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The incidence of antimicrobial resistance (AMR) in the environment is often overlooked and leads to serious health threats under the One Health paradigm. Infection with extended-spectrum β-lactamase (ESBL) producing bacteria in humans and animals has been widely examined, with the mode of transmission routes such as food, water, and contact with a contaminated environment. The purpose of this study was to determine the occurrence and molecular characteristics of resistant Salmonella enterica (S. enterica) (n = 59) and Escherichia coli (E. coli) (n = 392) isolated from produce commodities collected from fresh markets and supermarkets in Bangkok, Thailand. In this study, the S. enterica isolates exhibited the highest prevalence of resistance to tetracycline (11.9%) and streptomycin (8.5%), while the E. coli isolates were predominantly resistant to tetracycline (22.5%), ampicillin (21.4%), and sulfamethoxazole (11.5%). Among isolates of S. enterica (6.8%) and E. coli (15.3%) were determined as multidrug resistant (MDR). The prevalence of ESBL-producing isolates was 5.1% and 1.0% in S. enterica and E. coli, respectively. A minority of S. enterica isolates, where a single isolate exclusively carried blaCTX-M-55 (n = 1), and another isolate harbored both blaCTX-M-55 and blaTEM-1 (n = 1); similarly, a minority of E. coli isolates contained blaCTX-M-55 (n = 2) and blaCTX-M-15 (n = 1). QnrS (11.9%) and blaTEM (20.2%) were the most common resistant genes found in S. enterica and E. coli, respectively. Nine isolates resistant to ciprofloxacin contained point mutations in gyrA and parC. In addition, the odds of resistance to tetracycline among isolates of S. enterica were positively associated with the co-occurrence of ampicillin resistance and the presence of tetB (P = 0.001), while the E. coli isolates were positively associated with ampicillin resistance, streptomycin resistance, and the presence of tetA (P

The Produce Rule regulates food safety among produce growers in the United States. Policy exemptions (e.g., for farms with average produce sales under USD 30,000 over three years) pose a threat to local food safety, particularly given exempt growers' role in fresh produce production. Understanding exempt growers' perceptions of food safety practices and the factors influencing their voluntary adoption is crucial. A cross-sectional quantitative survey was conducted to assess growers' current engagement, their perceptions of agricultural water safety practices, and their influence on adoption decisions. The findings indicate inadequate engagement with agricultural water safety practices, with production water-related safety practices showing higher engagement compared to post-harvest water practices. Overall, growers reported favorable perceptions toward agricultural water safety practices. The perceived compatibility, relative advantage, and observability predicted adoption decisions among exempt growers, with the perceived compatibility being the strongest predictor. Policymakers and education professionals should design strategies and tailored educational interventions and messaging for exempt growers to emphasize the compatibility, relative advantage, and observability of agricultural water safety practices. Furthermore, it is recommended to explore policies and other mechanisms to increase the perceived relative advantage (immediacy of rewards and direct benefits) for exempt growers to promote voluntary adoption of these on-farm food safety practices. [ABSTRACT FROM AUTHOR]

In the U.S., baby spinach is mostly produced in Arizona (AZ) and California (CA). Characterizing the impact of growing region on the bacterial quality of baby spinach can inform quality management practices in industry. Between December 2021 and December 2022, baby spinach was sampled after harvest and packaging for microbiological testing, including shelf-life testing of packaged samples that were stored at 4°C. Samples were tested to (i) determine bacterial concentration, and (ii) obtain and identify bacterial isolates. Packaged samples from the Salinas, CA, area (n = 13), compared to those from the Yuma, AZ, area (n = 9), had a significantly higher bacterial concentration, on average, by 0.78 log10 CFU/g (P < 0.01, based on aerobic, mesophilic plate count data) or 0.67 log10 CFU/g (P < 0.01, based on psychrotolerant plate count data); the bacterial concentrations of harvest samples from the Yuma and Salinas areas were not significantly different. Our data also support that an increase in preharvest temperature is significantly associated with an increase in the bacterial concentration on harvested and packaged spinach. A Fisher's exact test and linear discriminant analysis (effect size), respectively, demonstrated that (i) the genera of 2,186 bacterial isolates were associated (P < 0.01) with growing region and (ii) Pseudomonas spp. and Exiguobacterium spp. were enriched in spinach from the Yuma and Salinas areas, respectively. Our findings provide preliminary evidence that growing region and preharvest temperature may impact the bacterial quality of spinach and thus could inform more targeted strategies to manage produce quality. [ABSTRACT FROM AUTHOR]

College students may have limited access to produce and may lack confidence in preparing it, but cooking videos can show how to make healthy dishes. The Cognitive Theory of Multimedia Learning suggests that learning is enhanced when visual and auditory information is presented considering cognitive load (e.g., highlighting important concepts, eliminating extraneous information, and keeping the video brief and conversational). The purpose of this project was to pilot test a food label for produce grown at an urban university and assess whether student confidence in preparing produce improved after using the label and QR code to view a recipe video developed using principles from the Cognitive Theory of Multimedia Learning. The video showed a student preparing a salad with ingredients available on campus. Students indicated the label was helpful and reported greater perceived confidence in preparing lettuce after viewing the label and video (mean confidence of 5.60 ± 1.40 before vs. 6.14 ± 0.89 after, p = 0.016, n = 28). Keeping the video short and providing ingredients and amounts onscreen as text were cited as helpful. Thus, a brief cooking video and interactive label may improve confidence in preparing produce available on campus. Future work should determine whether the label impacts produce consumption and if it varies depending on the type of produce used. [ABSTRACT FROM AUTHOR]

Background: Regular consumption of produce is a challenge for families with young children in low food access areas. Objective: A community partnership formed to evaluate feasibility of and interest in a child care center-based program for produce delivery from an online grocery vendor. Methods: Surveys were collected from caregivers across three child care centers, including produce program participants. Descriptive statistics summarize household characteristics and participants’ experience with the program. Results: Challenges related to online payment and difficulty planning delivery times led to implementation of a modified intervention. Survey results revealed factors related to food access and storage that may impact interest and feasibility of online grocery in some communities. Conclusions: Online grocery vendors may increase acces-sibility to fresh produce for families, but barriers to their use still exist. Trusted community partners such as child care centers may offer some of the resources needed for success.

Diarrheal disease is a leading cause of death in children in low- and moderate-income countries. Fresh produce, including fruits and vegetables, may harbor diarrheal disease-causing bacteria including strains of Salmonella enterica and Escherichia coli. This study aimed to determine the prevalence and antibiotic resistance profiles of S. enterica and E. coli isolated from produce samples (n = 207) obtained from retail markets in northern Botswana in Chobe District of Botswana in 2022. Samples were enriched in the appropriate selective media: Brilliant Green Bile Broth for E. coli and Rappaport Vassiliadis Broth for S. enterica. E. coli were confirmed by PCR detecting the phoA gene, and classified as potentially pathogenic through screening for the eae, stx, and stx2 and estIb genes. S. enterica isolates were confirmed using invA primers. Isolates were evaluated for resistance to ampicillin, amoxicillin-clavulanic acid, chloramphenicol, cefotaxime, doxycycline, streptomycin, sulfamethoxazole, and tetracycline antibiotic using the Kirby-Bauer Disk Diffusion method. E. coli was isolated from 15.5% of produce samples (n = 207). The gene eae was detected from 1.5% of samples, while stx1, stx2, and estIb were not detected. Resistance to one or more antibiotics was common (72%) with the majority of the resistant E. coli (n = 32) isolated from fruits (22%) and greens (18%) compared to other types of vegetables. Multidrug resistance (MDR, resistant to 3 or more antibiotics) was identified in 18% of samples. S. enterica was isolated from 3.4% of produce samples (7, n = 207). Resistance was uncommon among the S. enterica isolates (1/7). Overall prevalence of diarrheagenic S. enterica and E. coli was low; however, their presence and that of MDR E. coli in foods commonly consumed raw increases the risk to vulnerable populations. Strategies to reduce contamination of fresh produce and public education on washing and cooking some types of produce may be useful to reduce disease.

Listeria monocytogenes is a foodborne pathogen that lives in nature as a saprophyte. Two of the three most common serotypes that cause foodborne listeriosis are 1/2a and 4b. Within serotype 4b, there is a variant called 4bv-1. In the last decade, several produce-related outbreaks (linked to leafy salad, caramel apples, and stone fruit) were linked to 4bv-1 strains, specifically those of Sequence Type 382. This study assessed the fitness of ST 382 strains on lettuce leaf sections to determine if they are more fit on produce than strains of other serotypes. Strains of serotypes 1/2a, 4b, and ST 382 were inoculated as mixtures onto lettuce and incubated at 4 °C for 7 days or 25 °C for 24 h. Thirty L. monocytogenes colonies resulting from the growth on each lettuce piece were characterized for serotype by multiplex PCR, and the percentages of each serotype recovered were compared. In the individual mixtures with three strains, none of the ST 382 strains showed better fitness for growth on lettuce at either 4 °C or 25 °C. Overall, ST 382 strains showed better recovery from lettuce sections grown at 4 °C than at 25 °C. Statistical analysis of the recovery of twelve strains tested in competition experiments indicated that ST 382 strains were less fit for lettuce growth when competing against the other serotypes. The data indicate that ST 382 strains do not have a competitive fitness advantage on cut lettuce sections.

IntroductionThe impact of water quality on the survival of human norovirus (NoV) was determined in irrigation water field run-off (tail water) and well water from a representative Central Coast vegetable production site in the Salinas Valley, California.MethodsTail water, well water, and ultrapure water samples were inoculated separately with two surrogate viruses for human NoV-Tulane virus (TV) and murine norovirus (MNV)-to achieve a titer of 1×105 plaque forming units (PFU)/ml. Samples were stored at 11, 19, and 24°C for 28 days. Additionally, inoculated water was applied to soil collected from a vegetable production site in the Salinas Valley or to the surface of growing romaine lettuce leaves, and virus infectivity was evaluated for 28 days in a growth chamber.ResultsVirus survival was similar for water stored at 11, 19, and 24°C and there was no difference in infectivity based on water quality. After 28 days, a maximum 1.5 log reduction was observed for both TV and MNV. TV decreased by 1.97-2.26 log and MNV decreased by 1.28- 1.48 logs after 28 days in soil; infectivity was not influenced by water type. Infectious TV and MNV were recovered from lettuce surfaces for up to 7 and 10 days after inoculation, respectively. Across the experiments there was no significant impact of water quality on the stability of the human NoV surrogates.DiscussionOverall, the human NoV surrogates were highly stable in water with a less than 1.5 log reduction over 28 days and no difference observed based on the water quality. In soil, the titer of TV declined by approximately 2 logs over 28 days, while MNV declined by 1 log during the same time interval, suggesting surrogate-specific inactivation dynamics in the soil tested in this study. A 5-log reduction in MNV (day 10 post inoculation) and TV (day 14 post inoculation) was observed on lettuce leaves, and the inactivation kinetics were not significantly impacted by the quality of water used. These results suggest that human NoV would be highly stable in water, and the quality of the water (e.g., nutrient content, salinity, and turbidity) does not significantly impact viral infectivity.

Foodborne pathogens can cause gastrointestinal infections in consumers and in some cases can even lead to outbreaks. In the last decade, it has been observed that some zoonotic pathogenic bacteria can use plants as secondary hosts. Contamination with foodborne bacteria becomes relevant in foods that are regularly eaten raw, such as lettuce, cilantro, fenugreek, rocket leaves, basil, and so forth, and some fruits such as tomatoes, melons, and green peppers; because the elimination of these pathogenic bacteria is difficult to achieve with conventional sanitization processes. Contamination of produce can occur throughout the entire production chain. In farmlands, pathogenic bacteria can contaminate the seed, mainly when contaminated water is used for irrigation. Later, bacteria can reach other plant tissues such as the stems, leaves, and fruits. Another form of contamination is when the produce is in contact with feces from domestic, production, or wild animals. Additionally, poor handling practices during harvest, packaging, distribution, and sale can contaminate produce. Studies have shown that foodborne pathogens can adhere to produce, sometimes forming a biofilm, and can also be internalized into the plant or fruit, which protects them from sanitation processes. For this reason, in this text we address three biocontrol strategies such as bacteria, lytic bacteriophages, and some fungi, as an alternative approach for the control of both foodborne and plant pathogens. Additionally, the use of these biological agents can represent an advantage for the development of the plant, making them a good strategy to favor yield. [ABSTRACT FROM AUTHOR]

Recent cyclosporiasis outbreaks associated with fresh produce grown in the United States highlight the need to better understand Cyclospora cayetanensis prevalence in U.S. agricultural environments. In this study, C. cayetanensis occurrence was assessed in municipal wastewater sludge, on-farm portable toilets, irrigation pond water, and spent packing house dump tank water in a Southeastern Georgia growing region over two years. Detection of the C. cayetanensis 18S rRNA qPCR gene target in pond samples was 0%, 28%, and 42% (N = 217) depending on the detection definition used, and ≤1% in dump tank samples (N = 46). However, no qPCR detections were confirmed by sequencing, suggesting false detection occurred due to cross-reactions. C. cayetanensis qPCR detections were confirmed in 9% of wastewater sludge samples (N = 76). The human-specific fecal markers HF183 and crAssphage were detected in 33% and 6% of pond samples, respectively, and 4% and 0% of dump tank samples, respectively. Despite community Cyclospora shedding and evidence of human fecal contamination in irrigation water, there was no correlation between C. cayetanensis and HF183 qPCR detections, further supporting that 18S gene target qPCR amplifications were due to cross−reactions. When evaluating C. cayetanensis qPCR environmental detection data, the impact of assay specificity and detection criteria should be considered. Moreover, additional sequence-based testing may be needed to appropriately interpret Cyclospora qPCR environmental data.

Cyclospora cayetanensis is a coccidian parasite of the phylum Apicomplexa that causes cyclosporiasis, a human-specific gastrointestinal disease. Unlike most enteric pathogens, C. cayetanensis does not infect via direct fecal–oral transmission between humans because shed oocysts must be exposed to environmental triggers prior to becoming infectious. The development of specific and sensitive detection methods for C. cayetanensis is crucial to effectively address data gaps and provide regulatory support during outbreak investigations. In this study, new more specific molecular markers for the detection of C. cayetanensis were developed based on updated genomic databases of Apicomplexa mitochondrial sequences. Novel alternative reagents and supplies, as well as optimization protocols, were tested in spiked produce and agricultural water samples. The selected Mit1C primers and probe combined showed at least 13 mismatches to other related species. The new optimized qualitative real-time PCR assay with modifications to sample processing and replacement of discontinued items produced results comparable to the previously validated methods. In conclusion, the new optimized qualitative Mit1C real-time PCR assay demonstrated an increase in its specificity in comparison to other detection methods previously published, while it showed to be robust and as sensitive as the previously validated method at the FDA. This study has also expanded the array of PCR reagents that can be used to detect C. cayetanensis in produce and agricultural water samples and provided several improvements to the method for detection in agricultural water including replacements for discontinued items and a new dialysis filter for water filtration.

Limited data exist on the environmental factors that impact pathogen prevalence in the soil. The prevalence of foodborne pathogens, Salmonella and Listeria monocytogenes, and the prevalence and concentration of generic E. coli in Florida’s agricultural soils were evaluated to understand the potential risk of microbial contamination at the preharvest level. For all organisms but L. monocytogenes, a longitudinal field study was performed in three geographically distributed agricultural areas across Florida. At each location, 20 unique 5 by 5 m field sampling sites were selected, and soil was collected and evaluated for Salmonella presence (25 g) and E. coli and coliform concentrations (5 g). Complementary data collected from October 2021 to April 2022 included: weather; adjacent land use; soil properties, including macro- and micro-nutrients; and field management practices. The overall Salmonella and generic E. coli prevalence was 0.418% (1/239) and 11.3% (27/239), respectively; with mean E. coli concentrations in positive samples of 1.56 log CFU/g. Farm A had the highest prevalence of generic E. coli, 22.8% (18/79); followed by Farm B, 10% (8/80); and Farm C 1.25% (1/80). A significant relationship (p

At the beginning of the COVID-19 Pandemic, many warned that the resilience of the global, industrial food system would be tested. We conducted regular interviews in 2020 with key actors at the Ontario Food Terminal, North America’s third largest produce wholesale market, to better understand urban food system resilience in the first year of the Pandemic. How major wholesale marketplaces, such as the Ontario Food Terminal, fare during emergencies is key to understanding urban food system resilience, as these institutions connect farms to cities. Widescale interruptions to the supply of fresh produce did not take place at the Terminal despite challenges. We present data from the frontlines, documenting the challenges participants faced and their adaptive capacity. We find that food system resilience was rooted in pre-existing relationships, the adaptability of actors in produce supply chains, and worker stress and effort. We caution that, even though the system displayed resiliency, this does not mean that it is inherently resilient. We highlight vulnerabilities in the status quo and raise a red flag around the future ability of the system to withstand shocks. We conclude that, because the system resilience we document depends on people, the well-being of humans in the system is key to resilience of the food system itself.

College students may have limited access to produce and may lack confidence in preparing it, but cooking videos can show how to make healthy dishes. The Cognitive Theory of Multimedia Learning suggests that learning is enhanced when visual and auditory information is presented considering cognitive load (e.g., highlighting important concepts, eliminating extraneous information, and keeping the video brief and conversational). The purpose of this project was to pilot test a food label for produce grown at an urban university and assess whether student confidence in preparing produce improved after using the label and QR code to view a recipe video developed using principles from the Cognitive Theory of Multimedia Learning. The video showed a student preparing a salad with ingredients available on campus. Students indicated the label was helpful and reported greater perceived confidence in preparing lettuce after viewing the label and video (mean confidence of 5.60 ± 1.40 before vs. 6.14 ± 0.89 after, p = 0.016, n = 28). Keeping the video short and providing ingredients and amounts onscreen as text were cited as helpful. Thus, a brief cooking video and interactive label may improve confidence in preparing produce available on campus. Future work should determine whether the label impacts produce consumption and if it varies depending on the type of produce used.

Wild birds pose a difficult food safety risk to manage because they can avoid traditional wildlife mitigation strategies, such as fences. Birds often use agricultural fields and structures as foraging and nesting areas, which can lead to defecation on crops and subsequent transfer of foodborne pathogens. To assess the food safety risk associated with these events, wild bird feces were collected from produce fields across the southeastern United States during the 2021 and 2022 growing seasons. In total 773 fecal samples were collected from 45 farms across Florida, Georgia, South Carolina, and Tennessee, and 2.1% (n = 16) of samples were Salmonella-positive. Importantly, 75% of Salmonella were isolated from moist feces, showing reduced Salmonella viability when feces dry out. 16S microbiome analysis showed that presence of culturable Salmonella in moist feces correlated to a higher proportion of the Enterobacteriaceae family. From the Salmonellapositive samples, 62.5% (10/16) contained multi-serovar Salmonella populations. Overall, 13 serovars were detected, including six most commonly attributed to human illness (Enteriditis, Newport, Typhimurium, Infantis, Saintpaul, and Muenchen). PCR screening identified an additional 59 Salmonella-positive fecal samples, which were distributed across moist (n = 44) and dried feces (n = 15). On-farm point counts and molecular identification from fecal samples identified 57 bird species, including for 10 Salmonella-positive fecal samples. Overall, there was a low prevalence of Salmonella in fecal samples, especially in dried feces, and we found no evidence of Salmonella transmission to proximal foliage or produce. Fecal samples collected in farms close together shared highly related isolates by whole genome sequencing and also had highly similar Salmonella populations with comparable relative frequencies of the same serovars, suggesting the birds acquired Salmonella from a common source. [ABSTRACT FROM AUTHOR]

The incidence of antimicrobial resistance (AMR) in the environment is often overlooked and leads to serious health threats under the One Health paradigm. Infection with extended-spectrum β-lactamase (ESBL) producing bacteria in humans and animals has been widely examined, with the mode of transmission routes such as food, water, and contact with a contaminated environment. The purpose of this study was to determine the occurrence and molecular characteristics of resistant Salmonella enterica (S. enterica) (n = 59) and Escherichia coli (E. coli) (n = 392) isolated from produce commodities collected from fresh markets and supermarkets in Bangkok, Thailand. In this study, the S. enterica isolates exhibited the highest prevalence of resistance to tetracycline (11.9%) and streptomycin (8.5%), while the E. coli isolates were predominantly resistant to tetracycline (22.5%), ampicillin (21.4%), and sulfamethoxazole (11.5%). Among isolates of S. enterica (6.8%) and E. coli (15.3%) were determined as multidrug resistant (MDR). The prevalence of ESBL-producing isolates was 5.1% and 1.0% in S. enterica and E. coli, respectively. A minority of S. enterica isolates, where a single isolate exclusively carried blaCTX-M-55 (n = 1), and another isolate harbored both blaCTX-M-55 and blaTEM-1 (n = 1); similarly, a minority of E. coli isolates contained blaCTX-M-55 (n = 2) and blaCTX-M-15 (n = 1). QnrS (11.9%) and blaTEM (20.2%) were the most common resistant genes found in S. enterica and E. coli, respectively. Nine isolates resistant to ciprofloxacin contained point mutations in gyrA and parC. In addition, the odds of resistance to tetracycline among isolates of S. enterica were positively associated with the co-occurrence of ampicillin resistance and the presence of tetB (P = 0.001), while the E. coli isolates were positively associated with ampicillin resistance, streptomycin resistance, and the presence of tetA (P

An increase in acute gastroenteritis occurred in Chanthaburi Province, Thailand, during December 2021‒January 2022. Of the norovirus genotypes we identified in hospitalized patients and produce from local markets, genotype GII.3[P25] accounted for one third. We found no traceable link between patients and produce but found evidence of potential viral intake.

Whole genome sequencing (WGS) is a powerful tool that may be used to assist in identifying Listeria contamination sources and movement within environments, and to assess persistence. This study investigated sites in a produce packinghouse where Listeria had been historically isolated; and aimed to characterize dispersal patterns and identify cases of transient and resident Listeria. Environmental swab samples (n = 402) were collected from 67 sites at two time-points on three separate visits. Each sample was tested for Listeria, and Listeria isolates were characterized by partial sigB sequencing to determine species and allelic type (AT). Representative isolates from the three most common L. monocytogenes ATs (n = 79) were further characterized by WGS. Of the 144 Listeria species positive samples (35.8%), L. monocytogenes was the most prevalent species. L. monocytogenes was often coisolated with another species of Listeria. WGS identified cases of sporadic and continued reintroduction of L. monocytogenes from the cold storages into the packinghouse and demonstrated cases of L. monocytogenes persistence over 2 years in cold storages, drains, and on a forklift. Nine distinct clusters were found in this study. Two clusters showed evidence of persistence. Isolates in these two clusters (N = 11, with one historical isolate) were obtained predominantly and over multiple samplings from cold storages, with sporadic movement to sites in the packing area, suggesting residence in cold storages with opportunistic dispersal within the packinghouse. The other seven clusters demonstrated evidence of transient Listeria, as isolation was sporadic over time and space during the packing season. Our data provide important insights into likely L. monocytogenes harborage points and transfer in a packinghouse, which is key to root cause analysis. While results support Listeria spp. as a suitable indicator organism for environmental monitoring surveys, findings were unable to establish a specific species as an index organism for L. monocytogenes. Findings also suggest long-term persistence with substantial SNP diversification, which may assist in identifying potential contamination sources and implementing control measures.

Wild birds pose a difficult food safety risk to manage because they can avoid traditional wildlife mitigation strategies, such as fences. Birds often use agricultural fields and structures as foraging and nesting areas, which can lead to defecation on crops and subsequent transfer of foodborne pathogens. To assess the food safety risk associated with these events, wild bird feces were collected from produce fields across the southeastern United States during the 2021 and 2022 growing seasons. In total 773 fecal samples were collected from 45 farms across Florida, Georgia, South Carolina, and Tennessee, and 2.1% (n = 16) of samples were Salmonella-positive. Importantly, 75% of Salmonella were isolated from moist feces, showing reduced Salmonella viability when feces dry out. 16S microbiome analysis showed that presence of culturable Salmonella in moist feces correlated to a higher proportion of the Enterobacteriaceae family. From the Salmonella-positive samples, 62.5% (10/16) contained multi-serovar Salmonella populations. Overall, 13 serovars were detected, including six most commonly attributed to human illness (Enteriditis, Newport, Typhimurium, Infantis, Saintpaul, and Muenchen). PCR screening identified an additional 59 Salmonella-positive fecal samples, which were distributed across moist (n = 44) and dried feces (n = 15). On-farm point counts and molecular identification from fecal samples identified 57 bird species, including for 10 Salmonella-positive fecal samples. Overall, there was a low prevalence of Salmonella in fecal samples, especially in dried feces, and we found no evidence of Salmonella transmission to proximal foliage or produce. Fecal samples collected in farms close together shared highly related isolates by whole genome sequencing and also had highly similar Salmonella populations with comparable relative frequencies of the same serovars, suggesting the birds acquired Salmonella from a common source.

Current California agricultural practices strive to comanage food safety and habitat conservation on farmland. However, the ecology of foodborne pathogens in wild bird populations, especially those avian species residing in proximity to fresh produce production fields, is not fully understood. In this repeated cross-sectional study, avifauna within agricultural lands in California were sampled over 1 year. Feces, oral swabs, and foot/feather swabs were cultured for zoonotic Salmonella spp., Escherichia coli O157:H7, and non-O157 Shiga toxin-producing E. coli (STEC) and characterized by serotyping and pulsed-field gel electrophoresis. Of 60 avian species sampled, 8 species (13.3%, bird groups of sparrows, icterids, geese, wrens, and kinglets) were positive for at least one of these foodborne pathogens. At the individual bird level, the detection of foodborne pathogens was infrequent in feces (n = 583; 0.5% Salmonella, 0.34% E. coli O157:H7, and 0.5% non-O157 STEC) and in feet/feathers (n = 401; 0.5% non-O157 STEC), and it was absent from oral swabs (n = 353). Several subtypes of public health importance were identified, including Salmonella enterica serotype Newport, E. coli O157:H7, and STEC serogroups O103 and O26. In late summer and autumn, the same STEC subtype was episodically found in several individuals of the same and different avian species, suggesting a common source of contamination in the environment. Sympatric free-range cattle shared subtypes of STEC O26 and O163 with wild geese. A limited rate of positive detection in wild birds provides insights into broad risk profile for contamination considerations but cannot preclude or predict risk on an individual farm.IMPORTANCE The shedding dynamics of foodborne pathogens by wild birds on farmland are not well characterized. This yearlong study sampled wild birds for foodborne pathogens within agricultural lands in northern California. There was a low prevalence of Salmonella spp., Escherichia coli O157:H7, and non-O157 Shiga-toxin producing E. coli (prevalence, 0.34% to 0.50%) identified in bird populations in this study. However, pathogens of public health importance (such as Salmonella Newport, E. coli O157:H7, and STEC O103 and O26) were identified in fecal samples, and two birds carried STEC on their feet or feathers. Identical pathogen strains were shared episodically among birds and between wild geese and free-range cattle. This result suggests a common source of contamination in the environment and potential transmission between species. These findings can be used to assess the risk posed by bird intrusions in produce fields and enhance policy decisions toward the comanagement of food safety and farmland habitat conservation.

Abstract Background The global community has battled the spread of SAR-CoV-2 for almost 2 years, and the projection is that the virus may be recurrent like the seasonal flu. The SARS-CoV-2 pandemic disrupted activities within the food supply chain that cost billions of dollars globally. This has heightened concerns about fomite spread of the virus through surfaces. There is an urgent need to understand the risk portends by this virus along the produce supply chain with conditions (low temperature and high relative humidity) conducive to extended survival of the virus. Main body Pre-dating SARS-CoV-2 are other types of coronaviruses that had lower infection and mortality rates. There are some similarities between the former and the new coronavirus, especially with regards to transmission modes and their survivability on surfaces. There is evidence of other coronaviruses’ survival on surfaces for weeks. Currently, there are limited evidence-based studies to enlighten us on how the virus is transmitted within the produce supply chain. A few studies claim that the virus could spread through the cold supply chains. However, these are not sufficient to make a conclusive inference about the deadly SARS-CoV-2. Conclusions This paper provides a succinct review of the literature on current understanding of the transmission, survivability, and risk SARS-CoV-2 portend to humans within the produce supply chain and calls for more evidence-based research to allay or alert us of the potential risk of fomite transmission of SARS-CoV-2. The paper also highlights examples of conventional and novel non-thermal inactivation and sanitation methods applicable to this type of virus.

Introduction: The impact of water quality on the survival of human norovirus (NoV) was determined in irrigation water field run-off (tail water) and well water from a representative Central Coast vegetable production site in the Salinas Valley, California. Methods: Tail water, well water, and ultrapure water samples were inoculated separately with two surrogate viruses for human NoV--Tulane virus (TV) and murine norovirus (MNV)--to achieve a titer of 1×105 plaque forming units (PFU)/ml. Samples were stored at 11, 19, and 24°C for 28 days. Additionally, inoculated water was applied to soil collected from a vegetable production site in the Salinas Valley or to the surface of growing romaine lettuce leaves, and virus infectivity was evaluated for 28 days in a growth chamber. Results: Virus survivalwas similar for water stored at 11, 19, and 24°C and there was no difference in infectivity based on water quality. After 28 days, a maximum1.5 log reduction was observed for both TV and MNV. TV decreased by 1.97-2.26 log and MNV decreased by 1.28-1.48 logs after 28 days in soil; infectivitywas not influenced bywater type. Infectious TV and MNV were recovered from lettuce surfaces for upto 7 and 10 days after inoculation, respectively. Across the experiments there was no significant impact of water quality on the stability of the human NoV surrogates. Discussion: Overall, the human NoV surrogates were highly stable in water with a less than 1.5 log reduction over 28 days and no difference observed based on the water quality. In soil, the titer of TV declined by approximately 2 logs over 28 days, while MNV declined by 1 log during the same time interval, suggesting surrogatespecific inactivation dynamics in the soil tested in this study. A 5-log reduction in MNV (day 10 post inoculation) and TV (day 14 post inoculation) was observed on lettuce leaves, and the inactivation kinetics were not significantly impacted by the quality of water used. These results suggest that human NoV would be highly stable in water, and the quality of the water (e.g., nutrient content, salinity, and turbidity) does not significantly impact viral infectivity. [ABSTRACT FROM AUTHOR]

Listeriosis outbreaks were associated with contaminated fruits and vegetables, including cantaloupe, apples, and celery. Grape seed extract (GSE) is a natural antimicrobial with potential for reducing Listeria monocytogenes contamination in food. This study assessed the effectiveness of GSE to reduce L. monocytogenes on fresh produce and the impact of food matrices on its antilisterial activity. GSE showed MIC values of 30–35 μg/mL against four Listeria strains used in this study. A total of 100 g portions of cantaloupe, apples, and celery were inoculated with L. monocytogenes and treated with 100–1000 μg/mL of GSE for 5 or 15 min. Results were analyzed using Rstudio and a Tukey's test. Treated produce had significantly lower L. monocytogenes counts than the control samples (p-value < 0.05). The inhibition was significantly higher on apples and lowest on cantaloupe. Moreover, a 15 min treatment was found to be more effective than a 5 min treatment in reducing L. monocytogenes on all produce types. The reduction in L. monocytogenes levels varied between 0.61 and 2.5 log10 CFU reduction, depending on the treatment concentration, duration, and produce matrix. These findings suggest that GSE is an effective antilisterial treatment for fresh produce, with varying levels of effectiveness depending on the food matrix and treatment time. [ABSTRACT FROM AUTHOR]

This study investigated the inhibitory activity of organic solutions containing 5, 10, 15, 20 and 30% (w/v) sodium chloride and citric acid solution and 15:10, 15:15, 15:20 and 15:30% (w/v) sodium chloride (NaCl) combined with citric acid (CA) solution (salt/acid solution) for 10 min against microorganisms isolated from trimmed young coconut: Bacillus cereus, B. subtilis, Staphylococcus aureus, S. epidermidis, Enterobacter aerogenes, Serratia marcescens, Candida tropicalis, Lodderromyces elongisporus, Aspergillus aculeatus and Penicillium citrinum. Commercial antimicrobial agents such as potassium metabisulfite and sodium hypochlorite (NaOCl) were used as the controls. Results showed that 30% (w/v) NaCl solution displayed antimicrobial properties against all microorganisms, with s reduction range of 0.00–1.49 log CFU/mL. Treatment of 30% (w/v) CA solution inhibited all microorganisms in the reduction range of 1.50–8.43 log CFU/mL, while 15:20% (w/v) salt/acid solution was the minimum concentration that showed a similar antimicrobial effect with NaOCl and strong antimicrobial effect against Gram-negative bacteria. The mode of action of this solution against selected strains including B. cereus, E. aerogenes and C. tropicalis was also determined by scanning electron microscopy and transmission electron microscopy. B. cereus and E. aerogenes revealed degradation and detachment of the outer layer of the cell wall and cytoplasm membrane, while cytoplasmic inclusion in treated C. tropicalis cells changed to larger vacuoles and rough cell walls. The results suggested that a 15:20% (w/v) salt/acid solution could be used as an alternative antimicrobial agent to eliminate microorganisms on fresh produce. [ABSTRACT FROM AUTHOR]

The foodborne pathogen Listeria monocytogenes generally infects immunocompromised individuals, such as cancer patients, more frequently and with higher morbidity and mortality than the general population. Because of the anticipated risk associated with L. monocytogenes and other pathogens in produce, immunocompromised individuals are often placed on neutropenic diets that exclude fresh produce, though these risks have not been quantified. Therefore, this study developed a data-driven risk model for listeriosis in cancer patients who consume ready-to-eat (RTE) salads, consisting of leafy greens, cucumbers, and tomatoes, as influenced by kitchen-scale treatments and storage practices. Monte Carlo simulations were used to model the risk of invasive listeriosis during one chemotherapy cycle. Refrigerating all salad components decreased the median risk by approximately one-half log. For refrigerated salads with no treatment, the predicted median risk was ≤ 4.3 × 10−08. When salad ingredients were surface blanched with greens rinsed, the predicted risk decreased to 5.4 × 10−10. Predicted risk was lowest (1.4 × 10−13) for a blanched “salad” consisting of solely cucumbers and tomatoes. Interestingly, rinsing, as recommended by FDA, only decreased the median risk by 1 log. A sensitivity analysis revealed that the highly variable dose-response parameter k strongly influenced risk, indicating that reducing uncertainty in this variable may improve model accuracy. Overall, this study demonstrates that kitchen-scale pathogen reduction approaches have high risk reduction efficacy and could be considered as an alternative to diets that exclude produce when making risk management decisions.

Produce (e.g. fruit and vegetable) varies in quality when it is picked from plants. Then a produce wholesaler has two sales strategies: (1) unsorted selling (US), i.e. selling the produce to the market at one price and (2) sorted selling (SS), i.e. pre-sorting the produce into different grades by quality and selling them at different prices. When the SS strategy is adopted, the demands of the different grades are uncertain and substitutable. In this paper, we study the joint optimal purchase and sales strategy for selling mixed quality produce to customers with heterogeneous quality preferences. We first derive the optimal purchase policies under the two strategies and then identify the conditions under which the SS or US strategy is optimal. We show that the optimal sales strategy is determined by the marginal purchase and sorting cost and the fixed investment cost, the overall quality level of the produce, and the substitution rate for the high-quality and low-quality produce. This finding differs from the conventional wisdom that the SS strategy is optimal in the produce retail business. Finally, we conduct numerical studies based on real-world data to generate managerial insights from the analytical findings. [ABSTRACT FROM AUTHOR]

This randomized controlled trial characterized the transfer of E. coli from animal feces and/or furrow water onto adjacent heads of lettuce during foliar irrigation, and the subsequent survival of bacteria on the adaxial surface of lettuce leaves. Two experiments were conducted in Salinas Valley, California: (1) to quantify the transfer of indicator E. coli from chicken and rabbit fecal deposits placed in furrows to surrounding lettuce heads on raised beds, and (2) to quantify the survival of inoculated E. coli on Romaine lettuce over 10 days. E. coli was recovered from 97% (174/180) of lettuce heads to a maximal distance of 162.56 cm (5.33 ft) from feces. Distance from sprinklers to feces, cumulative foliar irrigation, and lettuce being located downwind of the fecal deposit were positively associated, while distance from fecal deposit to lettuce was negatively associated with E. coli transference. E. coli exhibited decimal reduction times of 2.2 and 2.5 days when applied on the adaxial surface of leaves within a chicken or rabbit fecal slurry, respectively. Foliar irrigation can transfer E. coli from feces located in a furrow onto adjacent heads of lettuce, likely due to the kinetic energy of irrigation droplets impacting the fecal surface and/or impacting furrow water contaminated with feces, with the magnitude of E. coli enumerated per head of lettuce influenced by the distance between lettuce and the fecal deposit, cumulative application of foliar irrigation, wind aspect of lettuce relative to feces, and time since final irrigation. Extending the time period between foliar irrigation and harvest, along with a 152.4 cm (5 ft) no-harvest buffer zone when animal fecal material is present, may substantially reduce the level of bacterial contamination on harvested lettuce.

Field trials were conducted in July-August and October 2012 to quantify the inactivation rate of Escherichia coli O157:H7 when mixed with fecal slurry and applied to romaine lettuce leaves. Lettuce was grown under commercial conditions in Salinas Valley, California. One-half milliliter of rabbit, chicken, or pig fecal slurry, containing an average of 4.05 × 107 CFU E. coli O157:H7 (C0), was inoculated onto the upper (adaxial) surface of a lower leaf on 288 heads of lettuce per trial immediately following a 2.5 h irrigation event. To estimate the bacterial inactivation rate as a function of time, fecal matrix, irrigation and seasonal climate effects, sets of lettuce heads (n = 28) were sampled each day over 10 days and the concentration of E. coli O157:H7 (Ct) determined. E. coli O157:H7 was detected on 100% of heads during the 10-day duration, with concentrations ranging from ≤340 MPN/head (∼5-log reduction) to >3.45 × 1012 MPN/head (∼5-log growth). Relative to C0, on day 10 (Ct = 12) we observed an overall 2.6-log and 3.2-log mean reduction of E. coli O157:H7 in July and October, respectively. However, we observed relative maximum concentrations due to bacterial growth on day 6 (maximum Ct = 8) apparently stimulated by foliar irrigation on day 5. From this maximum there was a mean 5.3-log and 5.1-log reduction by day 10 (Ct = 12) for the July and October trials, respectively. This study provides insight into the inactivation and growth kinetics of E. coli O157:H7 on romaine lettuce leaves under natural field conditions. This study provides evidence that harvesting within 24 h post irrigation has the potential to increase the concentration of E. coli O157:H7 contamination, if present on heads of romaine lettuce; foliar irrigation can temporarily stimulate substantial regrowth of E. coli O157:H7.

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.

With the spread of global markets for modern technical education and the diversity of programs for the requirements of the local and global market for information and communication technology, the universities began to race among themselves to earn their academic reputation. In addition, they want to enhance their technological development by developing IMT systems with integrated technology as the security and fastest response with the speed of providing the required service and sure information and linking it The network and using social networking programs with wireless networks which in turn is a driver of the emerging economies of technical education. All of these facilities opened the way to expand the number of students and solve the problem of accumulation, collection and analysis of data by storing it with large, expanded and automatically interconnected databases between university places and departments to provide services adapted to the desire of demand. This research dealt with a sample from of the academic's opinions and students. The sample is 319 questionnaires. It concluded that each of the infrastructure, devices, Internet of things, smart classrooms, and administrative database, with the presence of the fifth-generation network and its equipment, have a statistically significant correlation with technical education technology. [ABSTRACT FROM AUTHOR]

Persistent Listeria monocytogenes contamination may occur in a packinghouse if the pathogen successfully infiltrates the facility and reaches a harborage site, where it may be difficult to remove and may contaminate produce within the facility. There is a need for simulation-based decision support tools that can predict which equipment sites are more likely to undergo persistent contamination and simulate potential corrective actions to prevent this contamination. Thus, we adapted for longer term simulation two existing applications of an agent-based model of Listeria spp. hourly contamination dynamics in produce packinghouses. Next, we developed a novel approach to identify and analyze persistent and transient Listeria contamination patterns on simulated agents representing equipment sites and employees. Testing of corrective actions showed that methods that involved targeted, facility-specific, risk-based sanitation were the most effective in reducing both the likelihood and duration of persistent contamination. Generic approaches to controlling Listeria (e.g., more concentrated sanitizers) are unlikely to be successful and suggest that use of sanitation schedules produced through facility-specific root cause analysis and hygienic design are key in reducing persistence. Hourly Listeria contamination patterns also suggest that transient contamination may be mistaken for persistent contamination, depending on the frequency of environmental sampling. Likewise, as concentrations of Listeria on most contaminated agents were predicted to be very low, there is also a possibility to mistake persistence for transient contamination of sites, or even miss it outright, due to false-negative environmental Listeria monitoring results. These findings support that agent-based models may be valuable decision support tools, aiding in the identification of contamination patterns within packinghouses and assessing the viability of specific corrective actions. [ABSTRACT FROM AUTHOR]

Objective: Produce sold as plastic-wrapped packs of two to four individual items (i.e., produce micro-packs) that are low cost and placed at checkout may appeal to shoppers with budget constraints and provide a second chance to purchase items available elsewhere in the store. This study examined the impact of an intervention that placed produce micro-packs at checkout and promoted them in grocery stores across New Mexico, USA. Design: This quasi-experimental study placed produce micro-packs at checkout end-caps in thirteen stores (group 1), with eight stores serving as controls (group 2) from 1 July 2019 through 31 January 2020 (first phase). The intervention was extended to group 2 stores from 1 February 2020 through 30 June 2020 (second phase). Cashiers were directed to upsell the micro-packs to Special Supplemental Nutrition Program for Women, Infants, and Children recipients who had unspent cash value benefits for produce purchases. Setting: Twenty-one grocery stores across New Mexico. Participants: Twenty-one produce items sold as micro-packs in stores from July 2019 through June 2020. Results: A random effects model showed that the daily sales of micro-packs increased by 47 % during each intervention period. Group 2 stores had lower sales than group 1 stores during the first phase of the intervention. Once extended to group 2 stores, sales of micro-packs in those stores increased and sales in group 1 stores continued at the higher level. Conclusions: Placing produce micro-packs at checkout may increase produce sales and support health promotion efforts by public and private stakeholders. [ABSTRACT FROM AUTHOR]

Nearly half of foodborne illnesses are linked to produce and nuts, and water used for produce post-harvest activities can contribute to contamination. Surface water serves as an economical source for agricultural activities; however, exposure to the environment increases microbial risks and impacts its physicochemical characteristics. In this study, peroxyacetic acid (PAA) and chlorine (Cl) were evaluated as treatments for simulated surface water to determine their efficacy at achieving 'no detectable generic Escherichia coli' in 100 mL. Simulated surface water was prepared to turbidities of 2 and 100 NTU, adjusted to pH 6.5 or 8.4, equilibrated to 32 or 12 °C, inoculated with 5 logs per mL of non-pathogenic (generic) E. coli, and treated with Cl 25 ± 2 ppm, PAA 75 ± 5 ppm, or sterile water control (W). Dey-Engley neutralization was followed by enumeration on E. coli/Coliform Petrifilm at times (t) 0 to 2880 min (48 h) post-treatment. When not detected, treatments were further evaluated through enrichment in 2X Brain Heart Infusion (BHI) broth. Enrichments were streaked on MacConkey agar (MAC) to confirm E. coli absence. All Cl and PAA treated samples were below the test limit of detection (<5 CFU/mL), and E. coli was not detected in 5 mL enrichments even at t = 0 (shortly after treatment). These data suggest that Cl and PAA interventions may be effective for treating surface water for post-harvest uses. [ABSTRACT FROM AUTHOR]

Under conditions of blocked main hubs, frozen container traffic, high container freight rates, difficult deliveries and a number of other shocks accompanying the Covid-19 crisis, businesses were faced with a number of challenges to which they had to adapt. In this regard, the factors that influence the decision not to produce, but to purchase parts and components under the new conditions, have also aroused interest. This aspect is related to the make-or-buy strategic decision, where a choice is made between producing materials or purchasing them from an external supplier. The aim of this article is both to highlight the importance of a group of factors that influence the decision of companies to buy parts and components, rather than produce them independently, and to make an attempt to compare the importance of these factors in different periods, namely before and during the Covid - 19 crisis. The assertion is that the degree of importance of the factors that are taken into account in the decision not to manufacture but to buy parts and components is different in each individual situation. [ABSTRACT FROM AUTHOR]

The contamination of ready-to-eat produce with Listeria monocytogenes (LM) can often be traced back to environmental sources in processing facilities and packinghouses. To provide an improved understanding of Listeria sources and transmission in produce operations, we performed whole-genome sequencing (WGS) of LM (n = 169) and other Listeria spp. (n = 107) obtained from 13 produce packinghouses and three fresh-cut produce facilities. Overall, a low proportion of LM isolates (9/169) had inlA premature stop codons, and a large proportion (83/169) had either or both of the LIPI-3 or LIPI-4 operons, which have been associated with hypervirulence. The further analysis of the WGS data by operation showed a reisolation (at least 2 months apart) of highly related isolates (,10 hqSNP differences) in 7/16 operations. Two operations had highly related strains reisolated from samples that were collected at least 1 year apart. The identification of isolates collected during preproduction (i.e., following sanitation but before the start of production) that were highly related to isolates collected during production (i.e., after people or products have entered and begun moving through the operation) provided evidence that some strains were able to survive standard sanitation practices. The identification of closely related isolates (,20 hqSNPs differences) in different operations suggests that cross-contamination between facilities or introductions from common suppliers may also contribute to Listeria transmission. Overall, our data suggest that the majority of LM isolates collected from produce operations are fully virulent and that both persistence and reintroduction may lead to the repeat isolation of closely related Listeria in produce operations. IMPORTANCE Listeria monocytogenes is of particular concern to the produce industry due to its frequent presence in natural environments as well as its ability to survive in packinghouses and fresh-cut processing facilities over time. The use of whole-genome sequencing, which provides high discriminatory power for the characterization of Listeria isolates, along with detailed source data (isolation date and sample location) shows that the presence of Listeria in produce operations appears to be due to random and continued reintroduction as well as to the persistence of highly related strains in both packinghouses and fresh-cut facilities. These findings indicate the importance of using high-resolution characterization approaches for root cause analyses of Listeria contamination issues. In cases of repeat isolation of closely related Listeria in a given facility, both persistence and reintroduction need to be considered as possible root causes. [ABSTRACT FROM AUTHOR]

Grains are strategic crops in the study area (wheat, barley, maize) that located between two latitudes (33/46 35-34 28/27) and two longitudes (41 10/39 36/49 east), exactly in the western part of Anbar province and Iraq, being one of the areas of al-Qaim district of 95,358 dunums (2.7%) at a rate of the area of al-Qaim district (3530,000) dunums. The area bordered to the west by the Syrian region of Albu Kamal, to the east and north-east, to the south and south-west by the Alratba district, where the area under cultivation of wheat crops for years reached (2000-2010-2020), (7732), (7,730), (9,417) dunums and at a rate of (31%) , (31%), (37.8%) Dunum, quantity of Production for the same years was (32064) (30253) and (65919) and 25 tons. ، (23,5%) ، (51,4%), the area cultivated of the barley crop for the years (2000-2010-2020), (290), (442), (261) dunums and (26.7%), (43.4%) (29.7%) The amount of production for the same years (4.9%) (83,5%) ،(11,4%) Tons, the area under cultivation of maize crops for years (2000-2010-2020), (887), (885), (1037) dunums and (31.5%) ،(31,5%)، (36,9%), Dunum, and production for the same years reached (13,845), (4,425), (25925) tons and at a rate of (3.3%) .(10%), (58.6 %) ton.

Salmonella spp. are remarkably adaptable pathogens, and this adaptability allows these bacteria to thrive in a variety of environments and hosts. The mechanisms with which these pathogens establish within a niche amid the native microbiota remain poorly understood. Here, we aimed to uncover the mechanisms that enable Salmonella enterica serovar Typhimurium strain ATCC 14028 to benefit from the degradation of plant tissue by a soft rot plant pathogen, Pectobacterium carotovorum The hypothesis that in the soft rot, the liberation of starch (not utilized by P. carotovorum) makes this polymer available to Salmonella spp., thus allowing it to colonize soft rots, was tested first and proven null. To identify the functions involved in Salmonella soft rot colonization, we carried out transposon insertion sequencing coupled with the phenotypic characterization of the mutants. The data indicate that Salmonella spp. experience a metabolic shift in response to the changes in the environment brought on by Pectobacterium spp. and likely coordinated by the csrBC small regulatory RNA. While csrBC and flhD appear to be of importance in the soft rot, the global two-component system encoded by barA sirA (which controls csrBC and flhDC under laboratory conditions) does not appear to be necessary for the observed phenotype. Motility and the synthesis of nucleotides and amino acids play critical roles in the growth of Salmonella spp. in the soft rot.IMPORTANCE Outbreaks of produce-associated illness continue to be a food safety concern. Earlier studies demonstrated that the presence of phytopathogens on produce was a significant risk factor associated with increased Salmonella carriage on fruits and vegetables. Here, we genetically characterize some of the requirements for interactions between Salmonella and phytobacteria that allow Salmonella spp. to establish a niche within an alternate host (tomato). Pathways necessary for nucleotide synthesis, amino acid synthesis, and motility are identified as contributors to the persistence of Salmonella spp. in soft rots.

Abstract Background Lettuce is linked to recurrent outbreaks of Shiga toxin-producing Escherichia coli (STEC) infections, the seasonality of which remains unresolved. Infections have occurred largely from processed lettuce, which undergoes substantial physiological changes during storage. We investigated the microbiome and STEC O157:H7 (EcO157) colonization of fresh-cut lettuce of two cultivars with long and short shelf life harvested in the spring and fall in California and stored in modified atmosphere packaging (MAP) at cold and warm temperatures. Results Inoculated EcO157 declined significantly less on the cold-stored cultivar with short shelf life, while multiplying rapidly at 24 °C independently of cultivar. Metagenomic sequencing of the lettuce microbiome revealed that the pre-storage bacterial community was variable but dominated by species in the Erwiniaceae and Pseudomonadaceae. After cold storage, the microbiome composition differed between cultivars, with a greater relative abundance (RA) of Erwiniaceae and Yersiniaceae on the cultivar with short shelf life. Storage at 24 °C shifted the microbiome to higher RAs of Erwiniaceae and Enterobacteriaceae and lower RA of Pseudomonadaceae compared with 6 °C. Fall harvest followed by lettuce deterioration were identified by recursive partitioning as important factors associated with high EcO157 survival at 6 °C, whereas elevated package CO2 levels correlated with high EcO157 multiplication at 24 °C. EcO157 population change correlated with the lettuce microbiome during 6 °C storage, with fall microbiomes supporting the greatest EcO157 survival on both cultivars. Fall and spring microbiomes differed before and during storage at both temperatures. High representation of Pantoea agglomerans was a predictor of fall microbiomes, lettuce deterioration, and enhanced EcO157 survival at 6 °C. In contrast, higher RAs of Erwinia persicina, Rahnella aquatilis, and Serratia liquefaciens were biomarkers of spring microbiomes and lower EcO157 survival. Conclusions The microbiome of processed MAP lettuce evolves extensively during storage. Under temperature abuse, high CO2 promotes a lettuce microbiome enriched in taxa with anaerobic capability and EcO157 multiplication. In cold storage, our results strongly support a role for season and lettuce deterioration in EcO157 survival and microbiome composition, suggesting that the physiology and microbiomes of fall- and spring-harvested lettuce may contribute to the seasonality of STEC outbreaks associated with lettuce grown in coastal California.

Cyclospora cayetanensis, a coccidian apicomplexan parasite, causes large outbreaks of foodborne diarrheal disease globally. Tracking the source of C. cayetanensis oocyst contamination in food items is essential to reduce, even prevent outbreaks. We previously showed that a genotyping method based on mitochondrial single nucleotide polymorphism (SNP) profiles had discriminatory power in classifying C. cayetanensis clinical isolates. In food specimens, low level contamination by oocysts and difficulties in DNA extraction present significant challenges in genotyping method development. Here, we report the development of a highly sensitive, custom-designed, targeted sequencing method based on the Illumina AmpliSeq platform; our method was capable of consistently generating near-complete mitochondrial genome sequences of C. cayetanensis from foods with low levels of contamination. To simulate environmentally observed contamination levels in foods, we seeded various food matrices, such as fresh produce and prepared dishes, with known quantities of oocysts, and isolated genomic DNA from washed food samples. Using the Ampliseq Targeted Sequencing method, we obtained near-complete mitochondrial genome sequences of C. cayetanensis from food samples seeded with as low as five to ten oocysts and used the data in downstream analysis. The flexibility of the AmpliSeq platform could potentially allow for more genomic targets to be added to achieve higher discriminatory power. This level of sensitivity in capturing high resolution genome data from contaminated food samples is a critical milestone towards the potential development of a comprehensive genotyping method for C. cayetanensis.

Some aspects of the biology, digestive enzymes and organosomatic indices of Chrysichthys nigrodigitatus purchased from fishermen in Oyan dam was investigated. This study was aimed at providing information on the composition of food materials found in the gut and specific activities of selected enzymes as it affects the domestication of the species. 100 specimens of the species were examined for stomach contents, length-weight relationship, digestive enzyme assay and organosomatic indices, using standard methods. Food items observed were detritus (4%), fish part (12%), Insecta (13%). Sand was observed to be 11% of total stomach volume. The logarithmic equation for length-weight relationship 𝑙𝑛𝑊 = 2.68𝑙𝑛𝑙 − 3.79 indicated that an increase in length led to a corresponding increase in weight with ‘R’ = 0.611, calculated ‘r’=0.78, ‘a’=2.68 and ‘b’=3.79 indicating positive allometric growth pattern. Amylase exhibited high activity in the stomach, while lipase and proteinase in the stomach and posterior intestine. Specific activities of digestive enzymes showed significant differences (p<0.05). Viscerosomatic (2.92± 0.25), hepatosomatic (2.27± 0.22) indices and Fulton condition factor (1.93± 0.06) were recorded. Feed items present in the species confirm its overlapping feeding habit, indicating that the species is an omnivorous detritivore. This was also depicted in the activities of the different digestive enzymes. [ABSTRACT FROM AUTHOR]