Consumers' demand for fresh produce is rising due to dietary preferences and public health campaigns. Fresh produce is often consumed raw or minimally processed to retain nutrient content. If contaminated, fresh produce can become a vehicle for pathogen transmission and potentially cause outbreaks. Contamination may occur within the farm border, but some evidence suggests different sources on adjacent and nearby land may be off-farm contributors. Natural ecosystems (e.g., wildlife habitats, vegetation), agricultural activities (e.g., livestock operations, manure and compost application, wastewater discharge), residential and industrial activities (e.g., septic systems, waste discharge), and recreational uses (e.g., parks, campgrounds, golf courses) all represent potential contamination sources. Environmental pathogens from adjacent land can reach produce farms through diverse pathways influenced by meteorological drivers (e.g., rainfall, wind) or by human activities, equipment, and animal intrusion. These adjacent land inputs interact with each other and farming practices, creating complex contamination mechanisms. This paper explored microbial contamination sources from adjacent and nearby land and proposed a risk ranking model with implications for produce safety. It aimed to provide an overall understanding of the interaction between environmental and adjacent land sources as a basis for future risk assessment and management research.

Contamination of food contact surfaces by airborne transmission of pathogens from the environment has contributed to disease outbreaks. Therefore, this study evaluated the survival and recovery of aerosolized generic Escherichia coli and Enterococcus faecium from four food contact surfaces (food-grade silicone rubber, high-density polyethylene [HDPE] plastic, stainless steel, and waxed cardboard), after four contact times (10, 20, 40, and 60 min), two relative humidity (RH) levels (high: 80%-90%, low: 40%-50%), three distances from aerosolization source (0, 36.5, and 73 cm; E. coli only), and with and without airflow (E. coli only). ANOVA test with Tukey's HSD at alpha = 0.05 was used to determine how treatment combinations influenced recovery. At high humidity, E. coli recovery on all materials after 40 min was similar to 1.0 log lower than recovery after 10 min, and further reduced by 1.0 log at 60 min. At lower humidity, E. coli recovery on all materials was similar to 1.0 log lower at 10 and 20 min compared with high humidity. Distances exerted no significance, whereas airflow presence lowered E. coli recovery. E. coli survival on all materials declined from similar to 5.0 log CFU/coupon at 0 h to 3.5 log CFU/coupon at 6 h, and 2.0 log CFU/coupon at 24 h post-inoculation. E. coli recovery was significantly lower (p < 0.05) on waxed cardboard. Low RH and longer contact time reduced E. coli recovery but not E. faecium. E. faecium recovery was consistent across treatment combinations, with changes < 0.5 log CFU/coupon. The findings are relevant for the survival of bacteria on common food contact surfaces and the potential of transmission to food products.

Listeria monocytogenes is a foodborne pathogen with the ability to persist and form biofilm matrices in processing environments of food manufacturing facilities. Bacteriophages are bacterial viruses with host specific lethality. Published research on the application of phage to control Listeria sp. in manufacturing environments is limited. In this study, we have assessed the capacity of bacteriophage P100 (Listex (TM)) to reduce incidence of Listeria sp. in the ready-to-eat (RTE) environment of refrigerated (4 degrees C) and ambient (20 degrees C) temperature facilities using two different application strategies. A moderate application applied as a single treatment every 24 hr over three days (2 x 10(7) PFU/ml) and an intensified application applied once every 6 hr over a 24 hr period (1 x 10(8) PFU/ml). Environmental nonfood contact surface (NFCS) samples were collected and analyzed for the presence of Listeria sp. before and after treatment. When the moderate treatment protocol was applied the incidence of positives decreased from 51.3 to 17.5% in the 4 degrees C environment and from 67.5 to 23.1% in the 20 degrees C production area. For the intensified phage treatment method, the initial positive rate in the 4 degrees C environment ranged from 5 to 47.5%, with an overall 43% reduction in Listeria sp. In the 20 degrees C facility, initial environmental Listeria sp. ranged from 15 to 50%, with an overall reduction of 32% after treatment with phage P100. Data indicate the application of Listeria specific phage P100 in RTE food production environments by either the moderate or intensified application method can reduce incidence and be considered an additional intervention strategy for controlling this pathogen on NFCS.

EDITORIAL article Front. Public Health, 09 June 2023Sec. Infectious Diseases: Epidemiology and Prevention Volume 11 - 2023 | https://doi.org/10.3389/fpubh.2023.1221763

Campylobacter and pathogenic Escherichia coliillnesses have been attributed to the consumption of fresh produce. The leafy green, kale, is increasingly consumed raw. In comparison to other leafy greens, kale has a longer shelf-life. Due to the extended shelf-life of kale, it is warranted to examine the survival of pathogenic Campylobacter jejuni and E. coli O157:H7 inoculated on the surface of kale stored in a controlled environment at 4 ± 1.4°C, and average humidity of 95 ± 1.9% over a 23-day period. At predetermined time points (days 0, 1, 2, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21), inoculated kale was destructively sampled and the surviving bacteria determined by serial dilution and plating onto Tryptic soy agar, Charcoal cefoperozone deoxycholate agar, and Eosin methylene blue for total aerobic bacteria, C. jejuni, and E. coli O157:H7, respectively. Enrichment and PCR were used for detection when pathogens were not detected using serial dilution and plating. Aerobic heterotrophic bacteria increased over the 23-day period, in contrast, significant declines in the inoculated pathogens were observed. Inoculated E. coli O157:H7 survived longer on kale (up to 19 d); in comparison, C. jejuni was undetectable by day 13 using enrichment and PCR or plating. In conclusion, C. jejuni and E. coli O157:H7 declined on fresh kale over time when held at refrigerated temperatures but were still detected during the majority of the time when the kale would likely still be considered edible by consumers.

Aims This study investigated Salmonella concentrations following combinations of horticultural practices including anaerobic soil disinfestation (ASD), soil amendment type and irrigation regimen. Methods and Results Sandy-loam soil was inoculated with a five-serovar Salmonella cocktail (5.5 +/- 0.2 log CFU per gram) and subjected to one of six treatments: (i) no soil amendment, ASD (ASD control), (ii) no soil amendment, no-ASD (non-ASD control) and (iii-vi) soil amended with pelletized poultry litter, rye, rapeseed or hairy vetch with ASD. The effect of irrigation regimen was determined by collecting samples 3 and 7 days after irrigation. Twenty-five-gram soil samples were collected pre-ASD, post-soil saturation (i.e. ASD-process), and at 14 time-points post-ASD, and Salmonella levels enumerated. Log-linear models examined the effect of amendment type and irrigation regimen on Salmonella die-off during and post-ASD. During ASD, Salmonella concentrations significantly decreased in all treatments (range: -0.2 to -2.7 log CFU per gram), albeit the smallest decrease (-0.2 log CFU per gram observed in the pelletized poultry litter) was of negligible magnitude. Salmonella die-off rates varied by amendment with an average post-ASD rate of -0.05 log CFU per gram day (CI = -0.05, -0.04). Salmonella concentrations remained highest over the 42 days post-ASD in pelletized poultry litter, followed by rapeseed, and hairy vetch treatments. Findings suggested ASD was not able to eliminate Salmonella in soil, and certain soil amendments facilitated enhanced Salmonella survival. Salmonella serovar distribution differed by treatment with pelletized poultry litter supporting S. Newport survival, compared with other serovars. Irrigation appeared to assist Salmonella survival with concentrations being 0.14 log CFU per gram (CI = 0.05, 0.23) greater 3 days, compared with 7 days post-irrigation. Conclusions ASD does not eliminate Salmonella in soil, and may in fact, depending on the soil amendment used, facilitate Salmonella survival. Significance and Impact of the Study Synergistic and antagonistic effects on food safety hazards of implementing horticultural practices should be considered.

Salmonella enterica contamination of low water activity foods (LWAFs) has resulted in recalls of spices, herbs, and seeds and outbreaks of salmonellosis. To improve the safety of these ready-to-eat products, new treatment methods, including fumigation with chlorine dioxide (ClO2) or hydrogen peroxide (H2O2) gas are being explored, and effectiveness determined. To prevent overestimation of treatment effectiveness, it is vital that recovery methods should accurately quantify all viable cells, even those injured. This study evaluated different media and supplements for the recovery of multiple strains of S. enterica and Enterococcus faecium NRRL B2354, from ClO2 or H2O2 treated black peppercorns, dried basil leaves, and chia seeds. Also, this study aimed to compare the log reduction of these two microorganisms to evaluate E. faecium NRRL B2354, as a surrogate for S. enterica. On average, recovery of S. enterica was improved by 1 log CFU from ClO2 and H2O2 treated LWAFs when a non-selective but differential media containing tryptic soy agar with yeast extract, ammonium iron citrate and sodium thiosulfate (MTSAYE) was used, when compared to plating on XLD (p < 0.05). Furthermore, addition of sodium pyruvate, ferrous sulfate, or 3’3’-thiodiproionate supplements to MTSAYE did not show increased recovery of either S. enterica or E. faecium NRRL B2354 (p > 0.05). On each treatment and LWAF combination tested, there was no significant difference between the log reduction of S. enterica and E. faecium NRRL B2354, indicating its suitability as a surrogate under the test conditions.

Fresh produce may be contaminated by bacterial pathogens, including Listeria monocytogenes, during harvesting, packaging, or transporting. A low-intensity cavitation process with air being injected into water was studied to determine the microbubbles’ efficiency when detaching L. monocytogenes from stainless steel and the surface of fresh cucumber and avocado. Stainless steel coupons (1″ × 2″), cucumber, and avocado surfaces were inoculated with L. monocytogenes (LCDC strain). After 1, 24 or 48 h, loosely attached cells were washed off, and inoculated areas were targeted by microbubbles (~0.1–0.5 mm dia.) through a bubble diffuser (1.0 L air/min) for 1, 2, 5, or 10 min. For steel, L. monocytogenes (48 h drying) detachment peaked at 2.95 mean log reduction after 10 min of microbubbles when compared to a no-bubble treatment. After 48 h pathogen drying, cucumbers treated for 10 min showed a 1.78 mean log reduction of L. monocytogenes. For avocados, L. monocytogenes (24 h drying) detachment peaked at 1.65 log reduction after 10 min of microbubbles. Microbubble applications may be an effective, economical, and environmentally friendly way to remove L. monocytogenes, and possibly other bacterial pathogens, from food contact surfaces and the surfaces of whole, intact fresh produce.

Listeria monocytogenes is a Gram-positive foodborne pathogen responsible for the severe disease listeriosis and notorious for its ability to persist in food processing plants, leading to contamination of processed, ready-to-eat foods. L. monocytogenes persistence in various food processing environments (FPEs) has been extensively investigated by various subtyping tools, with increasing use of whole genome sequencing (WGS). However, major knowledge gaps remain. There is a need for facility-specific molecular signatures not only for adequate attribution of L. monocytogenes to a specific FPE but also for improved understanding of the ecology and evolution of L. monocytogenes in the food processing ecosystem. Furthermore, multiple strains can be recovered from a single FPE sample, but their diversity can be underestimated with common molecular subtyping tools. In this study we investigated a panel of 54 L. monocytogenes strains from four turkey processing plants in the United States. A combination of WGS and phenotypic assays was employed to assess strain persistence as well as identify facility-specific molecular signatures. Comparative analysis of allelic variation across the whole genome revealed that allelic profiles have the potential to be specific to individual processing plants. Certain allelic profiles remained associated with individual plants even when closely-related strains from other sources were included in the analysis. Furthermore, for certain sequence types (STs) based on the seven-locus multilocus sequence typing scheme, presence and location of premature stop codons in inlA, inlB length, prophage sequences, and the sequence content of a genomic hotspot could serve as plant-specific signatures. Interestingly, the analysis of different isolates from the same environmental sample revealed major differences not only in serotype and ST, but even in the sequence content of strains of the same ST. This study highlights the potential for WGS data to be deployed for identification of facility-specific signatures, thus facilitating the tracking of strain movement through the food chain. Furthermore, deployment of WGS for intra-sample strain analysis allows for a more complete environmental surveillance of L. monocytogenes in food processing facilities, reducing the risk of failing to detect strains that may be clinically relevant and potentially novel.

Study objective was to determine efficacy of a bacteriophage suspension against Listeria spp. when applied to three common types of materials used in food manufacturing facilities. Materials included two food contact materials (stainless steel and polyurethane thermoplastic belting) and one noncontact material (epoxy flooring). Coupons of each material were inoculated with a cocktail containing L. monocytogenes and L. innocua (4 to 5-log(10) CFU/cm(2)). Two phage concentrations and a control, 0, 2 x 10(7) and 1 x 10(8) PFU/cm(2) were evaluated. Treated samples were held at 4 or 20 degrees C for 1 and 3 hr to determine the effect of temperature and treatment time. Reductions in Listeria populations ranged from 1.27 to 3.33 log(10) CFU/cm(2) on stainless steel, from 1.17 to 2.76 log(10) CFU/cm(2) on polyurethane thermoplastic belting, and from 1.19 to 1.76 log(10) CFU/cm(2) on epoxy resin flooring. Higher phage concentration (1 x 10(8) PFU/cm(2)), longer treatment time (3 hr), and processing area temperature of 20 degrees C showed a greater (p <= .05) reduction of Listeria on the stainless-steel and polyurethane thermoplastic belting coupons. Overall, Listeria reduction by phage treatment occurred on all three materials tested, under all conditions.

Effective disinfection in healthcare facilities prevents healthcare-associated infections. This study evaluated the ability of Environmental Protection Agency-approved disinfectants (quaternary ammonium compound, QAC; sodium hypochlorite, and hydrogen peroxide) applied with 3 wiping substrates (microfiber, nonwoven, and cotton) to remove Staphylococcus aureus from Formica surfaces. All treatments reduced S aureus on Formica squares with the exception of QAC applied with cotton and QAC, nondisinfectant, and control applied with a nonwoven cloth. Sodium hypochlorite or hydrogen peroxide applied with cotton or microfiber, respectively, may be the best choice for disinfection of Formica surfaces in healthcare settings.

The use of prewetted disinfectant towelettes in health care settings proves challenging because they may dry quickly, reducing disinfection. This study examined the drying time of various commercial disinfectant towelettes and the efficacy of these towelettes over time to eliminate Staphylococcus aureus from glass surfaces. This study confirms that these disinfectants dry quickly. Further disinfection after drying time on glass is minimal, but dependent on the type of disinfectant. (c) 2019 Published by Elsevier Inc. on behalf of Association for Professionals in Infection Control and Epidemiology, Inc.

Dynamics of a bubble impacting and sliding a tilted surface has been investigated through experimental and computational methods. \textcolor{blue}{Specifically, shear stress generated on the wall has been calculated and compared with bacterium adhesion force in order to evaluate a potential sanitization function. In experiments, the bubble-wall interaction has been characterized for several different wall angles. We numerically solved a force balance including buoyancy, hydrodynamic inertia \& drag, lift and thin film force to determine the bubble motion. Results showed that the shear stress increases with the wall inclination. The maximum shear stress goes up to more than 300 Pa as a single bubble impacts and scrubs a tilted wall. We found that such a high shear stress is attributed to a rapid change in thin film curvature (flipping bubble/water interface) during the bouncing stage. Later, during the sliding stage, a smaller shear stress up to around 45 Pa is generated for a longer period of time. We also showed that the shear stress generated during the bouncing and sliding stages is high enough to remove bacteria from a surface as a potential method for removing bacteria from tilted surfaces.

Salmonella serotypes linked to tomato-associated outbreaks were evaluated for survival in soil and water over a 40-day period. Salmonella enterica serotypes Anatum, Baildon, Braenderup, Montevideo, Newport, and Javiana were inoculated separately into sterile soil and water, followed by plating onto TSAYE and XLT4 at 10-day intervals. Biofilm production by Salmonella serotypes was measured on both quartz particles (soil surrogate) and glass coverslips, and was evaluated using a crystal violet dye assay. Salmonella populations in soil and water over 40 days indicated no significant differences between Salmonella serotypes tested (p > 0.05). Over a 40-day period, there was a 1.84 ± 0.22 log CFU/g and 1.56 ± 0.54 CFU/mL decrease in populations of Salmonella in soil and water, respectively. Enumeration indicated that Salmonella population fluctuated in water but decreased linearly in soil. All serotypes tested produced the "red dry and rough" morphotype on Congo Red agar. Biofilm produced by all the Salmonella serotypes tested was significantly different on quartz particles than on glass coverslips (p < 0.0001), indicating that material and surface characteristics could affect biofilm development. The ability of Salmonella serotypes to persist in soil or water and attach to abiotic surfaces through biofilm formation affirms that contact surfaces, soil, water, and sediment should be considered as possible sources of cross-contamination in the farm environment.

Foodborne illnesses have been a heavy burden in the United States and globally. Many medicinal herbs have been cultivated in the US and many of which contain antimicrobial compounds with the potential to be used for food preservation. Methanol/water extracts of pomegranate peel (“PP”, Punica Granatum L.), Chinese gallnut (“CG”, Galla chinensis), Forsythia fruit (“FF”, Forsythia suspensa) and Baikal skullcap root (“BS”, Scutellaria baicalensis) were tested for antimicrobial activity using the agar diffusion assay on tryptic soy agar (TSA) and microdilution assay in tryptic soy broth (TSB). CG and PP extracts showed good to excellent inhibitory effect against Vibrio parahaemolyticus and Listeria monocytogenes in both assays, with a minimum inhibitory concentration (MIC) range from 0.04 to 5 mg/mL. BS had moderate inhibitory effects against V. parahaemolyticus with an MIC of 5 mg/mL in TSB, and against L. monocytogenes with an MIC of 20 mg/mL on TSA. CG was analyzed using LC-MS and fractionated using HPLC. The major components were identified as gallic acid, digallic acid, methyl gallate, and gallotannins (oligo-galloyl-D-glucose, nGG, n = 1~10). Six fractions (I - VI) were collected and their antibacterial activities were tested against L. monocytogenes, and V. parahaemolyticus both on TSA and in TSB. On TSA, fraction III, IV and V inhibited V. parahaemolyticus but no fraction inhibited L. monocytogenes. In TSB, all fractions inhibited V. parahaemolyticus and fractions II - V inhibited L. monocytogenes. Future studies are needed to investigate the effects of medicinal plants on food products.

Raw produce has frequently been identified as the source of bacterial pathogens that can cause human illnesses, including listeriosis and salmonellosis. Microbial pathogens may attach and form biofilms on raw fruit surfaces and can be difficult to remove. A cavitation process (injection of bubbles into water) was studied for its effectiveness for removal and inactivation of Listeria monocytogenes and Salmonella Newport from the surfaces of fresh Roma tomatoes and cantaloupes. Individual fruit were separately inoculated with each pathogen, then submerged in a water tank and treated with a bubble flow through an air stone using one airflow rate (0 – 14 liters/min.) for up to 60 sec. As airflow increased, pathogen reduction increased up to 1.2 log CFU/fruit greater than with water alone (no bubbles). Additional pathogen reduction in the tank water (organisms detached from the fruit) was observed with the bubble treatments. Therefore, these bubble streams can be used to enhance the detachment of bacteria from fruit surfaces and to inactivate a proportion of these detached microorganisms. Additionally, recoveries of Salmonella from inoculated Roma tomatoes and cantaloupe were determined for treatment water that contained 50 or 150 ppm sodium hypochlorite. Combining both cavitating bubbles and chlorine in the tank water resulted in greater efficacy of removing or inactivating S. Newport from the surface of Roma tomatoes and cantaloupe than with cavitation or chlorine alone. The physical force of a bubble stream on raw produce can effectively detach and inactivate surface bacteria, and has the potential to reduce antimicrobial chemical use and water use in post-harvest packing operations.

Cetylpyridinium chloride (CPC) solutions (0, 0.5, or 1.0%) were applied to cantaloupe ("Athena" and "Hale's Best Jumbo" cultivars) rind plugs, either before or after inoculation with a broth culture of Salmonella Michigan (109 CFU/mL) and held at 37°C for 1 or 24 hr. Rind plugs were diluted, shaken, and sonicated, and solutions were enumerated. Texture quality and color were evaluated over 14 days storage at 4°C after 0 and 1% CPC spray applications. A 0.5 or 1.0% (vol/vol) application of CPC after Salmonella reduced the pathogen levels between 2.34 log CFU/mL and 5.16 log CFU/mL in comparison to the control (p < .01). No differences were observed in the firmness and color of 1% CPC treated cantaloupes. Salmonella concentrations on cantaloupes, treated with 1.0% CPC, were lower after 1 hr storage as compared to 24 hr. And, Salmonella on "Athena" surfaces were more susceptible to CPC spray treatments than on "Hale's Best Jumbo."PRACTICAL APPLICATIONS:Cetylpyridinium chloride (CPC) is the active ingredient of some antiseptic oral mouth rinses, and has a broad antimicrobial spectrum with a rapid bactericidal effect on gram-positive pathogens. The spray application of CPC solutions to cantaloupe may reduce the level of Salmonella surface contamination during production from irrigation water and manure fertilizers and, during food processing by contaminated equipment and food handlers. Since the surfaces of cantaloupes are highly rough or irregular, bacteria can easily attach to these surfaces and become difficult to remove. Appropriate postharvest washing and sanitizing procedures are needed that can help control Salmonella and other pathogens on melons, especially on cantaloupes with nested surfaces. A direct surface spray application of CPC may be an alternative antimicrobial postharvest treatment to reduce pathogen contamination of cantaloupe melons, while providing an alternative to chlorine-based solutions.

A computer vision technique was used to determine the volume of raw agricultural products with an irregular shape. Thirty images of each rotated raw roma tomato, salad tomato, white button mushroom, and strawberry were collected. Volume determinations of these products using the optical imaging system were compared to volume measurements collected using a water displacement/buoyant force method. A high correlation was found between measurements from both methods. Furthermore, regression analyses of weight and volume measurements of each product set were used to develop equations to predict object volume from object weight. Weight measurement of raw produce can be used as a non-destructive method to estimate unit volume for sorting and grading purposes.

Dietary supplementation of probiotics is growing as a scientifically valid alternative to antibiotics for enhancement of overall animal health and productivity in aquaculture. Strains of Bacillus subtilis are regarded as attractive probiotic candidates to the fish farming industry; however, there is a limited number of studies focused on the use of specific strains probiotics in tilapia, and therefore complicating replication. The objective of this study was to examine the effect of the strains NZ86 (NRRL B-50136) and O14VRQ (NRRL B-67221) of B. subtilis on various parameters of the innate immunity in Nile tilapia (Oreochromis niloticus) in a 51-day feeding trial. Supplementation of tilapia with either strain resulted in significant increases (p < 0.05) in plasma lysozyme concentration of varying degrees throughout the trial. Meanwhile, alternative complement activity was significantly elevated (p < 0.05) only after feeding of the NZ86 strain after 14 and 51 days. Conversely, supplementation with O14VRQ resulted in a significant increase (p < 0.05) in the percent of neutrophils in the peripheral blood of tilapia by day 28. At the end of the trial, there was a trend towards increased phagocytic and respiratory burst activities observed in immune organ derived leukocytes. Feeding with either probiotic appeared to have an up-regulation on the gene expression of both pro-inflammatory cytokines in the intestine, yet only O14VRQ was significantly different than the control. Moreover, the occurrence of these results could be associated with supplementation of the probiotic strains, given that Bacillus bacteria were observed to populate the intestines of the dietary treatment groups. These results suggest the potential roles of these B. subtilis probiotic candidates to stimulate immune responses both locally and systemically in tilapia.

A multiyear survey of 31 ready-to-eat (RTE) food processing plants in the United States was conducted to determine the incidence of Listeria spp. in various RTE production environments. Samples were collected from 22 RTE plants regulated by the U.S. Department of Agriculture's Food Safety and Inspection Service (FSIS) and from 9 RTE food plants regulated by the U.S. Department of Health and Human Services' Food and Drug Administration (FDA). Only nonfood contact surfaces in the RTE manufacturing areas with exposed RTE product were sampled. Each sample was individually analyzed for the presence of Listeria spp. by using a PCR-based rapid assay. In total, 4,829 samples were collected from various locations, including freezers, equipment framework, floors, walls, wall-floor junctures, drains, floor mats, doors, and cleaning tools. Nine (29%) of the facilities had zero samples positive for Listeria spp. in the production environment, whereas 22 (71%) had one or more samples positive for Listeria spp. The total incidence of Listeria spp. in all RTE food plants was 4.5%. The positive rate in plants regulated by the FSIS ranged from 0 to 9.7%, whereas the positive rate in plants regulated by the FDA ranged from 1.2 to 36%.