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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.

The antinoroviral effect of copper ions is well known, yet most of this work has previously been conducted in copper and copper alloy surfaces, not copper ions in solution. In this work, we characterized the effects that Cu ions have on human norovirus capsids' and surrogates' integrity to explain empirical data, indicating virus inactivation by copper alloy surfaces, and as means of developing novel metal ion-based virucides. Comparatively high concentrations of Cu(II) ions (10 mM) had little effect on the infectivity of human norovirus surrogates, so we used sodium ascorbate as a reducing agent to generate unstable Cu(I) ions from solutions of copper bromide. We found that significantly lower concentrations of monovalent copper ions (∼0.1 mM) compared to divalent copper ions cause capsid protein damage that prevents human norovirus capsids from binding to cell receptors

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 log

Background Sodium intake in the USA exceeds recommendations. The replacement of added sodium chloride (NaCl) with potassium chloride (KCl) provides a potential strategy to reduce sodium intake. Objective The purpose of this study was to quantitatively estimate changes in intakes of sodium and potassium by the US population assuming use of potassium-based NaCl replacers in top dietary sodium sources. Methods Data collected in the What We Eat in America (WWEIA) component of the 2015–2016 and 2009–2010 NHANES were used to identify top-ranking sources of dietary sodium among the population aged 2 y and older based on contributions from food categories aligning with the FDA draft guidance for voluntary sodium reduction. Predicted nutrient intakes were estimated in models assuming total and feasible and practical (F&P) replacement of added NaCl with KCl in foods and ingredients within the top food sources of sodium. An expert elicitation was conducted to collect information on the F&P KCl replacement of added NaCl. Results Using 2015–2016 consumption data, the total replacement of added NaCl with KCl in the 18 top-ranking sources of dietary sodium results in a predicted sodium intake of 2004 mg/d from the replacement of 1406 mg/d sodium with 1870 mg/d potassium as KCl. Modeled F&P replacement predicted sodium intakes of 3117 mg/d (range of 2953 to 3255 mg/d) from the replacement of 294 mg/d sodium (155 to 457 mg/d) with 390 mg/d potassium (206 to 608 mg/d). Similar results are seen with 2009–2010 data. Conclusions The F&P replacement of NaCl with KCl in top-ranking sources of dietary sodium modeled in this study can result in decreased sodium to a level consistent with the short-term intake goal targeted by the FDA of 3000 mg/d, with the mean potassium intake remaining in the range recommended for the apparently healthy population.

Cantaloupe melons, which have been responsible of an increasing number of foodborne disease outbreaks, may become contaminated with microbial pathogens during production. However, little information is available on the microbial populations in the cantaloupe farm environment. The purpose of this work was to characterize the bacterial communities present on cantaloupe farms. Fruit, soil, and harvester hand rinsates were collected from two Mexican cantaloupe farms, each visited three times. Microbiome analysis was performed by sequencing 16sRNA and analyzed using qiime2 software. Correlations were determined between sample type and microbial populations. The α and β diversity analysis identified 2777 sequences across all samples. The soil samples had the highest number and diversity of unique species (from 130 to 1329 OTUs); cantaloupe (from 112 to 205 OTUs), and hands (from 67 to 151 OTUs) had similar diversity. Collectively, Proteobacteria was the most abundant phyla (from 42 to 95%), followed by Firmicutes (1-47%), Actinobacteria (< 1 to 23%), and Bacteroidetes (< 1 to 4.8%). The most abundant genera were Acinetobacter (20-58%), Pseudomonas (14.5%), Erwinia (13%), and Exiguobacterium (6.3%). Genera with potential to be pathogenic included Bacillus (4%), Salmonella (0.85%), Escherichia-Shigella (0.38%), Staphylococcus (0.32%), Listeria (0.29%), Clostridium (0.28%), and Cronobacter (0.27%), which were found at lower frequencies. This study provides information on the cantaloupe production microbiome, which can inform future research into critical food safety issues such as antimicrobial resistance, virulence, and genomic epidemiology.

Successful human norovirus (HuNoV) cultivation in stem cell‐derived human intestinal enteroids (HIE) was recently reported. The purpose of this study was to evaluate the anti‐HuNoV efficacy of two alcohol‐based commercial hand sanitizers and 60% ethanol by suspension assay using RNase‐RT‐qPCR, with subsequent validation of efficacy by HuNoV cultivation using the HIE model. In suspension, when evaluated by RNase‐RT‐qPCR, 60% ethanol resulted in less than one log10 reduction in HuNoV genome equivalent copies (GEC) regardless of contact time (30 or 60s) or soil load. The two commercial products outperformed 60% ethanol regardless of contact time or soil load, providing 2·2–3·2 log10 HuNoV GEC reductions by suspension assay. Product B could not be validated in the HIE model due to cytotoxicity. Following a 60s exposure, viral replication in the HIE model increased 1·9 ± 0·2 log10 HuNoV GEC for the neutralization (positive) control and increased 0·9 ± 0·2 log10 HuNoV GEC in challenged HIE after treatment with 60% ethanol. No HuNoV replication in HIE was observed after a 60 s exposure to Product A., Significance and Impact of the Study: The purpose of this study was to evaluate the efficacy of two alcohol‐based commercial hand sanitizers and 60% ethanol by suspension assay using RNase‐RT‐qPCR, with subsequent validation of efficacy by human norovirus (HuNoV) cultivation using stem cell‐derived human intestinal enteroids (HIE). When HuNoV was treated with hand sanitizers, a 2·2–3·2 log10 reduction was observed using RNase‐RTqPCR. Adsorption to HIE was inefficient, and replication was nonexistent at 72 h post infection, suggesting sanitizer treatment affects capsid binding and infectivity. Validating molecular data using an infectivity model is an important advancement towards more reliable characterization of anti‐HuNoV efficacy of sanitizers.

Essential food workers experience an elevated risk of SARS-CoV-2 infection due to prolonged occupational exposures (e.g., frequent close contact, enclosed spaces) in food production and processing areas, shared transportation (car or bus), and employer-provided shared housing. The purpose of this study was to evaluate the impact of combined food industry interventions and vaccination on reducing the daily cumulative risk of SARS-CoV-2 infection for produce workers. Six linked quantitative microbial risk assessment models were developed in R to simulate daily scenarios experienced by a worker. Standard industry interventions (2 m physical distancing, handwashing, surface disinfection, universal masking, increased ventilation) and two-dose mRNA vaccinations (86–99% efficacy) were modeled individually and jointly to assess risk reductions. The infection risk for an indoor (0.802, 95% Uncertainty Interval [UI]: 0.472–0.984) and outdoor (0.483, 95% UI: 0.255–0.821) worker was reduced to 0.018 (93% reduction) and 0.060 (87.5% reduction) after implementation of combined industry interventions. Upon integration of these interventions with vaccination, the infection risk for indoor (0.001, 95% UI: 0.0001–0.005) and outdoor (0.004, 95% UI: 0.001–0.016) workers was reduced by ≥99.1%. Food workers face considerable risk of occupationally-acquired SARS-CoV-2 infection without interventions; however, consistent implementation of key infection control measures paired with vaccination effectively mitigates these risks.SynopsisBundled interventions, particularly if they include vaccination, produce significant reductions (>99%) in SARS-CoV-2 infection risk for essential food workers.Graphical Abstract

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.

This study was conducted to test the effectiveness of a consumer poultry washing educational intervention that included video observation of meal preparation with participants who self-reported washing poultry. Treatment group participants received three e-mail messages containing information that the U.S. Department of Agriculture has used on social media sites (video and infographics) related to poultry preparation, including advising against washing chicken. Participants were observed cooking chicken thighs (inoculated with traceable nonpathogenic Escherichia coli strain DH5α) and preparing a salad to determine whether they washed the chicken and the extent of cross-contamination to the salad and areas of the kitchen. After meal preparation, participants responded to an interview about food handling behaviors, including questions about the intervention for treatment group participants. Three hundred people participated in the study (158 control, 142 treatment). The intervention effectively encouraged participants not to wash chicken before cooking; 93% of treatment group participants but only 39% of control group participants did not wash the chicken (P0.0001). The high levels of E. coli DH5α detected in the sink and on the salad lettuce suggest that microbes transferred to the sink from the chicken, packaging, or contaminated hands are a larger cause for concern than is splashing contaminated chicken fluids onto the counter. Among chicken washers, 26 and 30% of the lettuce from the prepared salad was contaminated for the control and treatment groups, respectively. For nonwashers, 31 and 15% of the lettuce was contaminated for the control and treatment groups, respectively. Hand-facilitated cross-contamination is suspected to be a factor in explaining this resulting lettuce cross-contamination. This study demonstrates the need to change the frame of "don't wash your poultry" messaging to instead focus on preventing contamination of sinks and continuing to emphasize the importance of hand washing and cleaning and sanitizing surfaces.

In-home pet food handling and food dish hygiene practices can have adverse health impacts for both humans and pets. Safe food and dish handling guidelines are not easily evidenced for pet owners. The study was designed to investigate dog owners’ feeding habits and evaluate the impact of the Food and Drug Association (FDA) hygiene protocols on dog food dish contamination. Procedures and surveys were approved by NCSU-IACUC and -IRB. Pet feeding and food dish hygiene data were collected from 417 dog owner surveys and 68 food dish swabs. Total aerobic cell counts (TAC) were performed on 68 dishes and randomly assigned into Group A (FDA pet food handling and dish hygiene guidelines), Group B (FDA pet and human food handling and dish hygiene guidelines), or Group C (no guidelines). Hygiene protocols were instituted in-home for 1 week, followed by a second TAC and follow-up survey. Survey from dog owners-households indicated: 4.7% were aware of FDA pet food handling and dish hygiene guidelines; 36% have individuals ≤ 13 years old and/or immunocompromised; 43% store dog food 0-5 feet from human food; 34% wash their hands after feeding; and 33% prepare their dog food on human food preparation surfaces. The hygiene protocols followed by Groups A and B resulted in significant decreases in food dish TAC (P160F) washing decreased TAC (p=0.005) over cold/lukewarm water. In the follow-up survey, 8% of Group A and B respondents reported likely to adhere to protocols long-term. This study suggests a need for pet food handling and dish hygiene guideline education to minimize bacterial contamination of dishes, especially in high-risk households.

In-home pet food handling and food dish hygiene practices can have adverse health impacts for both humans and pets. Safe food and dish handling guidelines are not easily evidenced for pet owners. The study was designed to investigate dog owners’ feeding habits and evaluate the impact of the Food and Drug Association (FDA) hygiene protocols on dog food dish contamination. Procedures and surveys were approved by North Carolina State University Institutional Animal Care and Use Committee and Institutional Review Board. Pet feeding and food dish hygiene data were collected from 417 dog owner surveys and 68 food dish swabs. Total aerobic plate counts (APC) were performed on 68 dishes and randomly assigned into Group A (FDA pet food handling and dish hygiene guidelines), Group B (FDA pet and human food handling and dish hygiene guidelines), or Group C (no guidelines). Hygiene protocols were instituted in-home for 1 week, followed by a second APC and follow-up survey. Survey from dog owners-households indicated: 4.7% were aware of FDA pet food handling and dish hygiene guidelines; 36% have individuals ≤ 13 years old and/or immunocompromised; 43% stored dog food 0–5 feet from human food; 34% washed their hands after feeding; and 33% prepared their dog food on human food preparation surfaces. The hygiene protocols followed by Groups A and B resulted in significant decreases in food dish APC (p160° F or 71.1°C) washing decreased APC (p

SummaryBackgroundCountries continue to debate the need for decontamination of cold-chain food packaging to reduce possible SARS-CoV-2 fomite transmission among workers. While laboratory-based studies demonstrate persistence of SARS-CoV-2 on surfaces, the likelihood of fomite-mediated transmission under real-life conditions is uncertain.MethodsUsing a quantitative risk assessment model, we simulated in a frozen food packaging facility 1) SARS-CoV-2 fomite-mediated infection risks following worker exposure to contaminated plastic packaging; and 2) reductions in these risks attributed to masking, handwashing, and vaccination.FindingsIn a representative facility with no specific interventions, SARS-CoV-2 infection risk to a susceptible worker from contact with contaminated packaging was 2·8 × 10−3per 1h-period (95%CI: 6·9 × 10−6, 2·4 × 10−2). Implementation of standard infection control measures, handwashing and masks (9·4 × 10−6risk per 1h-period, 95%CI: 2·3 × 10−8, 8·1 × 10−5), substantially reduced risk (99·7%). Vaccination of the susceptible worker (two doses Pfizer/Moderna, vaccine effectiveness: 86-99%) combined with handwashing and masking reduced risk to less than 1·0 × 10−6. Simulating increased infectiousness/transmissibility of new variants (2-, 10-fold viral shedding) among a fully vaccinated workforce, handwashing and masks continued to mitigate risk (2·0 × 10−6-1·1 × 10−5risk per 1h-period). Decontamination of packaging in addition to these interventions reduced infection risks to below the 1·0 × 10−6risk threshold.InterpretationFomite-mediated SARS-CoV-2 infection risks were very low under cold-chain conditions. Handwashing and masking provide significant protection to workers, especially when paired with vaccination.FundingU.S. Department of Agriculture

The SARS-CoV-2 global pandemic poses significant health risks to workers who are essential to maintaining the food supply chain. Using a quantitative risk assessment model, this study characterized the impact of risk reduction strategies for controlling SARS-CoV-2 transmission (droplet, aerosol, fomite-mediated) among front-line workers in a representative enclosed food manufacturing facility. We simulated: 1) individual and cumulative SARS-CoV-2 infection risks from close contact (droplet and aerosols at 1-3m), aerosol, and fomite-mediated exposures to a susceptible worker following exposure to an infected worker during an 8h-shift; and 2) the relative reduction in SARS-CoV-2 infection risk attributed to infection control interventions (physical distancing, mask use, ventilation, surface disinfection, hand hygiene). Without mitigation measures, the SARS-CoV-2 infection risk was largest for close contact (droplet and aerosol) at 1m (0.96, 95%CI: 0.67–1.0). In comparison, risk associated with fomite (0.26, 95%CI: 0.10–0.56) or aerosol exposure alone (0.05, 95%CI: 0.01–0.13) at 1m distance was substantially lower (73-95%). At 1m, droplet transmission predominated over aerosol and fomite-mediated transmission, however, this changed by 3m, with aerosols comprising the majority of the exposure dose. Increasing physical distancing reduced risk by 84% (1 to 2m) and 91% (1 to 3m). Universal mask use reduced infection risk by 52-88%, depending on mask type. Increasing ventilation (from 0.1 to 2-8 air changes/hour) resulted in risk reductions of 14-54% (1m) and 55-85% (2m). Combining these strategies, together with handwashing and surface disinfection, resulted in Significance StatementUsing mathematical modeling, we find that workers in enclosed food manufacturing facilities are at higher risk of SARS-CoV-2 infection from close contact transmission (exposure to large droplets and small aerosol particles) than fomite transmission. Thus, strategies protecting workers should prioritize close contact transmission pathways, such as physical distancing, universal mask use, and room air changes, with surface disinfection (reducing fomite transmission) and handwashing of secondary importance. Our work supports current international (EU-OSHA), domestic (FDA, OSHA), and food industry-standard guidance for managing COVID-19 transmission in essential workers in the food manufacturing sector. Although our model was designed for an indoor food manufacturing setting, it can be readily adapted to other indoor environments and infectious respiratory pathogens.

Cantaloupes contaminated with pathogens have led to many high-profile outbreaks and illnesses. Since bacterial virulence genes (VGs) can act in tandem with antibiotic-resistance and mobile genetic elements, there is a need to evaluate these gene reservoirs in fresh produce, such as cantaloupes. The goal of this study was to assess the distribution of antibiotic-resistance, virulence, and mobile genetic elements genes (MGEGs) in cantaloupe farm environments. A total of 200 samples from cantaloupe melons (

Countries continue to debate the need for decontamination of cold-chain food packaging to reduce possible severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) fomite transmission among frontline workers. While laboratory-based studies demonstrate persistence of SARS-CoV-2 on surfaces, the likelihood of fomite-mediated transmission under real-life conditions is uncertain. Using a quantitative microbial risk assessment model of a frozen food packaging facility, we simulated 1) SARS-CoV-2 fomite-mediated infection risks following worker exposure to contaminated plastic packaging; and 2) reductions in these risks from masking, handwashing, and vaccination. In a frozen food facility without interventions, SARS-CoV-2 infection risk to a susceptible worker from contact with contaminated packaging was 1.5 × 10−3 per 1h-period (5th – 95th percentile: 9.2 × 10−6, 1.2 × 10−2). Standard food industry infection control interventions, handwashing and masking, reduced risk (99.4%) to 8.5 × 10−6 risk per 1h-period (5th – 95th percentile: 2.8 × 10−8, 6.6 × 10−5). Vaccination of the susceptible worker (two doses Pfizer/Moderna, vaccine effectiveness: 86–99%) with handwashing and masking reduced risk to 5.2 × 10−7 risk per 1h-period (5th – 95th percentile: 1.8 × 10−9, 5.4 × 10−6). Simulating increased transmissibility of current and future variants (Delta, Omicron), (2-, 10-fold viral shedding) among a fully vaccinated workforce, handwashing and masking continued to mitigate risk (1.4 × 10−6 - 8.8 × 10−6 risk per 1h-period). Additional decontamination of frozen food plastic packaging reduced infection risks to 1.2 × 10−8 risk per 1h-period (5th – 95th percentile: 1.9 × 10−11, 9.5 × 10−8). Given that standard infection control interventions reduced risks well below 1 × 10−4 (World Health Organization water quality risk thresholds), additional packaging decontamination suggest no marginal benefit in risk reduction. Consequences of this decontamination may include increased chemical exposures to workers, food quality and hazard risks to consumers, and unnecessary added costs to governments and the global food industry.

Recent epidemiological evidence indicates that enteric viruses are the leading cause of foodborne disease in the U.S.A. and, indeed, worldwide. Certainly, advances in epidemiology and molecular biology have improved the ability to study this previously elusive group of foodborne pathogens. The purpose of this article is to review the agents, transmission routes, epidemiology, persistence, diagnosis, and detection of foodborne viruses and their diseases, with specific reference to the role that contemporary technologies have had in improving our understanding of this important group of emerging foodborne pathogens.

Human norovirus (NoV) is the leading cause of acute viral gastroenteritis and a major source of foodborne illness. Detection of NoV in food and environmental samples is typically performed using molecular techniques, including real-time reverse transcription polymerase chain reaction (RT-PCR) and less frequently, nested real-time PCR. In this study, we conducted a controlled comparison of two published NoV detection assays: a broadly reactive one-step real-time RT-PCR and a two-step nested real-time PCR assay. A 20% human fecal suspension containing a genogroup II human NoV was serially diluted, genome extracted, and subjected to amplification using the two assays compared via PCR Units. Additional amplicon confirmation was performed by dot blot hybridization using digoxigenin (DIG)-labeled oligonucleotide probes. Both assays displayed similar amplification standard curves/amplification efficiencies; however, the nested assay consistently detected one log10 lower virus. Dot blot hybridization improved the detection limit of the nested real-time PCR by one log10 NoV genome copies but impaired the detection limit of the one-step real-time RT-PCR by one log10 NoV genome copies. These results illustrate the complexities in designing and interpreting molecular techniques having a sufficient detection limit to detect low levels of viruses that might be anticipated in contaminated food and environmental samples.

Hand hygiene interventions are critical for reducing farmworker hand contamination and preventing the spread of produce-associated illness. Hand hygiene effectiveness may be produce-commodity specific, which could influence implementation strategies. This study’s goal was to determine if produce commodity influences the ability of handwashing with soap and water or two-step alcohol-based hand sanitizer (ABHS) interventions to reduce soil and bacteria on farmworker hands. Farmworkers (n = 326) harvested produce (cantaloupe, jalapeño, and tomato) for 30 to 90 minutes before engaging in handwashing, two-step ABHS (jalapeño and cantaloupe), or no hand hygiene. Hands were rinsed to measure amounts of soil (absorbance at 600 nm) and indicator bacteria (coliforms, Enterococcus sp., generic Escherichia coli, and Bacteroidales universal [AllBac] and human-specific [BFD] 16S rRNA gene markers). Without hand hygiene, bacterial concentrations (0.88 to 5.1 log(10) CFU/hand) on hands significantly differed by the produce commodity harvested. Moderate significant correlations (ρ = −0.41 to 0.56) between soil load and bacterial concentrations were observed. There were significant produce-commodity-specific differences in the ability of handwashing and two-step ABHS interventions to reduce soil (P

Our food systems depend on complex interactions between farmers and food producers, local and federal governments, and consumers. Underlying these interactions are economic, environmental, and societal factors that can impact the types of food available, access to food, affordability, and food safety. The recent SARS-CoV-2 global pandemic has affected multiple aspects of our food systems, from federal governments' decisions to limit food exports, to the ability of government agencies to inspect food and facilities to the ability of consumers to dine at restaurants. It has also provided opportunities for societies to take a close look at the vulnerabilities in our food systems and reinvent them to be more robust and resilient. For the most part, how these changes ultimately affect the safety and accessibility of food around the world remains to be seen.

The purpose of this study was to test the effectiveness of an intervention for consumer thermometer use by using a randomized experimental design and direct observation of meal preparation. The study was conducted in test kitchen facilities in two locations in North Carolina (one urban and one rural). Cameras recorded participants' actions at various locations throughout the kitchen and recorded the meal preparation from beginning to end. Before preparing the meal, a randomized treatment group watched a 3-min U.S. Department of Agriculture (USDA) food safety video "The Importance of Cooking to a Safe Internal Temperature and How to Use a Food Thermometer." Participants in the control and treatment groups were observed while cooking turkey burgers and preparing a salad to determine whether a thermometer was used to check the doneness of the turkey patties. Following meal preparation, all participants responded to a postobservation interview about food handling behaviors. Treatment group participants were also asked about the intervention. A total of 383 people participated in the study (201 in the control group and 182 in the treatment group). Participants who viewed the video were twice as likely to use a thermometer to check the doneness of the turkey patties compared with the participants who were not exposed to the video (75 versus 34%) and twice as likely to place the thermometer in the correct location (52 versus 23%). Sixty-seven percent of participants who watched the video reported that it influenced their behavior in the kitchen. This study demonstrates the importance of timing and framing of a behavioral intervention for thermometer use and highlights considerations for the development of additional messages (e.g., proper insertion). U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS) [GS-00F-354CA, AG3A94-D-16-0130] This work was funded by the U.S. Department of Agriculture, Food Safety and Inspection Service (FSIS) contract GS-00F-354CA, order AG3A94-D-16-0130. All views expressed here are those of the authors and not necessarily of the FSIS. Public domain – authored by a U.S. government employee

The most commonly used indicator of fecal contamination in fresh produce production and packing is Escherichia coli. In depth analysis of the prevalence and characteristics of naturally occurring E. coli strains in these environments is important because it can (1) serve as an indicator of sources of fecal contamination; and (2) provide information on strain pathogenicity, persistence, and other defining characteristics such as multidrug resistance. In this study, we analyzed 341 E. coli strains isolated from the jalapeno pepper, tomato and cantaloupe farm environments, in Northeast Mexico. Strains were isolated from produce, farmworkers' hands, soil and water. Pathotypes, genotypes, biofilm formation and antibiotic resistance were characterized. Phylogenetic subgroups and identification of diarrheagenic E. coli were determined by PCR; biofilm formation was quantified using a plate-based colorimetric method. Antibiotic resistance was analyzed by the Kirby Bauer diffusion disc method. Most isolates (N = 293, 86%) belonged to phylogenetic group A. Only four isolates (1.2%) were diarrheagenic: EPEC (N = 3) and ETEC (N = 1). Antibiotic resistance to tetracycline (23.2%) and ampicillin (19.9%) was high, and only 3.5% of the strains presented resistance to >5 antibiotics. Biofilms were produced by most strains (76%), among which 34.4% were categorized as high producers. The presence of antibiotic resistant E. coli strains that may contain gene markers for pathogenicity and which can form biofilms suggests potential health risks for consumers.

Consumption of cucumbers (Cucumis sativus var. sativus) has been linked to several foodborne outbreaks involving Salmonella enterica. The purpose of this work was to investigate the efficiency of colonization and internalization of S. enterica into cucumber plants by various routes of contamination. Produce-associated outbreak strains of Salmonella (a cocktail of serovars Javiana, Montevideo, Newport, Poona, and Typhimurium) were introduced to three cultivars of cucumber plants (two slicing cultivars and one pickling) via blossoms (ca. 6.4 log10 CFU/blossom, 4.5 log10 CFU/blossom, or 2.5 log10 CFU/blossom) or soil (ca. 8.3 log10 CFU/root zone) and were analyzed for prevalence of Salmonella contamination (internal and external) and serovar predominance in fruit and stems. Of the total slicing fruit harvested from Salmonella-inoculated blossoms (ca. 6.4, 4.5, or 2.5 log10 CFU/blossom), 83.9% (47/56), 81.4% (48/59) or 71.2% (84/118) were found colonized and 67.9% (38/56), 35.6% (21/59) or 22.0% (26/118) had Salmonella internalized into the fruit, respectively. S. Poona was the most prevalent serovar isolated on or in cucumber fruits at all inoculation levels. When soil was inoculated at 1 day post-transplant (dpt), 8% (10/120) of the plants were shown to translocate Salmonella to the lower stem 7 days post-inoculation (dpi). Results identified blossoms as an important route by which Salmonella internalized at a high percentage into cucumbers, and S. Poona, the same strain isolated from the 2015 outbreak of cucumbers imported from Mexico, was shown to be well-adapted to the blossom niche.

Cantaloupes have emerged as significant vehicles of widespread foodborne illness outbreaks caused by bacterial pathogens, including Salmonella. The purpose of this study was to investigate the efficiency of Salmonella colonization and internalization in cantaloupes by relevant routes of contamination. Cantaloupe plants (Cucumis melo ‘reticulatus’) from two cultivars ‘Athena’ (Eastern) and ‘Primo’ (Western) were grown from commercial seed. Plants were maintained in the NCSU BSL-3P phytotron greenhouse. Salmonella enterica (a cocktail of cantaloupe-associated outbreak serovars Javiana, Newport, Panama, Poona and Typhimurium) contamination was introduced via blossoms or soil at ca. 4.4 log10 CFU/blossom or 8.4 log10 CFU/root zone, respectively. Cantaloupes were analyzed for Salmonella by enrichment in accordance with modified FDA-BAM methods. Five randomly chosen colonies from each Salmonella-positive sample were typed using the Agilent 2100 bioanalyzer following multiplex PCR. Data were analyzed for prevalence of contamination and serovar predominance in fruit, stems and soil. Of the total cantaloupe fruit harvested from Salmonella-inoculated blossoms (n = 63), 89% (56/63) were externally contaminated and 73% (46/63) had Salmonella internalized into the fruit. Serovar Panama was the most commonly isolated from the surface of fruit while S. Panama and S. Poona were the most prevalent inside the fruit. When soil was inoculated with Salmonella at one day post-transplant, 13% (8/60) of the plants were shown to translocate the organism to the lower stem (ca. 4 cm) by 7 days post-inoculation (dpi). We observed Salmonella persistence in the soil up to 60 dpi with S. Newport being the predominant serovar at 10 and 20 dpi. These data demonstrate that contaminated soil and blossoms can lead to Salmonella internalization into the plant or fruit at a relatively high frequency.

A single stranded (ss) DNA aptamer, specific to members of Listeria genus, was used to develop a two-site binding sandwich assay for capture and detection of L. monocytogenes. Antibody-immobilized immunomagnetic beads were used to capture L. monocytogenes, followed by their exposure to the aptamer detector. Detection was achieved by amplification of cell-bound aptamers by qPCR. The lower limit of detection for the combined assay was 2.5 CFU L. monocytogenes in 500 μl buffer. This is juxtaposed to a detection limit of 2.4 log10 CFU in 500 μl buffer for immunomagnetic separation coupled with qPCR detection of L. monocytogenes targeting the hly gene. When applied to turkey deli meat, subjected to 24 h of non-selective enrichment, the two-site binding sandwich assay showed positive results at initial inoculum levels of 1–2 log10 CFU per 25 g sample. Because of its lower limit of detection, the assay reported here could be useful for detection of L. monocytogenes in foods and environmental samples.

In the agricultural setting, core global food safety elements, such as hand hygiene and worker furlough, should reduce the risk of norovirus contamination on fresh produce. However, the effect of these practices has not been characterized. Using a quantitative microbial risk model, we evaluated the individual and combined effect of farm-based hand hygiene and worker furlough practices on the maximum risk of norovirus infection from three produce commodities (open leaf lettuce, vine tomatoes, and raspberries). Specifically, we tested two scenarios where a harvester’s and packer’s norovirus infection status was: 1) assumed positive; or 2) assigned based on community norovirus prevalence estimates. In the first scenario with a norovirus-positive harvester and packer, none of the individual interventions modeled reduced produce contamination to below the norovirus infectious dose. However, combined interventions, particularly high handwashing compliance (100%) and efficacy (6 log(10) virus removal achieved using soap and water for 30 seconds), reduced produce contamination to

Viral enteric disease imposes a considerable public health and economic burden globally in both humans and livestock. Because enteric viruses are highly transmissible and resistant to numerous control strategies, making early in-field or point-of-care detection is important. There are problems with ligand-based detection strategies (e.g., sensitivity, false positive/negatives) for virus detection. Traditional amplification-based strategies are sensitive, but not as portable or rapid. Recombinase polymerase amplification is a new isothermal technique that utilizes bacterial genome repair enzymes to rapidly amplify target sequences. This report reviews the use of recombinase polymerase amplification for virus detection, showing that the method has favorable fundamental properties supporting its promise for rapid point-of-care detection of enteric viruses.

More efficient sampling and detection methods of pathogens on fresh produce are needed. The purpose of this study was to compare a novel rinse-membrane filtration method (RMFM) to a more traditional sponge rubbing or stomaching method in processing jalapeno peppers and cantaloupe samples for detection of Escherichia coli, Salmonella enterica, and Listeria monocytogenes. For jalapeno peppers inoculated with 106, 104, and 102 CFU of each pathogen and cantaloupes inoculated at 106 and 104 CFU, all pathogens were detected in all (100%) samples by RMFM at a 10-mL filtration volume, as well as by the stomacher and sponge rubbing methods. However, for cantaloupe inoculated at 102 CFU, detection differed by pathogen: S. enterica (20% RMFM, 60% stomacher, and 20% sponge), L. monocytogenes (40% RMFM, 60% stomacher, and 20% sponge), and E. coli O157:H7 (100% RMFM, 75% stomacher, and 75% sponge). When RMFM was compared with the other methods, in accordance with guidelines in the International Organization for Standardization 16140:2003 protocol, it produced values >95% in relative accuracy, relative specificity, and relative sensitivity. Overall, the RMFM performed similar to or better than the homogenization and sponge surface rubbing methods and is a good alternative for processing large numbers of produce samples for bacterial pathogen detection.

Background The purpose of this study was to relate experimentally measured log10 human norovirus reductions for a nonresidual (60% ethanol) and a residual (quaternary ammonium-based) hand sanitizer to infection risk reductions. Methods Human norovirus log10 reductions on hands for both sanitizers were experimentally measured using the ASTM International Standard E1838-10 method, with modification. Scenarios included product application to: (1) inoculated fingerpads with 30- and 60-second contact times, and (2) hands followed by inoculation with human norovirus immediately and 4 hours later. Hand sanitizer efficacies were used in a mathematical model estimating norovirus infection risk from a single hand-to-fomite contact under low and high environmental contamination conditions. Results The largest log10 reductions for the residual and nonresidual hand sanitizers were for a 60-second contact time, reducing infection risk by approximately 99% and 85%, respectively. Four hours after application, the residual hand sanitizer reduced infection risks by 78.5% under high contamination conditions, whereas the nonresidual hand sanitizer offered no reduction. Discussion Log10 virus and infection risk reductions were consistently greater for the residual hand sanitizer under all scenarios. Further data describing residual hand sanitizer efficacy with additional contamination or tactile events are needed. Conclusions Residual antinoroviral hand sanitizers may reduce infection risks for up to 4 hours.

Objective Research suggests human norovirus binding to histo-blood group antigen (HBGA)-like molecules on enteric bacteria may enhance viral pathogenesis; however, the properties of these bacterial ligands are not well known. Previous work identified, but did not characterize, seven norovirus-binding bacteria. To further examine this bacteria–virus binding interaction, enteric bacteria were analyzed via Western blot with anti-HBGA antibodies and lectins targeting HBGA-associated sugar components. Virus overlay assays using capsids from six different human norovirus strains further identified responsible ligands and strain dependent binding properties. Results Each bacterial species possessed varying degrees of HBGA-like activity, and lectin binding further elucidated potential sugar residues involved (N-acetyl-galactosamine, α-d-galactose or α-l-fucose). Both GI and GII norovirus capsids bound specific bacterial ligand sizes, and generally corresponded to anti-HBGA Western blot patterns. A 35-kDa band reacted with all HBGA antibodies, bound all six of the noroviruses tested, and had a high affinity for the lectins. Collectively, this work characterizes the varying carbohydrate residues potentially responsible for norovirus–bacteria interactions and provides a basis for future ligand identification.

Detection of enteric virus contamination of shellfish is limited by current methodology, which is time-consuming, tedious, and lacking in sensitivity due to reliance on cell culture infectivity. Alternative detection methods based on nucleic acid amplification have been hampered by high sample volumes and the presence of enzymatic inhibitors. The goal of this study was to develop methods to purify and concentrate intact virions from oyster extracts to a volume and quality compatible with viral genomic nucleic acid amplification by reverse transcriptase-polymerase chain reaction (RT-PCR). Fifty-gram oyster samples were homogenized and processed by standard adsorption-elution-precipitation methodology and then seeded with 10 5 PFU of poliovirus 1 (PV 1 ) or hepatitis A virus (HAV). Seeded viruses were concentrated by fluorocarbon extraction, polyethylene glycol (PEG) precipitation, chloroform extraction, and cetyltrimethyl ammonium bromide (CTAB) precipitation to a volume of 100 μl with removal of RT-PCR inhibitors. Virus recovery after elution of PEG precipitates was 50% for PV1 and 15 to 20% for HAV as evaluted by cell culture infectivity. The CTAB precipitation step yielded a concentrated sample which was directly compatible with RT-PCR reactions and capable of detecting about 100 placque=forming units (PFU) of PVI or HAV. When 50-g oyster extracts were seeded and processed by the entire concentration and purification scheme, direct RT-PCR detection of viral genomic RNA was possible at initial inoculum levels of 10 4 PFU of HAV and 10 3 PFU of PVI, with recoveries of 1 to 5% of seeded viruses.

Aims Human norovirus is a major public health burden and is resistant to numerous sanitizers and disinfectants. In this study, we tested the efficacy of an antimicrobial product containing a blend of silver ions and citric acid (silver dihydrogen citrate; SDC) against GI.6 and GII.4 HuNoV. Methods and Results Pure® hard surface disinfectant (Pure Bioscience, El Cajon, CA) was evaluated using ASTM International virucidal suspension and stainless steel carrier assays. The effect of SDC (or citrate alone) exposure on viral integrity was evaluated using RT-qPCR, transmission electron microscopy, sodium dodecyl sulphate-polyacrylamide gel electrophoresis/Western blot analysis and a receptor-binding assay. Suspension assays showed a 4·0 log10 reduction in RNA copy number within 5 min, while carrier assays showed a 2·0–3·0 log10 reduction in 30 min. Incorporating a simulated soil load into the sample matrix significantly reduced product efficacy. Treated particles displayed deformation and aggregation, a 50% reduction in viral capsid protein band intensity, and an 80% reduction in histo-blood group antigen receptor-binding ability. Conclusions Our results suggest that SDC acts exclusively on the viral capsid, and shows efficacy against HuNoV when used on precleaned surfaces. Significance and Impact of the Study This study sheds light on the mechanisms and efficacy of a novel antimicrobial against HuNoV. Our results suggest: (i) silver ions exclusively target the viral capsid, and not the RNA genome; (ii) citrate is not crucial for HuNoV capsid deformation.

According to the U.S. Food and Drug Administration 2013 Model Food Code, it is the duty of a food establishment to disclose and remind consumers of risk when ordering undercooked food such as ground beef. The purpose of this study was to explore actual risk communication behaviors of food establishment servers. Secret shoppers visited 265 restaurants in seven geographic locations across the United States, ordered medium rare burgers, and collected and coded risk information from chain and independent restaurant menus and from server responses. The majority of servers reported an unreliable method of doneness (77%) or other incorrect information (66%) related to burger doneness and safety. These results indicate major gaps in server knowledge and risk communication, and the current risk communication language in the Model Food Code does not sufficiently fill these gaps. The question is "should servers even be acting as risk communicators?" There are numerous challenges associated with this practice, including high turnover rates, limited education, and the high stress environment based on pleasing a customer. If servers are designated as risk communicators, food establishment staff should be adequately trained and provided with consumer advisory messages that are accurate, audience appropriate, and delivered in a professional manner so that customers can make informed food safety decisions.

The need for improved pathogen separation and concentration methods to reduce time-to-detection for foodborne pathogens is well recognized. Apolipoprotein H (ApoH) is an acute phase human plasma protein that has been previously shown to interact with viruses, lipopolysaccharides (LPS) and bacterial proteins. The purpose of this study was to determine if ApoH was capable of binding and efficiently capturing two representative human norovirus strains (GI.1 and GII.4), a cultivable surrogate, and four bacterial pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus). Experiments were carried out using an ApoH-conjugated magnetic bead-based capture followed by pathogen detection using nucleic acid amplification. For all three viruses studied, >10% capture efficiency (