Bacteriophage MS2 Research Articles

Modelling ribosome kinetics and translational control on dynamic mRNA

The control of protein synthesis and the overall levels of various proteins in the cell is critical for achieving homoeostasis. Regulation of protein levels can occur at the transcriptional level, where the total number of messenger RNAs in the overall transcriptome are controlled, or at the translational level, where interactions of proteins and ribosomes with the messenger RNA determine protein translational efficiency. Although transcriptional control of mRNA levels is the most commonly used regulatory control mechanism in cells, positive-sense single-stranded RNA viruses often utilise translational control mechanisms to regulate their proteins in the host cell. Here I detail a computational method for stochastically simulating protein synthesis on a dynamic messenger RNA using the Gillespie algorithm, where the mRNA is allowed to co-translationally fold in response to ribosome movement. Applying the model to the test case of the bacteriophage MS2 virus, I show that the models ability to accurately reproduce experimental measurements of coat protein production and translational repression of the viral RNA dependant RNA polymerase at high coat protein concentrations. The computational techniques reported here open up the potential to examine the infection dynamics of a ssRNA virus in a host cell at the level of the genomic RNA, as well as examine general translation control mechanisms present in polycistronic mRNAs.

Bacterial RNA virus MS2 exposure increases the expression of cancer progression genes in the LNCaP prostate cancer cell line.

Bacteriophages effectively counteract diverse bacterial infections, and their ability to treat most types of cancer has been explored using phage engineering or phage-virus hybrid platforms. In the present study, it was demonstrated that the bacteriophage MS2 can affect the expression of genes associated with the proliferation and survival of LNCaP prostate epithelial cells. LNCaP cells were exposed to bacteriophage MS2 at a concentration of 1×107 plaque forming units/ml for 24-48 h. After exposure, various cellular parameters, including cell viability, morphology, and changes in gene expression, were examined. MS2 affected cell viability adversely, reducing viability by 25% in the first 4 h of treatment; however, cell viability recovered within 24-48 h. Similarly, the AKT, androgen receptor, integrin α5, integrin β1, MAPK1, MAPK3, STAT3, and peroxisome proliferator-activated receptor-γ coactivator 1α genes, which are involved in various normal cellular processes and tumor progression, were significantly upregulated, whereas the expression levels of HSP90, ITGB5, ITGB3, HSP27, ITGAV, and PI3K genes were unchanged. Therefore, based on viability and gene expression changes, bacteriophage MS2 severely impaired LNCaP cells by reducing anchorage-dependent survival and androgen signaling. A caveolin-mediated endocytosis mechanism for MS2-mediated signaling in prostate cancer cells was proposed based on reports involving bacteriophages T4, M13, and MS2, and their interactions with LNCaP and PC3 cell lines.

CRISPR/Cas12a-Assisted Dual Visualized Detection of SARS-CoV-2 on Frozen Shrimps

Given the possibility that food contaminated with SARS-CoV-2 might become an infection source, there is an urgent need for us to develop a rapid and accurate nucleic acid detection method for SARS-CoV-2 in food to ensure food safety. Here, we propose a sensitive, specific, and reliable molecular detection method for SARS-CoV-2. It has a mechanism to control amplicon contamination. Swabs from spiked frozen shrimps were used as detection samples, which were processed by heating at 95 °C for 30 s. These preprocessed samples served as the templates for subsequent amplification. A colorimetric LAMP reaction was carried out to amplify both the SARS-CoV-2 target and the MS2 phage simultaneously in one tube. MS2 phage was detected by colorimetric LAMP as the internal control, while SARS-CoV-2 was detected with a CRISPR/Cas12a system. The fluorescence results could be visually detected with an ultraviolet lamp. Meanwhile, uracil was incorporated during the LAMP reaction to provide an amplicon contamination proof mechanism. This test could detect as low as 20 copies of SARS-CoV-2 in one reaction. Additionally, the detection could be finished in 45 min. The test only needs a heating block and an ultraviolet lamp, which shows the potential for field detection.

Antimicrobial and Virucidal Potential of Morpholinium-Based Ionic Liquids.

Witnessed by the ongoing spread of antimicrobial resistant bacteria as well as the recent global pandemic of the SARS-CoV-2 virus, the development of new disinfection strategies is of great importance, and novel substance classes as effective antimicrobials and virucides are urgently needed. Ionic liquids (ILs), low-melting salts, have been already recognized as efficient antimicrobial agents with prospects for antiviral potential. In this study, we examined the antiviral activity of 12 morpholinium based herbicidal ionic liquids with a tripartite test system, including enzyme inhibition tests, virucidal activity determination against five model viruses and activity against five bacterial species. The antimicrobial and enzymatic tests confirmed that the inhibiting activity of ILs corresponds with the number of long alkyl side chains and that [Dec2Mor]+ based ILs are promising candidates as novel antimicrobials. The virucidal tests showed that ILs antiviral activity depends on the type and structure of the virus, revealing enveloped Phi6 phage as highly susceptible to the ILs action, while the non-enveloped phages PRD1 and MS2 proved completely resistant to ionic liquids. Furthermore, a comparison of results obtained for P100 and P001 phages demonstrated for the first time that the susceptibility of viruses to ionic liquids can be dependent on differences in the phage tail structure.

Grating-like DBD plasma for air disinfection: Dose and dose-response characteristics

Atmospheric pressure dielectric barrier discharge (DBD) plasma is an emerging technique for effective bioaerosol decontamination and is promising to be used in indoor environments to reduce infections. However, fundamental knowledge of the dose and dose-response characteristics of plasma-based disinfection technology is very limited. By examining the single-pass removal efficiency of S. lentus aerosol by in-duct grating-like DBD plasma reactors with varied discharge setups (gap distance, electrode size, number of discharge layers, frequency, dielectric material), it was found that the specific input energy (SIE) could be served as the dose for disinfection, and the efficiency was exponentially dependent on SIE in most cases. The corresponding susceptibility constants (Z values) were obtained hereinafter. Humidity was a prominent factor boosting the efficiency with a Z value of 0.36 L/J at relative humidity (RH) of 20% and 1.68 L/J at RH of 60%. MS2 phage showed a much higher efficiency of 2.66–3.08 log10 of reduction than those of S. lentus (38–85%) and E. coli (42%–95%) under the same condition. Using SIE as the dose, the performance of plasma reactors in the literature was compared and evaluated. This work provides a theoretical and engineering basis for air disinfection by plasma-based technology.

Efficacy of Triethylene Glycol (Grignard Pure) Against Bacteriophage MS2 in the Air and on Surfaces

Efficacy of Triethylene Glycol (Grignard Pure) Against Bacteriophage MS2 in the Air and on Surfaces

Digital PCR: A Partitioning-Based Application for Detection and Surveillance of SARS-CoV-2 from Sewage Samples.

The wastewater-based surveillance of SARS-CoV-2 has emerged as a potential tool for cost-effective, simple, and long-term monitoring of the pandemic. Since the COVID-19 pandemic, several developed countries have incorporated the national wastewater surveillance program into their national policies related to pandemic management. Various research groups have utilized the approach ofreal-time quantitative reverse transcriptionPCR (RT-qPCR) for the quantification of SARS-CoV-2 from environmental samples like sewage water. However, detection and quantification using RT-qPCR relies on standards and is known to have lesser tolerance to inhibitors present in the sample. Unlike RT-qPCR, digital PCR (dPCR) offers an absolute and sensitive quantification without a need reference and offershigher tolerance to inhibitors present in the wastewater samples. Additionally, the accuracy of detection increases with the presence of rare target copies in the sample. The methodology herein presented comprises the detection and quantification of SARS-CoV-2 from sewer shed samples using the dPCR approach. The main features of the process include virus concentration and absolute quantification of the virus surpassing the substantial presence of inhibitors in the sample. This chapter presents the optimized PEG and NaCl-based protocol for virus concentration followed by nucleic acid extraction and quantification using CDC-approved N1+N2 assay. The protocol uses MS2 bacteriophage as a process recovery or internal control.The methodology herein described highlights the importance of digital PCR technologies for environmental surveillance of important emerging pathogens or pandemics.

Application of NaCl in Cold Atmospheric Plasma Jet and Plasma-Activated Solution to Enhance Virus Inactivation

Viruses are serious pathogenic contaminants that gravely threaten human health. There is an urgent need for environment-friendly and novel disinfection methods to inactivate various viruses. In this study, we applied the technique of cold atmospheric plasma (CAP) for virus disinfection. Particularly, we utilized the cold atmospheric plasma jet (CAPJ) and the plasma-activated solution (PAS) approaches for the treatment of bacteriophage MS2, and we added NaCl in the treatment system to enhance the disinfection efficiency. The inactivation effects under different CAPJ-PAS conditions using various plasma working gases were investigated. Also, the involved inactivation mechanisms due to respective active species were scrutinized. Our results showed that the addition of NaCl could considerably enhance the virus inactivation efficiency in the air/oxygen-CAPJ-PAS treatment, and during the direct CAPJ treatment, the primary inactivation factors were attributed to ·OH, <sup>1</sup>O<sub>2</sub>, ONOOH and ClO<sup>-</sup>, while for the indirect PAS treatment, the major inactivation contributors stemmed from <sup>1</sup>O<sub>2</sub> and ClO<sup>-</sup>. This work therefore demonstrates the usefulness of NaCl in the CAPJ-PAS treatment in the virus inactivation and offers new perspectives on the application of CAP technology in disinfection of viruses.

Improved extracellular vesicle-based mRNA delivery for familial hypercholesterolemia treatment.

Extracellular vesicle (EV)-based low-density lipoprotein receptor (Ldlr) mRNA delivery showed excellent therapeutic effects in treating familial hypercholesterolemia (FH). Nevertheless, the loading inefficiency of EV-based mRNA delivery presents a significant challenge. Recently, RNA-binding proteins (RBPs) have been fused to EV membrane proteins for selectively encapsulating targeted RNAs to promote loading efficiency. However, the strong interaction between therapeutic RNAs and RBPs prevents RNA release from endosomes to the cytosol in the recipient cells. In this study, an improved strategy was developed for efficient encapsulation of Ldlr mRNA into EVs in donor cells and controllable release in recipient cells. Methods: The MS2 bacteriophage coat protein (CD9-MCP) fusion protein, Ldlr mRNA, and a customized MS2 containing RNA aptamer base-pair matched with Ldlr mRNA were expressed in donor cells. Cells receiving the above therapeutic EVs were simultaneously treated with EVs containing "Ldlr releaser" with a sequence similar to the recognition sites in Ldlr mRNA. Therapeutic effects were analyzed in Ldlr-/- mice receiving EV treatments via the tail vein. Results: In vitro experiments demonstrated improved loading efficiency of Ldlr mRNA in EVs via MS2-MCP interaction. Treatment of "Ldlr releaser" competitively interacted with MS2 aptamer with higher affinity and released Ldlr mRNA from CD9-MCP for efficient translation. When the combinatory EVs were delivered into recipient hepatocytes, the robust LDLR expression afforded therapeutic benefits in Ldlr-/- mice. Conclusion: We proposed an EV-based mRNA delivery strategy for enhanced encapsulation of therapeutic mRNAs in EVs and RNA release into the cytosol for translation in recipient cells with great potential for gene therapy.

An actinometric method to characterize performance of reflecting UVC reactors used for water treatment

The applicability of chemical actinometry to characterize the fluence in UV reactors with reflections, non-parallel light, and variable water transmittance is limited due to the unknown effective path length or hydraulic shortcuts within the reactor. In this study, the effects of reflection and transmittance on actinometry were examined and a new, optimized and easy method for determining fluence was developed. KI/KIO3 and uridine actinometry experiments were carried out under controlled conditions using a collimated beam apparatus and a completely mixed batch reactor with or without diffuse reflection and compared to biodosimetry results. Whereas optically opaque actinometers such as KI/KIO3 are not directly capable of predicting the fluence of reflecting reactors, the results of uridine actinometry are influenced by reflection and transmission. To precisely predict the fluence rate in UV reactors with uridine, knowledge about the effective optical path length of the light is needed. Here, an existing method to mathematically calculate the optical path length was adopted and optimized for uridine actinometry. Results for average fluence were validated by biodosimetry using MS2 phages under different degrees of reflection and transmission. It could be shown that by modifying the bottom of the reactor with diffusely reflecting polytetrafluoroethylene foil, the fluence rate was increased by a factor of approximately 2.6 and the path length by factor of 2.4. When only half of the bottom was covered with reflective foil, fluence rate increased by a factor of 1.8 and path length by 1.8. Although this new approach cannot replace biodosimetry, to predict the fluence distribution received by microorganisms, it can provide means to characterize more complex reactor designs, validate results of advanced reactor modeling, and quantify fluence for non-parallel irradiation and reflective light, especially for the application of high fluence (e.g., advanced oxidation processes), where biodosimetry may be too sensitive. Further, comparing the fluence obtained with actinometry to the results of biodosimetry might qualitatively indicate hydraulic short cuts or unideal fluence distributions for flow-through reactors.

Ions play different roles in virus removal caused by different NOMs in UF process: Removal efficiency and mechanism analysis

In this study, we investigated the effect of different compositions of aquatic natural organic matter (NOM) and ions on virus removal by ultrafiltration (UF). MS2 bacteriophage was used as a surrogate. Humic acid (HA) improved the MS2 removal rate from 1.95 ± 0.09 LRV to 2.40 ± 0.03 LRV at the HA dosage of 9 mg/L through the combined mechanisms of size exclusion, electrostatic repulsion and hydrophobicity. MS2 removal rate further increased to 3.10 ± 0.05 LRV by 10 mmol/L Na+ dosage and 3.19 ± 0.12 LRV by Ca2+ 1 mmol/L in the HA-containing UF system. Size exclusion turned into the dominant virus removal mechanism according to the results of the fouling model fitting and the weakening of electrostatic repulsion and hydrophobicity. The complexation of Ca2+ also played a role in MS2 removal based on the analysis of interaction force. MS2 removal rate by bovine serum albumin (BSA) was poor, which was 2.07 ± 0.06 LRV at the BSA dosage of 9 mg/L. Hydrophobicity was greatly reduced and the dominant virus removal mechanisms were size exclusion and electrostatic repulsion. 10 mmol/L Na+ in the presence of BSA deteriorated MS2 removal rate to 2.02 ± 0.07 LRV by the weakening of electrostatic repulsion, hydrophobicity and size exclusion. Electrostatic repulsion severely decreased by 1 mmol/L Ca2+ and the enhanced adsorption barrier represented competitive adsorption of Ca2+ by BSA and MS2 contributed for MS2 removal further decline (1.99 ± 0.05 LRV). Complex components in water will have different effects on virus removal due to their properties and interactions. This study can provide references for selecting more efficient water treatment methods according to the different compositions of raw water in actual water treatment applications during the UF process. Moreover, the retention of virus by UF can be predicted based on our study results.

Estimation of Bacteriophage MS2 Inactivation Parameters During Microwave Heating of Frozen Strawberries

Frozen berries have been repeatedly linked to acute gastroenteritis caused by norovirus, the most common cause of foodborne illness in the United States. Many guidelines recommend that frozen berries be microwaved for at least 2 min, but it is unclear if this thermal treatment is effective at inactivating norovirus. The objective of this study was to model the effect of microwave heating at varying power levels on the survival of bacteriophage MS2, a norovirus surrogate, when inoculated onto frozen strawberries. Bacteriophage MS2 was inoculated onto the surface of frozen strawberries with a starting concentration of approximately 10 log PFU/g. Samples (either 3 or 5 whole strawberries) were heated in a 1300-Watt domestic research microwave oven (frequency of 2450 MHz) at power levels of 30, 50, 70, and 100% (full power), for times ranging from 15 to 300 s to determine inactivation. Temperatures at berry surfaces were monitored during heating using fiberoptic thermometry. All experiments were conducted in triplicate. The primary model for thermal inactivation was a log-linear model of logN vs. time. The secondary model was for a D-value decreasing linearly with temperature and an added term that was path-dependent on the thermal history. Parameters in the model were estimated using dynamic temperature history at the surface of the berry, via nonlinear regression using all data simultaneously. The root mean square error was ∼0.5 PFU/g out of a total 6-log reduction. Log reductions of 1.1 ± 0.4, 1.5 ± 0.5, 3.1 ± 0.1, and 3.8 ± 0.2 log PFU/g were observed for 30, 50, 70, and 100% microwave power levels when three berries were heated for 60 s. D-values were 21.4 ± 1.95 s and 10.6 ± 1.1 s at 10 and 60°C, respectively. This work demonstrates an approach to estimate inactivation parameters for viruses from dynamic temperature data during microwave heating. These findings will be useful in predicting the safety effect of microwave heating of berries in the home or food service.

Antimicrobial Lipstick: Bio-Based Composition against Viruses, Bacteria, and Fungi.

The COVID-19 pandemic has speeded up the race to find materials that could help limit or avoid the spread of SARS-CoV-2, while infections by multidrug-resistant bacteria and fungi are now becoming a serious threat. In this study, we developed a novel bio-based lipstick containing cranberry extract, a substance able to inactivate a broad range of microorganisms: enveloped viruses such as bacteriophage Φ6, a surrogate of SARS-CoV-2; non-enveloped viruses including bacteriophage MS2; multidrug-resistant bacteria like methicillin-resistant Staphylococcus aureus, Escherichia coli, and Mycobacterium smegmatis, a surrogate of Mycobacterium tuberculosis; and the Candida albicans fungus. The proposed antimicrobial lipstick offers a new form of protection against a broad range of microorganisms, including enveloped and non-enveloped viruses, bacteria, and fungi, in the current COVID-19 pandemic and microbial-resistant era.

Antimicrobial properties of a multi-component alloy

High traffic touch surfaces such as doorknobs, countertops, and handrails can be transmission points for the spread of pathogens, emphasizing the need to develop materials that actively self-sanitize. Metals are frequently used for these surfaces due to their durability, but many metals also possess antimicrobial properties which function through a variety of mechanisms. This work investigates metallic alloys comprised of several metals which individually possess antimicrobial properties, with the target of achieving broad-spectrum, rapid sanitation through synergistic activity. An entropy-motivated stabilization paradigm is proposed to prepare scalable alloys of copper, silver, nickel and cobalt. Using combinatorial sputtering, thin-film alloys were prepared on 100 mm wafers with ≈50% compositional grading of each element across the wafer. The films were then annealed and investigated for alloy stability. Antimicrobial activity testing was performed on both the as-grown alloys and the annealed films using four microorganisms—Phi6, MS2, Bacillus subtilis and Escherichia coli—as surrogates for human viral and bacterial pathogens. Testing showed that after 30 s of contact with some of the test alloys, Phi6, an enveloped, single-stranded RNA bacteriophage that serves as a SARS-CoV-2 surrogate, was reduced up to 6.9 orders of magnitude (> 99.9999%). Additionally, the non-enveloped, double-stranded DNA bacteriophage MS2, and the Gram-negative E. coli and Gram-positive B. subtilis bacterial strains showed a 5.0, 6.4, and 5.7 log reduction in activity after 30, 20 and 10 min, respectively. Antimicrobial activity in the alloy samples showed a strong dependence on the composition, with the log reduction scaling directly with the Cu content. Concentration of Cu by phase separation after annealing improved activity in some of the samples. The results motivate a variety of themes which can be leveraged to design ideal antimicrobial surfaces.

Isoporous catalytic ceramic membranes for ultrafast contaminants elimination through boosting confined radicals

Catalytic membrane based oxidation-filtration processes (AOP-CM), a derivative concept of membrane process that combines physical separation and chemical oxidation, offers a high-efficient water purification strategy. However, the application of AOP-CM was still hampered by the low heterogeneous AOPs efficiency of catalytic membranes. In order to improve the heterogeneous AOP efficiency, an isoporous AlOx/La2CoMnO6-δ ceramic membrane (IAPCM) with nano-confinement characteristics was prepared via sol–gel based block copolymer self-assembly route. Benefiting from the well-designed pore structure, IAPCM exhibited excellent pure water permeance (313 L·m−2·h−1·bar−1) and size-exclusion performance (complete rejection of MS2 phages with a diameter of ∼ 20 nm). With the addition of peroxymonosulfate (PMS), IAPCM achieved ultrafast degradation of organic micropollutants (e.g. atrazine, carbamazepine and sulfamethazine) at 0.5 bar (equivalent to a retention time of 4.3 × 10−4 s). Finite-Element analysis confirmed that high-concentration radical fields were generated in the confined nanoscale-pores within the isoporous La2CoMnO6-δ layer. The boosted mass transfer rates and high-concentration radical fields induced ultrafast degradation of micropollutants in IAPCM based oxidation-filtration system. This work highlights the significance of pore structure design for high-performance AOP-CM processes.

Direct and Indirect Cationization of Cellulose Nanocrystals: Structure-Properties Relationship and Virus Capture Activity.

Due to increasing public concern over hygiene, there have been many studies investigating antimicrobial and antiviral agents recently. With the aim of developing biobased virucidal/virus capture agents, we report a chemical modification of the cellulose nanocrystals (CNCs) surface with poly(2-dimethylamino) ethyl acrylate) methyl chloride quaternary salt (Q-PDMAEA) to introduce the positively charged functional groups. The surface of CNCs was modified through direct and indirect graft polymerization. Subsequently, the direct and indirect cationization effect on the degree of functionalization, thermal stability, crystallinity, and antiviral activity of CNCs was investigated. Indirect cationization produced the highest degree of polymer grafting, increasing particle size and thermal stability. Further, the modified CNCs were tested for their ability to capture nonenveloped bacteriophages PhiX174 (ΦX174) and MS2. We observed a significant (>4.19 log10) reduction in total viral load by specific functionalized CNCs. However, the activity depended on the structure of functional groups, surface charge density, and the type of virus under study. Overall, the direct and indirect cationization of CNC leads to biobased agents with immobilized cationic charge, with good virus capture activity. Such agents can be used for various applications including textiles, packaging, wastewater treatment, etc.

Evaluation of pathogen disinfection efficiency of electrochemical advanced oxidation to become a sustainable technology for water reuse

Water treatment and reuse is gaining acceptance as a strategy to fight against water contamination and scarcity, but it usually requires complex treatments to ensure safety. Consequently, the electrochemical advanced processes have emerged as an effective alternative for water remediation. The main objective here is to perform a systematic study that quantifies the efficiency of a laboratory-scale electrochemical system to inactivate bacteria, bacterial spores, protozoa, bacteriophages and viruses in synthetic water, as well as in urban wastewater once treated in a wetland for reuse in irrigation. A Ti|RuO2-based plate and Si|BDD thin-film were comparatively employed as the anode, which was combined with a stainless-steel cathode in an undivided cell operating at 12 V. Despite the low resulting current density (<15 mA/cm2), both anodes demonstrated the production of oxidants in wetland effluent water. The disinfection efficiency was high for the bacteriophage MS2 (T99 in less than 7.1 min) and bacteria (T99 in about 30 min as maximum), but limited for CBV5 and TuV, spores and amoebas (T99 in more than 300 min). MS2 presented a rapid exponential inactivation regardless of the anode and bacteria showed similar sigmoidal curves, whereas human viruses, spores and amoebas resulted in linear profiles. Due the different sensitivity of microorganisms, different models must be considered to predict their inactivation kinetics. On this basis, it can be concluded that evaluating the viral inactivation from inactivation profiles determined for bacteria or some bacteriophages may be misleading. Therefore, neither bacteria nor bacteriophages are suitable models for the disinfection of water containing enteric viruses. The electrochemical treatment added as a final disinfection step enhances the inactivation of microorganisms, which could contribute to safe water reuse for irrigation. Considering the calculated low energy consumption, decentralized water treatment units powered by photovoltaic modules might be a near reality.

Surrogate Selection for Foot-and-Mouth Disease Virus in Disinfectant Efficacy Tests by Simultaneous Comparison of Bacteriophage MS2 and Bovine Enterovirus Type 1.

In South Korea, testing disinfectants against foot-and-mouth disease virus (FMDV) that are contagious in livestock or that require special attention with respect to public hygiene can be manipulated only in high-level containment laboratories, which are not easily available. This causes difficulties in the approval procedure for disinfectants, such as a prolonged testing period. Additionally, the required biosafety level (BSL) in the case of FMDV has hindered its extensive studies. However, this drawback can be circumvented by using a surrogate virus to improve the performance of the efficacy testing procedure for disinfectants. Therefore, we studied bacteriophage MS2 (MS2) and bovine enterovirus type 1 (ECBO) with respect to disinfectant susceptibility for selecting a surrogate for FMDV according to the Animal and Plant Quarantine Agency (APQA) guidelines for efficacy testing of veterinary disinfectants. Effective concentrations of the active substances in disinfectants (potassium peroxymonosulfate, sodium dichloroisocyanurate, malic acid, citric acid, glutaraldehyde, and benzalkonium chloride) against FMDV, MS2, and ECBO were compared and, efficacies of eight APQA-listed commercial disinfectants used against FMDV were examined. The infectivity of FMDV and ECBO were confirmed by examination of cytopathic effects, and MS2 by plaque assay. The results reveal that the disinfectants are effective against MS2 and ECBO at higher concentrations than in FMDV, confirming their applicability as potential surrogates for FMDV in efficacy testing of veterinary disinfectants.

Plasma-generated reactive water mist for disinfection of N95 respirators laden with MS2 and T4 bacteriophage viruses

Due to the shortage of personal protective equipment (PPE) during the COVID-19 pandemic, the interest and demand for sterilization devices to reuse PPE has increased. For reuse of face masks, they must be effectively decontaminated of potential infectious agents without compromising its filtration ability during sterilization. In this study, we utilized an atmospheric pressure pulsed dielectric barrier discharge (DBD), combined with nebulized liquid microdroplets to generate plasma-activated mist (PAM). MS2 and T4 bacteriophages were used to conduct the decontamination tests on two types of N95 respirators. Results showed at least a 2-log reduction of MS2 and T4 on N95 respirators treated in one cycle with 7.8% hydrogen peroxide PAM and at least a 3-log reduction treated in 10% hydrogen peroxide PAM. In addition, it was found that there was no significant degradation in filtration efficiency of N95 respirators (3M 1860 and 1804) treated in 10% hydrogen peroxide PAM found after 20 cycles. In terms of re-useability of masks after treatment as determined, it was shown that the elastic straps of 3M 1804 were fragmented after 20 treatment cycles rendering them unusable, while the straps of 3M 1860 were not negatively affected even after 20 disinfection cycles.

Evaluation of disinfection efficacy of single UV-C, and UV-A followed by UV-C LED irradiation on Escherichia coli, B. spizizenii and MS2 bacteriophage, in water

Ultraviolet light-emitting diodes (UV LEDs) have shown ability to inactivate microorganisms and viruses in water. The unique characteristic of the UV-LEDs' diversity in wavelengths ranging from UV-C, UV-B, and UV-A, allows for wavelengths to be combined in different manners for polychromatic irradiation. Previous studies reported no synergy from simultaneous or sequential UV-C and UV-B as well as UV-C or UV-B followed by UV-A irradiation. However, synergy was reported for UV-A followed by UV-C or UV-B irradiation on various microorganisms. Nevertheless, no clear ground has been reached on whether to adopt single UV-C wavelengths or UV-A followed by UV-C LED, irradiation on inactivation of microorganisms and viruses in water. Therefore, this work evaluates the disinfection efficacy of single UV-C as well as UV-A followed by UV-C LED irradiation on Escherichia coli, Bacillus spizizenii spores and MS2 bacteriophage in water. The UV-C wavelengths were represented by 267 and 278 nm UV LEDs, and UV-A by 368 nm UV LEDs. In this study, E. coli was highly susceptible to UV radiation followed by B. spizizenii spores, and lastly MS2. Repair following UV inactivation was only observed in E. coli. The synergistic effect found in both E. coli, and B. spizizenii spores was attributed to the different inactivation mechanisms of the UV-C and UV-A wavelengths. In both single UV-C, and UV-A followed by UV-C LED irradiations, single 267 nm UV-C LED showed higher inactivation efficacy. Meanwhile, single 278 nm UV-C LED showed higher efficacy in terms of suppression of repair, and electrical energy consumption. Using single UV-C LEDs in a water disinfection system cuts down on related extra costs by avoiding combined wavelengths while still attaining better levels of microorganism inactivation, repair suppression and electrical energy consumption. These findings are applicable for the design and implementation of UV LED water disinfection systems.