Reduce Virus Infectivity Research Articles

Potassium ion channels as a molecular target to reduce virus infection and mortality of honey bee colonies

Declines in managed honey bee populations are multifactorial but closely associated with reduced virus immunocompetence and thus, mechanisms to enhance immune function are likely to reduce viral infection rates and increase colony viability. However, gaps in knowledge regarding physiological mechanisms or ‘druggable’ target sites to enhance bee immunocompetence has prevented therapeutics development to reduce virus infection. Our data bridge this knowledge gap by identifying ATP-sensitive inward rectifier potassium (KATP) channels as a pharmacologically tractable target for reducing virus-mediated mortality and viral replication in bees, as well as increasing an aspect of colony-level immunity. Bees infected with Israeli acute paralysis virus and provided KATP channel activators had similar mortality rates as uninfected bees. Furthermore, we show that generation of reactive oxygen species (ROS) and regulation of ROS concentrations through pharmacological activation of KATP channels can stimulate antiviral responses, highlighting a functional framework for physiological regulation of the bee immune system. Next, we tested the influence of pharmacological activation of KATP channels on infection of 6 viruses at the colony level in the field. Data strongly support that KATP channels are a field-relevant target site as colonies treated with pinacidil, a KATP channel activator, had reduced titers of seven bee-relevant viruses by up to 75-fold and reduced them to levels comparable to non-inoculated colonies. Together, these data indicate a functional linkage between KATP channels, ROS, and antiviral defense mechanisms in bees and define a toxicologically relevant pathway that can be used for novel therapeutics development to enhance bee health and colony sustainability in the field.

Immunogenicity and efficacy of a novel multi-patch SARS-CoV-2/COVID-19 vaccine candidate.

While there has been considerable progress in the development of vaccines against SARS-CoV-2, largely based on the S (spike) protein of the virus, less progress has been made with vaccines delivering different viral antigens with cross-reactive potential. In an effort to develop an immunogen with the capacity to induce broad antigen presentation, we have designed a multi-patch synthetic candidate containing dominant and persistent B cell epitopes from conserved regions of SARS-CoV-2 structural proteins associated with long-term immunity, termed CoV2-BMEP. Here we describe the characterization, immunogenicity and efficacy of CoV2-BMEP using two delivery platforms: nucleic acid DNA and attenuated modified vaccinia virus Ankara (MVA). In cultured cells, both vectors produced a main protein of about 37 kDa as well as heterogeneous proteins with size ranging between 25-37 kDa. In C57BL/6 mice, both homologous and heterologous prime/boost combination of vectors induced the activation of SARS-CoV-2-specific CD4 and CD8 T cell responses, with a more balanced CD8+ T cell response detected in lungs. The homologous MVA/MVA immunization regimen elicited the highest specific CD8+ T cell responses in spleen and detectable binding antibodies (bAbs) to S and N antigens of SARS-CoV-2. In SARS-CoV-2 susceptible k18-hACE2 Tg mice, two doses of MVA-CoV2-BMEP elicited S- and N-specific bAbs as well as cross-neutralizing antibodies against different variants of concern (VoC). After SARS-CoV-2 challenge, all animals in the control unvaccinated group succumbed to the infection while vaccinated animals with high titers of neutralizing antibodies were fully protected against mortality, correlating with a reduction of virus infection in the lungs and inhibition of the cytokine storm. These findings revealed a novel immunogen with the capacity to control SARS-CoV-2 infection, using a broader antigen presentation mechanism than the approved vaccines based solely on the S antigen.

The ICAM-1 ligand HRV-A89 is internalized independently of clathrin-mediated endocytosis and its capsid reaches late endosomes

The human rhinovirus (HRV) A2 is endocytosed by clathrin-mediated endocytosis (CME) bound to the classical LDL receptor and releases its RNA during its transport to late endosomes. Here it is shown that - presumably due to an effect on virus recycling - a low concentration of the CME inhibitor chlorpromazine present during virus internalization (30 min) did not reduce HRV-A2 infection, but strongly inhibited short-time (5 min) endocytosis of HRV-A2. Chlorpromazine had no effect on the colocalization of the ICAM-1 ligand HRV-A89 with early endosomes, excluding CME as the main endocytosis pathway of this virus. As published for HRV-A2 and HRV-A14, HRV-A89 partially colocalized with lysosome-associated membrane protein 2 and the microtubule inhibitor nocodazole did not reduce virus infection when present only during virus internalization. Together with previous work these data suggest that there are no principal differences between endocytosis pathways of ICAM-1-binding rhinoviruses in different cell types.

Effects of Essential Oils and Hydrolates on the Infectivity of Murine Norovirus.

The use of natural substances with antiviral properties might reduce foodborne viral diseases. In this study, we evaluated the virucidal effect of Citrus limon and Thymus serpyllum essential oils (EOs) and of Citrus Limon, Thymus serpyllum and Thymus vulgaris hydrolates on murine norovirus (MNV), a human norovirus surrogate. To assess the virucidal effect of these natural substances, the reduction in viral infectivity was estimated by comparing the TCID50/mL of untreated viral suspension and the viral suspension treated with hydrolates and EOs at different concentrations. The results showed a natural loss of infectivity of the untreated virus after 24 h of approx. 1 log. The EO (1%) of T. serpyllum, and hydrolates (1% and 2%) of T. serpyllum and T. vulgaris immediately caused a reduction in MNV infectivity of about 2 log but did not provide a further significant decrease after 24 h. Instead, the EO (1%) and hydrolate (1% and 2%) of C. limon exerted an immediate reduction in the viral infectivity of about 1.3 log and 1 log, respectively, followed by a further reduction in infectivity of 1 log after 24 h for the hydrolate. These results will allow for the implementation of a depuration treatment based on the use of these natural compounds.

Hydroxy-xanthones as promising antiviral agents: Synthesis and biological evaluation against human coronavirus OC43

A number of synthetic hydroxy-xanthones related to isolates from the plant genus Swertia (family Gentianaceae) were prepared and their antiviral activity assessed against human coronavirus OC43. Overall, the results of the initial screening of the test compounds in BHK-21 cell lines show promising biological activity, with a significant reduction in viral infectivity (p ≤ 0.05). In general, the addition of functionality around the xanthone core increases the biological activity of the compounds compared to xanthone itself. More detailed studies are needed to determine mechanism of action, but favourable property predictions make them interesting lead compounds for further development as potential treatments for coronavirus infections.

Ratooning increases production of sweetpotato seed vines multiplied in insect-proof net tunnels in Tanzania

Summary Insect-proof net tunnels can help reduce virus infection of clean virus-tested sweetpotato seed produced by decentralized seed producers. However, optimal management is required to maintain both quality and quantity of seed produced. This study investigated the effect of the ratoon cropping technique on vine production in net tunnels and open fields. Virus-tested planting material of two varieties, Kabode and Mataya, were grown in net tunnels and open fields. Each variety had 80 plants per plot, with 40 following the ratooning technique and 40 a replanting technique. The ratooned crop was harvested six times, comprising the initial harvest and five regrowths. This covered 14 months representing six generations of vine production. The number of vines, number of nodes per vine, and vine length were recorded. The number of plants showing virus symptoms was also recorded. The ratoon cropping technique produced more vines compared with the replanting technique in both net tunnels and open fields. Cv. Kabode produced more vines in open fields compared with net tunnels regardless of cropping technique. On the other hand, cv. Mataya produced relatively equal numbers of vines in net tunnels and open fields. Despite ratooning leading to more vine production compared with replanting, the technique led to higher virus incidences on plants grown in the open. This also varied with variety with the highest virus disease incidences being recorded on cv. Mataya. We recommend the ratoon cropping technique for sweetpotato vine production in net tunnels. Replanting technique should be adopted for vine production in the open fields because it acts as a key control strategy for virus infections even for susceptible varieties.

Examining the Effect of Organic Acids on Inactivation of Hepatitis E Virus

Infection with hepatitis E virus genotype 3 (HEV-3) is an emerging cause of illness in developed countries. In North America and Europe, HEV-3 has been increasingly detected in swine, and exposure to pigs and pork products is considered the primary source of infection. We have previously demonstrated the prevalence of the HEV-3 genome in commercial pork products in Canada. In this study, we investigated the application of citric acid and acetic acid to inactivate HEV-3 on food and on food contact surfaces. For this purpose, plastic, stainless steel, and pork pâté surfaces were inoculated with HEV-3 and were treated with acetic acid or citric acid at 1, 3, or 5%. The infectivity of posttreatment viral particles was determined by cell culture. A greater than 2-log reduction in viral infectivity was observed on plastic and stainless steel treated with the organic acids, but the treatment was less effective on HEV infectivity on pork pâté (average reductions of 0.47 log citric acid and 0.63 log acetic acid). Therefore, we conclude that citric acid and acetic acid have potential application to control HEV-3 on food contact surfaces but are not suitable for food.

The Presence of Virus Neutralizing Antibodies Is Highly Associated with Protection against Virulent Challenge in Domestic Pigs Immunized with ASFV live Attenuated Vaccine Candidates.

African swine fever virus (ASFV) is currently producing a pandemic affecting a large area of Eurasia, and more recently, the Dominican Republic in the Western Hemisphere. ASFV is a large and structurally complex virus with a large dsDNA genome encoding for more than 150 genes. Live attenuated virus strains can induce protection in domestic swine against disease produced by homologous virulent parental viruses. The roles of the different immune mechanisms induced by the attenuated strains in protection still need to be understood. In particular, the role of ASFV neutralizing antibody in protection still is an important controversial issue to be elucidated. Here we present the development of a novel methodology to detect virus neutralizing antibodies based on the reduction of virus infectivity in a Vero cell adapted ASFV strain. The described method was used to assess levels of virus neutralizing antibodies in domestic swine inoculated with live attenuated ASFV. Results demonstrated a high association between the presence of virus neutralizing antibodies and protection in 84 animals immunized with the recombinant vaccine candidates ASFV-G-Δ9GL/ΔUK or ASFV-G-ΔI177L. To our knowledge, this is the first report demonstrating an association between virus neutralizing antibodies and protection against virulent challenge in such a large number of experimental individuals.

Impact of Protein Nitration on Influenza Virus Infectivity and Immunogenicity.

Influenza viruses are deadly respiratory pathogens of special importance due to their long history of global pandemics. During influenza virus infections, the host responds by producing interferons, which activate interferon-stimulated genes (ISGs) inside target cells. One of these ISGs is inducible nitric oxide synthase (iNOS). iNOS produces nitric oxide (NO) from arginine and molecular oxygen inside the cell. NO can react with superoxide radicals to form reactive nitrogen species, principally peroxynitrite. While much work has been done studying the many roles of nitric oxide in influenza virus infections, the direct effect of peroxynitrite on influenza virus proteins has not been determined. Manipulations of NO, either by knocking out iNOS or chemically inhibiting NO, produced no change in virus titers in mouse models of influenza infection. However, peroxynitrite has a known antimicrobial effect on various bacteria and parasites, and the reason for its lack of antimicrobial effect on influenza virus titers in vivo remains unclear. Therefore, we wished to test the direct effect of nitration of influenza virus proteins. We examined the impact of nitration on virus infectivity, replication, and immunogenicity. We observed that the nitration of influenza A virus proteins decreased virus infectivity and replication ex vivo. We also determined that the nitration of influenza virus hemagglutinin protein can reduce antibody responses to native virus protein. However, our study also suggests that nitration of influenza virus proteins in vivo is likely not extensive enough to inhibit virus functions substantially. These findings will help clarify the role of peroxynitrite during influenza virus infections. IMPORTANCE Nitric oxide and peroxynitrite produced during microbial infections have diverse and seemingly paradoxical functions. While nitration of lung tissue during influenza virus infection has been observed in both mice and humans, the direct effect of protein nitration on influenza viruses has remained elusive. We addressed the impact of nitration of influenza virus proteins on virus infectivity, replication, and immunogenicity. We observed that ex vivo nitration of influenza virus proteins reduced virus infectivity and immunogenicity. However, we did not detect nitration of influenza virus hemagglutinin protein in vivo. These results contribute to our understanding of the roles of nitric oxide and peroxynitrite in influenza virus infections.

Major target for UV-induced complete loss of HIV-1 infectivity: A model study of single-stranded RNA enveloped viruses

Deep ultraviolet light (UV) is useful for the disinfection of microorganisms, including bacteria and viruses. Although genome damage by UV has been widely accepted, the adverse effects of UV on the activity and/or function of viral proteins including the envelope components are poorly documented. Worthy of note, the observed unfavorable UV-effects for viruses are only insufficiently analyzed in association with the reduction in viral infectivity. In this study, we aimed to clarify which component of virions affected by UV significantly correlates with the loss of viral infectivity using HIV-1 as a model for single-stranded RNA enveloped viruses. Using our UV irradiation apparatus at three wavelengths (265, 280, and 300 nm), we first quantitatively determined the UV power density and irradiation period of each wavelength required for a reduction in infectivity. A heat-treated sample as a control drastically reduced the virion-associated reverse transcriptase (RT) activity and Gag-p24 level. The UV-irradiated samples at the three wavelengths, completely lacking viral infectivity, showed p24 levels similar to those without irradiation. While the virion-associated RT activity was gradually decreased in a wavelength and power density dependent manner, this reduction did not explain the loss of viral infectivity by UV. Remarkably, virological assays revealed that the entry efficiency of the UV-irradiated virus samples at the three wavelengths is comparable to those without irradiation. Importantly, this result shows that, even the virions exposed to UV of various wavelengths at the lethal level, still maintain the function of their envelope composed of a host lipid bilayer and viral proteins. In sharp contrast, UV-induced genome damage shown by semiquantitative RT-PCR correlated well with the reduction in viral infectivity, indicating that it is a major determinant for virus inactivation by UV. The degree of damage was found to be distinct among the regions analyzed. This was probably due to the different nucleotide sequences in those genomic regions amplified by PCR. Our data clearly demonstrate a principal mechanism for viral inactivation by UV and provide information contributing to the improvement of UV-based disinfection technology for microorganisms.

Reducing Virus Infection Risk in Space Environments through Nutrient Supplementation.

Space exploration has brought many challenges to human physiology. In order to evaluate and reduce possible pathological reactions triggered by space environments, we conducted bioinformatics analyses on the methylation data of the Mars 520 mission and human transcriptome data in the experiment simulating gravity changes. The results suggest that gene expression levels and DNA methylation levels were changed under the conditions of isolation and gravity changes, and multiple viral infection-related pathways were found in the enrichment analysis results of changed genes including Epstein Barr virus (EBV) infection, Hepatitis B virus (HBV) infection, Herpes simplex virus (HSV) infection and Kaposi’s sarcoma-associated herpesvirus (KHSV) infection. In this study, we found that Epigallocatechin-3-gallate (EGCG) and vitamin D are helpful in reducing viral infection risk. In addition, the causal associations between nutrients and viral infections were calculated using Two sample Mendelian Randomization (2SMR) method, the results indicated that vitamin D can reduce EBV infection and HBV infection risk. In summary, our study suggests that space environments increase the risk of human viral infection, which may be reduced by supplementing EGCG and vitamin D. These results can be used to formulate medical plans for astronauts, which have practical application value for future space exploration.

Analysis of the effects of the mutation of selected genes of pedilanthus leaf curl virus on infectivity, symptoms and the maintenance of tobacco leaf curl betasatellite

Pedilanthus leaf curl virus (PeLCV) is a widely encountered monopartite begomovirus with a wide host range that usually associates with a betasatellite. The cloned PeLCV used was highly infectious to Nicotiana benthamiana plants by agroinoculation and symptoms were enhanced by co-inoculation with tobacco leaf curl betasatellite (TbLCB). The study investigated the effects of the mutation of the coat protein (CP), V2, C2 and C4 genes of PeLCV on infectivity, symptoms and maintenance of TbLCB in N. benthamiana. Mutation of the CP abolished infectivity of PeLCV, which was not rescued by co-inoculation with TbLCB. Mutation of the V2 gene severely reduced infectivity that, in a small number of plants, was rescued by TbLCB, resulting in plants with very mild symptoms. Mutation of the C2 or V4 genes led to a significant reduction in virus infectivity, but also to non-symptomatic infections. Co-inoculation of either mutant with TbLCB led to a greater number of plants infected but fewer plants ultimately maintained the betasatellite, compared with inoculation of the betasatellite with the wild-type virus, indicating that the C2 and C4 proteins play a role in the maintenance of the betasatellite. For the C2 mutation, but not the C4 mutation, the presence of the betasatellite also resulted in mild symptoms. The results here, in conjunction with earlier studies, highlight commonalities as well as differences for the effects of gene mutations and interactions with betasatellites of different begomoviruses, indicating that begomovirus--betasatellite interaction is highly nuanced.

Inhibitory mechanism of Ambroxol and Bromhexine Hydrochlorides as potent blockers of molecular interaction between SARS-CoV-2 spike protein and human angiotensin-converting Enzyme-2

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infects the host cells through interaction of its spike protein with human angiotensin-converting enzyme 2 (hACE-2). High binding affinity between the viral spike protein and host cells hACE-2 receptor has been reported to enhance the viral infection. Thus, the disruption of this molecular interaction will lead to reduction in viral infectivity. This study, therefore, aimed to analyze the inhibitory potentials of two mucolytic drugs; Ambroxol hydrochlorides (AMB) and Bromhexine hydrochlorides (BHH), to serve as potent blockers of these molecular interactions and alters the binding affinity/efficiency between the proteins employing computational techniques. The study examined the effects of binding of each drug at the receptor binding domain (RBD) of the spike protein and the exopeptidase site of hACE-2 on the binding affinity (ΔGbind) and molecular interactions between the two proteins. Binding affinity revealed that the binding of the two drugs at the RBD-ACE-2 site does not alter the binding affinity and molecular interaction between the proteins. However, the binding of AMB (−56.931 kcal/mol) and BHH (−46.354 kcal/mol) at the exopeptidase site of hACE-2, significantly reduced the binding affinities between the proteins compared to the unbound, ACE-2-RBD complex (−64.856 kcal/mol). The result further showed the two compounds have good affinity at the hACE-2 site, inferring they might be potent inhibitors of hACE-2. Residue interaction networks analysis further revealed the binding of the two drugs at the exopeptidase site of hACE-2 reduced the number of interacting amino residues, subsequently leading to loss of interactions between the two proteins, with BHH showing better reduction in the molecular interaction and binding affinity than AMB. The result of the structural analyses additionally, revealed that the binding of the drugs considerably influences the dynamic of the complexes when compared to the unbound complex. The findings from this study suggest the binding of the two drugs at the exopeptidase site reduces the binding effectiveness of the proteins than their binding at the RBD site, and consequently might inhibit viral attachment and entry.

Distinctive Roles of Furin and TMPRSS2 in SARS-CoV-2 Infectivity.

ABSTRACTThe spike protein (S) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directs infection of the lungs and other tissues following its binding to the angiotensin-converting enzyme 2 (ACE2) receptor. For effective infection, the S protein is cleaved at two sites: S1/S2 and S2′. The “priming” of the surface S protein at S1/S2 (PRRAR685↓) [the underlined basic amino acids refer to critical residues needed for the furin recognition] by furin has been shown to be important for SARS-CoV-2 infectivity in cells and small-animal models. In this study, for the first time we unambiguously identified by proteomics the fusion activation site S2′ as KPSKR815↓ (the underlined basic amino acids refer to critical residues needed for the furin recognition) and demonstrated that this cleavage was strongly enhanced by ACE2 engagement with the S protein. Novel pharmacological furin inhibitors (BOS inhibitors) effectively blocked endogenous S protein processing at both sites in HeLa cells, and SARS-CoV-2 infection of lung-derived Calu-3 cells was completely prevented by combined inhibitors of furin (BOS) and type II transmembrane serine protease 2 (TMPRSS2) (camostat). Quantitative analyses of cell-to-cell fusion and S protein processing revealed that ACE2 shedding by TMPRSS2 was required for TMPRSS2-mediated enhancement of fusion in the absence of S1/S2 priming. We further demonstrated that the collectrin dimerization domain of ACE2 was essential for the effect of TMPRSS2 on cell-to-cell fusion. Overall, our results indicate that furin and TMPRSS2 act synergistically in viral entry and infectivity, supporting the combination of furin and TMPRSS2 inhibitors as potent antivirals against SARS-CoV-2.IMPORTANCE SARS-CoV-2, the etiological agent of COVID-19, has so far resulted in >6.1 million deaths worldwide. The spike protein (S) of the virus directs infection of the lungs and other tissues by binding the angiotensin-converting enzyme 2 (ACE2) receptor. For effective infection, the S protein is cleaved at two sites: S1/S2 and S2′. Cleavage at S1/S2 induces a conformational change favoring the S protein recognition by ACE2. The S2′ cleavage is critical for triggering membrane fusion and virus entry into host cells. Our study highlights the complex dynamics of interaction between the S protein, ACE2, and the host proteases furin and TMPRSS2 during SARS-CoV-2 entry and suggests that the combination of a nontoxic furin inhibitor with a TMPRSS2 inhibitor significantly reduces viral entry in lung cells, as evidenced by an average synergistic ∼95% reduction of viral infection. This represents a powerful novel antiviral approach to reduce viral spread in individuals infected by SARS-CoV-2 or future related coronaviruses.

The Interaction of Hypericin with SARS-CoV-2 Reveals a Multimodal Antiviral Activity

Hypericin is a photosensitizing drug that is active against membrane-enveloped viruses and therefore constitutes a promising candidate for the treatment of SARS-CoV-2 infections. The antiviral efficacy of hypericin is largely determined by its affinity toward viral components and by the number of active molecules loaded on single viruses. Here we use an experimental approach to follow the interaction of hypericin with SARS-CoV-2, and we evaluate its antiviral efficacy, both in the dark and upon photoactivation. Binding to viral particles is directly visualized with fluorescence microscopy, and a strong affinity for the viral particles, most likely for the viral envelope, is measured spectroscopically. The loading of a maximum of approximately 30 molecules per viral particle is estimated, despite with marked heterogeneity among particles. Because of this interaction, nanomolar concentrations of photoactivated hypericin substantially reduce virus infectivity on Vero E6 cells, but a partial effect is also observed in dark conditions, suggesting multiple mechanisms of action for this drug.

SAMDC3 enhances resistance to Barley stripe mosaic virus by promoting the ubiquitination and proteasomal degradation of viral γb protein

Posttranslational modifications (PTMs) play important roles in virus-host interplay. We previously demonstrated that Barley stripe mosaic virus (BSMV) γb protein is phosphorylated by different host kinases to support or impede viral infection. However, whether and how other types of PTMs participate in BSMV infection remains to be explored. Here, we report that S-adenosylmethionine decarboxylase 3 (SAMDC3) from Nicotiana benthamiana or wheat (Triticum aestivum) interacts with γb. BSMV infection induced SAMDC3 expression. Overexpression of SAMDC3 led to the destabilization of γb and reduction in viral infectivity, whereas knocking out NbSAMDC3 increased susceptibility to BSMV. NbSAMDC3 positively regulated the 26S proteasome-mediated degradation of γb via its PEST domain. Further mechanistic studies revealed that γb can be ubiquitinated in planta and that NbSAMDC3 promotes the proteasomal degradation of γb by increasing γb ubiquitination. We also found evidence that ubiquitination occurs at nonlysine residues (Ser-133 and Cys-144) within γb. Together, our results provide a function for SAMDC3 in defence against BSMV infection through targeting of γb abundance, which contributes to our understanding of how a plant host deploys the ubiquitin-proteasome system to mount defences against viral infections.

New food safety challenges of viral contamination from a global perspective: Conventional, emerging, and novel methods of viral control.

Food- and waterborne viruses, such as human norovirus, hepatitis A virus, hepatitis E virus, rotaviruses, astroviruses, adenoviruses, and enteroviruses, are major contributors to all foodborne illnesses. Their small size, structure, and ability to clump and attach to inanimate surfaces make viruses challenging to reduce or eliminate, especially in the presence of inorganic or organic soils. Besides traditional wet and dry methods of disinfection using chemicals and heat, emerging physical nonthermal decontamination techniques (irradiation, ultraviolet, pulsed light, high hydrostatic pressure, cold atmospheric plasma, and pulsed electric field), novel virucidal surfaces, and bioactive compounds are examined for their potential to inactivate viruses on the surfaces of foods or food contact surfaces (tools, equipment, hands, etc.). Every disinfection technique is discussed based on its efficiency against viruses, specific advantages and disadvantages, and limitations. Structure, genomic organization, and molecular biology of different virus strains are reviewed, as they are key in determining these techniques effectiveness in controlling all or specific foodborne viruses. Selecting suitable viral decontamination techniques requires that their antiviral mechanism of action and ability to reduce virus infectivity must be taken into consideration. Furthermore, details about critical treatments parameters essential to control foodborne viruses in a food production environment are discussed, as they are also determinative in defining best disinfection and hygiene practices preventing viral infection after consuming a food product.

Photothermal inactivation of universal viral particles by localized surface plasmon resonance mediated heating filter membrane

This study introduces localized surface plasmon resonance (L-SPR) mediated heating filter membrane (HFM) for inactivating universal viral particles by using the photothermal effect of plasmonic metal nanoparticles (NPs). Plasmonic metal NPs were coated onto filter membrane via a conventional spray-coating method. The surface temperature of the HFM could be controlled to approximately 40–60 °C at room temperature, owing to the photothermal effect of the gold (Au) NPs coated on them, under irradiation by visible light-emitting diodes. Due to the photothermal effect of the HFMs, the virus titer of H1Npdm09 was reduced by > 99.9%, the full inactivation time being < 10 min, confirming the 50% tissue culture infective dose (TCID50) assay. Crystal violet staining showed that the infectious samples with photothermal inactivation lost their infectivity against Mardin-Darby Canine Kidney cells. Moreover, photothermal inactivation could also be applied to reduce the infectivity of SARS-CoV-2, showing reduction rate of 99%. We used quantitative reverse transcription polymerase chain reaction (qRT-PCR) techniques to confirm the existence of viral genes on the surface of the HFM. The results of the TCID50 assay, crystal violet staining method, and qRT-PCR showed that the effective and immediate reduction in viral infectivity possibly originated from the denaturation or deformation of membrane proteins and components. This study provides a new, simple, and effective method to inactivate viral infectivity, leading to its potential application in various fields of indoor air quality control and medical science.

Antiviral activity of chitosan nanoparticles for controlling plant-infecting viruses

Chitosan nanoparticles (ChiNPs) are a potentially effective means for controlling numerous plant diseases. This study firstly describes the antiviral capabilities of ChiNPs to control plant viral diseases compared to its bulk form. Bean yellow mosaic virus (BYMV) was used as a model plant virus affecting faba bean plants and many other legumes. The antiviral effectiveness of ChiNPs and chitosan were evaluated as a curative application method, using six dosage rates (50, 100, 200, 250, 300 and 400 mg/L). Results indicated that ChiNPs curatively applied 48 h post virus inoculation entirely inhibit the disease infectivity and viral accumulation content at 300 mg/L and 400 mg/L. The virus titre was greatly alleviated within the plant tissues by 7.71% up to100% depending on ChiNP dosage rates. However, chitosan used in its bulk-based material form revealed a relatively low to an intermediate reduction in virus infectivity by 6.67% up to 48.86%. Interestingly, ChiNPs affect the virus particle’s integrity by producing defective and incomplete BYMV viral particles, defeating their replication and accumulation content within the plant tissues. Simultaneously, ChiNP applications were appreciably shown to promote the pathogenesis-related (PR-1) gene and other defence-related factors. The mRNA of the PR-1 gene was markedly accumulated in treated plants, reaching its maximum at 400 mg/L with 16.22-fold relative expression change over the untreated control. Further, the total phenol dynamic curve was remarkably promoted for 30 days in response to ChiNP application, as compared to the untreated control. Our results provide the first report that chitosan-based nanomaterials have a superior effect in controlling plant viruses as an antiviral curing agent, suggesting that they may feasibly be involved in viral disease management strategies under field conditions without serious health concerns and environmental costs.Significance: Our findings show that chitosan nanoparticles have a powerful curing antiviral activity against BYMV disease. These findings open the door for the use of eco-friendly nano-based tools in controlling numerous plant viruses. The use of eco-friendly nano-based materials could result in a successful integrative control strategy for plant viruses under field conditions, negating the need for the conventional measure used to control most of the insect-transmitted plant viruses, that is insecticide application against vector insects.

Amino acid substitutions at the HIV-1 transframe region significantly impair virus infectivity.

A transframe region within HIV-1 Gag-Pol (referred to as p6* or p6pol), directly linked to the protease (PR) N-terminus, plays a pivotal role in modulating PR activation. To identify specific p6* residues involved in PR activation, we created a series of p6* mutants by making substitutions for conserved p6* residues. Our results indicate that some p6* mutants were defective in terms of virus infectivity, despite displaying a wild-type virus particle processing pattern. Mutations at p6* F8 reduced virus infectivity associated with insufficient virus processing, due in part to impaired PR maturation and RT packaging. Our data strongly suggest that conserved Phe (F) residues at position 8 of p6* are involved in the PR maturation process.