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This study aimed to investigate the probiogenomic features of artisanal bacteriocin-producing Enterococcus faecium BGPAS1-3 and the use of the improved pMALc5HisEk expression vector for overexpressing class II bacteriocins and the application of purified bacteriocin 31 in a milk model as a preservative against L. monocytogenes. The BGPAS1-3 strain was isolated from traditional fresh soft cheese manufactured in households on a small scale in rural locations surrounding Pale Mountain City in Bosnia and Herzegovina. The whole-genome sequencing approach and bioinformatics analyses revealed that the strain BGPAS1-3 was non-pathogenic to humans. The presence of bacteriocin operons suggested the ability of the isolate to suppress the growth of pathogens. Coding regions for three maturated bacteriocins (bacteriocin 31, bacteriocin 32, and enterocin P) produced by BGPAS1-3 were amplified and expressed in Escherichia coli ER2523 using the pMALc5HisEk system. All three bacteriocins were successfully overexpressed and purified after enterokinase cleavage but showed different antimicrobial activity. Bacteriocin 31 showed significantly stronger antimicrobial activity compared with bacteriocin 32. It was the only one that proved to be suitable for use as a food preservative against L. monocytogenes in a milk model.

Raw milk cheeses harbor complex microbial communities. Some of these microorganisms are technologically essential, but undesirable microorganisms can also be present. While most of the microbial dynamics and cross-talking studies involving interaction between food-derived bacteria have been carried out on agar plates in laboratory-controlled conditions, the present study evaluated the modulation of the resident microbiota and the changes of metabolite production directly in ripening raw milk cheese inoculated with Listeria innocua strains. Using a proxy of the pathogenic Listeria monocytogenes, we aimed to establish the key microbiota players and chemical signals that characterize Latteria raw milk cheese over 60 days of ripening time. The microbiota of both the control and Listeria-inoculated cheeses was analyzed using 16S rRNA targeted amplicon sequencing, while direct analysis in real time mass spectrometry (DART-HRMS) was applied to investigate the differences in the metabolic profiles of the cheeses. The diversity analysis showed the same microbial diversity trend in both the control cheese and the inoculated cheese, while the taxonomic analysis highlighted the most representative genera of bacteria in both the control and inoculated cheese: Lactobacillus and Streptococcus. On the other hand, the metabolic fingerprints revealed that the complex interactions between resident microbiota and L. innocua were governed by continuously changing chemical signals. Changes in the amounts of small organic acids, hydroxyl fatty acids, and antimicrobial compounds, including pyroglutamic acid, hydroxy-isocaproic acid, malic acid, phenyllactic acid, and lactic acid, were observed over time in the L. innocua-inoculated cheese. In cheese that was inoculated with L. innocua, Streptococcus was significantly correlated with the volatile compounds carboxylbenzaldheyde and cyclohexanecarboxylic acid, while Lactobacillus was positively correlated with some volatile and flavor compounds (cyclohexanecarboxylic acid, pyroxidal acid, aminobenzoic acid, and vanillic acid). Therefore, we determined the metabolic markers that characterize a raw milk cheese inoculated with L. innocua, the changes in these markers with the ripening time, and the positive correlation of flavor and volatile compounds with the resident microbiota. This multi-omics approach could suggest innovative food safety strategies based on the enhanced management of undesirable microorganisms by means of strain selection in raw matrices and the addition of specific antimicrobial metabolites to prevent the growth of undesirable microorganisms.

AimsRecent foodborne disease outbreaks have caused farmers to re-evaluate their practices. In particular, concern that soil amendments could introduce foodborne pathogens onto farms and promote their survival in soils has led farmers to reduce or eliminate the application of animal-based composts. However, organic amendments (such as composts and cover crops) could bolster food safety by increasing soil microbial diversity and activity, which can act as competitors or antagonists and reduce pathogen survival.Methods and resultsLeveraging a study of a 27-year experiment comparing organic and conventional soil management, we evaluate the impacts of composted poultry litter and cover crops on soil chemistry, bacterial communities and survival of Salmonella enterica and Listeria monocytogenes. We found that bacterial community composition strongly affected pathogen survival in soils. Specifically, organic soils managed with cover crops and composts hosted more macronutrients and bacterial communities that were better able to suppress Salmonella and Listeria. For example, after incubating soils for 10 days at 20°C, soils without composts retained fourfold to fivefold more Salmonella compared to compost-amended soils. However, treatment effects dissipated as bacterial communities converged over the growing season.ConclusionsOur results suggest that composts and cover crops may be used to build healthy soils without increasing foodborne pathogen survival.Significance and impact of the studyOur work suggests that animal-based composts do not promote pathogen survival and may even promote bacterial communities that suppress pathogens. Critically, proper composting techniques are known to reduce pathogen populations in biological soil amendments of animal origin, which can reduce the risks of introducing pathogens to farm fields in soil amendments. Thus, animal-based composts and cover crops may be a safe alternative to conventional fertilizers, both because of the known benefits of composts for soil health and because it may be possible to apply amendments in such a way that food-safety risks are mitigated rather than exacerbated.

Sous-vide is a cooking technique which is considered safe for food preparation, however, the recommended temperatures and times for preparing seafood and fish may not be sufficient to ensure safety, and additionally, thermal stress may cause changes in virulence and resistance. The objective of the present investigation was to determine the reduction of Listeria monocytogenes isolated from fish after sous-vide preparation. For this purpose flow cytometry was used. The impact of stress during the preparation of the fish-based dish using the sous-vide technique on the virulence potential and resistance profile was determined geno- and phenotypically. The study was conducted at four different temperatures, that are typical for fish preparation (42, 52, 57 and 60 °C) showing that none of them could be effective against L. monocytogenes. In addition, the investigation of changes in the virulence, phenotypic antibiotic resistance and gene expression showed that the stress during the preparation of sous-vide decrease virulence potential of the L. monocytogenes strains. It was observed that the greatest changes in antibiotic resistance profile were noted for tetracycline and notable changes were also observed for some strains in the case of fosfomycin and ciprofloxacin. Therefore it is important not to consider the sous-vide method as ensuring the microbiological quality of products, especially those with high contamination levels, and choose products of the best quality for processing.

ABSTRACT Since 2010, the genus Listeria has had the addition of 22 new species that more than tripled the number of species identified until 2010. Sixteen of these 22 new species are distantly related to the type species, Listeria monocytogenes, and several of these present phenotypes that distinguish them from classical Listeria species (L. monocytogenes, Listeria innocua, Listeria ivanovii, Listeria seeligeri, Listeria welshimeri, and Listeria grayi). These 22 newly described species also show that Listeria is more genetically diverse than previously estimated. While future studies and surveys are needed to clarify the distribution of these species, at least some of these species may not be widely spread, while other species may be frequently found spread to human-related settings (e.g., farms and processing facilities), and others may be adapted to specific environmental habitats. Here, we review the taxonomic, phylogenetic, and ecological characteristics of these new Listeria species identified since 2010 and re-iterate the suggestion of re-classification of some species into three new genera: Murraya, Mesolisteria, and Paenilisteria. We also provide a review of current detection issues and the relevance to food safety related to the identification of these new species. For example, several new non-pathogenic species could be misidentified as the pathogen L. monocytogenes, based on methods that do not target L. monocytogenes-specific virulence genes/factors, leading to unnecessary product recalls. Moreover, eight species in the proposed new genus Mesolisteria are not good indicators of environmental conditions that could allow L. monocytogenes to grow since Mesolisteria species are unable to grow at low temperatures.

IntroductionThis study aimed to determine the prevalence and virulome of Listeria in fresh produce distributed in urban communities.MethodsA total of 432 fresh produce samples were collected from farmer’s markets in Michigan and West Virginia, USA, resulting in 109 pooled samples. Listeria spp. were isolated and L. monocytogenes was subjected to genoserogrouping by PCR and genotyping by pulsed-field gel electrophoresis (PFGE). Multi-locus sequence typing (MLST) and core-genome multi-locus sequence typing (cgMLST) were conducted for clonal identification.ResultsForty-eight of 109 samples (44.0%) were contaminated with Listeria spp. L. monocytogenes serotype 1/2a and 4b were recovered from radishes, potatoes, and romaine lettuce. Four clonal complexes (CC) were identified and included hypervirulent CC1 (ST1) and CC4 (ST219) of lineage I as well as CC7 (ST7) and CC11 (ST451) of lineage II. Clones CC4 and CC7 were present in the same romaine lettuce sample. CC1 carried Listeria pathogenicity island LIPI-1 and LIPI-3 whereas CC4 contained LIPI-1, LIPI-3, and LIPI-4. CC7 and CC11 had LIPI-1 only.DiscussionDue to previous implication in outbreaks, L. monocytogenes hypervirulent clones in fresh produce pose a public health concern in urban communities.

This is a review of U.S. Food and Drug Administration (FDA) recalls of products that are for dogs and cats which took place from 2003 through 2022. It includes recalls for pet foods (food, treats, and chews), ingredients, supplements (vitamins and minerals), and drugs. There were 3,691 recalls during this period: 51% were Class I, 35% were Class II, and 14% were Class III. Food items and ingredients accounted for the majority or 68%, drugs for 27%, and supplements (vitamins and minerals) accounted for 5% of these recalls. Recalls that could be associated with dogs only accounted for 42%, with cats only 18%, and with multiple species 40%. The primary reasons for the recalls were biological contamination at 35%, chemical contamination at 32%, and cGMP violations at 8%. Almost 25% of the total recalls in the past 20 years were due to a melamine incident in 2007/2008 (73% of those were Class I). Salmonella recalls for the 20 years accounted for 23% of the total recalls (94 % of those were Class I). Although the recalls for vitamins and minerals accounted for only 5.6% percent of the total, 70% of those were Class I and 30% Class II. Pet food is a complex part of the processed food industry, and the processing of pet food is subject to at least 40 different federal regulations. To avoid recalls and be successful, pet food manufacturers need a robust food safety culture to meet all of these requirements to produce a safe product. In contrast, the melamine contamination (an adulteration event) in 2007/2008 which resulted in animal deaths and recalls is a prime example of the need for an effective and robust supplier approval program in order to avoid fraudulent suppliers in the future.

Abstract This report by the European Food Safety Authority and the European Centre for Disease Prevention and Control presents the results of the zoonoses monitoring and surveillance activities carried out in 2022 in 27 Member States (MSs), the United Kingdom (Northern Ireland) and 11 non‐MSs. Key statistics on zoonoses and zoonotic agents in humans, food, animals and feed are provided and interpreted historically. In 2022, the first and second most reported zoonoses in humans were campylobacteriosis and salmonellosis, respectively. The number of cases of campylobacteriosis and salmonellosis remained stable in comparison with 2021. Nineteen MSs and the United Kingdom (Northern Ireland) achieved all the established targets in poultry populations for the reduction of Salmonella prevalence for the relevant serovars. Salmonella samples from carcases of various animal species, and samples for Campylobacter quantification from broiler carcases, were more frequently positive when performed by the competent authorities than when own checks were conducted. Yersiniosis was the third most reported zoonosis in humans, followed by Shiga toxin‐producing Escherichia coli (STEC) and Listeria monocytogenes infections. L. monocytogenes and West Nile virus infections were the most severe zoonotic diseases, with the most hospitalisations and highest case fatality rates. In 2022, reporting showed an increase of more than 600% compared with 2021 in locally acquired cases of human West Nile virus infection, which is a mosquito‐borne disease. In the EU, the number of reported foodborne outbreaks and cases, hospitalisations and deaths was higher in 2022 than in 2021. The number of deaths from outbreaks was the highest ever reported in the EU in the last 10 years, mainly caused by L. monocytogenes and to a lesser degree by Salmonella. Salmonella and in particular S. Enteritidis remained the most frequently reported causative agent for foodborne outbreaks. Norovirus (and other calicivirus) was the agent associated with the highest number of outbreak human cases. This report also provides updates on brucellosis, Coxiella burnetii (Q fever), echinococcosis, rabies, toxoplasmosis, trichinellosis, infection with Mycobacterium tuberculosis complex (focusing on Mycobacterium bovis and Mycobacterium caprae) and tularaemia.

Proper activation of cytotoxic T cells via the T cell receptor and the costimulatory receptor CD28 is essential for adaptive immunity against viruses, intracellular bacteria, and cancers. Through biochemical analysis of RNA:protein interactions, we uncovered a non-coding RNA circuit regulating activation and differentiation of cytotoxic T cells composed of the long non-coding RNA Malat1 (Metastasis Associated Lung Adenocarcinoma Transcript 1) and the microRNA family miR-15/16. miR-15/16 is a widely and highly expressed tumor suppressor miRNA family important for cell proliferation and survival. miR-15/16 play important roles in T cell responses to viral infection, including the regulation of antigen-specific T cell expansion and memory. Comparative Argonaute-2 high-throughput sequencing of crosslinking immunoprecipitation (AHC) combined with gene expression profiling in normal and miR-15/16-deficient mouse T cells revealed a large network of hundreds of direct miR-15/16 target mRNAs, many with functional relevance for T cell activation, survival and memory formation. Among these targets, Malat1 contained the largest absolute magnitude miR-15/16-dependent AHC peak. This binding site was among the strongest lncRNA:miRNA interactions detected in the T cell transcriptome. We used CRISPR targeting with homology directed repair to generate mice with a 5-nucleotide mutation in the miR-15/16-binding site in Malat1. This mutation interrupted Malat1:miR-15/16 interaction, and enhanced the repression of other miR-15/16 target genes, including CD28. Interrupting Malat1 interaction with miR-15/16 decreased cytotoxic T cell activation, including the expression of interleukin 2 (IL-2) and a broader CD28-responsive gene program. Accordingly, Malat1 mutation diminished memory cell persistence in mice following LCMV Armstrong and Listeria monocytogenes infection. This study marks a significant advance in the study of long non-coding RNAs in the immune system by ascribing cell-intrinsic, sequence-specific in vivo function to Malat1. These findings have implications for T cell-mediated autoimmune diseases, antiviral and anti-tumor immunity, as well as lung adenocarcinoma and other malignancies where Malat1 is overexpressed.

ABSTRACTA high-fat (HF) diet reduces resistance to the foodborne pathogen Listeria monocytogenes. We demonstrate that short-term gavage with A. muciniphila increases resistance to oral and systemic L. monocytogenes infection in mice fed a HF diet. A. muciniphila reduced inflammation in the gut and liver of mice fed a high-fat diet prior to infection and reduced inflammatory cell infiltration in the ileum to levels similar to mice fed a low-fat (LF) diet. Akkermansia administration had minimal impacts upon the microbiota and microbial metabolites and did not affect individual taxa or impact the Bacteroidetes to Firmicutes ratio. In summary, A. muciniphila increased resistance to L. monocytogenes infection in mice fed a HF diet by moderating immune/physiological effects through specific interaction between A. muciniphila and the host gut.

The accuracy of Oxford Nanopore Technology (ONT) sequencing has significantly improved thanks to new flowcells, sequencing kits, and basecalling algorithms. However, novel modification types untrained in the basecalling models can seriously reduce the quality. Here we reports a set of ONT-sequenced genomes with unexpected low quality due to novel modification types. Demodification by whole-genome amplification significantly improved the quality but lost the epigenome. We also developed a reference-based method, Modpolish, for correcting modification-mediated errors while retaining the epigenome when a sufficient number of closely-related genomes is publicly available (default: top 20 genomes with at least 95% identity). Modpolish not only significantly improved the quality of in-house sequenced genomes but also public datasets sequenced by R9.4 and R10.4 (simplex). Our results suggested that novel modifications are prone to ONT systematic errors. Nevertheless, these errors are correctable by nucleotide demodification or Modpolish without prior knowledge of modifications. The authors report excessive amounts of modification-mediated errors in Listeria strains during Nanopore sequencing that are due to novel modifications that were untrained in the Nanopore basecalling model. A whole-genome amplification and a reference-based computational methods (Modpolish) can correct these modification-mediated errors without the need for demodification. [ABSTRACT FROM AUTHOR]

Foodborne illness outbreaks in the U.S. associated with consumption of both fresh and dried specialty mushrooms have recently occurred. Dried wood ear mushrooms were implicated in a salmonellosis outbreak in 2020, while fresh enoki mushrooms were associated with two listeriosis outbreaks in 2020 and 2023. These specialty mushrooms are commercially available in both their fresh and dried states. Due to the short shelf life of mushrooms, dehydration is a common method used in both industry and by consumers to extend the shelf life and preserve quality. Therefore, the aim of this study was to evaluate the use of dehydration on the inactivation kinetics of both Listeria monocytogenes and Salmonella enterica on enoki and wood ear mushrooms. Fresh mushrooms were inoculated with four strain cocktails of either L. monocytogenes or S. enterica and dried at ambient conditions for 10  min. Following drying of the inoculum, mushrooms were placed into food dehydrators preheated to 70, 80, or 90°C and treated for up to 24  h. At treatment intervals, mushrooms were removed from the dehydrators for pathogen enumeration. Inactivation kinetics for both pathogens were modeled using the Weibull, log-linear with tail, and log-linear with shoulder models. Pathogen reductions of >4 log CFU/g were achieved on both enoki and wood ear mushrooms during dehydration at 90°C after only 2–4  h. At 70 and 80°C, log reductions of >4 log CFU/g were observed on wood ear mushrooms after 4–8  h. On enoki mushrooms, a tailing effect was observed with residual populations (>2 log CFU/g) of L. monocytogenes and S. enterica remaining even after 24  h of treatment at both 70 and 80°C. This study emphasizes the need for an individualized dehydration strategy for each mushroom type to ensure the effectiveness of dehydration as a process to reduce pathogen populations. Results of this study will aid in informing proper time and temperature combinations for dehydration of specialty mushrooms to ensure product safety. [ABSTRACT FROM AUTHOR]

The present study was aimed to find the prevalence of Listeria monocytogenes in raw chicken meat purchased across the Nagpur city. Cultural examination of 100 raw poultry meat samples revealed 35.0 % prevalence of Listeria spp. Among them, L. grayi was predominant (15 %) followed by L. monocytogenes (12 %), L. welshimeri (5 %), L. innocua (2 %) and L. seeligeri (1 %). A typical β-haemolysis clear zone was exhibited by 18 listeriae isolates. Of these, 12 isolates with characteristic biochemical pattern and showing haemolytic zone. All 12 L. monocytogenes isolates expressed the PI PLC activity. Multiplex PCR study of Listeria spp. revealed that four L. monocytogenes isolates were positive for plcA, actA, hlyA and iap genes. Amplified products of three virulence-associated genes namely, actA, hlyA and iap were detected in five L. monocytogenes and three L. monocytogenes isolates were positive for hlyA and iap genes. L. grayi isolates revealed the amplified product of iap gene. Two isolate of L. innocua were positive for actA and iap genes, Five L. welshimeri isolates and one L. seeligeri isolate were positive for iap gene, lacking plcA, actA and hlyA genes. As actA and iap genes were detected in a two isolate of L. innocua. It is clear that actA and iap genes are shared by L. monocytogenes and L. innocua. It is concluded that genes in three different combinations were recorded in Listeria spp. and that prfA and hlyA detected by PCR distinctively in L. monocytogenes suggest the usefulness of PCR amplification from various food samples. [ABSTRACT FROM AUTHOR]

Typically, short‐ and long‐chain lipids from oils exhibit different antimicrobial activities and therefore have been used in agriculture and aquaculture, biomedical therapeutic and antibacterial fields. However, these fatty acids have limitations in terms of bioactive efficacy, thermostability and aqueous solubility. In this study, water‐soluble iso‐fatty acid arginate hydrochloride derivatives with antimicrobial properties were produced by introducing branched (iso‐) chain and other linear‐ (n‐) chain fatty acids to the "arginine" amino acid molecule. The two‐step synthetic route was straightforward and provided an efficient 88% and 76% product yields for ethyl n‐oleoyl arginate hydrochloride and ethyl iso‐oleoyl arginate hydrochloride, respectively. ATR‐FT‐IR, NMR, and LC‐MS‐Q‐TOF techniques were used to thoroughly characterize and confirm the products. These arginate products had strong antimicrobial activities against Listeria innucua, a Gram‐positive bacterium with minimum inhibitory concentrations and minimum bactericidal concentrations ranging from 1.8 µg mL−1 to 29.1 µg mL−1. Therefore, the study demonstrated the development of a novel class of antimicrobial compounds from iso‐fatty acids and arginates. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a facultative intracellular foodborne pathogen and the causative agent of the severe disease listeriosis. It is found ubiquitously in the environment and exhibits innate resistance to certain antimicrobials, but acquired antimicrobial resistance remains relatively uncommon. Given the potentially dire health outcomes associated with listeriosis, acquisition of antimicrobial resistance (AMR) by this pathogen is of considerable public health concern. AMR in L. monocytogenes has been surveyed frequently in strains of clinical and food origin, but much less commonly in wildlife. We analyzed 158 strains of L. monocytogenes and 27 of non-pathogenic Listeria spp. isolated from wild black bears (Ursus americanus) for resistance to a panel of antimicrobials. AMR was uncommon and noted mostly for tetracycline. Tetracycline resistance was more common in Listeria innocua than in L. monocytogenes. All tetracycline-resistant L. monocytogenes strains belonged to sequence type ST1039 and harbored the Tn916-like tet(M) transposon Tn916.1039 in a conserved chromosomal location. In contrast, three different tetracycline resistance elements, i.e., the tet(M) elements Tn5801.UAM and Tn5801.551 and the tet(S) element Tn6000.205, were ident ified among tetracycline-resistant strains of L. innocua. The greater prevalence and diversity of tetracycline resistance elements among bear-derived non-pathogenic Listeria strains suggest the potential of the latter to serve as reservoirs for retention and dive rsification of AMR determinants in this wildlife host and warrant their further monitoring and study. IMPORTANCE Listeria monocytogenes causes severe foodborne illness and is the only human pathogen in the genus Listeria. Previous surveys of AMR in Listeria focused on clinical sources and food or food processing environments, with AMR in strains from wildlife and other natural ecosystems remaining under-explored. We analyzed 185 sequenced strains from wild black bears (Ursus americanus) from the United States, including 158 and 27 L. monocytogenes and L. innocua, respectively. Tetracycline resistance was the most prevalent resistance trait. In L. monocytogenes, it was encountered exclusively in serotype 4b strains with the novel Tn916-like element Tn916.1039. In contrast, three distinct, novel tetracycline resistance elements (Tn5801.UAM, Tn5801.551, and Tn6000.205) were identified in L. innocua. Interestingly, Tn5801.551 was identical to elements in L. monocytogenes from a major foodborne outbreak in the United States in 2011. The findings suggest the importance of wildlife and non-pathogenic Listeria species as reservoir for resistance elements in Listeria. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the microbial aspect of some meat products by performing S. aureus count, detection of S. aureus enterotoxins as well as isolation and identification of some food poisoning bacteria. Therefore, fifty random samples of beef luncheon and kofta (25 of each) were equally collected from different localities in Gharbia governorate, then bacteriologically examined, where values of S. aureus count were 7.61x10²±0.54x10² in beef luncheon, 2.82x10³±0.39x10³ (cfu/g) in kofta samples. Meanwhile, in beef luncheon enterotoxin B and enterotoxin (A&D) were detected with a percentage of 4% of each, while in kofta samples enterotoxin A, enterotoxin D, and enterotoxin (A&C) were detected with a percentage of 8%, 4%, and 4%, respectively. Moreover, Salmonellae, E. coli, Klebsiella pneumoniae, and Listeria were detected at 16%, 20%, 20%, and 20% in the examined beef luncheon. while, in kofta samples 24 %, 36%, 28% and 16%, respectively. Meanwhile, E. coli identified as O26 : H11, O44 : H18, O78, O91 : H21, O103 : H2, O111 : H4, O127 : H6, O146 : H21 and O159 with various percentages. Salmonella spp. identified as S. Enteritidis, S. Typhimurium, S. Labadi, S. Infantis, and S. Molade. On the other side, Listeria species serologically identified as L. monocytogenes, L. ivanovii, L. innocuai, and L. welshimeri. Moreover, isolated serotypes of K. pneumonia as K1 and K2 were 16%, and 4% in luncheon samples. While 16% and 12% were in kofta samples. Furthermore, high virulent K. pneumonia (HVKP) and classic K. pneumonia (CKP) were 12% and 8% in luncheon, while were 20% and 8% in kofta samples, respectively. Achieved results in the current study proved that most of the examined meat products were contaminated with E. coli, Salmonella spp., S. aureus, Listeria spp. and K. pneumonia, this considered objectionable, as they render the product of inferior quality and unfit for consumption [ABSTRACT FROM AUTHOR]

Listeriosis, an invasive illness with a fatality rate between 20% and 30%, is caused by the ubiquitous bacterium Listeria monocytogenes. Human listeriosis has long been associated with foods. This is because the ubiquitous nature of the bacteria renders it a common food contaminant, posing a significant risk to the food processing sector. Although several sophisticated stress coping mechanisms have been identified as significant contributing factors toward the pathogen's persistence, a complete understanding of the mechanisms underlying persistence across various strains remains limited. Moreover, aside from genetic aspects that promote the ability to cope with stress, various environmental factors that exist in food manufacturing plants could also contribute to the persistence of the pathogen. The objective of this review is to provide insight into the challenges faced by the dairy industry because of the pathogens' environmental persistence. Additionally, it also aims to emphasize the diverse adaptation and response mechanisms utilized by L. monocytogenes in food manufacturing plants to evade environmental stressors. The persistence of L. monocytogenes in the food processing environment poses a serious threat to food safety and public health. The emergence of areas with high levels of L. monocytogenes contamination could facilitate Listeria transmission through aerosols, potentially leading to the recontamination of food, particularly from floors and drains, when sanitation is implemented alongside product manufacturing. Hence, to produce safe dairy products and reduce the frequency of outbreaks of listeriosis, it is crucial to understand the factors that contribute to the persistence of this pathogen and to implement efficient control strategies. [ABSTRACT FROM AUTHOR]

Collation of the current scope of literature related to population dynamics (i.e., growth, die‐off, survival) of foodborne pathogens on fresh produce can aid in informing future research directions and help stakeholders identify relevant research literature. A scoping review was conducted to gather and synthesize literature that investigates population dynamics of pathogenic and non‐pathogenic Listeria spp., Salmonella spp., and Escherichia coli on whole unprocessed fresh produce (defined as produce not having undergone chopping, cutting, homogenization, irradiation, or pasteurization). Literature sources were identified using an exhaustive search of research and industry reports published prior to September 23, 2021, followed by screening for relevance based on strict, a priori eligibility criteria. A total of 277 studies that met all eligibility criteria were subjected to an in‐depth qualitative review of various factors (e.g., produce commodities, study settings, inoculation methodologies) that affect population dynamics. Included studies represent investigations of population dynamics on produce before (i.e., pre‐harvest; n = 143) and after (i.e., post‐harvest; n = 144) harvest. Several knowledge gaps were identified, including the limited representation of (i) pre‐harvest studies that investigated population dynamics of Listeria spp. on produce (n = 13, 9% of pre‐harvest studies), (ii) pre‐harvest studies that were carried out on non‐sprouts produce types grown using hydroponic cultivation practices (n = 7, 5% of pre‐harvest studies), and (iii) post‐harvest studies that reported the relative humidity conditions under which experiments were carried out (n = 56, 39% of post‐harvest studies). These and other knowledge gaps summarized in this scoping review represent areas of research that can be investigated in future studies. [ABSTRACT FROM AUTHOR]

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Cancer receives enduring international attention due to its extremely high morbidity and mortality. Immunotherapy, which is generally expected to overcome the limits of traditional treatments, serves as a promising direction for patients with recurrent or metastatic malignancies. Bacteria-based vectors such as Listeria monocytogenes take advantage of their unique characteristics, including preferential infection of host antigen presenting cells, intracellular growth within immune cells, and intercellular dissemination, to further improve the efficacy and minimize off-target effects of tailed immune treatments. Listeria monocytogenes can reshape the tumor microenvironment to bolster the antitumor effects both through the enhancement of T cells activity and a decrease in the frequency and population of immunosuppressive cells. Modified Listeria monocytogenes has been employed as a tool to elicit immune responses against different tumor cells. Currently, Listeria monocytogenes vaccine alone is insufficient to treat all patients effectively, which can be addressed if combined with other treatments, such as immune checkpoint inhibitors, reactivated adoptive cell therapy, and radiotherapy. This review summarizes the recent advances in the molecular mechanisms underlying the involvement of Listeria monocytogenes vaccine in anti-tumor immunity, and discusses the most concerned issues for future research. [ABSTRACT FROM AUTHOR]

"Chouriço de carne" is a Portuguese ready-to-eat dry fermented sausage which relies on effective spontaneous fermentation to ensure its microbial safety and desired organoleptic properties. This study aimed to assess selected microbiological and physicochemical characteristics of artisanal chouriço produced by 14 different regional producers. Aerobic mesophilic bacteria, lactic acid bacteria, Staphylococcus aureus, Clostridium spp., Listeria spp. and Salmonella spp., in addition to pH, water activity and moisture, were evaluated for each of 70 samples. Principal Component Analysis of all these attributes was performed to build quality maps of the analyzed lots. The results showed great variability between sausages of different producers within the same geographic region, and S. aureus, Clostridium spp., Salmonella spp. and Listeria spp. were identified in sausages of several producers, highlighting the need for stricter microbiological control and standardization of production processes among artisanal producers of chouriço. [ABSTRACT FROM AUTHOR]

Many systemically administered cancer therapies exhibit dose-limiting toxicities that reduce their effectiveness. To increase efficacy, bacterial delivery platforms have been developed that improve safety and prolong treatment. Bacteria are a unique class of therapy that selectively colonizes most solid tumors. As delivery vehicles, bacteria have been genetically modified to express a range of therapies that match multiple cancer indications. In this review, we describe a modular "build-a-bug" method that focuses on five design characteristics: bacterial strain (chassis), therapeutic compound, delivery method, immune-modulating features, and genetic control circuits. We emphasize how fundamental research into gut microbe pathogenesis has created safe bacterial therapies, some of which have entered clinical trials. The genomes of gut microbes are fertile grounds for discovery of components to improve delivery and modulate host immune responses. Future work coupling these delivery vehicles with insights from gut microbes could lead to the next generation of microbial cancer therapy. Several genera of bacteria colonize tumors with high selectivity. New discoveries regarding gut pathogenesis, coupled with advances in synthetic biology, have enabled bacteria to be engineered as safe and effective cancer therapies. Raman et al. describe a "build-a-bug" design method that can be used to create any bacterial cancer therapy. [ABSTRACT FROM AUTHOR]

This study determined the prevalence, risk factors, and molecular characteristics of Listeria species detected in beef and beef products sampled in Mpumalanga province, South Africa. Four hundred beef and beef products were collected from 30 retail outlets in three districts (Bronkhorstspruit, Emalahleni, and Middelburg) within the province. Standard bacteriological and polymerase chain reaction (PCR) assays were used in the study. The overall prevalence of L. monocytogenes and other Listeria spp. in the samples was 8.3% (33/400) and 30% (120/400) (p <.05), respectively. For the five variables investigated, statistically significant effects were evident only for the region (p <.001) and type of product (p <.0001) for L. monocytogenes, the type of outlet (p =.011) and the type of product (p <.0001) for Listeria spp. Of the 20 types of beef and beef products tested, 15 (75%) and 17 (85%) were contaminated by L. monocytogenes and Listeria spp., respectively (p =.429). Among the four categories of products tested, the prevalence of L. monocytogenes was 7.3% (8/109), 10.6% (11/104), 7.5% (8/106), and 7.4% (6/81) for raw beef, ready‐to‐eat (RTE) products, milled beef, and offal & organs, respectively (p >.799). Among the 33 L. monocytogenes isolates, PCR genoserogroup IIa (42.4%, 1/2a‐3a) was most frequently detected. All (100%) of the isolates carried one or more of the eight virulence‐associated genes assessed, with genes inlC and inlJ detected in all the isolates. The overall prevalence of L. monocytogenes (8.3%) and the high frequency of virulent serogroups of L. monocytogenes commonly associated with human listeriosis pose a food safety risk to consumers of beef and beef‐based products contaminated by L. monocytogenes. [ABSTRACT FROM AUTHOR]

Key Clinical Message: Listeria can cause neurological symptoms in immunocompromised and older patients. Additionally, it is impossible to rule out meningitis by the absence of typical meningeal irritation signs. Therefore, patients with fever and neurological impairments should be rapidly examined for blood and cerebrospinal fluid cultures to rule out Listeria meningitis. A woman in her 90s developed fever, dysarthria, and transient disturbance of consciousness. Physical examination revealed no meningeal irritation signs. Listeria monocytogenes were detected in her blood culture the following day. Because of an increased number of cells in cerebrospinal fluid, she was diagnosed with Listeria meningitis. [ABSTRACT FROM AUTHOR]

Urine and tissues samples were collected for bacteriological examination from 50 apparently diseased sheep of different ages and sexes which were clinically suspected suffering from renal infection and from slaughtered animals. We were collected samples from many locations at Matrouh Governorate and sent for laboratory. Post mortem examination of the renal system of affected cases revealed different investigations. kidneys appeared swollen, its surface was mottled red. Some kidneys were pale and greatly enlarged. Cut section of the kidney showed areas of hemorrhage and abscess formation. The results indicated that 80% (40) sheep were positive for bacterial infection however 10 show negative results of bacterial renal infection. Bacteriological investigations revealed that the isolated organisms were: Escherichia coli (20%), Staphyolcoccus aureus (25%), Pseudomonas aeruginosa (5.0%), Streptococcus spp. (7.5%), Enterococcus faeclis (7.5%), Corynebaeterium spp. (17.5%), Klebseilla pneumoniae (2.5%). and. Listeria monocytogene (2.5%). Moreover, mixed infection found in 12.5% of the examined samples. A total of eight isolates of E coli from renal examined samples were serologically positive to O111, O104, O26, O113, O91, O103 and O126. PCR results showed that two strains for Listeria isolates were Listeria monocytogenes and some of them have InI B gene. Antibiogram was applied upon the isolated bacterial pathogens and found that Garamycine was the drug of choice for treatment of infected animals. [ABSTRACT FROM AUTHOR]

Bacteriophages are viruses that infect and kill bacteria. Currently, phage products are available for the control of the pathogen Listeria monocytogenes in food products in the United States. In this study, we explore whether experimental evolution can be used to generate phages with improved abilities to function under specific food-relevant conditions. Ultra-pasteurized oat and whole milk were chosen as test matrices as they represent different food groups, yet have similar physical traits and macronutrient composition. We showed that (i) wild-type phage LP-125 infection kinetics are different in the two matrices and (ii) LP-125 has a significantly higher burst size in oat milk. From this, we attempted to evolve LP-125 to have improved infection kinetics in whole milk. Ancestral LP-125 was passaged through 10 rounds of amplification in milk conditions. Plaquepurified DNA samples from milk-selected phages were isolated and sequenced, and mutations present in the isolated phages were identified. We found two nonsynonymous substitutions in LP125_108 and LP125_112 genes, which encode putative baseplate-associated glycerophosphoryl diester phosphodiesterase and baseplate protein, respectively. Protein structural modeling showed that the substituted amino acids in the mutant phages are predicted to localize to surface-exposed helices on the corresponding structures, which might affect the surface charge of proteins and their interaction with the bacterial cell. The phage containing the LP125_112 mutation adsorbed significantly faster than the ancestral phage in both oat and whole milk. Follow-up experiments suggest that fat content may be a key factor for the expression of the phenotype of this mutation. [ABSTRACT FROM AUTHOR]

Antimicrobial blue light (aBL) in the 400-470 nm wavelength range has been reported to kill multiple bacteria. This study assessed its potential for mitigating an important foodborne pathogen, Listeria monocytogenes (Lm), focusing on surface decontamination. Three wavelengths were tested, with gallic acid as a photosensitizing agent (Ps), against dried cells obtained from bacterial suspensions, and biofilms on stainless-steel (SS) coupons. Following aBL exposure, standard microbiological analysis of inoculated coupons was conducted to measure viability. Statistical analysis of variance was performed. Confocal laser scanning microscopy was used to observe the biofilm structures. Within 16 h of exposure at 405 nm, viable Lm dried cells and biofilms were reduced by approx. 3 log CFU/cm2 with doses of 2,672 J/cm2. Application of Ps resulted in an additional 1 log CFU/cm2 at 668 J/cm2, but its effect was not consistent. The highest dose (960 J/cm2) at 420 nm reduced viable counts on the biofilms by 1.9 log CFU/cm2. At 460 nm, after 800 J/cm2, biofilm counts were reduced by 1.6 log CFU/cm2. The effect of material composition on Lm viability was also investigated. Irradiation at 405 nm (668 J/cm2) of cells dried on polystyrene resulted in one of the largest viability reductions (4.0 log CFU/cm2), followed by high-density polyethylene (3.5 log CFU/cm2). Increasing the dose to 4,008 J/cm2 from 405 nm (24 h), improved its efficacy only on SS and polyvinyl chloride. Biofilm micrographs displayed a decrease in biofilm biomass due to the removal of biofilm portions from the surface and a shift from live to dead cells suggesting damage to biofilm cell membranes. These results suggest that aBL is a potential intervention to treat Lm contamination on typical material surfaces used in food production. [ABSTRACT FROM AUTHOR]

The high mortality rate associated with Listeria monocytogenes can be attributed to its ability to invade the body systemically and to activate inflammasomes. Both of these processes are facilitated by expressing a major virulence factor known as listeriolysin O, a 56 kDa pore‐forming protein encoded by the hly gene. Listeriolysin O plays a crucial role in the pathogenesis of the bacterium by facilitating the escape of the pathogen from the phagosome into the cytosol. This process is essential for the successful establishment of infection. In addition, listeriolysin O is known as an immunomodulator that activates host signal transduction. In addition to listeriolysin O, Listeria expresses a variety of bacterial ligands, such as lipoteichoic acid, nucleotide, and flagellin, that are recognized by host intracellular pattern‐recognition receptors including Nod‐like receptors, AIM2‐like receptors, and RIG‐I‐like receptors. This review introduces intracellular recognition of Listeria monocytogenes since recent studies have revealed that the activation of inflammasome exacerbates Gram‐positive bacteria infection. [ABSTRACT FROM AUTHOR]

Multiple recalls and outbreaks involving Listeria monocytogenes-contaminated apples have been linked to the post-harvest packing environment where this pathogen can persist in biofilms. Therefore, this study assessed L. monocytogenes survival on apples as affected by harvest year, apple cultivar, storage atmosphere, and growth conditions. Unwaxed Gala, Granny Smith, and Honeycrisp apples were dip-inoculated in an 8-strain L. monocytogenes cocktail of planktonic- or biofilm-grown cells (~6.5 log CFU/mL), dried, and then examined for numbers of L. monocytogenes during air or controlled atmosphere (CA) (1.5% O2, 1.5% CO2) storage at 2 °C. After 90 days, air or CA storage yielded similar L. monocytogenes survival (p > 0.05), regardless of harvest year. Populations gradually decreased with L. monocytogenes quantifiable in most samples after 7 months. Apple cultivar significantly impacted L. monocytogenes survival (p < 0.05) during both harvest years with greater reductions (p < 0.05) seen on Gala compared to Granny Smith and Honeycrisp. Biofilm-derived cells survived longer (p < 0.05) on L. monocytogenes-inoculated Gala and Honeycrisp apples compared to cells grown planktonically. These findings should aid in the development of improved L. monocytogenes intervention strategies for apple growers and packers. [ABSTRACT FROM AUTHOR]

In the ready-to-eat food industry, Listeria control is mandatory to ensure the food safety of the products since its presence could cause a disease called listeriosis. The objective of the present study was to carry out a challenge test to verify the efficiency of different combinations of natural antimicrobial ingredients against Listeria monocytogenes to be used in ready-to-eat foods. Six different formulations of cooked ham were prepared: a control formulation and five different formulations. An initial inoculation of 2 log cycles was used in the different products, and the growth of Listeria was monitored at different temperatures and times (4 °C for 17 w and 7 °C for 12 w). Control samples showed a progressive growth, reaching 5–6 log after 3 or 4 weeks. The rest of the samples showed constant counts of Listeria during the entire study. Only samples containing 100 ppm nitrite + 250 PPM ascorbic acid + 0.7% PRS-DV-5 did not control the growth of Listeria at 7 °C after 7 w of storage. The results obtained allowed us to classify the cooked ham prepared using natural ingredient combinations as a "Ready-to-eat food unable to support the growth of L. monocytogenes other than those intended for infants and for special medical purposes". [ABSTRACT FROM AUTHOR]

Seal populations in Canadian waters provide sustenance to coastal communities. There is potential for pathogenic and/or antimicrobial‐resistant bacteria to transfer to humans through inadvertent faecal contamination of seal products. The objective of this study was to investigate the occurrence and potential antimicrobial resistance of Salmonella spp., Escherichia coli and Listeria monocytogenes in faecal samples collected from grey seals (Halichoerus grypus) in the Gulf of St. Lawrence and from ringed seals (Pusa hispida) in Frobisher Bay and Eclipse Sound, Nunavut, Canada. Grey seals were harvested during commercial hunts or during scientific sampling; ringed seals were collected by Inuit hunters during subsistence harvests. Virulence genes defining pathogenic E. coli were identified by PCR, and antimicrobial susceptibility testing was performed on recovered isolates. In grey seals, E. coli was detected in 34/44 (77%) samples, and pathogenic E. coli (extraintestinal E. coli [ExPEC], enteropathogenic E. coli [EPEC] or ExPEC/EPEC) was detected in 13/44 (29%) samples. Non‐susceptibility to beta‐lactams and quinolones was observed in isolates from 18 grey seals. In ringed seals from Frobisher Bay, E. coli was detected in 4/45 (9%) samples; neither virulence genes nor antimicrobial resistance was detected in these isolates. In ringed seals from Eclipse Sound, E. coli was detected in 8/50 (16%) samples and pathogenic E. coli (ExPEC and ExPEC/EPEC) in 5/50 (10%) samples. One seal from Eclipse Sound had an E. coli isolate resistant to beta‐lactams. A monophasic Salmonella Typhimurium was recovered from 8/50 (16%) seals from Eclipse Sound. All Salmonella isolates were resistant to ampicillin, streptomycin, sulfisoxazole and tetracycline. L. monocytogenes was not detected in any sample. These findings suggest that seals may act as important sentinel species and as reservoirs or vectors for antimicrobial‐resistant and virulent E. coli and Salmonella species. Further characterization of these isolates would provide additional insights into the source and spread of antimicrobial resistance and virulence genes in these populations of free‐living seals. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is an important pathogen that can cause listeriosis. Despite the growing recognition of Listeria spp. as a foodborne and environmental pathogen, the understanding of its prevalence and characteristics of Listeria spp. in the marine environment remains unknown. In this study, we first investigated the genetic and phenotypic characteristics of Listeria species isolated in a coastal city in China. The findings revealed that the sequence type 87 (ST87) L. monocytogenes, a prevalent clinical and seafood strain in China, dominates in recreational beach sands and possesses a notable biofilm-forming capacity in seawater. The presence of ST87 L. monocytogenes in coastal environments indicates the potential health risks for both recreational activities and seafood consumption. Moreover, the ST121 isolates from sand had a versatile plasmid encoding multifunctional genes, including uvrX for UV resistance, gbuC for salt resistance, and npx for oxidative resistance and multiple transposases, which potentially aid in survival under natural environments. Black-headed gulls potentially facilitate the spread of L. monocytogenes, with similar ST35 strains found in gulls and beach sand. As a reservoir of microbes from marine environments and human/animal excrement, coastal sand would play an important role in the spread of L. monocytogenes and is an environmental risk for human listeriosis. [ABSTRACT FROM AUTHOR]

Listeria continues to be a persistent foodborne pathogen that is responsible for human cases of listeriosis when contaminated food products are consumed. Human subjects considered to be most susceptible include the elderly, immunocompromised, and pregnant women. Listeria is characterized as a saprophytic organism with the capability of responding and adapting to constantly changing environments because it possesses multiple stress response mechanisms to overcome varying temperatures, salt concentrations, and pH, among others. Primary foods and food products associated with listeriosis include dairy products and ready-to-eat meats such as turkey products. Historically, chicken eggs have not been identified as a primary source of Listeria, but the potential for contamination during egg production and processing does exist. Listeria species have been isolated from egg-processing plant equipment and are presumed to occur in egg-processing plant environments. Whether Listeria is consistently disseminated onto eggs beyond the egg-processing plant is a risk factor that remains to be determined. However, research has been conducted over the years to develop egg wash solutions that generate combinations of pH and other properties that would be considered inhibitory to Listeria. Even less is known regarding the association of Listeria with alternative egg production systems, but Listeria has been isolated from pasture flock broilers, so it is conceivable, given the nature of the outdoor environments, that layer birds under these conditions would also be exposed to Listeria and their eggs become contaminated. This review focuses on the possibility of Listeria occurring in conventional and alternative egg-laying production and processing systems. [ABSTRACT FROM AUTHOR]