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Listeria monocytogenes, an intra-cellular, Gram-positive, pathogenic bacterium, is one of the leading agents of foodborne illnesses. The morbidity of human listeriosis is low, but it has a high mortality rate of approximately 20% to 30%. L. monocytogenes is a psychotropic organism, making it a significant threat to ready-to-eat (RTE) meat product food safety. Listeria contamination is associated with the food processing environment or post-cooking cross-contamination events. The potential use of antimicrobials in packaging can reduce foodborne disease risk and spoilage. Novel antimicrobials can be advantageous for limiting Listeria and improving the shelf life of RTE meat. This review will discuss the Listeria occurrence in RTE meat products and potential natural antimicrobial additives for controlling Listeria. [ABSTRACT FROM AUTHOR]

The global food demand is expected to increase in the coming years, along with challenges around climate change and food security. Concomitantly, food safety risks, particularly those related to bacterial pathogens, may also increase. Thus, the food sector needs to innovate to rise to this challenge. Here, we discuss recent advancements in molecular techniques that can be deployed within various foodborne bacteria surveillance systems across food settings. To start with, we provide updates on nucleic acid‐based detection, with a focus on polymerase chain reaction (PCR)‐based technologies and loop‐mediated isothermal amplification (LAMP). These include descriptions of novel genetic markers for several foodborne bacteria and progresses in multiplex PCR and droplet digital PCR. The next section provides an overview of the development of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR‐associated (Cas) proteins systems, such as CRISPR‐Cas9, CRISPR‐Cas12a, and CRISPR‐Cas13a, as tools for enhanced sensitive and specific detection of foodborne pathogens. The final section describes utilizations of whole genome sequencing for accurate characterization of foodborne bacteria, ranging from epidemiological surveillance to model‐based predictions of bacterial phenotypic traits through genome‐wide association studies or machine learning. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes virulence factor InlB specifically interacts with the receptors c-Met and gC1q-R. Both receptors are present in non-professional and professional phagocytes, including macrophages. Phylogenetically defined InlB isoforms differently support invasion into non-professional phagocytes. This work deals with the effects of InlB isoforms on L. monocytogenes uptake and intracellular proliferation in human macrophages. Three isoforms of the receptor binding domain (idInlB) were derived from phylogenetically distinct L. monocytogenes strains belonging to the highly virulent CC1 (idInlBCC1), medium-virulence CC7 (idInlBCC7), and low-virulence CC9 (idInlBCC9) clonal complexes. The constant dissociation increased in the order idInlBCC1 << idInlBCC7 < idInlBCC9 for interactions with c-Met, and idInlBCC1 ≈ idInlBCC7 < idInlBCC9 for interactions with gC1q-R. The comparison of uptake and intracellular proliferation of isogenic recombinant strains which expressed full-length InlBs revealed that the strain expressing idInlBCC1 proliferated in macrophages twice as efficiently as other strains. Macrophage pretreatment with idInlBCC1 followed by recombinant L. monocytogenes infection disturbed macrophage functions decreasing pathogen uptake and improving its intracellular multiplication. Similar pretreatment with idInlBCC7 decreased bacterial uptake but also impaired intracellular multiplication. The obtained results demonstrated that InlB impaired macrophage functions in an idInlB isoform-dependent manner. These data suggest a novel InlB function in L. monocytogenes virulence. [ABSTRACT FROM AUTHOR]

The wild boar is an abundant game species with high reproduction rates. The management of the wild boar population by hunting contributes to the meat supply and can help to avoid a spillover of transmissible animal diseases to domestic pigs, thus compromising food security. By the same token, wild boar can carry foodborne zoonotic pathogens, impacting food safety. We reviewed literature from 2012–2022 on biological hazards, which are considered in European Union legislation and in international standards on animal health. We identified 15 viral, 10 bacterial, and 5 parasitic agents and selected those nine bacteria that are zoonotic and can be transmitted to humans via food. The prevalence of Campylobacter, Listeria monocytogenes, Salmonella, Shiga toxin-producing E. coli, and Yersinia enterocolitica on muscle surfaces or in muscle tissues of wild boar varied from 0 to ca. 70%. One experimental study reported the transmission and survival of Mycobacterium on wild boar meat. Brucella, Coxiella burnetii, Listeria monocytogenes, and Mycobacteria have been isolated from the liver and spleen. For Brucella, studies stressed the occupational exposure risk, but no indication of meat-borne transmission was evident. Furthermore, the transmission of C. burnetii is most likely via vectors (i.e., ticks). In the absence of more detailed data for the European Union, it is advisable to focus on the efficacy of current game meat inspection and food safety management systems. [ABSTRACT FROM AUTHOR]

Introduction: Listeriosis, caused by infection with Listeria monocytogenes (Lm), is a relatively rare but severe disease with one of the highest mortality rates among bacterial foodborne illnesses. A better understanding on the degree of Lm clustering, the temporal distribution of the clusters, and their association with the various food sources is expected to lead to improved source tracing and risk-based sampling. Methods: We investigated the genomic epidemiology of Lm in the Netherlands between 2010 and 2020 by analyzing whole-genome-sequencing (WGS) data of isolates from listerioss patients and food sources from nationwide integrated surveillance and monitoring. WGS data of 756 patient and 770 food/environmental isolates was assessed using core-genome multi-locus sequence typing (cgMLST) with Hamming distance as measure for pairwise distances. Associations of genotype with the epidemiological variables such as patient’s age and gender, and systematic use of specific drugs were tested by multinomial logistic regressions. Genetic differentiation of the Lm within and between food categories was calculated based on allele frequencies at the 1701 cgMLST loci in each food category. Results: We confirmed previous results that some clonal complexes (CCs) are overrepresented among clinical isolates but could not identify any epidemiological risk factors. The main findings of this study include the observation of a very weak attribution of Lm types to food categories and a much better attribution to the producer level. In addition, we identified a high degree of temporal persistence of food, patient and mixed clusters, with more than half of the clusters spanning over more than 1 year and up to 10  years. Discussion: Taken together this would indicate that identifying persistent contamination in food production settings, and producers that process a wide variety of raw food produce, could significantly contribute to lowering the Lm disease burden. [ABSTRACT FROM AUTHOR]

The contamination of fresh produce with foodborne pathogens has been an on-going concern with outbreaks linked to these commodities. Evaluation of farm practices, such as use of manure, irrigation water source, and other factors that could influence pathogen prevalence in the farming environment could lead to improved mitigation strategies to reduce the potential for contamination events. Soil, water, manure, and compost were sampled from farms in Ohio and Georgia to identify the prevalence of Salmonella, Listeria monocytogenes (Lm), Campylobacter, and Shiga-toxin-producing Escherichia coli (STEC), as well as Arcobacter, an emerging human pathogen. This study investigated agricultural practices to determine which influenced pathogen prevalence, i.e., the percent positive samples. These efforts identified a low prevalence of Salmonella, STEC, and Campylobacter in soil and water (< 10%), preventing statistical modeling of these pathogens. However, Lm and Arcobacter were found in soil (13 and 7%, respectively), manure (49 and 32%, respectively), and water samples (18 and 39%, respectively) at a comparatively higher prevalence, suggesting different dynamics are involved in their survival in the farm environment. Lm and Arcobacter prevalence data, soil chemical characteristics, as well as farm practices and weather, were analyzed using structural equation modeling to identify which factors play a role, directly or indirectly, on the prevalence of these pathogens. These analyses identified an association between pathogen prevalence and weather, as well as biological soil amendments of animal origin. Increasing air temperature increased Arcobacter and decreased Lm. Lm prevalence was found to be inversely correlated with the use of surface water for irrigation, despite a high Lm prevalence in surface water suggesting other factors may play a role. Furthermore, Lm prevalence increased when the microbiome’s Simpson’s Diversity Index decreased, which occurred as soil fertility increased, leading to an indirect positive effect for soil fertility on Lm prevalence. These results suggest that pathogen, environment, and farm management practices, in addition to produce commodities, all need to be considered when developing mitigation strategies. The prevalence of Arcobacter and Lm versus the other pathogens suggests that multiple mitigation strategies may need to be employed to control these pathogens. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a foodborne pathogen with the ability to persist and form biofilm matrices in processing environments of food manufacturing facilities. Bacteriophages are bacterial viruses with host specific lethality. Published research on the application of phage to control Listeria sp. in manufacturing environments is limited. In this study, we have assessed the capacity of bacteriophage P100 (Listex™) to reduce incidence of Listeria sp. in the ready‐to‐eat (RTE) environment of refrigerated (4°C) and ambient (20°C) temperature facilities using two different application strategies. A moderate application applied as a single treatment every 24 hr over three days (2 × 107 PFU/ml) and an intensified application applied once every 6 hr over a 24 hr period (1 × 108 PFU/ml). Environmental nonfood contact surface (NFCS) samples were collected and analyzed for the presence of Listeria sp. before and after treatment. When the moderate treatment protocol was applied the incidence of positives decreased from 51.3 to 17.5% in the 4°C environment and from 67.5 to 23.1% in the 20°C production area. For the intensified phage treatment method, the initial positive rate in the 4°C environment ranged from 5 to 47.5%, with an overall 43% reduction in Listeria sp. In the 20°C facility, initial environmental Listeria sp. ranged from 15 to 50%, with an overall reduction of 32% after treatment with phage P100. Data indicate the application of Listeria specific phage P100 in RTE food production environments by either the moderate or intensified application method can reduce incidence and be considered an additional intervention strategy for controlling this pathogen on NFCS. [ABSTRACT FROM AUTHOR]

Although wooden cutting boards have been used for centuries, debate remains on how best to coat/seal the boards for microbial protection. A growing body of the literature points to the hygroscopicity of wood—its ability to draw water and bacteria from its surface, deep into the wood, where the bacteria are trapped and die—as the wood attempts to even out its moisture content. There is little research as to how coatings affect this mechanism. In this study, four wood species were coated with either a hardening or nonhardening oil and then, were inoculated with either Listeria or Salmonella, then 'stamped' onto sterile agar plates at given time intervals to check microbial growth. The coated woods had significantly more recoverable bacteria on their surface than did the uncoated samples. There was no significant difference in performance between the oils. Remaining bacterial loads did vary significantly by wood species, with European beech having significantly less surface bacteria when inoculated with Salmonella, and the oak species having significantly less surface bacteria with Listeria. Multiple factors appeared to affect the ability of wood to move bacteria from its surface, including porosity of the wood, type of bacteria applied, and time. These results run counter to popular understanding of wood cutting board maintenance and will hopefully impact the way households and professional kitchens use and care for their food-contact wood in the future. [ABSTRACT FROM AUTHOR]

The hitherto implemented Listeria monocytogenes detection techniques are cumbersome or require expensive non-portable instrumentation, hindering their transposition into on-time surveillance systems. The current work proposes a novel integrated system resorting to loop-mediated isothermal amplification (LAMP), assisted by a bacteriophage P100–magnetic platform, coupled to an endpoint electrochemical technique, towards L. monocytogenes expeditious detection. Molybdophosphate-based optimization of the bacterial phagomagnetic separation protocol allowed the determination of the optimal parameters for its execution (pH 7, 25 °C, 32 µg of magnetic particles; 60.6% of specific capture efficiency). The novel LAMP method targeting prfA was highly specific, accomplishing 100% inclusivity (for 61 L. monocytogenes strains) and 100% exclusivity (towards 42 non-target Gram-positive and Gram-negative bacteria). As a proof-of-concept, the developed scheme was successfully validated in pasteurized milk spiked with L. monocytogenes. The phagomagnetic-based approach succeeded in the selective bacterial capture and ensuing lysis, triggering Listeria DNA leakage, which was efficiently LAMP amplified. Methylene blue-based electrochemical detection of LAMP amplicons was accomplished in 20 min with remarkable analytical sensitivity (1 CFU mL−1). Hence, the combined system presented an outstanding performance and robustness, providing a 2.5 h-swift, portable, cost-efficient detection scheme for decentralized on-field application. [ABSTRACT FROM AUTHOR]

Reported cases of listeriosis from food of non-animal origin (FNAO) are increasing. In order to assess the risk of exposure to Listeria monocytogenes from FNAO, the genetic characterization of the pathogen in FNAO products and in primary production and processing plants needs to be investigated. For this, 123 samples of fresh and frozen soft fruit and 407 samples of 39 plants in Bavaria, Germany that produce and process FNAO were investigated for Listeria contamination. As a result, 64 Listeria spp. isolates were detected using ISO 11290-1:2017. Environmental swabs and water and food samples were investigated. L. seeligeri (36/64, 56.25%) was the most frequently identified species, followed by L. monocytogenes (8/64, 12.50%), L. innocua (8/64, 12.50%), L. ivanovii (6/64, 9.38%), L. newyorkensis (5/64, 7.81%), and L. grayi (1/64, 1.56%). Those isolates were subsequently sequenced by whole-genome sequencing and subjected to pangenome analysis to retrieve data on the genotype, serotype, antimicrobial resistance (AMR), and virulence markers. Eight out of sixty-four Listeria spp. isolates were identified as L. monocytogenes. The serogroup analysis detected that 62.5% of the L. monocytogenes isolates belonged to serogroup IIa (1/2a and 3a) and 37.5% to serogroup IVb (4b, 4d, and 4e). Furthermore, the MLST (multilocus sequence typing) analysis of the eight detected L. monocytogenes isolates identified seven different sequence types (STs) and clonal complexes (CCs), i.e., ST1/CC1, ST2/CC2, ST6/CC6, ST7/CC7, ST21/CC21, ST504/CC475, and ST1413/CC739. The core genome MLST analysis also showed high allelic differences and suggests plant-specific isolates. Regarding the AMR, we detected phenotypic resistance against benzylpenicillin, fosfomycin, and moxifloxacin in all eight L. monocytogenes isolates. Moreover, virulence factors, such as prfA, hly, plcA, plcB, hpt, actA, inlA, inlB, and mpl, were identified in pathogenic and nonpathogenic Listeria species. The significance of L. monocytogenes in FNAO is growing and should receive increasing levels of attention. [ABSTRACT FROM AUTHOR]

The gram-positive bacterium Listeria monocytogenes is an important foodborne pathogen contaminating dairy products. Closely related to L. monocytogenes saprophytic Listeria spp. are also frequent contaminators of food and, particularly, dairy products. To distinguish L. monocytogenes from nonpathogenic Listeria spp. and other bacteria, a dot-immunoassay was developed. The immunoassay is based on the polyclonal antibody to the secreted form of the surface virulence-associated L. monocytogenes -specific InlB protein. To increase InlB production, bacteria were grown on the brain-heart infusion agar supplemented with 0.2% activated charcoal (BHIC agar). Direct plating of artificially contaminated raw milk samples on the BHIC agar followed by the dot-immunoassay allowed a rapid identification of L. monocytogenes in concentrations as little as 10 cfu/mL. Using the developed approach, preliminary results were obtained within 14 h, and the final results were obtained after 26 h. The dot-immunoassay was tested on L. monocytogenes strains belonging to different clonal complexes and phylogenetic lineages, Listeria spp., and other bacterial species. Results showed the exceptional specificity of the developed dot-immunoassay for the rapid identification of L. monocytogenes. [ABSTRACT FROM AUTHOR]

Biofilms are surface-associated microbial clusters embedded in extracellular polymeric substances. Biofilms formed on food-contact surfaces create challenges for the food industry due to their increased tolerance to antimicrobial agents and disinfectants. This study aimed to evaluate the effect of the biofilm maturation period on their resistance to gaseous ClO2. Listeria monocytogenes, Salmonella serotype Typhimurium, and Escherichia coli O157:H7 biofilms formed on stainless steel (SS) and high-density polyethylene (HDPE) surfaces were investigated. The total cell mass and protein content significantly increased (P  < .05) between the second and the fifth day of maturation, and the biofilms' resistance to gaseous ClO2 increased as they matured. Generally, the cell counts of 0-day-old L. monocytogenes, Salm. Typhimurium, and E. coli O157:H7 biofilms on SS and HDPE reduced below the detection limit (0.48 log CFU/cm2) within 5 min. The cell counts of 2-day-old biofilms of the three pathogens were reduced by 6.22 to over 7.52 log, while those of 5-day-old biofilms were reduced by 3.64 to over 6.34 log after 20 min of treatment with 30 ppmv of gaseous ClO2. Therefore, as resistance increases with biofilm maturation, daily gaseous ClO2 treatment would maximize the antimicrobial efficacy of the cleaning strategy against biofilms. [ABSTRACT FROM AUTHOR]

Objective: Endocarditis is a rare but serious infection caused by Listeria monocytogenes. Panax ginseng demonstrated multiple immunomodulatory effects in earlier studies. Ampicillin is known as an effective antibiotic in the treatment of this disease. Therefore, this study aimed to evaluate the effect of hydro-alcoholic extract of P. ginseng and ampicillin treatment in an animal model of Listeria monocytogenes-induced endocarditis. Materials and Methods: Thirty mice, 5-7 weeks old, were randomly divided into five groups (n=6) including Healthy Control, Infected, Ampicillin (20 mg/kg, subcutaneous) treatment, Ginseng (0.025 mg/kg, intraperitoneal) treatment, and Ginseng (0.025 mg/kg, intraperitoneal) +Ampicillin (15 mg/kg, subcutaneous) treatment groups. The concentration of cytokines in heart tissue, such as IL-1 (interleukine-1), IL-6, IL-8, and TNF-a (Tumor Necrosis Factor-a), was measured. Histopathological changes were evaluated in heart tissues. Results: The levels of cytokines were significantly decreased in the Ampicillin+Ginseng treated group compared to the other experimental groups. Microscopically, pathologic changes in heart tissue were concomitant with biochemical findings, which in the infected group, neutrophils and mononuclear cells infiltration in endocardial tissue, myocardial cell necrosis, and edema were detectable. The Ampicillin+Ginseng group showed no significant changes compared to the normal control group. Conclusion: This study showed that ginseng hydro-alcoholic extract plus ampicillin has better efficacy than the extract or antibiotic alone against experimental endocarditis caused by Listeriosis. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a widespread foodborne pathogen contaminating foods during their production or processing stages. Fresh meat is susceptible to such contamination if it is not properly preserved. Our study was conducted to reveal the level of contamination and prevalence of Listeria spp. present in livestock and poultry meat from Gansu province. A total of 1,387 samples were collected from five cities in Gansu Province according to standard sampling procedures, of which 174 samples (12.5%) were positive for Listeria species. Among them, 14 isolates of L. monocytogenes (1.0%), 150 isolates of Listeria innocua (10.8%), and ten isolates of Listeria welshimeri (0.7%) were identified by conventional bacteriological and molecular identification methods. All isolates were subjected to serological assays, antimicrobial susceptibility tests, growth curve assays, determination of biofilm-forming capacity, and cluster analysis of the 16S rRNA gene sequences. Four predominant serotypes of L. monocytogenes were identified, including 1/2a (35.7%, 5/14), 1/2b (14.3%, 2/14), 1/2c (42.9%, 6/14), and 4b (7.1%, 1/14). All L. monocytogenes isolates were resistant to tetracycline and cefoxitin. Most L. innocua isolates (63.6%, 14/22) and L. welshimeri (40%, 4/10) were resistant to tetracycline. The high biofilm-forming ability was observed among 1/2c and 1/2a serotype isolates. The cluster analysis of the 16S rRNA gene sequences revealed a close genetic relationship between the three Listeria species. This study fills the gap in the knowledge of livestock and poultry meat that carry Listeria in slaughterhouses and markets in Gansu Province. [ABSTRACT FROM AUTHOR]

Strain ST182Gu, isolated from fresh guava fruit, was identified as Enterococcus casseliflavus on the basis of biochemical tests, sugar fermentation reactions (API20Strip), PCR with genus-specific primers, and 16S rRNA sequencing. This appears to be the first documentation of the presence of this species in guava. E. casseliflavus ST182Gu was shown to produce a 4.8 kDa class IIa bacteriocin, active against various lactic acid bacteria including Enterococcus spp. and Streptococcus spp., and Staphylococcus aureus, and different serotypes of Listeria spp. The activity of the peptide was reduced by treatment with 0.1 mg/mL proteolytic enzymes, but not by α-amylase, catalase, lipase, and 1% (w/v) sodium dodecyl sulphate (SDS), Tween-20, Tween-80, urea, NaCl, and EDTA. No change in activity was recorded after adjustment to pH values of between 2.0 and 12.0 for 2 h, and after treatment at 100 °C for 120 min or 121°C for 20 min, compared with non-treated antimicrobial peptide. The mode of action against representative susceptible bacteria was shown to be bactericidal and associated with cell lysis and enzyme- and DNA-leakage. These susceptible bacteria, Listeria ivanovii subsp. ivanovii ATCC 19119, Listeria monocytogenes ATCC 15313, and Enterococcus faecalis ATCC 19443 differed however in their sensitivity to bacteriocin ST182Gu (6,553,600 AU/mL, 102,400 AU/mL, and 51,200 AU/mL, respectively). No significant differences were detected in cell growth and bacteriocin production when strain ST182Gu was grown in MRS broth at 26 °C, 30 °C, and 37 °C for 24 h. Bacteriocin ST182Gu recovery from the surface of the producer cells showed different activity, dependent of the applied test organisms (3200, 800 and 400 AU/mL, evaluated versus L. ivanovii subsp. ivanovii ATCC 19119, L. monocytogenes ATCC 15313 and E. faecalis ATCC 19443, respectively), however, with proportional values with the activity recorded in cell free supernatant versus same test microorganisms. When bacteriocin ST182Gu was combined with sublethal doses of ciprofloxacin, synergistic inhibition of L. ivanovii subsp. ivanovii ATCC 19119 was demonstrated. This increase in ciprofloxacin sensitivity may be due to the dissipation of the proton gradient in the cell membrane of the target organism associated with exposure to bacteriocin ST182Gu. Apart from reducing the MIC of classical therapeutic antibiotics, bacteriocins such as ST182Gu may also play an important role in the treatment of multidrug resistant strains. [ABSTRACT FROM AUTHOR]

Concerns regarding the role of antimicrobial resistance (AMR) in disease outbreaks are growing due to the excessive use of antibiotics. Moreover, consumers are demanding food products that are minimally processed and produced in a sustainable way, without the use of chemical preservatives or antibiotics. Grape seed extract (GSE) is isolated from wine industry waste and is an interesting source of natural antimicrobials, especially when aiming to increase sustainable processing. The aim of this study was to obtain a systematic understanding of the microbial inactivation efficacy/potential of GSE against Listeria monocytogenes (Gram-positive), Escherichia coli and Salmonella Typhimurium (Gram-negative) in an in vitro model system. More specifically, for L. monocytogenes, the effects of the initial inoculum concentration, bacterial growth phase and absence of the environmental stress response regulon (SigB) on the GSE microbial inactivation potential were investigated. In general, GSE was found to be highly effective at inactivating L. monocytogenes, with higher inactivation achieved for higher GSE concentrations and lower initial inoculum levels. Generally, stationary phase cells were more resistant/tolerant to GSE as compared to exponential phase cells (for the same inoculum level). Additionally, SigB appears to play an important role in the resistance of L. monocytogenes to GSE. The Gram-negative bacteria under study (E. coli and S. Typhimurium) were less susceptible to GSE as compared to L. monocytogenes. Our findings provide a quantitative and mechanistic understanding of the impact of GSE on the microbial dynamics of foodborne pathogens, assisting in the more systematic design of natural antimicrobial-based strategies for sustainable food safety. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a Gram-positive pathogen responsible for the severe foodborne disease listeriosis. A chromosomal hotspot between lmo0301 and lmo0305 has been noted to harbor diverse restriction modification (RM) systems. Here, we analyzed 872 L. monocytogenes genomes to better understand the prevalence and types of RM systems in this region, designated the immigration control region (ICR). Type I, II, III and IV RM systems were found in 86.1% of strains inside the ICR and in 22.5% of strains flanking the ICR. ICR content was completely conserved within the same multilocus sequence typing-based sequence type (ST), but the same RM system could be identified in diverse STs. The intra-ST conservation of ICR content suggests that this region may drive the emergence of new STs and promote clone stability. Sau3AI-like, LmoJ2 and LmoJ3 type II RM systems as well as type I EcoKI-like, and type IV AspBHI-like and mcrB-like systems accounted for all RM systems in the ICR. A Sau3AI-like type II RM system with specificity for GATC was harbored in the ICR of many STs, including all strains of the ancient, ubiquitous ST1. The extreme paucity of GATC recognition sites in lytic phages may reflect ancient adaptation of these phages to preempt resistance associated with the widely distributed Sau3AI-like systems. These findings indicate that the ICR has a high propensity for RM systems which are intraclonaly conserved and may impact bacteriophage susceptibility as well as ST emergence and stability. [ABSTRACT FROM AUTHOR]

Atmospheric cold plasma (ACP) treatment can reduce bacterial pathogens in foods. Additional reduction in bacterial cells during storage after ACP treatment was previously reported. The underlying mechanisms of bacterial inactivation during ACP treatment and post-treatment storage need to be understood. This study investigated the changes in the morpho-physiological status of Listeria monocytogenes on ham surfaces after post-ACP-treatment storage of 1 h, 24 h, and 7 days at 4 °C. The membrane integrity, intracellular oxidative stress, and esterase activity of L. monocytogenes were evaluated by flow cytometry. L. monocytogenes cells were under high oxidative stress conditions with slightly permeabilized membranes after 1 h of post-ACP-treatment storage according to the flow cytometry data. During the extended storage of 24 h, the percentage of cells with a slightly permeabilized membrane increased; subsequently, the percentage of cells with intact membranes decreased. The percentage of L. monocytogenes cells with intact membranes decreased to <5% with a treatment time of 10 min and after 7 days of post-treatment storage. In addition, the percentage of L. monocytogenes cells under oxidation stress decreased to <1%, whereas the percentage of cells with completely permeabilized membranes increased to more than 90% for samples treated with ACP for 10 min and 7 days of post-treatment storage. With increased ACP treatment time, for 1 h stored samples, the percentage of cells with active esterase and slightly permeabilized membranes increased. However, during the extended post-treatment storage of 7 days, the percentage of cells with active esterase and slightly permeabilized membranes decreased to below 1%. At the same time, the percentage of cells with permeabilized membrane increased to more than 92% with an increase in ACP treatment time of 10 min. In conclusion, the higher inactivation after 24 h and 7 days post-ACP-treatment storage compared to 1 h stored samples correlated with the loss of esterase activity and membrane integrity of L. monocytogenes cells. [ABSTRACT FROM AUTHOR]

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This work aimed to optimize the E-beam treatment of whole-boned dry-cured hams to reach the food safety objective for Listeria monocytogenes. According to the zero-tolerance criteria in the USA and China, 2.39 decimal reductions are required. Boned hams were grouped by their thickness in clusters A (≤ 7 cm) and B (> 7 cm). All of them were contaminated with Listeria innocua as a surrogate of the pathogen. The inoculum (approx. 107 cells) was placed in the space before occupied by the bone. Then, bilateral treatments at 1 and 2 kGy were applied to A and B clusters, respectively. Listerial load reductions higher than 2.39 log colony forming units (CFU) were observed in the innermost part of the pieces. The absorbed doses by the treated hams were < 3 kGy, avoiding the radiation over-exposure that could compromise their sensory quality. Therefore, taking into account the thickness of whole-bone ham pieces could optimize the sanitization effect of the E-beam treatment. [ABSTRACT FROM AUTHOR]

Little is known about the microbial communities found in distribution centers (DCs), especially in those storing and handling food. As many foodborne bacteria are known to establish residence in food facilities, it is reasonable to assume that DCs handling foods are also susceptible to pathogen colonization. To investigate the microbial communities within DCs, 16S amplicon sequencing was completed on 317 environmental surface sponge swabs collected in DCs (n = 18) across the United States. An additional 317 swabs were collected in parallel to determine if any viable Listeria species were also present at each sampling site. There were significant differences in median diversity measures (observed, Shannon, and Chao1) across individual DCs, and top genera across all reads were Carnobacterium_A, Psychrobacter, Pseudomonas_E, Leaf454, and Staphylococcus based on taxonomic classifications using the Genome Taxonomy Database. Of the 39 16S samples containing Listeria ASVs, four of these samples had corresponding Listeria positive microbiological samples. Data indicated a predominance of ASVs identified as cold-tolerant bacteria in environmental samples collected in DCs. Differential abundance analysis identified Carnobacterium_A, Psychrobacter, and Pseudomonas_E present at a significantly greater abundance in Listeria positive microbiological compared to those negative for Listeria. Additionally, microbiome composition varied significantly across groupings within variables (e.g., DC, season, general sampling location).

This review analyzes the seafood recalls registered by the United States Food and Drug Administration (USFDA) from October 2002 through March 2022. There were more than 2,400 recalls for seafood products over this 20-year period. Biological contamination was the listed root cause for about 40% of these recalls. Almost half were designated as Class I recalls, due to the high risk of the recalled seafood to cause disease or death. Independent of the recall classification, 74% of the recalls were due to violations of the Current Good Manufacturing Practices (cGMPs) regulations. The most common cause for these seafood recalls was due to undeclared allergens (34%). More than half of the undeclared allergen recalls were for undeclared milk and eggs. Recalls for Listeria monocytogenes accounted for 30% of all recalls and were all Class I. Finfish comprised 70% of the recall incidents, and salmon was the single most recalled species (22%). Improper cold smoking treatment that resulted in Listeria monocytogenes contamination was the most common reason reported for the salmon recalls.The goal of this review was to evaluate the main causes of food safety failures within the seafood manufacturing and distribution sectors. Human errors and failures to control food safety risks during the processing of food are the main driving factors for most reported recalls in the U.S. Properly applying the Hazard Analysis Critical Control Points (HACCP) approach and procedures are needed to identify the potential food safety risks. The key to reducing the risks of human error and loss of process control is the development and implementation of an effective food safety culture program at the manufacturing facility, which must require strong senior management support at corporate and enterprise levels.

Raw, ready-to-eat (RTE) seafood products, such as ceviche, poke, and sushi, have experienced growing demand globally; however, these products have the potential to be contaminated with foodborne pathogens. The objective of this study was to determine the prevalence of Escherichiacoli/coliforms, Salmonella, and Listeria in ceviche, poke, and sushi dishes sold at the retail level in Orange County, CA, USA. Additional organisms detected during testing were also considered in the results. A total of 105 raw, RTE samples of ceviche, poke, and sushi were collected from restaurants and grocery stores in Orange County, CA. Samples were tested for Salmonella and Listeria utilizing methods from the Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM). E. coli and total coliforms were enumerated utilizing 3 M Petrifilm plates. Overall, two samples (1.9%) were positive for generic E. coli, with a range of 5–35 CFU/g. Coliforms were detected in 85 samples (81%), with a range of 5–1710 CFU/g. The average coliform levels in ceviche samples (259 CFU/g) were significantly higher than the levels in sushi samples (95 CFU/g), according to a Kruskal-Wallis H test followed by the Dunn test (p

SUMMARY: Free-range poultry has become more popular due to consumer demands for improvements in animal welfare. The current study was conducted to determine the impact of paddock area stocking density on the egg and environmental microbiology of free-range laying flocks. A flock of brown egg layers was split and maintained at low (4.02 m2/bird) and high (2.01 m2/bird) paddock area stocking densities. Egg and environmental samples were taken every 7 to 9 wk between 20 and 52 weeks (wk) of age for the enumeration of aerobic bacteria, Enterobacteriaceae, yeast and mold as well as the presence of Campylobacter spp., Listeria spp., and Salmonella spp.At 20 wk, shells from high-density flocks had higher levels of aerobic bacteria (P < 0.02) and Enterobacteriaceae (P < 0.008) compared to low-density flocks. However, at wk 44, nestbox swabs from low-density flocks had higher levels of aerobic bacteria and Enterobacteriaceae. Nestbox swabs (P < 0.04) of high-density flocks (43.3%) had a higher prevalence of Campylobacter compared to low-density flocks (26.7%). Differences in prevalence of Listeria or Salmonella were not observed in this study.Previous studies had shown differences in range use and behavioral expression between low and high paddock area stocking density free-range hens. These differences could have impacts on egg and environmental microbiology. Further research is needed to investigate the relationship between hen behavior and egg and environmental microbiology of free-range laying flocks.

Cold-smoked salmon are ready-to-eat products that may support the growth of pathogenic Listeria monocytogenes during their long shelf-life. Consumption of such contaminated products can cause fatal listeriosis infections. Another challenge and potential risk associated with CS salmon is their high levels of sodium salt. Excess dietary intake is associated with serious health complications. In the present study, anti-listerial bacteriocin (nisin), P100 bacteriophages (Phageguard L, PGL) and fermentates (Verdad N6, P-NDV) were evaluated as commercial bio-preservation strategies for increased control of L. monocytogenes in standard (with NaCl) and sodium-reduced (NaCl partially replaced with KCl) CS salmon. Treatments of CS salmon with nisin (1 ppm) and PGL (5 × 107 pfu/cm2) separately yielded significant initial reductions in L. monocytogenes (up to 0.7 log) compared to untreated samples. Enhanced additive reductions were achieved through the combined treatments of nisin and PGL. Fermentates in the CS salmon inhibited the growth of Listeria but did not lead to its eradication. The lowest levels of L. monocytogenes during storage were observed in nisin- and PGL-treated CS salmon containing preservative fermentates and stored at 4 °C, while enhanced growth was observed during storage at an abusive temperature of 8 °C. Evaluation of industry-processed standard and sodium-replaced CS salmon confirmed significant effects with up to 1.7 log reductions in L. monocytogenes levels after 34 days of storage of PGL- and nisin-treated CS salmon-containing fermentates. No differences in total aerobic plate counts were observed between treated (PGL and nisin) or non-treated standard and sodium-reduced CS salmon at the end of storage. The microbiota was dominated by Photobacterium, but with a shift showing dominance of Lactococcus spp. and Vagococcus spp. in fermentate-containing samples. Similar and robust reductions in L. monocytogenes can be achieved in both standard and sodium-replaced CS salmon using the bio-preservation strategies of nisin, PGL and fermentates under various and relevant processing and storage conditions.