Antibiotic susceptibility and genomic analysis of ciprofloxacin-resistant and ESBLs-producing Escherichia coli in vegetables and their irrigation water and growing soil.

The rise of antibiotic resistance in Escherichia coli has become a major global public health concern. While there is extensive research on antibiotic-resistant E. coli from human and animal sources, studies on vegetables and their environments are limited. This study investigated the prevalence and characteristics of ciprofloxacin-resistant (CIPR) E. coli in 13 types of edible raw vegetables, along with their irrigation water and soil in Shaanxi, China. Of 349 samples collected (157 vegetables, 59 water, and 133 soil), a total of 48 positive samples were detected, with one CIPRE. coli strain isolated from each sample being selected for further analyses. A striking observation was its high prevalence in irrigation water at 44.1 %, markedly exceeding that in vegetables (12.0 %) and soil (4.5 %). The susceptibility of Forty-eight CIPRE. coli isolates was evaluated using the disc diffusion method for 18 different antibiotics, all these isolates were not only resistant to the tested fluoroquinolones antibiotics (levofloxacin, nalidixic acid), but also displayed a multi-drug resistance (MDR) pattern. Twenty-eight (58.3 %) of 48 CIPRE. coli isolates exhibited extended spectrum ß-lactamases (ESBLs) (CIPR-ESBLs) producing phenotype. Subsequently, whole-genome sequencing was performed on these 28 isolates. We identified 12 serotypes and STs each, with O101: H9 (35.7 %, 10/28) and ST10 (21.4 %, 6/28) being the most common. Further classification placed these isolates into five phylogenetic groups: A (57.1 %, 16/28), B1 (32.1 %, 9/28), D (3.6 %, 1/28), B2 (3.6 %,1/28), and F (3.6 %,1/28). Notelly, Identical ST types, serotypes and phylogroups were found in certain CIPR-ESBLs-producing E. coli from both vegetables and adjacent irrigation water. Genomic analysis of the 28 CIPR-ESBLs-producing E. coli isolates unveiled 73 resistance genes, associated with 13 amino acid mutations in resistance-determining regions (QRDRs) and resistance to 12 types of antibiotics. Each isolate was confirmed to carry both ESBLs and fluoroquinolone resistance genes, with the Ser83Ala mutation in GyrA (96.4 %, 27/28) being the most prevalent. A detailed analysis of Mobile Genetic Elements (MGEs) revealed that IncFIB and IncFII plasmid subtypes were most prevalent in 60.7 % and 67.9 % of isolates, respectively, with 75 % containing over 10 insertion sequences (IS) each. Furthermore, we observed that certain ESBL and PMQR genes were located on plasmids or in proximity to insertion sequences. In conclusion, our research highlights the widespread presence of CIPRE. coli in irrigation water and thoroughly examines the genetic characteristics of CIPR-ESBLs-producing E. coli strains, underlining the need for ongoing monitoring and management to reduce multidrug-resistant bacteria in vegetables and their environment.

Prevalence and phylogenetic traits of nitrite-producing bacteria in raw ingredients and processed baby foods: Potential sources of foodborne infant methemoglobinemia.

Nitrite, which has been mainly regarded as a chemical hazard, can induce infant methemoglobinemia. As for nitrite as a product of microbial metabolism, the contribution of the oral or gut microbiome has mostly received attention, whereas the role of nitrite-producing bacteria (NPBs) in food has been less elucidated. In this study, mesophilic NPBs were isolated from food samples (n = 320) composed of raw ingredients for weaning foods (n = 160; beetroot, broccoli, carrot, lettuce, rice powder, spinach, sweet potato, and honey) and processed baby foods (n = 160; cereal snack, cheese, yogurt, powdered infant formula, sorghum syrup, vegetable fruit juice, and weaning food). The phylogenetic diversity of the NPB strains was analyzed via 16S rRNA sequencing. All 15 food items harbored NPBs, with a prevalence of 71.9 % and 34.4 % for the raw ingredients and processed foods, respectively. The NPBs isolated from the foods were identified as Actinomycetota (Actinomycetes), Bacteroidota (Flavobacteriia, Sphingobacteriia), Bacillota (Bacilli), or Pseudomonadota (Alpha-, Beta-, and Gammaproteobacteria). Among the raw and processed foods, beetroot (85.0 %) and powdered infant formula (70.0 %) showed had the highest NPB prevalence (P > 0.05). Bacillota predominated in both types of food. The contamination source of Pseudomonadota, which was another major phylum present in the raw ingredients, was presumed to be the soil and endophytes in the seeds, whereas that of Bacillota was the manufacturing equipment used with the raw ingredients. Common species for probiotics, such as Lacticaseibacillus, Leuconostoc, Enterococcus, and Bacillus, were isolated and identified as NPBs. To our knowledge, this is the first study to reveal the taxonomical diversity and omnipresence of NPBs in food for babies. The results of this study highlight the importance of food-mediated microbiological risks of infant methemoglobinemia which are yet underrecognized.

Unlocking sustainable solutions: Nanocellulose innovations for enhancing the shelf life of fruits and vegetables - A comprehensive review.

Regarding food security and waste reduction, preserving fruits and vegetables is a vital problem. This comprehensive study examines the innovative potential of coatings and packaging made of nanocellulose to extend the shelf life of perishable foods. The distinctive merits of nanocellulose, which is prepared from renewable sources, include exceptional gas barrier performance, moisture retention, and antibacterial activity. As a result of these merits, it is a good option for reducing food spoilage factors such as oxidation, desiccation, and microbiological contamination. Nanocellulose not only enhances food preservation but also complies with industry-wide environmental objectives. This review explores the many facets of nanocellulose technology, from its essential characteristics to its use in the preservation of fruits and vegetables. Furthermore, it deals with vital issues including scalability, cost-effectiveness, and regulatory constraints. While the use of nanocellulose in food preservation offers fascinating potential, it also wants to be cautiously careful to assure affordability, effectiveness, and safety. To fully use the potential of nanocellulose and advance the sustainability plan in the food business, collaboration between scientists, regulatory bodies, and industry stakeholders is important as we stand on the cusp of a revolutionary era in food preservation.

Analysis on Foodborne Pathogen Contamination of Food Samples in Longnan City from 2013 to 2022.

The objective was to determine the prevalence of foodborne pathogens in food in Longnan City, Gansu Province, China. In this research, we conducted tests on baked foods, catering foods, meat, and fruits and vegetables sold in supermarkets, farmers' markets, restaurants, retail stores, street stalls, and school canteens from 2013 to 2022. We analyzed the variety of foodborne pathogens (Salmonella, Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, and diarrheagenic Escherichia coli) in different sites and food types. Once foodborne pathogens were detected in the sample, it was deemed unqualified. The total detection rates of foodborne pathogens were 1.559%, 3.349%, 1.980%, 1.040%, 3.383%, and 1.303% in food from supermarkets, farmers' markets, restaurants, retail stores, street stalls, and school canteens, respectively. No pathogenic bacteria were detected in baked foods. Salmonella, S. aureus, L. monocytogenes, B. cereus, and diarrheagenic E. coli were detected in catering foods, among which B. cereus had the highest detection rate. Salmonella was the most common pathogenic bacteria detected in meat, while the detection rate of pathogenic bacteria in fruits and vegetables was low, with only one positive sample for diarrheagenic E. coli. Among the six sites, street stalls (3.382%) and farmers' markets (3.349%) had higher detection rates of pathogens. In general, the detection rate of pathogens from 2013 to 2022 was not high, but there were also some hidden dangers. Catering food is vulnerable to pathogen contamination, and street stalls and farmers' markets are the main sites of pollution. According to the above findings, the regulatory authorities should continue to strengthen supervision, guarantee food safety through early warning, and reduce the risk of food contamination.

Emergence of colistin-resistant Enterobacter cloacae and Raoultella ornithinolytica carrying the phosphoethanolamine transferase gene, mcr-9, derived from vegetables in Japan.

IMPORTANCE: Plasmid-mediated mobile colistin-resistance genes have been recognized as a global threat because they jeopardize the efficacy of colistin in therapeutic practice. Here, we described the genetic features of two mcr-9.1-carrying Gram-negative bacteria with a colistin-resistant phenotype derived from vegetables in Japan. The colistin-resistant mcr-9.1, which has never been detected in vegetables, was located on a large plasmid in Enterobacter cloacae CST17-2 and Raoultella ornithinolytica CST129-1, suggesting a high chance of horizontal gene transfer. To the best of our knowledge, this is the first report of mcr-9 in R. ornithinolytica. This study indicates that fresh vegetables might be a potential source for the transmission of mcr-9 genes encoding resistance to frontline (colistin) and clinically relevant antimicrobials. The study also provides additional consideration for colistin use and the relevance of routine surveillance in epidemiological perspective to curb the continuous spread of mcr alleles.

Endophytic bacterial communities in ungerminated and germinated seeds of commercial vegetables.

Chile is a prominent seed exporter globally, but the seed microbiome of vegetables (46% of seeds) and its role in the early stages of plant growth have remained largely unexplored. Here, we employed DNA metabarcoding analysis to investigate the composition and putative functions of endophytic bacterial communities in ungerminated and germinated seeds of the commercial vegetables Apiaceae (parsley and carrot), Asteraceae (lettuce), Brassicaceae (cabbage and broccoli), and Solanaceae (tomato). Bacterial quantification showed 104 to 108 copies of the 16S rRNA gene per gram of ungerminated and germinated seeds. Alpha diversity analysis (e.g., Chao1, Shannon, and Simpson indices) did not indicate significant differences (Kruskal-Wallis test) between ungerminated and germinated seeds, except for Solanaceae. However, beta diversity (PCoA) analysis showed distinctions (Adonis test) between ungerminated and germinated seeds, except Apiaceae. Pseudomonadota and Bacillota were identified as the dominant and specialist taxa in both ungerminated and germinated seed samples. Chemoheterotrophy and fermentation were predicted as the main microbial functional groups in the endophytic bacterial community. Notably, a considerable number of the 143 isolated endophytic strains displayed plant growth-promoting traits (10 to 64%) and biocontrol activity (74% to 82%) against plant pathogens (Xanthomonas and Pseudomonas). This study revealed the high variability in the abundance, diversity, composition, and functionality of endophytic bacteria between ungerminated and germinated seeds in globally commercialized vegetables. Furthermore, potential beneficial endophytic bacteria contained in their seed microbiomes that may contribute to the microbiome of the early stages, development, growth and progeny of vegetables were found.

Growth models for Salmonella, E. coli O157:H7 and L. monocytogenes give different predictions for pathogen growth in cut leafy greens transportation, but are consistent in identifying higher risk conditions.

Leafy greens are frequently implicated in foodborne disease outbreaks and cut-leafy greens are a food that requires time and temperature control for safety. Predictive microbiology uses mathematical models to predict the growth of bacteria based on environmental conditions. The objective of our study was to compare published square root growth models for Salmonella (n = 6), pathogenic E. coli (n = 6) and Listeria monocytogenes (n = 4) using real world transport temperature data. Data from trucks transporting fresh-cut leafy greens during cross-country shipments were used as temperature inputs to the models. Bacterial growth was computed using the temperatures from each probe in every truck over the duration of transit, which resulted in 12-18 growth predictions per truck for each model. Each model generally gave significantly different predictions than other models for the same organism. The exception was for the two Salmonella models predicting the least growth and the two Salmonella models predicting the most growth which gave predictions that were not significantly different. Although different models tended to give different predictions, their ability to rank risk by truck was generally consistent across models. While absolute risk might be dependent upon choice of model, relative risk is independent of model choice.

LysP53 activity against Salmonella and its application in decontamination of Salmonella on fresh romaine lettuce.

Salmonella is a zoonotic pathogen that is commonly associated with foodborne disease outbreaks. This study found that a newly identified Gram-negative lysin LysP53 had good activity against a wide range of Salmonella, including Salmonella Newington, Salmonella Typhimurium, and Salmonella Dublin. Without the help of an outer membrane permeabilizer, 4 µM LysP53 could reduce 97.6% of planktonic Salmonella Enteritidis and 90% of the bacteria in biofilms. Moreover, LysP53 was highly thermostable because it maintained >90% activity even after exposure to temperatures up to 95 °C. Although high concentrations of salts could reduce the activity, LysP53 was found safe for oral gavage of mice without affecting body weights and cytokines in sera and able to reduce 90% of Salmonella Enteritidis loads on fresh romaine lettuce after 30 min of treatment. Because of its good activity against a wide range of bacteria, thermal stability, safe for oral administration, LysP53 could be used as a biocontrol agent for reducing bacterial loads in fresh vegetable food. KEY POINTS: • Lysin LysP53 has high bactericidal activity against Salmonella. • LysP53 is thermostable even at high temperature of up to 95 °C. • LysP53 can be used for topical decontamination of Salmonella on vegetables.

Ranking food products based on estimating and combining their microbiological, chemical and nutritional risks: Method and application to Ready-To-Eat dishes sold in France.

Microbiological (M), chemical (C), and nutritional (N) risks associated with food products are usually assessed and managed independently by experts in public services or food companies. This can render difficult the comparison of food products in term of overall risk for the consumer. The objective of this study was to suggest a relatively simple method to (i) classify food products based on their M, C and N risks, and (ii) aggregate these risks and rank the food products accordingly. The method was developed and applied to 17 ready-to-eat (RTE) dishes available on the French market. With regard to food safety, the individual M and C risks were characterized considering likelihood and severity as recommended by the Codex Alimentarius. With regard to nutrition/health, the N risk was estimated based on the tendency of the dish to contribute to nutrient adequacy and to a healthy eating pattern. Finally, the outranking method PROMETHEE was applied to aggregate the three M, C, N risks and rank the food dishes. Food products were ranked relatively to each other, not in absolute terms. When we attributed the same weight to M, C and N risks, the RTE dish "Duck Parmentier" had the highest risk score while "Papillote of chicken, potatoes and small vegetables" and "Vegetarian plate vegetables and quinoa" had the lowest. However, this overall ranking changed according to the weight assigned to individual M, C and N risks, at least for food products whose scores varied according to risk types, such as"sushi discovery" (high M and C risks, low N risk). Since the risk ranking method developed here was built with assumptions and hypotheses related to the specific case study, more applications are needed to assess whether it can be generic. Nevertheless, this method is well grounded, objective, transparent, relatively fast and easy to set up. It might lead to further development of decision tools, particularly for consumers. This study paves the way towards food product multi-risk ranking.

Quantitative proteomics analysis of bacteriocin-like substance producing Lactococcus spp. in simulated vegetable and fruit media.

Lactic acid bacteria (LAB) and their metabolites such as bacteriocins have gained considerable interest in terms of their bio-preservative properties to improve food safety and quality. In this study, a quantitative proteomic investigation employing stable isotope labeling by peptide demethylation was carried out to investigate changes in intracellular proteins of bacteriocin-like substance (BLS) producing Lactococcus spp. 7.17 grown in vegetable or fruit juice culture media at 10 °C for 0, 3 or 7 days. In total, 1053 proteins in vegetable medium and 1113 in fruit medium were identified and quantified. Proteins that changed more than two- fold were identified as increased or decreased ones and grouped into four clusters. Those increased proteins were involved in the events of low temperature and ROS stress responses, DNA processing, transcription and translation, central carbon metabolism, fatty acid and phospholipid metabolism, amino acid and cell wall biosynthesis. Key proteins in relation to BLS producing property were also identified suggesting that at least one bacteriocin IIa production system exists in Lactococcus spp. 7.17. These findings provide insights into protein changes of L. lactis at low temperature and lay foundations for further investigations on BLS producing LAB using targeted quantitative proteomic approaches. SIGNIFICANCE OF THE RESEARCH: The inhibitory effects of Lactococcus spp. 7.17 on Listeria innocua in fruit and vegetable juice culture media were confirmed. Using a quantitative proteomic approach employing stable isotope labeling by peptide demethylation, 99 or 113 significantly changed proteins of Lactococcus spp. 7.17 grown in vegetable or fruit juice medium were determined, respectively. The significant change in protein abundance suggested an adaptive mechanism of Lactococcus spp. to culture condition at low temperatures. This research provides insights on protein changes of Lactococcus spp. which has potential application in fresh and fresh-cut fruit and vegetables at low temperature.

Impacts of agroecological practices on soil microbial communities in experimental open-field vegetable cropping systems.

This study aimed to determine the impact of different agroecological practices on the composition and diversity of edaphic bacterial and fungal communities. We designed two experimental agroecological vegetable cropping systems and analyzed their effects on soil microbial communities by pyrosequencing the 16S and 18S ribosomal RNA genes. Our results highlighted modifications to the Operational Taxonomic Units in both experimental systems compared with bare soil, particularly for the phyla Actinobacteria, Ascomycota, Bacteroidetes, and Mucoromycota. Multidimensional scaling plots based on beta diversity showed a clear distinction between the two experimental systems for fungi, whereas differences were observed between bare soil and the two experimental systems for bacteria. Overall, the agroecological systems enhanced soil microbial diversity. We showed a distinction between the two experimental systems and bare soil, correlated with the high total N and total P contents in the agroecological systems. Both experimental systems promoted soil enrichment with certain essential minerals. The agroecological systems had a positive impact on soil microbial communities, particularly by promoting the development of beneficial soil bacteria like Actinobacteria. In the two experimental systems, changes in the quality and quantity of organic matter (i.e. mulch, vermicompost, plant diversity) could have modified the abundance and diversity of microbial communities.

Fresh vegetables and fruit as a source of Salmonella bacteria.

INTRODUCTION: Increased consumer awareness of the health aspects of the diet has influenced the increased consumption of fruit and vegetables. Due to the fact that these products are mainly consumed raw and are usually not subjected to processes that reduce their microbiological contamination, they become a source of infection and transmit pathogens causing food poisoning in humans. Salmonella bacteria are a serious treat to human health and remain a serious problem in many parts of the world. OBJECTIVE: The aim of this study was to review the current state of knowledge regarding the prevalence of Salmonella bacteria on fresh fruit and vegetables. Attention is also given to the mechanisms by which these bacteria adapt to colonize plants. Methods that can prevent contamination of plant products by the bacteria are also analyzed. REVIEW METHODS: The review was based on data obtained from scientific articles published in the Science Direct and Pub Med database between 2007 - 2022, found with the use of the following keywords: Salmonella, fruit, vegetables, food contamination. BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: Data from the literature report that fresh fruit and vegetables are a source of Salmonella contamination through contact with soil, manure, compost, water or staff. SUMMARY: Actions targeting salmonellosis prevention should be undertaken by both the public and private sectors. Government regulations and stricter measures put in place can provide a framework that guides both domestic production and international imports. Periodic training of workers dealing with food is also important. Attention should be directed mainly to production control and less to the testing of final products. Education leading to increased awareness of salmonellosis should be indispensable.

Are Fermented Foods Effective against Inflammatory Diseases?

Fermented foods have been used over the centuries in various parts of the world. These foods are rich in nutrients and are produced naturally using various biological tools like bacteria and fungi. Fermentation of edible foods has been rooted in ancient cultures to keep food for preservation and storage for a long period of time with desired or enhanced nutritional values. Inflammatory diseases like rheumatoid arthritis, osteoarthritis, and chronic inflammatory pain are chronic disorders that are difficult to treat, and current treatments for these disorders fail due to various adverse effects of prescribed medications over a long period of time. Fermented foods containing probiotic bacteria and fungi can enhance the immune system, improve gastrointestinal health, and lower the risk of developing various inflammatory diseases. Foods prepared from vegetables by fermentation, like kimchi, sauerkraut, soy-based foods, or turmeric, lack proper clinical and translational experimental studies. The current review has focused on the effectiveness of various fermented foods or drinks used over centuries against inflammation, arthritis, and oxidative stress. We also described potential limitations on the efficacies or usages of these fermented products to provide an overarching picture of the research field.

Salmonella and other Enterobacteriaceae in conventional and organic vegetables grown in Brazilian farms.

This study aimed to assess the microbiological profile of conventional and organic vegetables grown in Brazilian farms through the detection of Salmonella and other Enterobacteriaceae. A total of 200 samples (100 conventional and 100 organic), including leafy greens, spices/herbs, and other unusual vegetables, were submitted to the enumeration of Enterobacteriaceae by plating on VRBG agar. Moreover, colonies of Enterobacteriaceae were randomly selected and submitted to identification by MALDI-TOF MS. Samples were also tested for Salmonella, using culture-based and PCR-based enrichment methods. The mean counts of Enterobacteriaceae in conventional and organic vegetables were 5.1 ± 1.5 and 5.4 ± 1.4 log CFU/g, respectively (P > 0.05). A total of 18 genera (including 38 species) of Enterobacteriaceae were identified, and the most frequent ones found in samples from both farming systems were Enterobacter (76%) and Pantoea (68%). Salmonella was identified in 17 samples (8.5%): nine (4.5%) in conventional and eight (4.0%) in organic vegetables. These results indicate that the farming system had no impact on the Enterobacteriaceae populations and rates of Salmonella and revealed unsatisfactory microbiological safety of some samples, mainly due to the presence of Salmonella. These findings highlight the need for control measures during vegetable production, regardless of the farming system, to reduce microbial contamination and the risks of foodborne illnesses.

Profiling the composition and metabolic functions of microbial community in pellicle-forming radish paocai.

Pellicle formation is an obvious indicator of spoilage and is followed by a loss of flavor in a variety of fermented vegetables. In this study, the pellicle-forming microorganisms were isolated using culture-dependent approaches, then a comparative analysis between the pellicle-forming (PF) radish paocai and normal fermented paocai in the diversity and function of microbial community was conducted by metagenome sequencing. Based on a pairwise t-test and OPLS-DA analysis, diallyl sulfide, (z)-1-allyl-2-(prop-1-en-1-yl) disulfane, and terpineol were considered to be the main components responsible for the unpleasant flavor of PF paocai. Yarrowia spp., Enterobacter spp., and Pichia spp. were the main pellicle-forming microorganisms. All 17 isolated Enterobacter strains showed pectinase-producing and cellulase-producing abilities, and 3 isolated Pichia strains showed gas-producing capacity. According to LEfSe analysis based on metagenomes, unclassified_g__Citrobacter and Yarrowia lipolytica were the uppermost biomarkers that distinguished the PF paocai from normal paocai. Unclassified_g__Lactobacillus and Lactobacillus plantarum were found to be actively engaged in starch and sucrose metabolism, cysteine and methionine metabolism, galactose metabolism, fructose and mannose metabolism, lysine biosynthesis, fatty acid biosynthesis, and arginine biosynthesis, all of which contributed to the flavor formation of paocai. Combining the results of metagenome sequencing with the data obtained based on the culture-dependent method, we could deduce that the growth of Yarrowia lipolytica first promoted the increase of pH and the formation of pellicle, which provided a suitable niche for the growth of some harmful bacteria such as Enterobacter, Citrobacter, and Serratia. These hazardous bacteria then worked in concert to induce the odorous stench and texture softening of paocai, as well as more pellicle formation.

Microbiological food safety considerations in shelf-life extension of fresh fruits and vegetables.

There are a number of opportunities for reducing loss and waste, and extending shelf life of fresh produce that go beyond cold chain optimization. For example, plant genotype (including ripening-related genes), presence of phytopathogens, maturity at harvest, and environmental conditions close to the harvest time, storage conditions, and postharvest treatments (washing, cutting, and waxing) all impact both shelf life of produce and food safety outcomes. Therefore, loss can be reduced and shelf life of fresh produce can be extended with plant breeding to manipulate ripening-related traits, or with pre- and postharvest treatments delaying senescence and decay. Food safety considerations of these applications are discussed.

Ready-To-Eat Rocket Salads as Potential Reservoir of Bacteria for the Human Microbiome.

Reportedly, Western-type diets may induce the loss of key microbial taxa within the gastrointestinal microbiota, promoting the onset of noncommunicable diseases. It was hypothesized that the consumption of raw vegetables could contribute to the maintenance of the intestinal microbial community structure. In this context, we explored bacteria associated with commercial rocket salads produced through different farming practices: traditional (conventional, organic, and integrated) and vertical farming. Viable counts of mesophilic bacteria and lactic acid bacteria (LAB) were performed on plate count agar (PCA) and de Man-Rogosa-Sharpe (MRS) agar at pH 5.7, whereas metataxonomics through 16S rRNA gene sequencing was used to profile total bacteria associated with rocket salads. We found that rocket salads from vertical farming had much fewer viable bacteria and had a bacterial community structure markedly different from that of rocket salads from traditional farming. Furthermore, although α- and ß-diversity analyses did not differentiate rocket samples according to farming techniques, several bacterial taxa distinguished organic and integrated from conventional farming salads, suggesting that farming practices could affect the taxonomic composition of rocket bacterial communities. LAB were isolated from only traditional farming samples and belonged to different species, which were variably distributed among samples and could be partly associated with farming practices. Finally, the INFOGEST protocol for in vitro simulation of gastrointestinal digestion revealed that several taxonomically different rocket-associated bacteria (particularly LAB) could survive gastrointestinal transit. This study suggests that commercial ready-to-eat rocket salads harbor live bacteria that possess the ability to survive gastrointestinal transit, potentially contributing to the taxonomic structure of the human gut microbiota. IMPORTANCE Western-type diets are composed of foods with a reduced amount of naturally occurring microorganisms. It was hypothesized that a microbe-depleted diet can favor the alteration of the human intestinal microbial ecosystem, therefore contributing to the onset of chronic metabolic and immune diseases currently recognized as the most significant causes of death in the developed world. Here, we studied the microorganisms that are associated with commercial ready-to-eat rocket salads produced through different farming practices. We showed that rocket salad (a widely consumed vegetal food frequently eaten raw) may be a source of lactic acid bacteria and other microbes that can survive gastrointestinal transit, potentially increasing the biodiversity of the intestinal microbiota. This deduction may be valid for virtually all vegetal foods that are consumed raw.

Use of Vegetable Waste as a Culture Medium Ingredient Improves the Antimicrobial and Immunomodulatory Activities of Lactiplantibacillus plantarum WiKim0125 Isolated from Kimchi.

Lactic acid bacteria (LAB) isolated from kimchi (a traditional Korean dish typically made of fermented cabbage) can provide various health benefits, including anti-obesity, antioxidant, anti-inflammatory, anticancer, and antimicrobial effects. In this study, we examined the antimicrobial and immunomodulatory effects of Lactiplantibacillus plantarum WiKim0125 cultured in de Man, Rogosa, and Sharpe (MRS) medium containing vegetable waste. Live bacterial cells were eliminated via supernatant filtration or heat treatment. The cell-free supernatant (CFS) obtained from culture broth containing kimchi cabbage waste (KCW), cabbage waste (CW), or onion waste (OW) showed significantly higher antimicrobial activity against skin pathogens (Propionibacterium acnes and Staphylococcus aureus) and foodborne pathogens (Escherichia coli and Salmonella typhimurium), with inhibition zones ranging between 4.4 and 8.5 mm, compared to that in conventional MRS medium (4.0-7.3 mm). In lipopolysaccharide-stimulated RAW264.7 cells, both supernatant and heat-inactivated Lb. plantarum WiKim0125 from culture media containing KCW and CW suppressed the production of inflammatory cytokines (72.8% and 49.6%, respectively) and nitric oxide (62.2% and 66.7%, respectively) without affecting cell viability. These results indicate that vegetable waste can potentially increase the antimicrobial and immunoregulatory potency of LAB while presenting a molecular basis for applying postbiotics to health products.

Agroecosystem exploration for Antimicrobial Resistance in Ahmedabad, India: A Study Protocol.

Introduction: Antimicrobial resistance (AMR) has emerged as one of the leading threats to public health. AMR possesses a multidimensional challenge that has social, economic, and environmental dimensions that encompass the food production system, influencing human and animal health. The One Health approach highlights the inextricable linkage and interdependence between the health of people, animal, agriculture, and the environment. Antibiotic use in any of these areas can potentially impact the health of others. There is a dearth of evidence on AMR from the natural environment, such as the plant-based agriculture sector. Antibiotics, antibiotic-resistant bacteria (ARB), and related AMR genes (ARGs) are assumed to present in the natural environment and disseminate resistance to fresh produce/vegetables and thus to human health upon consumption. Therefore, this study aims to investigate the role of vegetables in the spread of AMR through an agroecosystem exploration in Ahmedabad, India. Protocol: The present study will be executed in Ahmedabad, located in Gujarat state in the Western part of India, by adopting a mixed-method approach. First, a systematic review will be conducted to document the prevalence of ARB and ARGs on fresh produce in South Asia. Second, agriculture farmland surveys will be used to collect the general farming practices and the data on common vegetables consumed raw by the households in Ahmedabad. Third, vegetable and soil samples will be collected from the selected agriculture farms and analyzed for the presence or absence of ARB and ARGs using standard microbiological and molecular methods. Discussion: The analysis will help to understand the spread of ARB/ARGs through the agroecosystem. This is anticipated to provide an insight into the current state of ARB/ARGs contamination of fresh produce/vegetables and will assist in identifying the relevant strategies for effectively controlling and preventing the spread of AMR.

What Is in the Salad? Escherichia coli and Antibiotic Resistance in Lettuce Irrigated with Various Water Sources in Ghana.

INTRODUCTION: Safety of the environment in which vegetables are grown, marketed and consumed is paramount as most are eaten raw. Irrigation sources include open drains and streams, which are often contaminated with human and animal waste due to poor sanitation infrastructure. In irrigated vegetable farms using such sources in Ghana, we assessed Escherichia coli counts, antibiotic resistance patterns and resistant genes on irrigated lettuce. METHODS: A cross-sectional study was conducted between January-May 2022, involving five major vegetable farms in Ghana. RESULTS: Escherichia coli was found in all 25 composite lettuce samples analyzed. Counts expressed in CFU/g ranged from 186 to 3000, with the highest counts found in lettuce irrigated from open drains (1670) and tap water using hose pipes (3000). Among all bacterial isolates, resistance ranged between 49% and 70% for the Watch group of antibiotics, 59% for the Reserved group and 82% were multidrug-resistant. Of 125 isolates, 60 (48%) were extended-spectrum beta-lactamase-producing, of which five (8%) had the blaTEM-resistant gene. CONCLUSIONS: Lettuce was contaminated with Escherichia coli with high levels of antibiotic resistance. We call on the Ghana Ministry of Food and Agriculture, Food and Drugs Authority and other stakeholders to support farmers to implement measures for improving vegetable safety.