Search Results (150)

Surrogate viruses provide a safe and scalable alternative for evaluating disinfectant efficacy when access to high-risk pathogens is restricted. This study evaluated the potential of Modified Vaccinia Ankara (MVA) virus, which can be handled under BSL-1/2 conditions, as a surrogate for avian influenza virus (AIV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV). A total of 64 commercially available disinfectants—classified into four major chemical groups: quaternary ammonium compounds, oxidizing agents, PPMS-based formulations, and organic acids—were tested in suspension assays using a ≥4 log reduction as the efficacy criterion. MVA showed the strongest predictive performance for FMDV (r = 0.671, AUC = 0.83), supporting its use for both binary classification and approximate quantitative prediction. Although its correlation with ASFV was weaker (r = 0.175), the classification performance remained moderate (AUC = 0.78), indicating conditional applicability. While MVA exhibited no meaningful correlation with AIV, its higher chemical resistance meant that disinfectants effective against MVA were consistently effective against AIV. These results support the use of MVA as a conservative exclusion tool for fragile viruses. Overall, the findings demonstrate that MVA can serve as a practical surrogate virus for disinfectant efficacy testing against FMDV, ASFV, and AIV, with application strategies tailored to each virus’s characteristics. Full article

Fluoropyrimidine derivatives can cause severe toxicity in patients with DPD deficiency. Regulatory agencies, such as the European Medicines Agency (EMA), recommend pre-emptive genotyping of the HapB3 haplotype, along with other variants. Historically, the two main HapB3 variants, the benign c.1236G>A and the pathogenic c.1129-5923C>G, have been assumed to be in complete linkage disequilibrium. Recent findings contradict this assumption, questioning the reliability of the HapB3 analysis through c.1236G>A, which could directly impact patient safety. The aim of this study is to assess the linkage disequilibrium between the c.1236G>A and c.1129-5923C>G variants, with the ultimate goal of revising genotyping guidelines. A total of 46 patients already heterozygous for the c.1236G>A variant have been carefully reviewed for the c.1129-5923C>G variant. From the 46 patients analyzed, 45 maintain complete linkage disequilibrium between both variants. However, there is one patient where this linkage disequilibrium is not complete, being heterozygous for c.1236G>A and homozygous for c.1129-5923C>G. These findings challenge the validity of c.1236G>A as a surrogate marker for pathogenic variant c.1129-5923C>G. This article highlights the need for a review of the recommendations of the EMA and suggests laboratories to analyze both variants, or at least the pathogenic one, to ensure accurate therapeutic decisions. Full article

Background: It is still challenging to develop effective vaccines against tumorigenic human gammaherpesviruses such as Epstein–Barr virus (EBV). A major obstacle is the lack of a small animal model that reproduces the natural infection course of human gammaherpesviruses to allow for proper assessment of vaccine efficacy. Murine gammaherpesvirus 68 (MHV68) is a natural pathogen of wild rodents and laboratory mice and therefore can be used as a surrogate for human gammaherpesviruses to evaluate vaccination strategies. Methods: In this study, two mRNA vaccine candidates were generated, one encoding a fusion protein of the MHV68 gH with the gL (gHgL-mRNA) and the other expressing the MHV68 gB protein (gB-mRNA). The immunogenicity and protective efficacy of the mRNA vaccine candidates were evaluated in a mouse model of MHV68 infection. Results: The gHgL-mRNA but not the gB-mRNA candidate vaccine was able to induce neutralizing antibodies in mice, whereas both vaccines could elicit antigen-specific T-cell responses. Following MHV68 intranasal inoculation, complete blocking of the establishment of viral latency was observed in some mice immunized with individual gHgL-mRNA or gB-mRNA vaccines. Notably, co-immunization with the two mRNA vaccines appeared to be more effective than individual vaccines, achieving sterile immunity in 50% of the vaccinated mice. Conclusions: This study demonstrates that immunization with mRNA platform-based subunit vaccines is indeed capable of preventing MHV68 latent infection, thus validating a safe and efficacious vaccination strategy that may be applicable to human gammaherpesviruses. Full article

Recent outbreaks of highly pathogenic human RNA viruses from probable zoonotic origin have highlighted the relevance of epidemic preparedness as a society. However, research in vaccinology and virology, as well as epidemiologic surveillance, is often constrained by the biological risk that live virus experimentation entails. These also involve expensive costs, time-consuming procedures, and advanced personnel expertise, hampering market access for many drugs. Most of these drawbacks can be circumvented with the use of pseudotyped viruses, which are surrogate, non-pathogenic recombinant viral particles bearing the surface envelope protein of a virus of interest. Pseudotyped viruses significantly expand the research potential in virology, enabling the study of non-culturable or highly infectious pathogens in a safer environment. Most are derived from lentiviral vectors, which confer a series of advantages due to their superior efficiency. During the past decade, many studies employing pseudotyped viruses have evaluated the efficacy of vaccines or monoclonal antibodies for relevant pathogens such as HIV-1, Ebolavirus, Influenza virus, or SARS-CoV-2. In this review, we aim to provide an overview of the applications of pseudotyped viruses when evaluating the neutralization capacity of exposed individuals, or candidate vaccines and antivirals in both preclinical models and clinical trials, to further help develop effective countermeasures against emerging neutralization-escape phenotypes. Full article

Background: Neisseria meningitidis B is one of the main causative pathogens of meningitis and other forms of severe meningococcal disease. In the past decade, meningococcal B vaccines have been developed to address this infection and its sequelae. Objective: This article aims to present an example of how the EU regulatory framework allowed the early authorisation of two life-saving vaccines initially based on immunogenicity surrogates of clinical evidence. This was subsequently followed by post-marketing surveillance providing real-world evidence to support their safety profile and impact on the paediatric population in the EU. Methods: We review the evidence supporting the initial regulatory approval of the vaccines, the confirmatory data demonstrating vaccine effectiveness post-authorisation, and the real-world impact of these vaccines on the paediatric population. Results: Two vaccines were approved in the EU for active immunisation to prevent IMD caused by MenB (4CMenB in 2013 and MenB-fHBP in 2017). Both marketing authorisations were based on immunogenicity data (efficacy studies were not feasible due to the rarity of the disease) and safety data generated from pre-authorisation studies. Additional pharmacovigilance activities to further investigate the safety profile and effectiveness studies were requested to be conducted after approval. Both the effectiveness and safety profile of the vaccines were confirmed by these data. Conclusions: This paper illustrates that the EU medicines regulatory framework and safety monitoring system are robust. By supplementing the initial evidence with post-authorisation studies, further effectiveness and safety data enabled regulators to confirm the positive benefit–risk of the vaccines without delaying their access to the people who need them. Full article

Research on highly pathogenic biosafety level 4 (BSL-4) viruses that are classified as Select Agents involves transferring inactivated materials to lower containment levels for further analysis. Compliance with Select Agent and BSL-4 safety regulations necessitates the validation and verification of inactivation procedures. To streamline this process, it would be beneficial to use surrogate BSL-2 viruses for inactivation studies. This not only simplifies BSL-4 work but also enables the testing and validation of inactivation procedures in research facilities that lack access to high-containment laboratories yet may receive samples containing highly pathogenic viruses that require efficient and complete inactivation. In this study, we used Hazara virus (HAZV) as a surrogate virus for Crimean–Congo hemorrhagic fever virus to show the efficacy of various inactivation methods. We demonstrate the successful inactivation of HAZV using TRIzol/TRIzol LS and aldehyde fixation. Importantly, the parameters of the aldehyde inactivation of cell pellets differed from those of the monolayers, highlighting the importance of inactivation validation. As part of this study, we also defined specific criteria that must be met by a BSL-2 virus to be used as a surrogate for a closely related BSL-4 virus. Defining these criteria helps identify suitable nonpathogenic surrogates for developing inactivation procedures for highly pathogenic viruses. Full article

Indoors, the infection risk of diseases transmitted through the airborne route is estimated from indoor carbon dioxide (CO₂) levels. However, the approaches to assess this risk do not account for the airborne concentration of pathogens, among other limitations. In this study, we analyzed the relationship between airborne SARS-CoV-2 levels and environmental parameters. Bioaerosols were sampled (n = 40) in hospital corridors of two wards differing in the COVID-19 severity of the admitted patients. SARS-CoV-2 levels were quantified using droplet digital PCR. SARS-CoV-2 was detected in 60% of the total air samples. The ward where the mildly ill patients were admitted had a higher occupancy, transit of people in the corridor, and CO₂ levels, but there were no significant differences in SARS-CoV-2 detection between wards. The mean CO₂ concentration in the positive samples was 569 ± 35.6 ppm. Considering all samples, the CO₂ levels in the corridor were positively correlated with patient door openings but inversely correlated with SARS-CoV-2 levels. In conclusion, airborne SARS-CoV-2 can be detected indoors with optimal ventilation, and its levels do not scale with CO₂ concentration in hospital corridors. Therefore, CO₂ assessment should not be interpreted as a surrogate of airborne viral presence in all indoor spaces. Full article

The production of hydroponic fresh produce presents unique food safety and intervention challenges. A systematic approach was used to map and characterize the evidence on hydroponic food safety. Quantitative data describing the effectiveness of intervention studies were extracted, synthesized, and assessed for quality. A search of electronic databases yielded 131 relevant papers related to hydroponic food safety. Thirty-two studies focusing on food safety interventions reported 53 different interventions using chemical (n = 39), physical (n = 10), multiple-hurdle (n = 2), and biological (n = 2) approaches. Human pathogen indicators and surrogates were most often studied (n = 19), while pathogenic strains like Salmonella spp. (n = 9), Shiga toxin-producing Escherichia coli (STEC) (n = 5), Listeria monocytogenes (n = 2), and viruses (Hepatitis A virus (HAV), n = 1; norovirus (NoV), n = 1) were studied less frequently. Of fourteen articles (43.8%) investigating pre-harvest interventions, most (42.9%) did not specify the hydroponic system type. Gaps remain in the available evidence regarding the efficacy of interventions for controlling human pathogens in near-commercial hydroponic systems. The quality assessment revealed a significant lack of detailed reporting on methods and outcomes, making it difficult to translate the findings into practical recommendations for the industry; therefore, this review provides recommendations for the scientific community to improve future research design and reporting in this field. Full article

Viral hemorrhagic septicemia virus (VHSV) is a major pathogen in Paralichthys olivaceus (olive flounder) aquaculture, necessitating effective vaccines and evaluation methods. This study assessed immune responses following intraperitoneal administration of an inactivated VHSV vaccine and identified surrogate markers of vaccine efficacy. Fish were sampled weekly over 8 weeks. Antigen-specific antibody titers were measured by ELISA, and the expression of seven immune-related genes (CD4, CD8, CD28, IgM, Mx, IFN, and IL-1β) was analyzed in the spleen, kidney, liver, and gill by qPCR. The vaccinated group showed 84% relative survival, while unvaccinated fish experienced 100% mortality. Antibody titers peaked at 4 weeks post-vaccination, aligning with elevated IgM expression in the spleen and kidney. CD4 (kidney) and CD28 (spleen) expression strongly correlated with antibody titers (r = 0.854 and 0.796, respectively), whereas IL-1β, IFN, and Mx showed moderate associations in specific tissues. These findings indicate that CD4, CD28, and IgM may serve as molecular indicators of humoral immune responses specifically following inactivated VHSV vaccination in olive flounder (Paralichthys olivaceus). Combining gene expression profiling with antibody analysis offers a non-lethal, practical approach to evaluating vaccine efficacy in aquaculture. Full article

Norovirus, a foodborne pathogen, causes a significant economic and health burden globally. Although detection methods exist, they are expensive and non-field deployable. A flow-based dielectrophoretic biosensor was designed for the detection of foodborne pathogenic viruses and was tested using bacteriophage MS2 as a norovirus surrogate. The flow-based MS2 sensor comprises a concentrator and a detector. The concentrator is an interdigitated electrode array designed to impart dielectrophoretic effects to manipulate viral particles toward the detector in a fluidic channel. The detector is made of a silver electrode conjugated with anti-MS2 IgG to allow for antibody–antigen biorecognition events and is supplied with the electrical current for the purpose of measurement. Serially diluted MS2 suspensions were continuously injected into the fluidic channel at 0.1 mL/min. A cyclic voltammogram indicated that current measurements from single-walled carbon nanotube (SWCNT)-coated electrodes increased compared to uncoated electrodes. Additionally, a drop in the current measurements after antibody immobilization and MS2 capture was observed with the developed electrodes. Antibody immobilization at the biorecognition site provided greater current changes with the antibody-MS2 complexes vs. the assays without antibodies. The electric field applied to the fluidic channel at 10 V_pp and 1 MHz contributed to an increase in current changes in response to MS2 bound on the detector and was dependent on the MS2 concentrations in the sample. The developed biosensor was able to detect MS2 with a sensitivity of 10² PFU/mL within 15 min. Overall, this work demonstrates a proof of concept for a rapid and field-deployable strategy to detect foodborne pathogens. Full article

We have reviewed the primary literature on the virucidal efficacy of microbicidal active ingredients, formulated microbicides, and physical inactivation approaches (heat, irradiation) for hemorrhagic fever viruses (HFVs) (arenaviruses, filoviruses, flaviviruses, hantaviruses, nairoviruses, and phenuiviruses), and for two non-typical HFV paramyxoviruses. As each of these HFVs are large, lipid-enveloped RNA viruses, their susceptibilities to virucidal agents are informed by the so-called hierarchy of susceptibility of pathogens to microbicides. The unique susceptibility of lipid-enveloped viruses to most classes of microbicides is based on the common mechanisms of action of envelope-disrupting microbicides. Despite this, due to the relatively great lethality of these viruses, it is prudent (where possible) to confirm the expected efficacies of inactivation approaches in testing involving the HFVs themselves (as opposed to less lethal surrogate viruses) using field-relevant methods. Empirical data for virucidal activities of microbicidal active ingredients, formulated microbicides, and physical inactivation approaches, such as heat, ultraviolet light, and gamma irradiation, that were collected specifically for HFVs have been reviewed and summarized in this paper. These empirical data for surface and hand hygiene approaches, liquid inactivation approaches, and approaches for rendering diagnostic samples safe to handle inform non-pharmaceutical interventions intended to mitigate transmission risk associated with these HFVs. Full article

SARS-CoV-2 is classified as a containment level 3 (CL3) pathogen, limiting research access and antiviral testing. To address this, we developed a non-infectious viral surrogate system using reverse genetics to generate sub-genomic replicons. These replicons contained the nsp1 mutations K164A and H165A and had the spike, membrane, ORF6, and ORF7a coding sequences replaced with various reporter and selectable marker genes. Replicons based on the ancestral Wuhan Hu-1 strain and the Delta variant of concern were replication-competent in multiple cell lines, as assessed by Renilla luciferase activity, fluorescence, immunofluorescence staining, and single-molecule fluorescent in situ hybridization. Antiviral assays using transient replicon expression showed that remdesivir effectively inhibited both replicon and viral replication. Ritonavir and cobicistat inhibited Delta variant replicons similarly to wild-type virus but did not inhibit Wuhan Hu-1 replicon replication. To further investigate the impact of nsp1 mutations, we generated a recombinant SARS-CoV-2 virus carrying the K164A and H165A mutations. The virus exhibited attenuated replication across a range of mammalian cell lines, was restricted by the type I interferon response, and showed reduced cytopathic effects. These findings highlight the utility of sub-genomic replicons as reliable CL2-compatible surrogates for studying SARS-CoV-2 replication and drug activity mechanisms. Full article

This paper describes an experimental investigation on the spread of a virus in a business jet cabin and the potential of ionization to reduce the pathogen load. In contrast to priorly investigated recirculation air cleaning, ionization can act directly in the cabin by introducing ions into the supply air. Tests were performed by emitting a surrogate virus through a breathing head in a business jet mock-up. The results allow for the conclusion that ionization technology, along with increased airflow, is a well-suited tool to sanitize cabins. Additionally, the effect of ionization on particles was investigated where it became obvious that the presence of particles reduces the ion level; however, the presence of ions hardly impact particles. Full article

Global water scarcity and antimicrobial resistance (AMR) represent two escalating crises that urgently demand integrated and effective solutions. While wastewater reuse is increasingly promoted as a strategy to alleviate water scarcity, conventional treatment processes often fail to eliminate persistent contaminants and antibiotic-resistant microorganisms. Cold plasma (CP), a non-thermal advanced oxidation process, has demonstrated the strong potential to simultaneously inactivate pathogens and degrade micropollutants. CP generates a diverse mix of reactive oxygen and nitrogen species (ROS and RNS), as well as UV photons and charged particles, capable of breaking down complex contaminants and inducing irreversible damage to microbial cells. Laboratory studies have reported bacterial log reductions ranging from 1 to >8–9 log₁₀, with Gram-negative species such as E. coli and Pseudomonas aeruginosa showing higher susceptibility than Gram-positive bacteria. The inactivation of endospores and mixed-species biofilms has also been achieved under optimized CP conditions. Viral inactivation studies, including MS2 bacteriophage and norovirus surrogates, have demonstrated reductions >99.99%, with exposure times as short as 0.12 s. CP has further shown the capacity to degrade antibiotic residues such as ciprofloxacin and sulfamethoxazole by >90% and to reduce ARGs (e.g., bla, sul, and tet) in hospital wastewater. This perspective critically examines the mechanisms and current applications of CP in wastewater treatment, identifies the operational and scalability challenges, and outlines a research agenda for integrating CP into future water reuse frameworks targeting AMR mitigation and sustainable water management. Full article

Most donor human milk (HM) banks use Holder pasteurization (HoP) to ensure microbiological safety, although it can degrade essential bioactive factors for newborns. This study evaluates the innovative ultra-high-pressure homogenization (UHPH) technology as a potential alternative. Listeria innocua, Staphylococcus carnosus, Franconibacter helveticus (formerly named Cronobacter helveticus) and Escherichia coli strains were used as surrogates for common HM pathogens according to European Milk Bank Association (EMBA) guidelines, to evaluate the efficacy of new technologies. A reconstituted powder milk formula inoculated with these strains was used to determine the most efficient conditions (those to achieve a lethality of ≥5 Log), applying treatments from 150 to 300 MPa. These treatments were later validated using inoculated HM with the same strains. Immunoglobulin (sIgA, IgG, IgM) retention was also evaluated and compared with HoP. Results showed that UHPH treatments at 200 MPa achieved a lethality > 5 Log for all strains, except for St. carnosus, which required 250 MPa for complete inactivation in HM. Unlike HoP, UHPH at 200 and 250 MPa did not significantly reduce the basal concentration of sIgA, IgG, or IgM compared with raw HM. These findings suggest UHPH as a promising alternative to HoP, maintaining both microbiological safety and immunological quality. Full article