Indirect contact with contaminated surfaces is a potential transmission route for COVID-19. Therefore, it is necessary to investigate convenient and inexpensive surface sanitization methods, such as HOCl, against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The SARS-CoV-2 surrogate, Phi6 (~ 7 log PFU/mL), was prepared in artificial saliva and tripartite matrices, spot inoculated on coupons of either stainless steel or vinyl, and allowed to dry. The coupons were sprayed with either 500 ppm or 1000 ppm HOCl, and remained on the surface for 0 s (control), 5 s, 30 s, or 60 s. Samples were enumerated via the double agar overlay assay. Statistical analysis was completed in R using a generalized linear model with Quasipoisson error approximations. Time, concentration, surface type, and inoculum matrix were all significant contributors to log reduction at P = 0.05. Significant three-way interactions were observed for 1000 ppm, vinyl, and 60 s ( P = 0.03) and 1000 ppm, tripartite, and 60 s ( P = 0.0121). A significant two-way interaction between vinyl and 60 s was also observed ( P = 0.0168). Overall, increased HOCl concentration and exposure time led to increased Phi6 reduction. Notably, the highest estimated mean log reduction was 3.31 (95% CI 3.14, 3.49) for stainless steel at 60 s and 1000 ppm HOCl in artificial saliva, indicating that this method of sanitization may not adequately reduce enveloped viruses to below infective thresholds.

Shiga toxin-producing Escherichia coli (STEC) bacteria continue to impact the food industry. Environmental sampling of potential sources of contamination is important to aid epidemiologic efforts in tracking foodborne illnesses throughout the United States. Here, the draft genome sequences of 110 STEC isolates from bovine manure collected in Florida and Texas are reported.

The present study evaluates the efficacy of a batch wash ozone sanitation system (BWOSS) and spray wash ozone sanitation system (SWOSS) against Listeria monocytogenes (two strains) and Salmonella enterica subsp. enterica (three serovars) inoculated on the surface of carrots, sweet potatoes, and butternut squash, commonly used in raw meat-based diets (RMBDs) marketed for companion animals such as dogs and cats. Produce either remained at room temperature for 2 h or were frozen at -20 degrees C and then tempered overnight at 4 degrees C to mimic the preprocessing steps of a raw pet food processing operation ('freeze-temper') prior to ozone treatment. Two ozone concentrations (0 and 5 ppm) were applied for either 20 s or 60 s for BWOSS and 20 s for SWOSS. Based on an ANOVA, BWOSS data showed no significant difference (P > 0.05) in microbial reduction between 0 and 5 ppm ozone concentration across all treatment durations for each produce type. BWOSS resulted in mean microbial reductions of up to 1.56 log CFU/mL depending on the treatment time and produce type. SWOSS data were analyzed using a generalized linear model with Quasipoisson errors. Freeze-tempered produce treated with SWOSS had a higher bacterial log reduction at 5 ppm ozone compared to 0 ppm ozone (P = 0.0013) whereas room temperature produce treated with SWOSS did not show any significant difference in microbial reduction between ozone concentrations. The potential to mitigate microbial cross-contamination was also investigated during SWOSS treatment. The results indicate that 5 ppm ozone decreased pathogens in the rinsate and proximal surfaces by 0.63-1.66 log CFU/mL greater than no ozone depending on the pathogen and sample. Overall, data from this study indicate that SWOSS would be more effective compared to BWOSS in reducing the microbial load present on the surface of root tubers and squash subjected to freezing and thawing and has the potential to mitigate cross-contamination within RMDB manufacturing environments.

Determining the prevalence and persistence of viruses outside the human host aids our ability to characterize exposure risk across multiple transmission pathways. Since 2020, the Coronavirus Disease 2019 pandemic has resulted in a surge of research regarding severe acute respiratory syndrome-coronavirus-type 2 (SARS-CoV-2) and its potential to spread via direct and indirect contact transmission routes. Here, the authors discuss the current state of the science concerning SARS-CoV-2 transmission via contaminated surfaces and its persistence on environmental surfaces. This review aims to provide the reader with an overview of the currently published SARS-CoV-2 persistence studies, factors impacting persistence, guidelines for performing persistence studies, limitation of current data, and future directions for assessing SARS-CoV-2 persistence on fomites.

AIMS:This study aimed to determine the extent of Phi6 (Φ6) transfer between skin and surfaces relevant to consumer-facing environments based on inoculum matrix, surface type and contact time.METHODS AND RESULTS:Φ6 transfer rates were determined from skin-to-fomite and fomite-to-skin influenced by inoculum matrix (artificial saliva and tripartite), surface type (aluminium, plastic, stainless steel, touchscreen, vinyl and wood) and contact time (5 and 10 s). Significant differences in estimated means were observed based on surface type (both transfer directions), inoculum matrix (skin-to-fomite) and contact time (both transfer directions). During a sequential transfer experiment from fomite-to-skin, the maximum number of consecutive transfer events observed was 3.33 ± 1.19, 2.33 ± 1.20 and 1.67 ± 1.21 for plastic, touchscreen and vinyl, respectively.CONCLUSIONS:Contact time significantly impacted Φ6 transfer rates, which may be attributed to skin absorption dynamics. Surface type should be considered for assessing Φ6 transfer rates.SIGNIFICANCE AND IMPACT OF THE STUDY:Although the persistence of Φ6 on fomites has been characterized, limited data are available regarding the transfer of Φ6 among skin and fomites. Determining Φ6 transfer rates for surfaces in consumer-facing environments based on these factors is needed to better inform future virus transmission mitigation strategies.

The process of washing tomatoes in dump (flume) tanks has been identified as a potential source of cross-contamination. This study's objective was to assess the potential for Salmonella enterica cross-contamination at various inoculation levels in the presence of free chlorine (HOCl) and organic matter. Uninoculated tomatoes were introduced into a laboratory-based model flume containing tomatoes inoculated with a cocktail of five rifampin-resistant S. enterica serovars at 104, 106, or 108 CFU per tomato in water containing 0 or 25 mg/L HOCl and 0 or 300 mg/L chemical oxygen demand (COD). Uninoculated tomatoes exposed to the inoculated tomatoes were removed from the water after 5, 30, 60, and 120 s and placed in bags containing tryptic soy broth supplemented with rifampin and 0.1% sodium thiosulfate. Following incubation, enrichment cultures were plated on tryptic soy agar supplemented with rifampin and xylose lysine deoxycholate agar to determine the presence of Salmonella. HOCl and pH were measured before and after each trial. The HOCl in water containing 300 mg/L COD significantly declined (P < 0.05) by the end of each 120-s trial, most likely due to the increased demand for the oxidant. Higher inoculum levels and lower HOCl concentrations were significant factors (P < 0.05) that contributed to increased cross-contamination. At 25 mg/L HOCl, no Salmonella was recovered under all conditions from uninoculated tomatoes exposed to tomatoes inoculated at 104 CFU per tomato. When the inoculum was increased to 106 and 108 CFU per tomato, cross-contamination was observed, independent of COD levels. The results from this study indicate that the currently required sanitizer concentration (e.g., 100 or 150 mg/L) for flume water may be higher than necessary and warrants reevaluation.

Phi 6 (Φ6) bacteriophage is a proposed surrogate to study pathogenic enveloped viruses including SARS-CoV-2-the causative agent of COVID-19-based on structural similarities, BSL-1 status, and ease of use. To determine the role of virus-contaminated hands in disease transmission, an enhanced understanding of buffer and method performance for Φ6 recovery needs to be determined. Four buffer types and three methodologies were investigated for the recovery of Φ6 from human fingerpads over a 30 min duration. Phosphate buffered saline (PBS), PBS + 0.1 % Tween, 0.1 M glycine + 3% beef extract, and viral transport medium were evaluated as buffers for recovery of Φ6 via a dish, modified glove juice, and vigorous swabbing method. Φ6 concentrations on fingerpads were determined at 0-, 5-, 10-, and 30-min post-inoculation. While there were observed differences in virus recovery across buffer and method types depending on the time point, log PFU recovery based on buffer type or methodology was not significantly different at any time point (P > 0.05). The results presented in this study will allow for future work on Φ6 persistence, transfer between hands and surfaces, and efficacy of hand hygiene methods to be performed using a well-characterized and validated recovery method.

An important consideration in virus contact transmission is the transfer rate between hands and surfaces, which is driven by several factors, including virus persistence on inanimate surfaces. This research characterized Φ6 persistence on surfaces commonly encountered in public settings based on various factors.

Microgreens are an emerging salad crop with properties similar to those of sprouted seeds and lettuce. This study aimed to determine bacterial pathogen persistence during microgreen cultivation and transfer from soil-free cultivation matrix (SFCM) to mature microgreens. Salmonella enterica subsp. enterica ser. Javiana and Listeria monocytogenes were inoculated onto biostrate mats as well as peat SFCM and sampled (day 0). Next, sunflower and pea shoot seeds were planted (day 0) and grown in a controlled environment until the microgreen harvest (day 10). On day 10, SFCM and microgreens were sampled to determine the pathogen levels in the SFCM and the pathogen transfer to microgreens during production. Salmonella Javiana log CFU/g were significantly higher than L. monocytogenes in SFCM on day 10 in both planted and unplanted regions (p < 0.05). Significant differences in pathogen transfer (log CFU/g) were observed between the pea shoot and sunflower microgreens, regardless of the pathogen or SFCM type (p < 0.05). Meanwhile, pathogen transfer to the pea shoot and sunflower microgreens from the biostrate was 1.53 (95% CI: −0.75–3.81) and 5.29 (95% CI: 3.01–7.57) mean log CFU/g, respectively, and transfer from the peat was 0.00 (95% CI: −2.28–2.28) and 2.64 (95% CI: 0.36–4.92) mean log CFU/g, respectively. Results demonstrate that pathogen transfer to microgreens during production is influenced by SFCM and microgreen variety.

This research investigated the influence of soil microbiota on Escherichia coli O157:H7 survival in soil rinse and artificial soil. Additionally, the influence of selected soil bacteria on E. coli O157:H7 in soil environments was determined.

Monitoring and maintenance of water quality in dump tanks or flume systems is crucial to maintaining proper sanitizer levels to prevent pathogen cross-contamination during postharvest washing of tomatoes, but there is limited information on how organic matter influences sanitizer efficacy in the water. The main objective of this study was to monitor water quality in flume tanks and evaluate the efficacy of postharvest washing of tomatoes in commercial packinghouses. Flume tank water samples (n = 3) were collected on an hourly basis from three packinghouses in Florida and analyzed for pH, total dissolved solids (TDS), free chlorine, chemical oxygen demand (COD), oxidation-reduction potential, and turbidity. Additionally, three flume-water samples were collected and tested for total aerobic plate count (APC), total coliforms (TC), and Escherichia coli. Fresh tomatoes (n = 3), both before and after washing, were collected and analyzed for the same bacterial counts. Turbidity, COD, and TDS levels in flume water increased over time in all packinghouses. Correlations observed include COD and turbidity (r = 0.631), turbidity and TDS (r = 0.810), and oxidation-reduction potential and chlorine (r = 0.660). APC for water samples had an average range of 0.0 to 4.7 log CFU/mL and TC average range of 0.0 to 4.7 log CFU/mL. All water samples were negative for E. coli. The average APC for pre- and postflume tomatoes from the three packinghouses was 6.0 log CFU per tomato and ranged from 2.2 to 7.4 log CFU per tomato. The average TC count was <1.5 and 7.0 log CFU per tomato for pre- and postwash tomatoes, respectively. There was no significant effect (P > 0.05) of postharvest washing on the microbiological qualities of tomatoes. Water quality in flume tanks deteriorated over time in all packinghouses during a typical operational day of 4 to 8 h.

Aims This research was performed to investigate the influence of clay and humic acid onEscherichia coliO157 survival in model soils. Additionally, the influence of pH and humic acid onE. coliO157 in liquid culture was investigated. Methods and Results Artificial soil microcosms were prepared with sand, kaolinite, bentonite and humic acid. Artificial soil microcosms pH was adjusted (6 center dot 0-7 center dot 0) with aluminium sulphate beforeE. coliO157 inoculation. After 56 days of incubation at 30 degrees C, significant differences inE. coliO157 log CFU per gram were observed between 0 and 1000 ppm (P < 0 center dot 0001) and 0 and 5000 ppm (P < 0 center dot 0001) humic acid in 1 center dot 5% clay soils, but not in 7 center dot 5 or 15% clay soils. Significant differences (P < 0 center dot 05) inE. coliO157 log CFU per ml were observed in liquid culture influenced by humic acid concentrations after 8 h at 37 degrees C. Conclusions The developed model soils supportE. coliO157 populations over 28 days, and higher clay soils may aid inE. coliO157 survival. Significance and Impact of the Study These results provide insights into physicochemical properties of soil that may influenceE. coliO157 in the environment and help explainE. coliO157 survival in various soils and geographical regions.

While the soil microbiome may influence pathogen survival, determining the major contributors that reduce pathogen survival is inconclusive. This research was performed to determine the survival of E. coli O157 in autoclaved and natural (unautoclaved) sandy soils. Soils were inoculated with three different E. coli O157 strains (stx1+/stx2+, stx1-/stx2-, and stx1-/stx2+), and enumerated until extinction at 30°C. There was a significant difference in the survival of E. coli O157 based on soil treatment (autoclaved versus natural) at 30°C on days 1 (P = 0.00022), 3, (P = 2.53e-14), 7 (P = 5.59e-16), 14 (P = 1.072e-12), 30 (P = 7.18e-9), and 56 (P = 0.00029), with greater survival in autoclaved soils. The time to extinction (two consecutive negative enrichments) for all three strains was 169 and 84 days for autoclaved and natural soils, respectively. A separate E. coli O157 trial supplemented with 16S rRNA gene sequencing of the soil microbiome was performed at 15°C and 30°C on days 0, 7, 14, and 28 for each soil treatment. Greater species richness (Chao1, P = 2.2e-16) and diversity (Shannon, P = 2.2e-16) was observed in natural soils in comparison with autoclaved soils. Weighted UniFrac (beta-diversity) showed a clear distinction between soil treatments (P = 0.001). The greatest reduction of E. coli O157 was observed in natural soils at 30°C, and several bacterial taxa positively correlated (relative abundance) with time (day 0 to 28) in these soils (P < 0.05), suggesting that the presence of those bacteria might cause the reduction of E. coli O157. Taken together, a clear distinction in E. coli O157 survival, was observed between autoclaved and natural soils along with corresponding differences in microbial diversity in soil treatments. This research provides further insights into the bacterial taxa that may influence E. coli O157 in soils.

Fresh produce outbreaks due to Shiga toxin-producing Escherichia coli (STEC) continue to occur in the United States (US). Manure-amended soils can pose a public health risk when used for growing raw agricultural commodities. Knowing the prevalence and concentration of STEC in untreated biological soil amendments of animal origin (BSAAO) is important to help guide the most appropriate pre-harvest interval(s) following application to limit risks from these soil amendments. Bovine manure samples were collected from 12 farms in Florida, including samples from piles, lagoons, barns, and screened solids. Two methods were used to detect stx1/2 and rfbE genes in samples. A prevalence rate of 9% for stx1 and/or stx2 and 19% for rfbE was observed from the 518 bovine manure samples evaluated. A most probable number (MPN) assay was performed on stx+ samples when applicable. The geometric mean for stx+ samples (n = 20) was 3.37 MPN g-1 (0.53 log MPN g-1) with a maximum value of 6,800 MPN g-1 (3.83 log MPN g-1). This research was part of a larger nationwide geographical study on the prevalence and concentration of STEC in bovine manure to help guide regulations on feasible pre-harvest intervals for the application of untreated BSAAO.

Microbial contamination of fresh produce (fresh fruits and vegetables) poses serious public health concerns worldwide. This study was conducted as a comprehensive analysis of biological hazards in the global fresh produce chain. Data about produce-related outbreaks and illness were collected from the annual reports and databases of foodborne outbreak surveillance systems in different regions and countries from 2010 to 2015. The global patterns of and regional differences in documented outbreaks and cases were analyzed, and produce commodities and pathogens of greatest concern were identified. Data on sporadic illnesses were also collected through a comprehensive literature review of case-control studies. We found 988 produce-related outbreaks (with known agents) and 45,723 cases in all regions and countries. The numbers of produce-related outbreaks per million person-years were approximately 0.76, 0.26, 0.25, 0.13, 0.12, and 0.05 in New Zealand, Australia, the United States, the European Union, Canada, and Japan, respectively. The top three food categories and pathogens contributing to produce-related outbreaks were vegetables and nonfruits (i.e., food other than fruits; 27.0%), unspecified vegetables (12.2%), and vegetable row crops (11.7%) and norovirus (42.4%), Salmonella enterica (19.9%), and Staphylococcus aureus (7.9%), respectively. Produce consumption was identified as a protective factor, a risk factor, and either a protective or risk factor for sporadic illnesses in 11, 5, and 5 studies, respectively, among 21 case-control studies. Risks associated with produce consumption in the United States and the European Union have been linked to various factors such as irrigation water, cross-contamination, storage time and temperature abuse, infected food handlers, and unprocessed contaminated ingredients. The results of the current study indicate the complexity of produce products consumed across the globe and the difficulty in tracing illnesses back to specific food ingredients.

Contamination of food products by pathogenic microorganisms continues to be a major public health and food industry concern. Non-typhoidal Salmonella species have led to numerous outbreaks associated with various foods. A wide variety of methods have been applied and introduced for treatment of fresh foods to eliminate pathogenic as well as spoilage microorganisms. Salmonella can become exposed to elevated temperatures while associated with hosts such as poultry. In addition, heat treatment is also applied at various stages of processing to retain the shelf life of food products. Despite this, these microorganisms may overcome exposure to such treatments through the efficient expression of stress response mechanisms and result in illness following consumption. Thermal stress induces a range of destructive exposures to bacterial cells such as protein damage and DNA damage caused by reactive oxygen species. In this study, we chose three genes (∆recD, ∆STM14_5307, and ∆aroD) associated with conditionally essential genes required for different aspects of optimal growth at 42 °C and evaluated the responses of wild type and mutant Salmonella Typhimurium strains to uncover potential mechanisms that may enable survival and resistance under thermal stress. The RecBCD complex that initiates repair of double-stranded DNA breaks through homologous recombination. STM14_5307 is a transcriptional regulator involved in stationary phase growth and inositol metabolism. The gene aroD is involved in metabolism and stationary phase growth. These strains were characterized via high throughput phenotypic profiling in response to two different growth temperatures (37 °C (human host temperature) and 42 °C (poultry host temperature)). The ∆aroD strain exhibited the highest sensitivity to the various temperatures followed by the ∆recD and ∆STM14_5307 strains, respectively. Achieving more understanding of the molecular mechanisms of heat survival may lead to the development of more effective strategies to limit Salmonella in food products through thermal treatment by developing interventions that specifically target the pathways these genes are involved in.

Commensal Escherichia coll are commonly utilized for investigating the genetic and biochemical requirements of microorganisms, and have served in a wide variety of applications. Pathogenic E. coli known as Shiga toxin (Stx)-producing E. coli (STEC) are associated with various food products including ground beef. These pathogens are present in a wide range of environments, and have caused numerous foodborne outbreaks and recalls. These outbreaks and the increased awareness of STEC have led to certain STEC serotypes to be declared adulterants in non-intact raw meat. Various STEC detection methods have been investigated, and numerous cultural and molecular-based detection methods continue to be modified to meet regulatory requirements. However, STEC serotypes may possess certain characteristics that lead to bias in the likelihood of a certain serotype being detected in an assay. Understanding the characteristics of these STEC serotypes will provide means for optimizing the detection platforms, and as a result limit foodborne illness and recalls caused by STEC due to enhanced cultural and molecular detection capabilities. (C) 2015 Elsevier Ltd. All rights reserved.

The purpose of this study was to assess the microbial community in ground beef purchased from local stores in the north central area of South Korea. A total of 31 ground beef samples showed 3.77 +/- 5.16 x 10(5) CFU/g on aerobic plate counts, and PCR-based denaturing gradient gel electrophoresis was performed to assess the microbial community, which revealed a diverse profile among the products. Four of these samples were selected for further analyses via 454 pyrosequencing. Pseudomonas was the most frequently identified genus in terms of relative abundance (4.54-73.8%), and other bacteria such as Lactobacillus, Acinetobacter, and Carnobacterium were also identified in the ground beef. The presence of bacteria can affect the spoilage of ground beef products depending on the microbial populations, and understanding these populations may help limit spoilage and/or foodborne illness. This study illustrates the microbial diversity occurring among Korean ground beef products, even within a local region, and provides a molecular approach for potential ground beef microbial quality assessment.

The purpose of this study was to characterize the microbial community in ground pork using molecular approaches. Forty six ground pork products were purchased from local stores in the north central area of South Korea. Aerobic plate counts varied 4.23 ± 5.14 × 10(5) CFU/g with the range between 5.00 × 10(3) and 1.85 × 10(6) CFU/g for ground pork samples. Four ground meat samples were further processed for metagenomic analysis. Pseudomonas species was the most relative abundant with a wide range occurring (1.72 to 77.7%) as part of the microbial genera in ground pork. Bacteria such as Carnobacterium, Yersinia, Photobacterium were also identified in ground pork. Despite the prominence of certain genera across all samples there was still extensive microbial diversity among ground pork products that originated from different slaughter houses and were processed in different markets. Such diversity indicates that designing interventions to extend shelf life may be hampered by the extensive variability in the microbial consortia associated with pork products. However, this diversity may be useful for developing microbial traceability signatures unique to a slaughter house or a particular market.