions, the impact of fractional orders on the infection dynamics is thoroughly examined. Additionally, Banach's fix point theorm is used to investigates the existence, uniqueness, and stability properties of the solutions, providing a deeper understanding of the key parameters that affect the spread and persistence of the infection.

Knowledge and utilization of Integrated Child Development Services among antenatal women during COVID-19 pandemic.

The coronavirus disease 2019 (COVID-19) pandemic has affected the healthcare delivery system in both developed and developing countries. Many antenatal women skipped their regular antenatal check-ups due to fear of viral contamination and lack of transport facilities. Integrated Child Development Services (ICDS), which plays a major role in antenatal care, also suffered a great deal during the pandemic. 1. To assess knowledge and utilization of ICDS services by antenatal women during the COVID-19 pandemic. 2. To determine factors influencing the knowledge and utilization of ICDS services by antenatal women. 3. To determine the barriers to utilization of ICDS services during the pandemic. This study was conducted as a descriptive study in a tertiary care center. Antenatal women whose pregnancy period coincided with the second wave of the COVID-19 pandemic were interviewed using a structured questionnaire. The sample size was calculated as 198. Data were entered in Microsoft Excel and analyzed using Statistical Package for Social Sciences (SPSS). Around 61% of the study population had adequate knowledge about ICDS services. The proportion of antenatal women utilizing ICDS services reduced from 87.8% to 69.6% during the pandemic. Only a few of the mothers were followed up by the Anganwadi workers during the pandemic. The main barriers to not utilizing ICDS services were fear of the spread of the pathogen and lack of awareness about the availability of services during the pandemic. There has been a reduction in the utilization of ICDS services by antenatal women during the COVID-19 pandemic. Measures should be taken by the government to ensure undisrupted care for antenatal women during such hard times.

Identification of infectious viruses for risk-based virus testing of CHO unprocessed bulk using next-generation sequencing.

It is important to increase manufacturing speed to make medicines more widely available. One bottleneck for CHO-based drug substance release is the in vitro viral (IVV) cell-based assay on unprocessed bulk. To increase process speed, we evaluate the suitability of replacing the IVV cell-based assay with next-generation sequencing (NGS). First, we outline how NGS is currently used in the pharmaceutical industry, and how it may apply to CHO virus testing. Second, we examine CHO virus contamination history. Since prior virus contaminants can replicate in the production bioreactor, we perform a literature search and classify 159 viruses as high, medium, low, or unknown risk based on their ability to infect CHO cells. Overall, the risk of virus contamination during the CHO manufacturing process is low. Only six viruses were reported to have contaminated CHO bioprocesses over the past several decades, and were primarily caused by fetal bovine serum or cell culture components. These virus contamination events can be mitigated through limitation and control of raw materials, combined with virus testing and virus clearance technologies. The list of CHO infectious viruses provides a starting framework for virus safety risk assessment and NGS development. Furthermore, ICH Q5A (R2) includes NGS as a molecular method for adventitious agent testing, paving a path forward for modernizing CHO virus testing.

Cytopathic Effect Detection and Clonal Selection using Deep Learning.

In biotechnology, microscopic cell imaging is often used to identify and analyze cell morphology and cell state for a variety of applications. For example, microscopy can be used to detect the presence of cytopathic effects (CPE) in cell culture samples to determine virus contamination. Another application of microscopy is to verify clonality during cell line development. Conventionally, inspection of these microscopy images is performed manually by human analysts. This is both tedious and time consuming. In this paper, we propose using supervised deep learning algorithms to automate the cell detection processes mentioned above. The proposed algorithms utilize image processing techniques and convolutional neural networks (CNN) to detect the presence of CPE and to verify the clonality in cell line development. We train and test the algorithms on image data which have been collected and labeled by domain experts. Our experiments have shown promising results in terms of both accuracy and speed. Deep learning algorithms achieve high accuracy (more than 95%) on both CPE detection and clonal selection applications, resulting in a highly efficient and cost-effective automation process.

Factors Determining Adherence to Hand Antisepsis by Healthcare Workers during Pandemic Infection Spread (as exemplified by COVID­-19)

Relevance. The healthcare personnel’s adherence to hand antisepsis is а pressing problem during any epidemic outbreak. In the context of high biological hazards, the possibility of performing proper antiseptic procedures is reduced while nosocomial pathogen transmission and healthcare-associated infection (HAI) rate growth are promoted. Aims. Studying the factors determining the healthcare workers’ adherence to hand antisepsis during a pandemic infection spread (as exemplified by COVID-19). Materials and methods. The study was conducted during the entire COVID-19 pandemic. To estimate the rate of adherence rate by the healthcare personnel to hand hygiene, we conducted an anonymous questionnaire survey of the staff in infectious disease hospitals (2021–2022) as well as assessed the viral and bacterial contamination rates of the personnel’s hands and the hospital environment items (2021–2023). Epidemiological (descriptive-evaluation), bacteriological (conventional culture, AMR determination using a VITEK2 analyzer), molecular biological (RT-PCR, Sanger sequencing) and statistical (a questionnaire survey, building machine learning models) methods were used in the study. Results. Based on the results of the survey, the main factors influencing the formation of attitudes towards antiseptics among the staff of medical organizations with medical and non-medical education were identified. It was found that the pandemic spread of infections negatively affects the level of contamination of the hands of infectious diseases hospital staff, which amounted to 35.3% of doctors, 28.8% of nurses, and 25% of hospital cleaners. The spectrum of isolated pathogens included viruses (SARS-CoV-2) and opportunistic microorganisms (Enterococcus faecalis, Enterococcus faecium, Pantoea agglomerans, Acinetobacter baumannii, Klebsiella pneumoniae, Staphylococcus aureus, Escherichia coli), including those with a wide profile of antimicrobial resistance. Conclusion. During the COVID-19 epidemic, the rate of adherence to hand antisepsis in the infectious disease hospital was 82.3% in the staff with a medical background and 71.2% in the staff without one. The results confirm the significant role of staff hands in the in-hospital transmission of dangerous pathogens and indicate the need for systematic monitoring of the level of knowledge on antiseptics among all employees of a medical organization, regardless of specialization and level of education.