Comparative Removal of Poliovirus, Rotavirus SA11 and MS2 Coliphage by Point-of-Use Devices used to Treat Drinking Water and Water Disinfectants: A Review.

Test protocols have been developed to test water treatment devices/systems for use for treating drinking water that are used at the individual and home level to ensure the removal of waterborne viruses. Current test procedures call for the use of poliovirus type 1 and/or rotavirus SA11. Recently we suggested that selected coliphages could be used as surrogates for poliovirus for testing of point-of-use (POU) water treatment devices, however, rotavirus was not used in those studies. The purpose of this review was to compare studies of POU devices which were tested with poliovirus type 1, simian rotavirus SA11 and coliphage MS2 to determine if the behavior of rotavirus SA11 was significantly different. In addition, an attempt was made to compare the relative resistance of these viruses by various disinfectants used to treat drinking water. In all cases SA11 was removed to an equal or greater degree than poliovirus. SA11 was found to be less resistant to halogens, although one study found it to be more resistance to chloramines than poliovirus and MS2. Based on this review, use of coliphages for testing POU devices appear justified. Additionally, data on chloramines for these viruses would be useful to determine if rotavirus is more resistant than poliovirus and MS2.

Mechanisms of action of microbicides commonly used in infection prevention and control.

SUMMARYUnderstanding how commonly used chemical microbicides affect pathogenic microorganisms is important for formulation of microbicides. This review focuses on the mechanism(s) of action of chemical microbicides commonly used in infection prevention and control. Contrary to the typical site-specific mode of action of antibiotics, microbicides often act via multiple targets, causing rapid and irreversible damage to microbes. In the case of viruses, the envelope or protein capsid is usually the primary structural target, resulting in loss of envelope integrity or denaturation of proteins in the capsid, causing loss of the receptor-binding domain for host cell receptors, and/or breakdown of other viral proteins or nucleic acids. However, for certain virucidal microbicides, the nucleic acid may be a significant site of action. The region of primary damage to the protein or nucleic acid is site-specific and may vary with the virus type. Due to their greater complexity and metabolism, bacteria and fungi offer more targets. The rapid and irreversible damage to microbes may result from solubilization of lipid components and denaturation of enzymes involved in the transport of nutrients. Formulation of microbicidal actives that attack multiple sites on microbes, or control of the pH, addition of preservatives or potentiators, and so on, can increase the spectrum of action against pathogens and reduce both the concentrations and times needed to achieve microbicidal activity against the target pathogens.

Eye infection risks from Pseudomonas aeruginosa via hand soap and eye drops.

Eye infections from bacterial contamination of bulk-refillable liquid soap dispensers and artificial tear eye drops continue to occur, resulting in adverse health outcomes that include impaired vision or eye enucleation. Pseudomonas aeruginosa (P. aeruginosa), a common cause of eye infections, can grow in eye drop containers and refillable soap dispensers to high numbers. To assess the risk of eye infection, a quantitative microbial risk assessment for P. aeruginosa was conducted to predict the probability of an eye infection for two potential exposure scenarios: (i) individuals using bacteria-contaminated eye drops and (ii) contact lens wearers washing their hands with bacteria-contaminated liquid soap prior to placing the lens. The median risk of an eye infection using contaminated eye drops and hand soap for both single and multiple exposure events (per day) ranged from 10-1 to 10-4, with contaminated eye drops having the greater risk. The concentration of P. aeruginosa was identified as the parameter contributing the greatest variance on eye infection risk; therefore, the prevalence and level of bacterial contamination of the product would have the greatest influence on health risk. Using eye drops in a single-use container or with preservatives can mitigate bacterial growth, and using non-refillable soap dispensers is recommended to reduce contamination of hand soap. Given the opportunistic nature of P. aeruginosa and its ability to thrive in unique environments, additional safeguards to mitigate bacterial growth and exposure are warranted.IMPORTANCEPseudomonas aeruginosa (P. aeruginosa) is a pathogen that can persist in a variety of unusual environments and continues to pose a significant risk for public health. This quantitative microbial risk assessment (QMRA) estimates the potential human health risks, specifically for eye infections, associated with exposure to P. aeruginosa in bacteria-contaminated artificial tear eye drops and hand soap. This study applies the risk assessment framework of QMRA to evaluate eye infection risks through both consumer products. The study examines the prevalence of this pathogen in eye drops and soap, as well as the critical need to implement measures that will mitigate bacterial exposure (e.g., single-use soap dispensers and eye drops with preservatives). Additionally, limitations and challenges are discussed, including the need to incorporate data regarding consumer practices, which may improve exposure assessments and health risk estimates.

Human adenovirus-associated health risk in the recreational waters of the Yal-ku lagoon in the Mexican Caribbean.

The study objective was to evaluate human faecal contamination impacts in the Yal-ku lagoon in the Mexican Caribbean and to estimate adenovirus infection and illness risks associated with recreational exposure during water activities. A total of 20 water samples (10 from each site × two sites) (50 L) were collected monthly over a period of 12 months from two selected sampling sites in the swimming area of the Yal-ku lagoon. The occurrence of faecal-associated viruses was explored, and human adenovirus (HAdV) and pepper mild mottle virus (PMMoV) concentrations were quantified. A quantitative microbial risk assessment (QMRA) model was used to estimate exposure and subsequent adenovirus infection and illness risk for 1 h of swimming or snorkelling. Somatic and F + -specific coliphages occurred in 100% of the samples. Both HAdV and PMMoV were detected at a 60% frequency thereby indicating persistent faecal inputs. PMMoV concentrations (44-370 GC/L) were relatively lower than the concentrations of HAdV (64-1,000 GC/L). Estimated mean adenovirus risks were greater for snorkelling than for swimming by roughly one to two orders of magnitude and estimated mean illness risks for snorkelling were >32/1,000. Human faecal contamination is frequent in the Yal-ku lagoon, which is associated with human gastrointestinal illness.

Transmission of Viruses from Restroom Use: A Quantitative Microbial Risk Assessment.

Restroom use has been implicated in a number of viral outbreaks. In this study, we apply quantitative microbial risk assessment to quantify the risk of viral transmission by contaminated restroom fomites. We estimate risk from high-touch fomite surfaces (entrance/exit door, toilet seat) for three viruses of interest (SARS-CoV-2, adenovirus, norovirus) through eight exposure scenarios involving differing user behaviors, and the use of hand sanitizer following each scenario. We assessed the impacts of several sequences of fomite contacts in the restroom, reflecting the variability of human behavior, on infection risks for these viruses. Touching of the toilet seat was assumed to model adjustment of the seat (open vs. closed), a common touch point in single-user restrooms (home, small business, hospital). A Monte Carlo simulation was conducted for each exposure scenario (10,000 simulations each). Norovirus resulted in the highest probability of infection for all exposure scenarios with fomite surfaces. Post-restroom automatic-dispensing hand sanitizer use reduced the probability of infection for each virus by up to 99.75%. Handwashing within the restroom, an important risk-reduction intervention, was not found to be as effective as use of a non-touch hand sanitizer dispenser for reducing risk to near or below 1/1,000,000, a commonly used risk threshold for comparison.

Antifungal activity and mechanism of action of natural product derivates as potential environmental disinfectants.

There have been a considerable number of antifungal studies that evaluated natural products (NPs), such as medicinal plants and their secondary metabolites, (phenolic compounds, alkaloids), essential oils, and propolis extracts. These studies have investigated natural antifungal substances for use as food preservatives, medicinal agents, or in agriculture as green pesticides because they represent an option of safe, low-impact, and environmentally friendly antifungal compounds; however, few have studied these NPs as an alternative to disinfection/sanitation for indoor air or environmental surfaces. This review summarizes recent studies on NPs as potential fungal disinfectants in different environments and provides information on the mechanisms of inactivation of these products by fungi. The explored mechanisms show that these NPs can interfere with ATP synthesis and Ca++ and K+ ion flow, mainly damaging the cell membrane and cell wall of fungi, respectively. Another mechanism is the reactive oxygen species effect that damages mitochondria and membranes. Inhibition of the overexpression of the efflux pump is another mechanism that involves damage to fungal proteins. Many NPs appear to have potential as indoor environmental disinfectants. ONE-SENTENCE SUMMARY: This review shows the latest advances in natural antifungals applied to different indoor environments. Fungi have generated increased tolerance to the mechanisms of traditional antifungals, so this review also explores the various mechanisms of action of various natural products to facilitate the implementation of technology.

Environmental dissemination of respiratory viruses: dynamic interdependencies of respiratory droplets, aerosols, aerial particulates, environmental surfaces, and contribution of viral re-aerosolization.

During the recent pandemic of COVID-19 (SARS-CoV-2), influential public health agencies such as the World Health Organization (WHO) and the U.S. Centers for Disease Control and Prevention (CDC) have favored the view that SARS CoV-2 spreads predominantly via droplets. Many experts in aerobiology have openly opposed that stance, forcing a vigorous debate on the topic. In this review, we discuss the various proposed modes of viral transmission, stressing the interdependencies between droplet, aerosol, and fomite spread. Relative humidity and temperature prevailing determine the rates at which respiratory aerosols and droplets emitted from an expiratory event (sneezing, coughing, etc.) evaporate to form smaller droplets or aerosols, or experience hygroscopic growth. Gravitational settling of droplets may result in contamination of environmental surfaces (fomites). Depending upon human, animal and mechanical activities in the occupied space indoors, viruses deposited on environmental surfaces may be re-aerosolized (re-suspended) to contribute to aerosols, and can be conveyed on aerial particulate matter such as dust and allergens. The transmission of respiratory viruses may then best be viewed as resulting from dynamic virus spread from infected individuals to susceptible individuals by various physical states of active respiratory emissions, instead of the current paradigm that emphasizes separate dissemination by respiratory droplets, aerosols or by contaminated fomites. To achieve the optimum outcome in terms of risk mitigation and infection prevention and control (IPAC) during seasonal infection peaks, outbreaks, and pandemics, this holistic view emphasizes the importance of dealing with all interdependent transmission modalities, rather than focusing on one modality.

Antiviral Natural Products, Their Mechanisms of Action and Potential Applications as Sanitizers and Disinfectants.

Plant extracts, natural products and plant oils contain natural virucidal actives that can be used to replace active ingredients in commercial sanitizers and disinfectants. This review focuses on the virucidal mechanisms of natural substances that may exhibit potential for indoor air and fomite disinfection. Review of scientific studies indicates: (1) most natural product studies use crude extracts and do not isolate or identify exact active antiviral substances; (2) many natural product studies contain unclear explanations of virucidal mechanisms of action; (3) natural product evaluations of virucidal activity should include methods that validate efficacy under standardized disinfectant testing procedures (e.g., carrier tests on applicable surfaces or activity against aerosolized viruses, etc.). The development of natural product disinfectants requires a better understanding of the mechanisms of action (MOA), chemical profiles, compound specificities, activity spectra, and the chemical formulations required for maximum activity. Combinations of natural antiviral substances and possibly the addition of synthetic compounds might be needed to increase inactivation of a broader spectrum of viruses, thereby providing the required efficacy for surface and air disinfection.

Evaluation of nuclear and mitochondrial phylogenetics for the subtyping of Cyclospora cayetanensis.

Cyclospora cayetanensis is an enteric coccidian parasite responsible for gastrointestinal disease transmitted through contaminated food and water. It has been documented in several countries, mostly with low-socioeconomic levels, although major outbreaks have hit developed countries. Detection methods based on oocyst morphology, staining, and molecular testing have been developed. However, the current MLST panel offers an opportunity for enhancement, as amplification of all molecular markers remains unfeasible in the majority of samples. This study aims to address this challenge by evaluating two approaches for analyzing the genetic diversity of C. cayetanensis and identifying reliable markers for subtyping: core homologous genes and mitochondrial genome analysis. A pangenome was constructed using 36 complete genomes of C. cayetanensis, and a haplotype network and phylogenetic analysis were conducted using 33 mitochondrial genomes. Through the analysis of the pangenome, 47 potential markers were identified, emphasizing the need for more sequence data to achieve comprehensive characterization. Additionally, the analysis of mitochondrial genomes revealed 19 single-nucleotide variations that can serve as characteristic markers for subtyping this parasite. These findings not only contribute to the selection of molecular markers for C. cayetanensis subtyping, but they also drive the knowledge toward the potential development of a comprehensive genotyping method for this parasite.

Inactivation of SARS-CoV-2 in Water by Chlorination.

Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) is present in both respiratory secretions and feces, creating its potential for transmission by swimming pools. Recreational water activity is known to be at increased risk of respiratory infections and respiratory viruses have caused been detected and have caused outbreaks in swimming pools. However, little is known regarding the chlorine inactivation of SARS-CoV-2 in water typical of swimming pools in the USA. In this study, the inactivation of SARS-CoV-2 Isolate hCoV-19/USA-WA1/2020 was observed in water by chlorination. All experiments were conducted within a BSL-3 laboratory at room temperature. Our results show that the virus was reduced by 3.5 log (> 99.9%) after 30 s of 2.05-mg/L free chlorine contact and greater than 4.17 log (limit of detection) (> 99.99%) within 2 min.

Evaluating infection risks and importance of hand hygiene during the household laundry process using a quantitative microbial risk assessment approach.

BACKGROUND: Contaminated laundry contributes to infectious disease spread in residential and home health care settings. The objectives were to (1) evaluate pathogen transmission risks for individuals doing laundry, and (2) compare hand hygiene timing to reduce risks. METHODS: A quantitative microbial risk assessment using experimental data from a laundry washing effectiveness study was applied to estimate infection risks from SARS-CoV-2, rotavirus, norovirus, nontyphoidal Salmonella, and Escherichia coli in 4 laundry scenarios: 1 baseline scenario (no hand hygiene event) and 3 hand hygiene scenarios (scenario 1: after moving dirty clothes to the washing machine, scenario 2: after moving washed clothes to the dryer, and scenario 3: hand hygiene events following scenario 1 and 2). RESULTS: The average infection risks for the baseline scenario were all greater than 2 common risk thresholds (1.0×10-6and 1.0×10-4). For all organisms, scenario 1 yielded greater risk reductions (39.95%-99.86%) than scenario 2 (1.35%-55.25%). Scenario 3 further reduced risk, achieving 1.0×10-6(SARS-CoV-2) and 1.0×10-4risk thresholds (norovirus and E. coli). CONCLUSIONS: The modeled results suggest individuals should reduce hand-to-facial orifice (eyes, nose, and mouth) contacts and conduct proper hand hygiene when handling contaminated garments. More empirical data are needed to confirm the estimated risks. DATA AVAILABILITY STATEMENT: The data and code that support the findings of this study can be retrieved via a Creative Commons Zero v1.0 Universal license in GitHub at https://github.com/yhjung1231/Laundry-QMRAproject-2022.git DOI: http://doi.org/10.5281/zenodo.7122065.

A new workflow for assigning removal credits to assess overall performance of managed aquifer recharge (MAR).

Pathogen removal in managed aquifer recharge (MAR) systems is dependent upon numerous operational, physicochemical water quality, and biological parameters. Due to the site-specific conditions affecting these parameters, guidelines for specifying pathogen removal have historically taken rather precautionary and conservative approaches in order to protect groundwater quality and public health. A literature review of regulated pathogens in MAR applications was conducted and compared to up-and-coming indicators and surrogates for pathogen assessment, all of which can be gathered into a toolbox from which regulators and operators alike can select appropriate pathogens for monitoring and optimization of MAR practices. Combined with improved knowledge of pathogen fate and transport obtained through lab- and pilot-scale studies and supported by modeling, this foundation can be used to select appropriate, site-specific pathogens for regarding a more efficient pathogen retention, ultimately protecting public health and reducing costs. This paper outlines a new 10 step-wise workflow for moving towards determining robust removal credits for pathogens based on risk management principles. This approach is tailored to local conditions while reducing overly conservative regulatory restrictions or insufficient safety contingencies. The workflow is intended to help enable the full potential of MAR as more planned water reuse systems are implemented in the coming years.

Survival of SARS-CoV-2 in wastewater.

The ongoing pandemic of Coronavirus disease 2019 (COVID-19) has affected >600 million people with >6 million deaths. Although Severe Acute Respiratory Syndrome-Coronavirus-2 (SARS-CoV-2), the etiologic agent of COVID-19, is transmitted via respiratory droplets or direct contact, isolation of viable SARS-CoV-2 in feces has been reported. Therefore, there is a need for understanding the persistence of SARS-CoV-2 and emerging variants in wastewater. In this study, the survival of SARS-CoV-2 isolate hCoV-19/USA-WA1/2020 was observed in three wastewater matrices - filtered and unfiltered raw wastewater, and secondary effluent. All experiments were conducted within a BSL-3 laboratory at room temperature. The time required for inactivation of 90 % (T90) of SARS-CoV-2 was 10.4, 10.8, and 18.3 h for unfiltered raw, filtered raw, and secondary effluent, respectively. Progressive decline in infectivity of the virus following first order kinetics was noted in these wastewater matrices. To the best of our knowledge, this is the first study to describe the survival of SARS-CoV-2 in secondary effluent.

In vitro antiviral effect of Mexican and Brazilian propolis and phenolic compounds against human coronavirus 229E.

Propolis is a resinous substance collected by bees (Apis mellifera). It is used for its biological properties. This natural product is available as a safe therapeutic option. Herein, we report the antiviral effects of brown propolis extract from Mexico and green and red propolis extracts from Brazil, as well as their phenolic compounds (quercetin, caffeic acid, and rutin) in preventing infection of MRC-5 cells by HCoV-229E. Normal human fibroblast lung cells (MRC-5) were used to determine the cytotoxicity of the compounds. All samples studied showed antiviral activity. Green and brown propolis extracts, and quercetin exhibited the best EC50 values with values of 19.080, 11.240, and 77.208 µg/mL against HCoV-229E, respectively, and with TC50 of 62.19, 29.192, and 298 µg/mL on MRC-5 cells, respectively. These results are the first in vitro study of the effects of propolis on HCoV-229E and provide the basis for the development of natural formulations against other coronavirus strains.

UV Inactivation of Common Pathogens and Surrogates Under 222 nm Irradiation from KrCl* Excimer Lamps.

Germicidal ultraviolet (UV) devices have been widely used for pathogen disinfection in water, air, and on food and surfaces. Emerging UV technologies, like the krypton chloride (KrCl*) excimer emitting at 222 nm, are rapidly gaining popularity due to their minimal adverse effects on skin and eyes compared with conventional UV lamps emitting at 254 nm, opening opportunities for UV disinfection in occupied public spaces. In this study, inactivation of seven bacteria and five viruses, including waterborne, foodborne and respiratory pathogens, was determined in a thin-film aqueous solution using a filtered KrCl* excimer emitting primarily at 222 nm. Our results show that the KrCl* excimer can effectively inactivate all tested bacteria and viruses, with most microorganisms achieving more than 4-log (99.99%) reduction with a UV dose of 10 mJ cm-2 . Compared with conventional UV lamps, the KrCl* excimer lamp exhibited better disinfection performance for viruses but was slightly less effective for bacteria. The relationships between UV sensitivities at 222 and 254 nm for bacteria and viruses were evaluated using regression analysis, resulting in factors that could be used to estimate the KrCl* excimer disinfection performance from well-documented UV kinetics using conventional 254 nm UV lamps. This study provides fundamental information for pathogen disinfection when employing KrCl* excimers.

Continuously active disinfectant inactivates severe acute respiratory coronavirus virus 2 (SARS-CoV-2) and human coronavirus 229E two days after the disinfectant was applied and following wear exposures.

The surface environment in rooms of coronavirus disease 2019 (COVID-19) patients may be persistently contaminated despite disinfection. A continuously active disinfectant demonstrated excellent sustained antiviral activity following a 48-hour period of wear and abrasion exposures with reinoculations. Reductions of >4-log10 were achieved within a 1-minute contact time for severe acute respiratory coronavirus virus 2 (SARS-CoV-2) and the human coronavirus, 229E.

Transfer efficiency of an enveloped virus, human coronavirus 229E, from various hard surface fomites to finger pads of the hands.

Respiratory viruses can be transmitted by fomite contact, but no data currently exist on the transfer of enveloped viruses. The transfer efficiency of human coronavirus from various hard surfaces ranged from 0.46% to 49.0%. This information can be used to model the fomite transmission of enveloped viruses.

Impacts of lid closure during toilet flushing and of toilet bowl cleaning on viral contamination of surfaces in United States restrooms.

BACKGROUND: Viral aerosols generated during toilet flushing represent a potential route of pathogen transmission. The goal of this study was to determine the impact of toilet lid closure prior to flushing on the generation of viral aerosols and cross-contamination of restroom fomites. METHODS: A surrogate for human enteric viruses (bacteriophage MS2) was added to household and public toilet bowls and flushed. The resulting viral contamination of the toilet and other restroom surfaces was then determined. RESULTS: After flushing the inoculated toilets, toilet seat bottoms averaged >107 PFU/100 cm2. Viral contamination of restroom surfaces did not depend on toilet lid position (up or down). After toilet bowls were cleaned using a bowl brush with or without a commercial product (hydrochloric acid), a >4 log10 (>99.99%) reduction in contamination of the toilet bowl water was observed versus no product. Bowl brush contamination was reduced by 1.6 log10 (97.64%) when the product was used versus no product. CONCLUSIONS: These results demonstrate that closing the toilet lid prior to flushing does not mitigate the risk of contaminating bathroom surfaces and that disinfection of all restroom surfaces (ie, toilet rim, floors) may be necessary after flushing or after toilet brush used for the reduction of virus cross-contamination.

Variations in Bacterial Communities and Antibiotic Resistance Genes Across Diverse Recycled and Surface Water Irrigation Sources in the Mid-Atlantic and Southwest United States: A CONSERVE Two-Year Field Study.

Reduced availability of agricultural water has spurred increased interest in using recycled irrigation water for U.S. food crop production. However, there are significant knowledge gaps concerning the microbiological quality of these water sources. To address these gaps, we used 16S rRNA gene and metagenomic sequencing to characterize taxonomic and functional variations (e.g., antimicrobial resistance) in bacterial communities across diverse recycled and surface water irrigation sources. We collected 1 L water samples (n = 410) between 2016 and 2018 from the Mid-Atlantic (12 sites) and Southwest (10 sites) U.S. Samples were filtered, and DNA was extracted. The V3-V4 regions of the 16S rRNA gene were then PCR amplified and sequenced. Metagenomic sequencing was also performed to characterize antibiotic, metal, and biocide resistance genes. Bacterial alpha and beta diversities were significantly different (p < 0.001) across water types and seasons. Pathogenic bacteria, such as Salmonella enterica, Staphylococcus aureus, and Aeromonas hydrophilia were observed across sample types. The most common antibiotic resistance genes identified coded against macrolides/lincosamides/streptogramins, aminoglycosides, rifampin and elfamycins, and their read counts fluctuated across seasons. We also observed multi-metal and multi-biocide resistance across all water types. To our knowledge, this is the most comprehensive longitudinal study to date of U.S. recycled water and surface water used for irrigation. Our findings improve understanding of the potential differences in the risk of exposure to bacterial pathogens and antibiotic resistance genes originating from diverse irrigation water sources across seasons and U.S. regions.

Minding the matrix: The importance of inoculum suspensions on finger transfer efficiency of virus.

AIMS: The aim of this study was to determine how the transfer efficiency of MS-2 coliphage from the toilet seat to hands and fingertip to lip differs according to the suspension of the inoculum. METHODS AND RESULTS: Hands were sampled after lifting a toilet seat which was inoculated with MS-2 on the underneath side. MS-2 was suspended in a spectrum of proteinaceous and non-proteinaceous solutions. Transfer efficiencies were greatest with the ASTM tripartite soil load (3.02% ± 4.03) and lowest with phosphate-buffered saline (PBS) (1.10% ± 0.81) for hand-to-toilet seat contacts. Finger-to-lip transfer rates were significantly different (p < 0.05) depending on suspension matrix, with PBS yielding the highest transfer (52.53% ± 4.48%) and tryptose soy broth (TSB) the lowest (23.15% ± 24.27%). Quantitative microbial risk assessment was used to estimate the probability of infection from adenovirus and norovirus from finger contact with a toilet seat. CONCLUSIONS: The greatest transfer as well as the largest variation of transfer were measured for finger-to-lip contacts as opposed to toilet seat-to-finger contacts. These factors influence the estimation of the probability of infection from micro-activity, that is, toilet seat adjustment. SIGNIFICANCE AND IMPACT: Viruses may be transferred from various human excreta with differing transfer efficiencies, depending on the protein content.