The Efficacy of Commercial Surface Sanitizers against Norovirus on Formica Surfaces with and without Inclusion of a Wiping Step.

Commonly used surface sanitizers often lack activity against human noroviruses (hNoVs). The impact of inactivation versus removal when these products are applied via wiping is poorly characterized. The purpose of this work was to assess the anti-hNoV efficacy of various surface sanitizer chemistries, as applied to a laminate material commonly used for restaurant tabletops, using standard surface assays (ASTM E1053-11) and a newly developed wiping protocol. Four commercially available products with different active ingredient(s) (i.e., ethanol [EtOH], acid + anionic surfactant [AAS], quaternary ammonium compound [QAC], and sodium hypochlorite [NaOCl]) and a water control were evaluated against hNoV GII.4 Sydney, hNoV GI.6, and the cultivable surrogate Tulane virus (TuV). Virus concentration was evaluated using RNase-reverse transcriptase (RT)-quantitative PCR (qPCR) (hNoV) and infectivity assay (TuV). Only the EtOH-based product significantly reduced virus concentration (>3.5 log10 reduction [LR]) by surface assay, with all other products producing ≤0.5 LR. The inclusion of a wiping step enhanced the efficacy of all products, producing complete virus elimination for the EtOH-based product and 1.6 to 3.8 LR for the other chemistries. For hNoVs, no detectable residual virus could be recovered from paper towels used to wipe the EtOH-based product, while high concentrations of virus could be recovered from the used paper towel and the wiped coupon (1.5 to 2.5 log10 lower genome equivalent copies [GEC] compared to control) for the QAC- and AAS-based products and for water. These results illustrate the variability in anti-hNoV activity of representative surface sanitizers and highlights the value of wiping, the efficacy of which appears to be driven by a combination of virus inactivation and removal. IMPORTANCE Human noroviruses (hNoVs) are the leading cause of acute gastroenteritis and food-borne disease worldwide. Noroviruses are difficult to inactivate, being recalcitrant to sanitizers and disinfectants commonly used by the retail food sector. This comparative study demonstrates the variability in anti-hNoV activity of representative surface sanitizers, even those allowed to make label claims based on the cultivable surrogate, feline calicivirus (FCV). It also highlights the importance of wiping in the process of sanitization, which significantly improves product efficacy through the action of physical removal of surface microbes. There is a need for more and better product formulations with demonstrated efficacy against hNoVs, which will likely necessitate the use of alternative cultivable surrogates, such as Tulane virus (TuV). These findings help food safety professionals make informed decisions on sanitizing product selection and application methods in order to reduce the risk of hNoV contamination and transmission in their facilities.

Efficacy of an alcohol-based surface disinfectant formulation against human norovirus.

AIM: To evaluate the anti-noroviral efficacy of PURELL® surface sanitizer and disinfectant spray (PSS, an alcohol-based formulation) using human norovirus GII.4 Sydney [hNoV, by RT-qPCR and human intestinal enteroid (HIE) infectivity assay] and its cultivable surrogate, Tulane virus (TuV, infectivity assay), compared to sodium hypochlorite (NaOCl) solutions. METHODS AND RESULTS: PSS efficacy was evaluated in suspension and on surfaces [stainless steel (SS)] using ASTM methods. Results were expressed as log10 reduction (LR) of genome equivalent copy number (GEC, for hNoV, assayed by RT-qPCR) and plaque forming units (PFU, for TuV, per infectivity assay). In suspension, PSS achieved a 2.9 ± 0.04 LR hNoV GEC irrespective of contact time (30 or 60 s) and soil load (2.5% or 5%). Under all treatment conditions, infectious TuV could not be recovered following exposure to PSS, corresponding to the assay limit of detection (3.1-5.2 log10 PFU). Infectious hNoV could not be detected in the HIE model after exposure to PSS. On SS and 2.5% soil, PSS produced a 3.1 ± 0.1 LR hNoV GEC, comparable to 500 ppm NaOCl for 60 s. With 5.0% soil, PSS produced a 2.5 ± 0.2 LR hNoV GEC, which was similar to 1000-5000 ppm NaOCl for 60 s. CONCLUSIONS: PSS showed high anti-hNoV efficacy by RT-qPCR and in in vitro (TuV) and ex vivo (HIE) infectivity assays and performed similar to 1000-5000 ppm NaOCl for a 60-s contact time on SS with added soil. SIGNIFICANCE AND IMPACT OF STUDY: hNoV remains a significant cause of morbidity globally, partly due to its resistance to numerous surface disinfectants. RT-qPCR results from this study indicate PSS efficacy against hNoV is comparable to NaOCl efficacy. Infectivity assays leveraging TuV and the HIE model for hNoV support and confirm loss of virus infectivity. Collectively, these results indicate the product's ability to inactivate hNoV quickly, which could be beneficial in settings having elevated risk for hNoV transmission.

Laboratory evidence of norwalk virus contamination on the hands of infected individuals.

Human norovirus (NoV) outbreak investigations suggest that the hands of infected individuals play an important role in NoV transmission. However, there is no experimental evidence documenting the likelihood and degree of NoV contamination on hands. As part of a clinical trial designed to evaluate the efficacy of high-pressure processing for Norwalk virus (NV) inactivation in oysters, 159 hand rinse samples were collected from 6 infected and 6 uninfected subjects. NV was concentrated from the samples by polyethylene glycol precipitation, followed by RNA extraction using an automated guanidinium isothiocyanate-silica method. NV RNA was detected and quantified using multiple NV-specific reverse transcription-quantitative PCR (RT-qPCR) assays. A total of 25.4% (18/71) of the hand rinse samples collected from 6 infected volunteers were presumptively positive for NV, with an average of 3.86 log10 genomic equivalent copies (GEC) per hand. Dot blot hybridization of PCR products obtained using a different primer set, and DNA sequencing of selected amplicons, provided further confirmation of the presence of NV in the hand rinses. NV contamination was also detected in two hand rinse samples obtained from one uninfected subject. These findings provide definitive evidence of NV contamination on the hands of infected subjects observed under controlled clinical research conditions. Such data support the need for better hand hygiene strategies to prevent NoV transmission.

The role of hands in cross-contamination of kitchen surfaces during meal preparation.

BACKGROUND: Foodborne pathogen transmission during food preparation is a common occurrence, and cross-contamination can be a contributing factor. Behaviors that lead to cross-contamination during meal preparation have not been well characterized. The study objective was to determine how hands and food handling behaviors (with a focus on handwashing and touch-based events) affect the risk of cross-contamination of kitchen surfaces and foods during meal preparation. METHODS: Data from a prior study in which participants were observed preparing turkey burgers inoculated with bacteriophage MS2 and a salad provided the data for analysis. Cross-contamination was assessed using environmental sampling data. Behavioral coding was performed for handwashing and touch-based behaviors. Cross-contamination risk was defined as the likelihood (number of contaminated surfaces) and degree (contaminant concentration) of MS2 on surfaces. Statistical analyses were performed in R, SPSS, and SigmaPlot. RESULTS: The significantly reduced risk of cross-contamination (P.ß<.ß.0001) was observed for participants who attempted handwashing or completed more handwashing steps. Scrubbing hands for 5.ßseconds, on average, reduced the risk of cross-contamination (P.ß<.ß.05). Cross-contamination regression models created using the most significant predictor variables showed that increased handwashing attempts, completion of more handwashing steps, and average scrub times>5.ßseconds all decreased the risk of cross-contamination (P.ß<.ß.05). CONCLUSIONS: This analysis can be used in future risk assessment modeling and for informing education and outreach to reduce pathogen transmission during food preparation.

Comparative Assessment of the Efficacy of Commercial Hand Sanitizers Against Human Norovirus Evaluated by an in vivo Fingerpad Method.

Human noroviruses (hNoV) are the leading cause of acute non-bacterial gastroenteritis worldwide and contaminated hands play a significant role in the spread of disease. Some hand sanitizers claim to interrupt hNoV transmission, but their antiviral efficacy on human hands is poorly characterized. The purpose of this work was to characterize the efficacy of representative commercial hand sanitizers against hNoV using an in vivo fingerpad method (ASTM E1838-17). Eight products [seven ethanol-based and one benzalkonium chloride (BAK)-based], and a benchmark 60% ethanol solution, were each evaluated on 10 human volunteers using the epidemic GII.4 hNoV strain. Virus titers before and after treatment were evaluated by RT-qPCR preceded by RNase treatment; product efficacy was characterized by log10 reduction (LR) in hNoV genome equivalent copies after treatment. The benchmark treatment produced a 1.7 ± 0.5 LR, compared with Product A (containing 85% ethanol) which produced a 3.3 ± 0.3 LR and was the most efficacious (p < 0.05). Product B (containing 70% ethanol), while less efficacious than Product A (p < 0.05), performed better than the benchmark with a LR of 2.4 ± 0.4. Five of the other ethanol-based products (labeled ethanol concentration ranges of 62-80%) showed similar efficacy to the 60% ethanol benchmark with LR ranging from 1.3 to 2.0 (p > 0.05). Product H (0.1% BAK) was less effective than the benchmark with a LR of 0.3 ± 0.2 (p < 0.05). None of the products screened were able to completely eliminate hNoV (maximum assay resolution 5.0 LR). Product performance was variable and appears driven by overall formulation. There remains a need for more hand sanitizer formulations having greater activity against hNoV, a virus that is comparatively recalcitrant relative to other pathogens of concern in community, healthcare, and food preparation environments.

Cross-Contamination to Surfaces in Consumer Kitchens with MS2 as a Tracer Organism in Ground Turkey Patties.

ABSTRACT: It is estimated that one in five cases of foodborne illnesses is acquired in the home. However, how pathogens move throughout a kitchen environment when consumers are preparing food is not well characterized. The purpose of this study was to determine the prevalence and degree of cross-contamination across a variety of kitchen surfaces during a consumer meal preparation event. Consumers (n = 371) prepared a meal consisting of turkey patties containing the bacteriophage MS2 as a tracer organism and a ready-to-eat lettuce salad. Half were shown a video on proper thermometer use before the trial. After meal preparation, environmental sampling and detection were performed to assess cross-contamination with MS2. For most surfaces, positivity did not exceed 20%, with the exception of spice containers, for which 48% of the samples showed evidence of MS2 cross-contamination. Spice containers also had the highest MS2 concentrations, at a mean exceeding 6 log viral genome equivalent copies per surface. The high level of MS2 on spice containers drove the significant differences between surfaces, suggesting the significance of spice containers as a vehicle for cross-contamination, despite the absence of previous reports to this effect. The thermometer safety intervention did not affect cross-contamination. The efficiency of MS2 transfer, when expressed as a percentage, was relatively low, ranging from an average of 0.002 to 0.07%. Quantitative risk assessment work using these data would aid in further understanding the significance of cross-contamination frequency and efficiency. Overall, these data will help create more targeted consumer messaging to better influence consumer cross-contamination behaviors.

Cross-Contamination on Atypical Surfaces and Venues in Food Service Environments.

ABSTRACT: Cross-contamination of raw food to other surfaces, hands, and foods is a serious issue in food service. With individuals eating more meals away from home, contracting a foodborne illness from a food service establishment is an increasing concern. However, most studies have concentrated on hands or food contact surfaces and neglected atypical and unusual surfaces (surfaces that are not typically identified as a source of cross-contamination) and venues. This review was conducted to identify atypically cross-contaminated surfaces and atypical venues where cross-contamination could occur that have not been examined thoroughly in the literature. Most surfaces that could be at risk for cross-contamination are frequently touched, are rarely cleaned and sanitized, and can support the persistence and/or growth of foodborne pathogens. These surfaces include menus, spice and condiment containers, aprons and coveralls, mobile devices and tablets, and money. Venues that are explored, such as temporary events, mobile vendors, and markets, are usually limited in space or infrastructure, have low compliance with proper hand washing, and provide the opportunity for raw and ready-to-eat foods to come into contact with one another. These factors create an environment in which cross-contamination can occur and potentially impact food safety. A more comprehensive cleaning and sanitizing regime encompassing these surfaces and venues could help mitigate cross-contamination. This review highlights key surfaces and venues that have the potential to be cross-contaminated and have been underestimated or not fully investigated. These knowledge gaps indicate where further work is needed to fully understand the role of these surfaces and venues in cross-contamination and how it can be prevented.

Characterisation of curli production, cell surface hydrophobicity, autoaggregation and attachment behaviour of Escherichia coli O157.

Escherichia coli O157 are an important group of foodborne pathogens with the ability to attach to materials commonly used in food processing environments such as slightly hydrophilic stainless steel. The aim of this study was to characterise six E. coli isolates, including five E. coli O157, for curli production, autoaggregation, hydrophobicity and attachment to highly hydrophilic glass and hydrophobic Teflon. Curli production and autoaggregation were determined using absorbance assays; hydrophobicity by bacterial adherence to hydrocarbons, hydrophobic interaction chromatography and contact angle measurements; and attachment using epifluorescence microscopy. Curli production varied between strains and for some strains correlated with autoaggregation. Curli production correlated with decreased hydrophobicity for two strains. Four of the six isolates increased attachment to glass, but decreased attachment to Teflon with increased curli production. In contrast, one of the six isolates decreased attachment to glass, but increased attachment to Teflon with increasing curli production. Curli production by the remaining isolate did not correlate with hydrophobicity or attachment. Attachment of some E. coli, including E. coli O157, to abiotic surfaces may be influenced by curli production, autoaggregation and hydrophobicity. However, for other strains, a variety of factors may be of greater influence on these properties and ability to attach to abiotic surfaces. This study highlights the complexity of bacterial surface properties and their relationship with bacterial attachment.

Comparative analysis of attachment of Shiga-toxigenic Escherichia coli and Salmonella strains to cultured HT-29 and Caco-2 cell lines.

The ability of Escherichia coli and Salmonella isolates to attach to Caco-2 and HT-29 cell monolayers was measured. All isolates displayed a greater ability to attach to Caco-2 cells than HT-29 cells, and overall E. coli isolates attached better to both cell lines than Salmonella isolates. Bacteria that were considered to be pathogenic displayed no greater ability to attach to cell lines than those that were not considered to be pathogenic. Additionally, no correlation was found between cell line attachment and previously determined hydrophobicity results.