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The original notifications (n=1355) managed by the Rapid Alert System for Food and Feed of the Tuscany region in the seven-year period 2015-2021 were analyzed. 68.9% of them were classified as alert notifications, and they mostly originated (56.3%) from official controls on the market, followed by the company's own checks (29.1%). Italy was the most represented notifying country (73.3%) and the most represented country of origin of the notified products (64.5%). Out of the 28 considered food categories, "fish and products thereof" (F), "cereals and bakery products" (C), "bivalve mollusks and products thereof" (B), and "meat and meat products (other than poultry)" (M) were the most notified. F were especially notified for the presence of heavy metals (45.7%), C for undeclared allergens (32.1%), B for microbial contaminants (Escherichia coli, 49.6%), and M for pathogenic microorganisms (55.9%), mostly Salmonella spp. and Listeria monocytogenes. This study provides an updated framework for the main food safety issues at the regional level. The Rapid Alert System for Food and Feed should be periodically analyzed to evaluate historical trends and emerging or poorly known hazards. This is especially important for the planning of official control activities and the prevention of risks through the food supply chain. [ABSTRACT FROM AUTHOR]

This article explores the role of Akkermansia muciniphila, a type of gut bacteria, in preventing Listeria monocytogenes infection in mice. The study found that mice treated with A. muciniphila had reduced bacterial loads and improved intestinal barrier function compared to control mice. A. muciniphila was also found to increase levels of linoleic acid in the intestines, which strengthened the epithelial barrier and protected against L. monocytogenes infection. These findings suggest that A. muciniphila or linoleic acid-based strategies could be potential approaches for preventing listeriosis. [Extracted from the article]

Current knowledge about effects of disturbance on the fate of invaders in complex microbial ecosystems is still in its infancy. In order to investigate this issue, we compared the fate of Klebsiella pneumoniae (Kp) and Listeria monocytogenes (Lm) in soil microcosms. We then used environmental disturbances (freeze–thaw or heat cycles) to compare the fate of both invaders and manipulate soil microbial diversity. Population dynamics of the two pathogens was assessed over 50 days of invasion while microbial diversity was measured at times 0, 20 and 40 days. The outcome of invasion was strain-dependent and the response of the two invaders to disturbance differed. Resistance to Kp invasion was higher under the conditions where resident microbial diversity was the highest while a significant drop of diversity was linked to a higher persistence. In contrast, Lm faced stronger resistance to invasion in heat-treated microcosms where diversity was the lowest. Our results show that diversity is not a universal proxy of resistance to microbial invasion, indicating the need to properly assess other intrinsic properties of the invader, such as its metabolic repertoire, or the array of interactions between the invader and resident communities. [ABSTRACT FROM AUTHOR]

Among 15 strains of Listeria monocytogenes tested, a synergy between amoxicillin and ceftriaxone was observed in 14 (93%) according to minimal inhibitory concentration strips and 12 (80%) per the checkerboard methods, as well as for 2 of the 3 strains tested by the time-killing curve. This association may be an alternative treatment for listeriosis in the future. [ABSTRACT FROM AUTHOR]

This study involved the evaluation of the effect of vacuum pasteurization on physicochemical characteristics (pH, total soluble solids, titratable acidity, chroma, tone, IO, vitamin C, 5-hydroxymethylfurfural), microbiological properties (Staphylococcus aureus, Listeria monocytogenes, Escherichia coli, total coliforms, total mesophilic aerobes, molds and yeasts) and sensory characteristics of orange and carrot nectar. The thermal treatments were designed based on the thermal lethality of two heat-resistant microorganisms typical of the product (Neosartorya fischeri and Zygosaccaromyces bailii). The evaluation was carried out on raw nectar and pasteurized nectar. The shelf life was estimated to be 30 days (6 °C). The most favorable results were obtained by applying a heat treatment at 88 °C for 32.68 min, managing to retain 85.87% of vitamin C and a microbiological stability of 12 days (6 ± 0.6 °C) with regard to total mesophilic aerobes. Likewise, the tasters established that this treatment resulted in the best flavor, texture and acceptability characteristics. [ABSTRACT FROM AUTHOR]

Simple Summary: As a crucial component of the innate immune system, inflammasomes are activated by cellular infection or stress, which leads to the secretion of proinflammatory cytokines IL-1β and IL-18. In order to avert harmful inflammatory responses and maintain immune homeostasis, the precise control mechanisms of inflammasome activation need to be further defined. Previously, researchers have characterized the bovine SP110c isoform (bSP110c) as a new trigger of inflammasome activation in macrophages stimulated with bacterial lipopolysaccharide; however, the exact molecular mechanism for negative regulation of bSP110c-induced inflammasome remains unknown. In this study, the researchers demonstrated that bovine DDX3X is a novel bSP110c binding protein that restricts the bSP110c-mediated inflammatory response through the negative regulation of the NLRP3 inflammasome. These findings support a new cooperative mechanism between bSP110c and bovine DDX3X to balance inflammasome activation and advance our understanding of the function of the SP110c protein in the bovine innate immune system. The inflammasome is a vital part of the host's innate immunity activated by cellular infection or stress. Our previous research identified the bovine SP110c isoform (bSP110c) as a novel activator of the inflammasome that promoted the secretion of proinflammatory cytokines IL-1β and IL-18 in macrophages infected with Listeria monocytogenes or stimulated with lipopolysaccharide (LPS). However, the exact molecular mechanism for inhibiting bSP110c-induced inflammasome activation requires further clarification. Here, the researchers identified bovine DDX3X (bDDX3X) as an NLRP3-associated protein and an inhibitor of the bSP110c-induced inflammasome in the human THP1 macrophage cell line. Immunoprecipitation showed that bDDX3X interacted with the bSP110c CARD domain via its helicase domain. The co-expression of bSP110c and bDDX3X in THP1 macrophages significantly prevented the bSP110c-induced activation of inflammasomes. In addition, both bDDX3X and bSP110c interacted with bovine NLRP3 (bNLRP3), and bDDX3X enhanced the interaction between bSP110c and bNLRP3. The expression of bDDX3X in nigericin-stimulated THP1 macrophages significantly suppressed NLRP3 inflammasome activation, ASC speck formation, and pyroptosis. These findings demonstrate that bDDX3X negatively regulates the bSP110c-mediated inflammatory response by restricting the activation of the NLRP3 inflammasome. This discovery unveils a novel regulatory mechanism involving bDDX3X and bSP110c in coordinating inflammasome activation and subsequent cell-fate decisions in LPS-treated macrophages and, in turn, constitutes a step forward toward the implementation of marker-assisted selection in breeding programs aimed at utilizing cattle's immune defenses. [ABSTRACT FROM AUTHOR]

The essential oils from Thymus vulgaris, Salvia officinalis, and Ocimum basilicum were extracted by hydrodistillation, characterized by gas chromatography/mass spectrometry, and quantified by gas chromatography/flame ionization detector. The principal constituents were thymol, ρ‐cymene and carvacrol (T. vulgaris); camphor, β‐pinene, and 1,8‐cineole (S. officinalis); and (E)‐anethole, linalool, and 1,8‐cineole (O. basilicum). The essential oil from T. vulgaris was the most effective, forming inhibition halos of 46.16 ± 0.16 and 26.38 ± 0.33 mm, respectively, for Salmonella choleraesuis and Listeria monocytogenes. This essential oil was also more effective against S. choleraesuis, with a minimum inhibitory concentration of 8.85 mg mL−1, and a minimum inhibitory concentration of 17.71 mg mL−1 for L. monocytogenes. No bactericidal activity against S. choleraesuis and L. monocytogenes was observed for the essential oils from S. officinalis, and O. basilicum. Scanning electron micrographs showed that the addition of essential oils left the bacterial cells damaged and deformed. Significant 2,2‐diphenyl‐1‐picrylhydrazyl free radical scavenging capacity and lipid substrate protection were observed in the β‐carotene bleaching assay for the essential oil from T. vulgaris, with IC50 of 231.13 ± 0.53 and 15.25 ± 0.38 μg mL−1, respectively. A dose‐dependent relationship between antioxidant activity and concentrations was observed in the tests. Toxicities of LC50 = 1.24, 3.51 and 1.19 mg mL−1 against Drosophila suzukii flies, respectively, were observed for the essential oils from T. vulgaris, S. officinalis, and O. basilicum. Results suggest that essential oils can be promising antioxidant agents, insecticides, and inhibitors of pathogenic bacteria. [ABSTRACT FROM AUTHOR]

Fresh apples are vulnerable to Listeria monocytogenes (L. monocytogenes) contamination during handling and storage causing subsequent foodborne outbreaks. Gaseous chlorine dioxide (ClO2) is an effective antimicrobial agent against L. monocytogenes and could augment food safety. This study investigates the stress and virulence response of L. monocytogenes on apples exposed to a sublethal concentration of ClO2. L. monocytogenes was inoculated on the surface of apples and exposed to gaseous ClO2 for 1 h. Gene expression analysis via Real‐Time quantitative polymerase chain reaction (RT‐qPCR) revealed minimal changes in gene expression compared to control samples. However, a surface‐dependent response was observed, with slight upregulation of relevant virulence (hly) and stress (clpC) genes on more hydrophilic surfaces. These findings indicate that gaseous ClO2 treatment can provide a biostatic effect against L. monocytogenes on fresh produce, shedding light on its antimicrobial mechanism and potential for enhancing apple safety. [ABSTRACT FROM AUTHOR]

The current foodborne pathogen detection methods, such as culture-based methods, polymerase chain reaction, and optical and electrochemical biosensors with nucleic acid, have high sensitivity and selectivity. However, they are slow, expensive, and require well-trained operators. In this study, we utilized a 3D printer to develop a novel chip with an aptamer-based nanointerferometer capable of identifying four distinct foodborne pathogens: Listeria monocytogenes, Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus. The aptamer sensor on the chip achieved a limit-of-detection of 10 colony forming unit (CFU)/ml. With its high sensitivity and specificity, this chip offers a cost-effective platform for distinguishing and screening different foodborne pathogens. [ABSTRACT FROM AUTHOR]

A one-shot CO2 laser-based strategy to generate conductive reduced graphene oxide (rGO) decorated with nanoceria (nCe) is proposed. The 2D/0D rGO-nCe films, integrated as catalytic sensing layers in paper-based sensors, were employed for on-site monitoring of indoor fogging treatments against Listeria monocytogenes (Lm), a ubiquitous pathogenic bacterium. The rGO-nCe laser-assisted synthesis was optimized to preserve the rGO film morphological and electron-transfer features and simultaneously integrate catalytic nCe. The films were characterized by microscopical (SEM), spectroscopical (EDX, Raman, and FTIR), and electrochemical techniques. The most performing film was integrated into a nitrocellulose substrate, and the complete sensor was assembled via a combination of xurography and stencil printing. The rGO-nCe sensor's catalytic activity was proved toward the detection of H2O2, obtaining sensitive determination (LOD = 0.3 µM) and an extended linear range (0.5–1500 µM). Eventually, the rGO-nCe sensor was challenged for the real-time continuous monitoring of hydrogen peroxide aerosol during no-touch fogging treatment conducted following the EU's recommendation for biocidal product use. Treatment effectiveness was proved toward three Lm strains characterized by different origins, i.e., type strain ATCC 7644, clinical strain 338, and food strain 641/6II. The sensor allows for discrimination and quantification treatments at different environmental biocidal amounts and fogging times, and correlates with the microbiological inhibition, promoting the proposed sensor as a useful tool to modulate and monitor no-touch treatments. [ABSTRACT FROM AUTHOR]

Between 2012 and 2024, a prolonged outbreak of 73 cases of L. monocytogenes ST173 infections ('My2' ‐cluster) has been ongoing in Belgium (5), Czechia (1), Germany (39), Finland (2), Italy (1), the Netherlands (20), and the United Kingdom (UK) (5). Fourteen deaths have been recorded as associated with this outbreak. Males aged over 60 years represent the most affected population group. Most patients with available information from case interviews report consuming various fish products before illness. Among 73 cases, an ongoing 'My2' sub‐cluster 1 was identified with 38 cases reported in six EU countries and the UK. The 'My2' sub‐cluster 1 includes 83 non‐human isolates, of which 48 are food isolates and six are environmental isolates with traceability data corresponding to 37 fish products (different typologies) and 12 fish manufacturers (located in six EU countries and one third country). The epidemiological and traceability information did not allow identification of a single common point or food business operator in the traceability of all products. Five countries reported the implementation of control measures. The whole genome analysis (WGS) analysis, the tracing evidence, and the detection in fish products between 2017 and 2024 suggest that the strain has been geographically spread in Europe for several years, most likely originated from a past single source high up in the production chain, and has been established in different fish processing plants. Overall, the evidence supports the hypothesis of fish products as the vehicle of infections. Given the detection of the outbreak strain in multiple types of fish products and its wide circulation in the EU/EEA fish production chain, new cases are likely to be reported. Corrective measures should be implemented in the plants where the contamination is detected, and further investigations should be carried out to identify the point of entry and prevent possible re‐contamination. [ABSTRACT FROM AUTHOR]

An enhanced fluorescent DNA probe based on aggregation-induced emission fluorogens (AIEgens) was developed, and a new method for sensitive detection of Listeria monocytogenes (LM) was established based on recombinant polymerase amplification (RPA) combined with CRISPR/Cas12a. The AIEgens enhanced probe contained both single-stranded and double-stranded DNA sequence, where a Black Hole Quencher group was labeled on the 5' end of probe, and the cationic AIEgens dramatically enhanced the fluorescent intensity of DNA probe via binding with the dsDNA. The fluorescent signal of the developed probe was turned on by the trans-cleavage of Cas12a protein, which was trigged by the target DNA fragment of LM. Using the AlEgens enhanced probes as a signal output, the sensitivity of CRISPR/Cas12a for the dsDNA fragment was achieved at 0.37 pmol·L-1,40 times higher than that of traditional fluorescent reporter. Combined with recombinase polymerase amplification, the sensitivity of the proposed CRISPR/Cas12a system was achieved at 3×10¹ CFU mL-1 for the LM determination. Intra-and inter-assays were evaluated by determining the artificially contaminated LM milk and flammulina mushroom samples. and the average recoveries ranged from 91.14% to 108.47% with the coefficient of variation less than 12.71%, indicating an accepted accuracy and precision. An AIEgens enhanced fluorescent DNA probe was constructed to improve the detection efficiency of CRISPR/Cas system, the proposed CRISPR/Cas system achieved highly sensitivity for the foodborne pathogenic detection. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes, a prominent foodborne pathogen responsible for zoonotic infections, owes a significant portion of its virulence to the presence of the phospholipase PlcB. In this study, we performed an in-depth examination of the intricate relationship between L. monocytogenes PlcB and host cell mitochondria, unveiling a novel participant in bacterial survival: the mitochondrial carboxylase propionyl-coenzyme A carboxylase (PCCA). Our investigation uncovered previously unexplored levels of interaction and colocalization between PCCA and PlcB within host cells, with particular emphasis on the amino acids 504-508 of PCCA, which play a pivotal role in this partnership. To assess the effect of PCCA expression on L. monocytogenes proliferation, PCCA expression levels were manipulated by siRNA-si-PCCA or pCMV-N-HA-PCCA plasmid transfection. Our findings demonstrated a clear inverse correlation between PCCA expression levels and the proliferation of L. monocytogenes. Furthermore, the effect of L. monocytogenes infection on PCCA expression was investigated by assessing PCCA mRNA and protein expression in HeLa cells infected with L. monocytogenes. These results indicate that L. monocytogenes infection did not significantly alter PCCA expression. These findings led us to propose that PCCA represents a novel participant in L. monocytogenes survival, and its abundance has a detrimental impact on bacterial proliferation. This suggests that L. monocytogenes may employ PlcB-PCCA interactions to maintain stable PCCA expression, representing a unique pro-survival strategy distinct from that of other intracellular bacterial pathogens. [ABSTRACT FROM AUTHOR]

The foodborne pathogen Listeria monocytogenes is differentiated into four distinct lineages which differ in their virulence. It remains unknown, however, whether the four lineages also differ with respect to their ability to persist in food processing facilities, their resistance to high pressure, a preservation method that is used commercially for Listeria control on ready-to-eat meats, and their ability to form biofilms. This study aimed to determine differences in the pressure resistance and biofilm formation of 59 isolates of L. monocytogenes representing lineages I and II. Furthermore, the genetic similarity of 9 isolates of L. monocytogenes that were obtained from a meat processing facility over a period of 1 year and of 20 isolates of L. monocytogenes from food processing facilities was analyzed to assess whether the ability of the lineages of L. monocytogenes to persist in these facilities differs. Analysis of 386 genomes with respect to the source of isolation revealed that genomes of lineage II are over-represented in meat isolates when compared with clinical isolates. Of the 38 strains of Lm. monocytogenes that persisted in food processing facilities (this study or published studies), 31 were assigned to lineage II. Isolates of lineage I were more resistant to treatments at 400 to 600 MPa. The thickness of biofilms did not differ between lineages. In conclusion, strains of lineage II are more likely to persist in food processing facilities while strains of lineage I are more resistant to high pressure. [ABSTRACT FROM AUTHOR]

Although various types of mRNA-based vaccines have been explored, the optimal conditions for induction of both humoral and cellular immunity remain rather unknown. In this study, mRNA vaccines of nucleoside-modified mRNA in lipoplexes (LPXs) or lipid nanoparticles (LNPs) were evaluated after administration in mice through different routes, assessing mRNA delivery, tolerability and immunogenicity. In addition, we investigated whether mRNA vaccines could benefit from the inclusion of the adjuvant alpha-galactosylceramide (αGC), an invariant Natural Killer T (iNKT) cell ligand. Intramuscular (IM) vaccination with ovalbumin (OVA)-encoding mRNA encapsulated in LNPs adjuvanted with αGC showed the highest antibody- and CD8+ T cell responses. Furthermore, we observed that addition of signal peptides and endocytic sorting signals of either LAMP1 or HLA-B7 in the OVA-encoding mRNA sequence further enhanced CD8+ T cell activation although reducing the induction of IgG antibody responses. Moreover, mRNA LNPs with the ionizable lipidoid C12–200 exhibited higher pro-inflammatory- and reactogenic activity compared to mRNA LNPs with SM-102, correlating with increased T cell activation and antitumor potential. We also observed that αGC could further enhance the cellular immunity of clinically relevant mRNA LNP vaccines, thereby promoting therapeutic antitumor potential. Finally, a Listeria monocytogenes mRNA LNP vaccine supplemented with αGC showed synergistic protective effects against listeriosis, highlighting a key advantage of co-activating iNKT cells in antibacterial mRNA vaccines. Taken together, our study offers multiple insights for optimizing the design of mRNA vaccines for disease applications, such as cancer and intracellular bacterial infections. [Display omitted] [ABSTRACT FROM AUTHOR]

Microbial biofilm formation represents a serious problem for both food industry and households. Natural biofilms are formed mostly by multiple species, and show resistance against most of the usual sanitizers. In this study, the effects of cinnamon (Cinnamomum zeylanicum), marjoram (Origanum majorana) and thyme (Thymus vulgaris) essential oils (EOs) and their main components (cinnamaldehyde, terpinene-4-ol, and thymol) were investigated on four-species biofilms of Escherichia coli, Listeria monocytogenes, Pseudomonas putida and Staphylococcus aureus. Minimum bactericide concentration (MBC) and killing time were determined by means of the microdilution method. MBC of the investigated EOs and components was between 0.5 mg/mL (cinnamaldehyde) to 25 mg/mL (terpinene-4-ol). Killing times for the four-species suspension were 5 or 10 min, time spans usable in the food industry. For eradication of the mixed-population biofilm from stainless steel (SS), polypropylene (PP), tile and wood surfaces, EO- or EO component-based disinfectant solutions were developed, and their effects were compared to a peracetic acid-based industrial sanitizer (HC-DPE). Total eradication of biofilms (99.9%) was achieved, with solutions containing cinnamon and thyme EO and EO components, from SS and PP, but not from tile or wood surfaces. Apparently, cinnamon EO, terpinene-4-ol and thymol have better disinfectant activity than HC-DPE. [ABSTRACT FROM AUTHOR]

All bacterial strains studied retained the viability and ability to form both mono- and polycultural biofilms under conditions of long-term culturing in artificial seawater at 6°C and without addition of nutrients. Bacillus sp. and Pseudomonas japonica presumably stimulated the growth and reproduction of the pathogenic bacteria Listeria monocytogenes and Yersinia pseudotuberculosis. Preserved cell viability in a monoculture biofilm for a long period without adding a food source can indicate allolysis. At the same time, in a polycultural biofilm, the metabolites secreted by saprotrophic strains can stimulate the growth of L. monocytogenes and Y. pseudotuberculosis. [ABSTRACT FROM AUTHOR]

Introduction: Listeriosis is a severe food-borne disease caused by Listeria monocytogenes infection. The data of listeriosis in Xi'an population are limited. The aim of this study is to evaluate the clinical features and fatality risk factors for listeriosis in three tertiary-care hospitals in Xi'an, China Methods: The characteristics of demographic data, underlying diseases, clinical manifestations, laboratory indicators, cranial imaging examination, antibiotics therapeutic schemes, and clinical outcomes were collected between 2011 and 2023. Logistic regression analysis was performed. Results: Seventy-one etiologically confirmed listeriosis patients were enrolled, including 12 neonatal and 59 non-neonatal cases. The majority of neonatal listeriosis presented as preterm (50%) and fetal distress (75%). The main clinical manifestations of non-neonatal listeriosis included fever (88%), headache (32%), disorder of consciousness (25%), vomiting (17%), abdominal pain (12%), and convulsions (8%). The fatality rate in neonatal cases was higher than in non-neonatal listeriosis (42 vs. 17%). Although no deaths were reported in maternal listeriosis, only two of 23 patients had an uneventful obstetrical outcome. Five maternal listeriosis delivered culture-positive neonates, three of whom decreased within 1 week post-gestation due to severe complications. Twenty-eight cases were neurolisteriosis and 43 cases were bacteremia. Neurolisteriosis had a higher fatality rate compared with bacteremia listeriosis (36 vs. 12%). The main neuroradiological images were cerebral edema/hydrocephalus, intracranial infection, and cerebral hernia. Listeria monocytogenes showed extremely low resistance to ampicillin (two isolates) and penicillin (one isolate). The fatality risk factors were the involvement of the central nervous system, hyperbilirubinemia, and hyponatremia for all enrolled subjects. Hyperuricemia contributed to the elevation of fatality risk in non-neonatal listeriosis. Conclusions: When the patients suffered with symptoms of fever and central nervous system infection, they should be alert to the possibility of listeriosis. Early administration of ampicillin- or penicillin-based therapy might be beneficial for recovery of listeriosis. [ABSTRACT FROM AUTHOR]

Lactic acid bacteria (LAB) can produce peptides known as bacteriocins with antagonistic activity against foodborne pathogens. The potential of LAB isolated from the surface of jalapeno peppers to produce bacteriocins with antagonistic activity against Listeria monocytogenes, Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella Typhimurium was evaluated. Previously isolated LAB strains were reactivated, and their cell-free supernatants (CFSs) were evaluated. Out of 390 reactivated strains, 60 produced bacteriocin-like inhibitory substances (BLIS) since their antagonistic activity was lost after proteases addition. Subsequently, 16 BLIS showed heat resistance (HR-BLIS), retaining their bioactivity after heat treatment (121 °C for 15 min). By 16S rRNA gene sequencing and antibiotic susceptibility tests, LAB strains producing HR-BLIS were identified as Enterococcus lactis. Four HR-BLIS exhibited a minimum inhibitory concentration (MIC) of 80 mg/mL against L. monocytogenes. MIC and minimum bactericidal concentration (MBC) of HR-BLIS-67 for S. aureus (MIC = 80 mg/mL; MBC = 320 mg/mL), S. Typhimurium (MIC = 150 mg/mL; MBC = 250 mg/mL), and E. coli O157:H7 (MIC = 250 mg/mL; MBC = 400 mg/mL) were determined. LAB isolated from the surface of jalapeno pepper produced HR-BLIS (possibly enterocin) that exhibited broad-spectrum antagonistic activity against foodborne pathogens; therefore, they are a promising source of natural antimicrobials to ensure food safety. [ABSTRACT FROM AUTHOR]

The objectives of this paper were to investigate the existence of Listeria monocytogenes (L. monocytogenes) in different types of cheese (Mozzarella, Thalaga, and Romy cheeses, 40 samples each) sold in Assiut dairy shops and to evalute antibacterial properties of chitosan and its nanoparticles against isolated strains. L. monocytogenes was isolated in higher percentage from Mozzarella cheese (25%) followed by Thalaga and Romy cheese samples. L. monocytogenes isolates were validated by PCR utilising the 16S rRNA gene. Zeta-sizer and TEM were used to characterise nano-chitosan. The minimum inhibitory concentrations of chitosan and nano-chitosan on L. monocytogenes were detected by MIC test. Nano-chitosan (3.125%) could prevent organism growth in cheese as early as the third week. The findings revealed that 3.125% nano-chitosan had a sufficient effect on L. monocytogenes while also providing good consumer satisfaction; therefore, nano-chitosan should be utilised as a natural food additive in Egyptian dairy products. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes, the causative agent of listeriosis, is a relevant pathogen in dry fermented sausages (DFSs), and the application of antilisterial starter cultures is an effective intervention strategy to control the pathogen during DFS production. The effect of factors in relation to DFS formulation and production, NaCl (0–40 g/L), Mn (0.08–0.32 g/L), glucose (0–40 g/L) and temperature (3–37 °C), on the behaviour of L. monocytogenes when cocultured with Latilactobacillus sakei 23K (non-bacteriocinogenic) and CTC494 (bacteriocinogenic) strains was studied through a central composite design in meat simulation media. L. sakei and L. monocytogenes counts, pH, lactic acid production and bacteriocin activity were determined in mono and coculture. The pH decrease and lactic acid production were highly influenced by glucose, while production of sakacin K by L. sakei CTC494 was observed at moderate (10 and 20 °C), but not at the lowest (3 °C) and highest (37 °C), temperatures. Coculture growth had no effect on the acidification and bacteriocin production but inhibited and inactivated L. monocytogenes when L. sakei 23K entered the early stationary phase and when L. sakei CTC494 produced sakacin K. Optimal conditions for achieving a 5-log units reduction of L. monocytogenes were at 20 °C, 20 g/L of NaCl, 0.20 g/L of Mn and 40 g/L of glucose, those highlighting the importance of considering product formulation and fermentation conditions for bioprotective starter cultures application. [ABSTRACT FROM AUTHOR]

Listeria (L.) monocytogenes is an opportunistic foodborne pathogen that causes listeriosis in humans and animals, reaching up to 30% case mortality. There are only a few reports in Mexico about the L. monocytogenes strains found in various foods. The aim of this study was to determine the prevalence of L. monocytogenes, serogroups, virulence genes, and antimicrobial resistance in different foods from Reynosa, Tamaulipas, Mexico. L. monocytogenes strains were characterized by microbiological and molecular methods. Susceptibility to 12 antibiotics was determined according to CLSI and EUCAST. A total of 300 samples of seafood, pasteurized and raw milk, cheese, beef, and chicken were collected from supermarkets and retail markets. The presence of L. monocytogenes was detected in 5.6% of the samples. Most strains belonged to serogroups 4b, 4d, and 4e (68.4%). All strains presented a minimum of four virulence genes; the most common were actA, hly, and plcB (92.1%). A high percentage of antimicrobial susceptibility was observed, with resistance only to STX-TMP (78.9%), STR (26.3%), MEM (21.0%), and E (2.6%). These results show that the foods in Reynosa, Tamaulipas, are a reservoir of L. monocytogenes and represent a potential health risk. [ABSTRACT FROM AUTHOR]

Loop-mediated isothermal amplification, LAMP, is nowadays the most popular isothermal nucleic acid amplification technique, and as such, several commercial, ready-to-use master mixes have flourished. Unfortunately, independent studies to determine their performance are limited. The current study performed an independent evaluation of the existing ready-to-use commercial LAMP master mixes WarmStart® LAMP Kit, LavaLAMP™ DNA Master Mix, Saphir Bst Turbo GreenMaster, OptiGene Fast Master Mix ISO-004, and SynLAMP Mix. To reduce bias, three different genes, namely ttr (Salmonella spp.), rfbE (E. coli O157), and hly (Listeria monocytogenes), were targeted. The comparison was based on amplification speed, performance with decreasing DNA concentrations, and the effect of five typical LAMP reaction additives (betaine, DMSO, pullulan, TMAC, and GuHCl). Significant differences were observed among the different master mixes. OptiGene provided the fastest amplification and showed less detrimental effects associated with the supplements evaluated. Out of the chemicals tested, pullulan provided the best results in terms of amplification speed. It is noteworthy that the different additives impacted the master mixes differently. Overall, the current study provides insights into the performance of commercial LAMP master mixes, which can be of value for the scientific community to better select appropriate reagents when developing new methods. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is an important foodborne pathogen implicated in outbreaks related to soft cheese, indicating the importance of its control. In the current study, the antimicrobial efficacy of octanoic acid (OA) and lauric arginate ester (LAE), individually or in combination, against L. monocytogenes in Domiati cheese was evaluated during storage at 4°C for 35 days. Data revealed that the population of L. monocytogenes in untreated Domiati cheese gradually increased during refrigerated storage. The application of 400ppm OA, 200ppm LAE, and 200ppm LAE+200ppm OA reduced the inoculated L. monocytogenes in Domiati cheese with 1.72±0.11, 2.14±0.01, and 2.63±0.08 log CFU/g reduction respectively, after 35 days of refrigerated storage. Moreover, the combination of 200ppm LAE+400ppm OA was the most effective treatment, leading to a 3.31±0.03 log CFU/g reduction of L. monocytogenes by the end of the refrigerated storage period. This study is a guide for the practical application of octanoic acid and lauric arginate during the manufacturing of Domiati cheese to control L. monocytogenes during the refrigerated storage. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes, along with various other pathogenic bacteria, may show resistance against a broad spectrum of antibiotics. Evaluating the extent of resistance in harmful microorganisms like Listeria monocytogenes holds significant importance in crafting novel therapeutic strategies to mitigate or combat the rise of infections stemming from antibiotic-resistant bacteria. The present work aims to investigate the occurrence of antimicrobial resistance among Listeria monocytogenes strains in meat products (n = 173), seafood (n = 54), dairy products (n = 19), sauces (n = 2), confectionary products (n = 1), ready-to-eat rice dishes (n = 1), and food-processing environments (n = 19). A total of 269 Listeria monocytogenes strains belonging to eight different serovars were tested against 10 antimicrobials. In the classes of antibiotics, most of the strains were resistant antibiotics belonging to the family of β-lactams (92.94%). High proportions of L. monocytogenes isolates were resistant to oxacillin (88.48%), followed by fosfomycin (85.87%) and flumenique (78.44%). The lowest level of resistance was observed against gentamycin (1.49%). A total of 235 strains (n = 87.36%) showed a profile of multidrug resistance. In conclusion, a high occurrence of resistant and multidrug-resistant strains of Listeria monocytogenes was observed among the examined serotypes isolated from different food sources. This understanding enables the adoption of suitable measures to avert contamination and the spread of resistant bacteria via food. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes, a foodborne pathogen causing listeriosis, poses substantial societal, economic, and public health challenges due to its resistance, persistence, and biofilm formation in the food industry. Exploring subinhibitory concentrations of compounds to target virulence inhibition and increase susceptibility to adverse conditions presents a promising strategy to mitigate its impact of L. monocytogenes and unveils new potential applications. Thus, this study aims to explore the effect of linalool on virulence factors of L. monocytogenes and potential use in the reduction in its tolerance to stressful conditions. This action was analysed considering the use of two sub-inhibitory concentrations of linalool, 0.312 and 0.625 mg/mL. We found that even with the lowest tested concentrations, a 65% inhibition of violacein production by Chromobacterium violaceum, 55% inhibition in biofilm formation by L. monocytogenes and 62% reduction on haemolysis caused by this bacterium were observed. In addition to its impact on virulence factors, linalool diminished the tolerance to osmotic stress (up to 4.3 log reduction after 24 h with 12% NaCl), as well as to high (up to 3.8 log reduction after 15 min at 55 °C) and low temperatures (up to 4.6 log reduction after 84 days with 12% NaCl at 4 °C). Thus, this study paves the way to further investigation into the potential utilization of linalool to mitigate the threat posed by L. monocytogenes in the field of food safety and public health. [ABSTRACT FROM AUTHOR]

In recent years, there has been an increasing interest in antibacterial biopolymeric films. Among the different approaches for tuning the release kinetic of antibacterial compounds, the use of natural fillers allows for this purpose while optimizing the processability and the mechanical properties of the products. In this work, the effect of three different concentrations of biochar (BC) was investigated on the morphological, rheological, mechanical, and thermal properties of Polybutylene adipate terephthalate/Carvacrol/BC (PBAT/CV/BC) ternary biocomposites. The films were fabricated by means of melt mixing and compression molding and compared to PBAT/BC samples. The carvacrol kinetic release was evaluated as a function of the BC concentration in the ternary system. Results highlighted that BC allows tuning the properties of PBAT and of PBAT/CV samples and modifying the release kinetic of CV. Finally, the antimicrobial analysis revealed that PBAT/CV films exhibited excellent antimicrobial properties against Escherichia coli, Listeria monocytogenes, Salmonella Enteritidis, and Staphylococcus aureus. [ABSTRACT FROM AUTHOR]

Fresh cheese is characterized by a limited shelf life, which represents a major challenge in its production. Wild thyme (Thymus serpyllum L.) has an antimicrobial capacity demonstrated in numerous studies. The utilisation of its by-product obtained in the production of filter tea could improve fresh cheese technology by obtaining a product with additional functional value and protecting the environment by reducing industrial waste. Our study sought to explore how incorporating wild thyme, in the form of dry extract, supercritical fluid extract, and herbal ground, affects the microbiological composition of fresh cheese made with kombucha inoculum as the starter culture over a 30-day storage period. To assess antimicrobial efficacy, we deliberately exposed the samples we produced to common foodborne pathogens, namely Listeria monocytogenes and Staphylococcus aureus. The results showed that the total number of L. monocytogenes and S. aureus in each sample (produced with dry extract, supercritical fluid extract, and herbal ground) decreased significantly during the storage period. The decrease in L. monocytogenes count varied from 0.6 to 1.7 log CFU/g. The results suggest that a by-product from the production of wild thyme filter tea is suitable for the production of fresh cheese to improve its antimicrobial properties against L. monocytogenes and S. aureus. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a human pathogen that has the ability to cause listeriosis, a disease with possible fatal outcomes. The typical route of infection is ingestion of the bacteria with contaminated food. In this study, 13 virulence-associated genes were examined with PCR in the genomes of 153 L. monocytogenes isolates collected from meat products and processing environments in Poland. All isolates possessed genes from LIPI-1—hly, actA, plcA, plcB and mpl—as well as four internalins: inlA, inlB, inlC, inlJ. Invasion-associated protein iap, as well as genes prfA and sigB, encoding regulatory proteins, were also detected in all isolates. Gene flaA, encoding flagellin, was detected in 113 (74%) isolates. This was the only gene that was not detected in all isolates, as its presence is serotype-dependent. Gene actA showed polymorphism with longer and shorter variants in PCR amplicons. Two isolates were characterized by truncated inlB genes, lacking 141 bp in their sequence, which was confirmed by gene sequencing. All isolates were positive in hemolysis assays, proving the synthesis of functional PrfA and Hly proteins. Four genotypes of L. monocytogenes based on actA polymorphism and two genotypes based on inlB polymorphism were distinguished within the isolates' collection. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a ubiquitous bacterial pathogen that threatens the food chain and human health. In this study, whole-genome sequencing (WGS) was used for the genomic characterization of L. monocytogenes (n = 24) from beef and beef-based products. Multilocus Sequence Type (MLST) analysis revealed that ST204 of CC204 was the most common sequence type (ST). Other sequence types detected included ST1 and ST876 of CC1, ST5 of CC5, ST9 of CC9, ST88 of CC88, ST2 and ST1430 of CC2, and ST321 of CC321. Genes encoding for virulence factors included complete LIPI-1 (pfrA-hly-plcA-plcB-mpl-actA) from 54% (13/24) of the isolates of ST204, ST321, ST1430, and ST9 and internalin genes inlABC that were present in all the STs. All the L. monocytogenes STs carried four intrinsic/natural resistance genes, fosX, lin, norB, and mprF, conferring resistance to fosfomycin, lincosamide, quinolones, and cationic peptides, respectively. Plasmids pLGUG1 and J1776 were the most detected (54% each), followed by pLI100 (13%) and pLM5578 (7%). The prophage profile, vB_LmoS_188, was overrepresented amongst the isolates, followed by LP_101, LmoS_293_028989, LP_030_2_021539, A006, and LP_HM00113468. Listeria genomic island 2 (LGI-2) was found to be present in all the isolates, while Listeria genomic island 3 (LGI-3) was present in a subset of isolates (25%). The type VII secretion system was found in 42% of the isolates, and sortase A was present in all L. monocytogenes genomes. Mobile genetic elements and genomic islands did not harbor any virulence, resistance, or environmental adaptation genes that may benefit L. monocytogenes. All the STs did not carry genes that confer resistance to first-line antibiotics used for the treatment of listeriosis. The characterization of L. monocytogenes in our study highlighted the environmental resistance and virulence potential of L. monocytogenes and the risk posed to the public, as this bacterium is frequently found in food and food processing environments. [ABSTRACT FROM AUTHOR]

Extensive research has been conducted to identify key proteins governing stress responses, virulence, and antimicrobial resistance, as well as to elucidate their interactions within Listeria monocytogenes. While these proteins hold promise as potential targets for novel strategies to control L. monocytogenes, given their critical roles in regulating the pathogen's metabolism, additional analysis is needed to further assess their druggability—the chance of being effectively bound by small-molecule inhibitors. In this work, 535 binding pockets of 46 protein targets for known drugs (mainly antimicrobials) were first analyzed to extract 13 structural features (e.g., hydrophobicity) in a ligand–protein docking platform called Molsoft ICM Pro. The extracted features were used as inputs to develop a logistic regression model to assess the druggability of protein binding pockets, with a value of one if ligands can bind to the protein pocket. The developed druggability model was then used to evaluate 23 key proteins from L. monocytogenes that have been identified in the literature. The following proteins are predicted to be high-potential druggable targets: GroEL, FliH/FliI complex, FliG, FlhB, FlgL, FlgK, InlA, MogR, and PrfA. These findings serve as an initial point for future research to identify specific compounds that can inhibit druggable target proteins and to design experimental work to confirm their effectiveness as drug targets. [ABSTRACT FROM AUTHOR]

Current diagnostic methods for detecting foodborne pathogens are time-consuming, require sophisticated equipment, and have a low specificity and sensitivity. Magnetic nanoparticles (MNPs) and plasmonic/colorimetric biosensors like gold nanoparticles (GNPs) are cost-effective, high-throughput, precise, and rapid. This study aimed to validate the use of MNPs and GNPs for the early detection of Escherichia coli O157:H7, Salmonella enterica spp., Campylobacter jejuni, and Listeria monocytogenes in bovine fecal samples. The capture efficiency (CE) of the MNPs was determined by using Salmonella Typhimurium (ATCC_13311) adjusted at an original concentration of 1.5 × 108 CFU/mL. One (1) mL of this bacterial suspension was spiked into bovine fecal suspension (1 g of fecal sample in 9 mL PBS) and serially diluted ten-fold. DNA was extracted from Salmonella Typhimurium to determine the analytical specificity and sensitivity/LOD of the GNPs. The results showed that the CE of the MNPs ranged from 99% to 100% and could capture as little as 1 CFU/mL. The LOD of the GNPs biosensor was 2.9 µg/µL. The GNPs biosensor was also tested on DNA from 38 naturally obtained bovine fecal samples. Out of the 38 fecal samples tested, 81.6% (31/38) were positive for Salmonella enterica spp., 65.8% (25/38) for C. jejuni, 55.3% (21/38) for L. monocytogenes, and 50% (19/38) for E. coli O157:H7. We have demonstrated that MNP and GNP biosensors can detect pathogens or their DNA at low concentrations. Ensuring food safety throughout the supply chain is paramount, given that these pathogens may be present in cattle feces and contaminate beef during slaughter. [ABSTRACT FROM AUTHOR]

Copyright of Kocatepe Veterinary Journal / Kocatepe Veteriner Dergisi is the property of Afyon Kocatepe University, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The identification of the coproporphyrin‐dependent heme biosynthetic pathway, which is used almost exclusively by monoderm bacteria in 2015 by Dailey et al. triggered studies aimed at investigating the enzymes involved in this pathway that were originally assigned to the protoporphyrin‐dependent heme biosynthetic pathway. Here, we revisit the active site of coproporphyrin ferrochelatase by a biophysical and biochemical investigation using the physiological substrate coproporphyrin III, which in contrast to the previously used substrate protoporphyrin IX has four propionate substituents and no vinyl groups. In particular, we have compared the reactivity of wild‐type coproporphyrin ferrochelatase from the firmicute Listeria monocytogenes with those of variants, namely, His182Ala (H182A) and Glu263Gln (E263Q), involving two key active site residues. Interestingly, both variants are active only toward the physiological substrate coproporphyrin III but inactive toward protoporphyrin IX. In addition, E263 exchange impairs the final oxidation step from ferrous coproheme to ferric coproheme. The characteristics of the active site in the context of the residues involved and the substrate binding properties are discussed here using structural and functional means, providing a further contribution to the deciphering of this enigmatic reaction mechanism. [ABSTRACT FROM AUTHOR]

The microbiological quality of dairy products from small-scale producers in Serbia was analysed. A total of 302 dairy products [raw (n = 111) and pasteurized milk cheeses (n = 79) and kajmak (n = 112)], were collected and tested for the presence of pathogens, Listeria monocytogenes and Salmonella spp., and enumerated for Coagulase-positive staphylococci (CPS), Escherichia coli, and yeasts and moulds. None of the samples tested positive for Salmonella spp., while L. monocytogenes was recovered from one raw milk cheese and five kajmak samples. Raw milk cheese and kajmak also had higher levels of indicator microorganisms, namely E. coli and yeast and moulds. Molecular serotyping grouped L. monocytogenes isolates into serogroups 1 (1/2a and 3a) and 3 (1/2b, 3b, and 7). When exposed to eight antibiotics, L. monocytogenes isolates were mostly sensitive, with the exception of oxacillin and reduced susceptibility to clindamycin, penicillin G, and trimethoprim/sulfamethoxazole, emphasizing the importance of continuous surveillance for antimicrobial resistance. Samples that tested positive for Listeria spp. also had higher loads of indicator microorganisms, namely E. coli and yeast and moulds, suggesting lapses in hygiene practices during production. Collectively, these data emphasize the need for improved food safety and hygiene practices among small-scale dairy producers. This is crucial to reduce the microbial contamination and improve both the quality and safety of dairy products in the Serbian market. [ABSTRACT FROM AUTHOR]

We report a very rare case of Listeria multiple brain abscesses manifested as delirium, which represented diagnostic and therapeutic challenges overcome only by the close cooperation between Infectious Diseases and Neuroradiology, without which a satisfactory outcome would not be achieved. An elderly man presented with confusion and drowsiness with a background of type-II diabetes mellitus. Although computed tomography of the brain only showed frontal lobe oedema, contrast magnetic resonance (MR) imaging showed numerous irregular rim-enhancing lesions containing central diffusion restriction, suggesting multiple pyogenic cerebral abscesses of unclear aetiology. Thereafter, Listeria monocytogenes was isolated from blood cultures, suggesting this as the causative organism. Deemed unsuitable for neurosurgical drainage, the patient received medical management with a protracted course of antibiotics. This case was extremely challenging, due to 1) the impossibility of source control, 2) the small number of effective antibiotics available to treat this condition, and 3) the inevitable antibiotic side-effects, derived from long-term exposure. A successful outcome was only possible thanks to strict close multidisciplinary follow up, requiring frequent MR imaging and a judicious antibiotic choice, including monitoring of their side-effects. Due to the rarity of this condition, there is lack of guidance on its management, hence the importance of multidisciplinary involvement with very close imaging and antibiotic monitoring. [ABSTRACT FROM AUTHOR]

The study aims to identify antibacterial properties in unfertilized eggs of Scyliorhinus canicula and Scyliorhinus stellaris. Despite challenging marine conditions, these eggs retain their integrity for extended periods and remain unaffected by pathogens. The antibacterial activity was measured using minimum inhibitory and minimum bactericidal concentration analysis. The eggs of S. stellaris exhibited a slight inhibitory effect against Staphylococcus aureus and Listeria monocytogenes, whereas both species' eggs showed no activity against gram‐negative microorganisms. [ABSTRACT FROM AUTHOR]

The contamination of Listeria monocytogenes (L. monocytogenes) in dairy products poses a significant risk to human health. It is urgent to develop on‐site rapid strategy to reduce associated threats. We employed carbon dots (CDs) and silicon nanoparticles (SiNPs) as energy donors and acceptors respectively. The CDs and SiNPs were functionalised with vancomycin and aptamers correspondingly, leading to the establishment of ratiometric fluorescent sensor. With a portable smartphone‐integrated device, the ratios of blue/green values from the colour images depended on L. monocytogenes concentrations for quantification. The sensors were demonstrated for monitoring L. monocytogenes in dairy products within 30 min. [ABSTRACT FROM AUTHOR]

CdaA is the most widespread diadenylate cyclase in many bacterial species, including several multidrug‐resistant human pathogens. The enzymatic product of CdaA, cyclic di‐AMP, is a secondary messenger that is essential for the viability of many bacteria. Its absence in humans makes CdaA a very promising and attractive target for the development of new antibiotics. Here, the structural results are presented of a crystallographic fragment screen against CdaA from Listeria monocytogenes, a saprophytic Gram‐positive bacterium and an opportunistic food‐borne pathogen that can cause listeriosis in humans and animals. Two of the eight fragment molecules reported here were localized in the highly conserved ATP‐binding site. These fragments could serve as potential starting points for the development of antibiotics against several CdaA‐dependent bacterial species. [ABSTRACT FROM AUTHOR]

A fast and accurate identification of Listeria monocytogenes. A new quartz crystal microbalance (QCM) aptasensor was designed for the specific and rapid detection of L. monocytogenes. Before detection of the target bacterium from samples in the QCM aptasensor, a magnetic pre-enrichment system was used to eliminate any contaminant in the samples. The prepared magnetic system was characterized using ATR-FTIR, SEM, VSM, BET, and analytical methods. The saturation magnetization values of the Fe3O4, Fe3O4@PDA, and Fe3O4@PDA@DAPEG particles were 57.2, 40.8, and 36.4 emu/g, respectively. The same aptamer was also immobilized on the QCM crystal integrated into QCM flow cell and utilized to quantitatively detect L. monocytogenes cells from the samples. It was found that a specific aptamer-magnetic pre-concentration system efficiently captured L. monocytogenes cells in a short time (approximately 10 min). The Fe3O4@PDA@DA-PEG-Apt particles provided selective isolation of L. monocytogenes from the bacteria-spiked media up to 91.8%. The immobilized aptamer content of the magnetic particles was 5834 µg/g using 500 ng Apt/mL. The QCM aptasensor showed a very high range of analytical performance to the target bacterium from 1.0 × 102 and 1.0 × 107 CFU/mL. The limit of detection (LOD) and limit of quantitation (LOQ) were 148 and 448 CFU/mL, respectively, from the feeding of the QCM aptasensor flow cell with the eluent of the magnetic pre-concentration system. The reproducibility of the aptasensor was more than 95%. The aptasensor was very specific to L. monocytogenes compared to the other Listeria species (i.e., L. ivanovii, L. innocua, and L. seeligeri) or other tested bacteria such as Staphylococcus aureus, Escherichia coli, and Bacillus subtilis. The QCM aptasensor was regenerated with NaOH solution, and the system was reused many times. [ABSTRACT FROM AUTHOR]

A prolonged genomic cluster of 20 human Listeria monocytogenes ST1607 infections has been reported to the European Centre for Disease Prevention and Control, with cases identified in Denmark (17), Germany (1), and Italy (2) since 2019. The number of cases has sharply increased after 2021. The most recent case was reported in March 2024 in Denmark, indicating an ongoing risk of further infections. Of 20 listeriosis cases, five have died (one due to another cause than L. monocytogenes infection), indicating high severity of infection particularly among elderly people with underlying chronic conditions. The national investigations the traceability and genomic data analyses revealed the detection of five matching L. monocytogenes ST1607 isolates from four sliced smoked salmon products (four batches) produced by the Danish Processing Plant A between 2021 and 2024. These contaminated products were distributed to the three countries (Denmark, Germany, and Italy) reporting human cases to ECDC. The identification of the outbreak strain from the environment of the Danish Processing Plant A in 2023 and in the products since 2021 indicates the persistence of the L. monocytogenes ST1607 within the plant and suggests that the point(s) of the contamination in the plant have not been identified and controlled. Although control measures had been implemented at the plant and withdrawals and recalls of contaminated batches were performed, new batches of contaminated products could still be placed on the market until the site(s) at the plant and the root(s) of the contamination in the fish production chain are identified and properly controlled. Based on the available information from case interviews, microbiological and traceability analysis, the conclusion is that sliced smoked salmon products are the likely sources of infection. New cases could occur in EU/EEA countries, particularly among vulnerable people, until the root(s) of contaminations are identified and corrective measures are implemented. [ABSTRACT FROM AUTHOR]

This study aimed to generate low molecular weight peptides (LMWP) from fish collagen type I hydrolysed by increasing activity of Alcalase® from 0.0 to 12.0 (AU-A per 100 g) at 55 °C and 7.4 of pH for 3 h 40 min. The results showed that all enzyme activity levels caused 34–55 % reductions in protein recovery, 1.0–3.0 folds' increase in free α-amino groups and 1.7–3.2 folds' increase in carboxyl groups. Degree of hydrolysis ranged from 20 to 30 % with increasing enzyme activity. The number average molecular weight significantly reduced from 3200 g/mol in 0.0 AU-A per 100 g enzyme activity to 1151, 1398, 1175, 1040 and 1246 g/mol in 2.4, 4.8, 7.2, 9.6 and 12.0 AU-A per 100 g enzyme activities, respectively. Depending upon enzyme activity level, the produced LMWP with reactive carboxyl and amino end-groups exhibited 2.5- to 4.0-fold increases in antioxidant capacity and 1.0–3.5 log cfu/ml inhibition of four pathogen bacteria. Highest inhibition of 2.5 log cfu/ml in Escherichia coli was obtained from 2.4 AU-A per 100 g enzyme activity and 3.5 log cfu/ml in Listeria monocytogenes from 9.6 and 12.0 AU-A per 100 g enzyme activity levels. Infrared spectroscopy clearly identified reactive end-groups and showed remarkably differences in molar absorptivity of various molecular regions between non-enzyme and enzyme treated collagen type I molecule. A 9.6 and 12.0 AU-A per 100 g enzyme activity levels were found optimally effective to generate LMWP. In conclusion, LMWP exhibited high antioxidant and antibacterial activity due to increased functional reactive end-groups, and these bio-active peptides may have greater potentialities in various food and pharmaceutical applications. [ABSTRACT FROM AUTHOR]

Traditional and regional foods have been increasing in popularity among consumers in Poland for many years. The observed trend of searching for natural and authentic taste encourages many producers to craft products from raw milk, including Koryciński cheeses. The aim of this study was to assess the microbiological hazards resulting from the presence of pathogenic bacteria in Koryciński cheeses available in retail trade. The tests were carried out using accredited methods, including the detection of the presence of Salmonella spp., the enumeration of Listeria monocytogenes, the enumeration of coagulase-positive staphylococci, and the detection of staphylococcal enterotoxins in food when the number of coagulase-positive staphylococci in the sample exceeded the limit of 105 cfu/g. The research material consisted of 45 Koryciński cheeses. The tests conducted revealed that Salmonella spp. was not detected in any of the examined cheeses. However, coagulase-positive staphylococci were present in 68.9% of the samples. In as many as 15 tested cheeses, the level of S. aureus contamination was above 105 cfu/g; therefore, these samples were tested for the presence of staphylococcal enterotoxins. The presence of staphylococcal enterotoxins was found in one Koryciński cheese. In four cheeses, the number of L. monocytogenes exceeded the level of 102 cfu/g, the limit specified in Regulation 2073/2005 on microbiological criteria for foodstuffs. The obtained research results confirm the validity of monitoring the microbiological quality of Koryciński cheeses and the need to increase awareness of ensuring proper hygienic conditions of production, including the increased risk associated with unpasteurized milk products. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is the causative agent of listeriosis, a severe foodborne illness characterized by septicemia, meningitis, encephalitis, abortions, and occasional death in infants and immunocompromised individuals. L. monocytogenes is composed of four genetic lineages (I, II, III, and IV) and fourteen serotypes. The aim of the current study was to identify proteins that can serve as biomarkers for detection of genetic lineage III strains based on simple antibody-based methods. Liquid chromatography (LC) with electrospray ionization tandem mass spectrometry (ESI MS/MS) followed by bioinformatics and computational analysis were performed on three L. monocytogenes strains (NRRL B-33007, NRRL B-33014, and NRRL B-33077), which were used as reference strains for lineages I, II, and III, respectively. Results from ESI MS/MS revealed 42 unique proteins present in NRRL B-33077 and absent in NRRL B-33007 and NRRL B-33014 strains. BLAST analysis of the 42 proteins against a broader panel of >80 sequenced strains from lineages I and II revealed four proteins [TM2 domain-containing protein (NRRL B-33077_2770), DUF3916 domain-containing protein (NRRL B-33077_1897), DNA adenine methylase (NRRL B-33077_1926), and protein RhsA (NRRL B-33077_1129)] that have no homology with any sequenced strains in lineages I and II. The four genes that encode these proteins were expressed in Escherichia coli strain DE3 and purified. Polyclonal antibodies were prepared against purified recombinant proteins. ELISA using the polyclonal antibodies against 12 L. monocytogenes lineage I, II, and III isolates indicated that TM2 protein and DNA adenine methylase (Dam) detected all lineage III strains with no reaction to lineage I and II strains. In conclusion, two proteins including TM2 protein and Dam are potentially useful biomarkers for detection and differentiation of L. monocytogenes lineage III strains in clinical, environmental, and food processing facilities. Furthermore, these results validate the approach of using a combination of proteomics and bioinformatics to identify useful protein biomarkers. [ABSTRACT FROM AUTHOR]

We present an unusual case of Listeria monocytogenes rhomboencephalitis in a young, healthy patient. Although L. monocytogenes meningitis is usually associated with immunodeficiency, rhomboencephalitis is more commonly seen in immunocompetent patients. The wide differential for rhomboencephalitis can create a diagnostic challenge. Without prompt pathogen identification and appropriate antibiotic regimen, L. monocytogenes central nervous system infections can be fatal. Cerebro-Spinal Fluid (CSF) Polymerase Chain Reaction (PCR) aided a prompt diagnosis and adjustment of therapy to achieve a good patient outcome. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Food Safety & Quality is the property of Journal of Food Safety & Quality Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Listeria monocytogenes, a foodborne pathogen, exhibits high adaptability to adverse environmental conditions and is common in the food industry, especially in ready-to-eat foods. L. monocytogenes strains pose food safety challenges due to their ability to form biofilms, increased resistance to disinfectants, and long-term persistence in the environment. The aim of this study was to evaluate the presence and genetic diversity of L. monocytogenes in food and related environmental products collected from 2014 to 2022 and assess antibiotic susceptibility and biofilm formation abilities. L. monocytogenes was identified in 13 out of the 227 (6%) of samples, 7 from food products (meat preparation, cheeses, and raw milk) and 6 from food-processing environments (slaughterhouse-floor and catering establishments). All isolates exhibited high biofilm-forming capacity and antibiotic susceptibility testing showed resistance to several classes of antibiotics, especially trimethoprim-sulfamethoxazole and erythromycin. Genotyping and core-genome clustering identified eight sequence types and a cluster of three very closely related ST3 isolates (all from food), suggesting a common contamination source. Whole-genome sequencing (WGS) analysis revealed resistance genes conferring resistance to fosfomycin (fosX), lincosamides (lin), fluoroquinolones (norB), and tetracycline (tetM). In addition, the qacJ gene was also detected, conferring resistance to disinfecting agents and antiseptics. Virulence gene profiling revealed the presence of 92 associated genes associated with pathogenicity, adherence, and persistence. These findings underscore the presence of L. monocytogenes strains in food products and food-associated environments, demonstrating a high virulence of these strains associated with resistance genes to antibiotics, but also to disinfectants and antiseptics. Moreover, they emphasize the need for continuous surveillance, effective risk assessment, and rigorous control measures to minimize the public health risks associated to severe infections, particularly listeriosis outbreaks. A better understanding of the complex dynamics of pathogens in food products and their associated environments can help improve overall food safety and develop more effective strategies to prevent severe health consequences and economic losses. [ABSTRACT FROM AUTHOR]

Resident memory T (TRM) cells have been recently established as an important subset of memory T cells that provide early and essential protection against reinfection in the absence of circulating memory T cells. Recent findings showing that TRM expand in vivo after repeated antigenic stimulation indicate that these memory T cells are not terminally differentiated. This suggests an opportunity for in vitro TRM expansion to apply in an immunotherapy setting. However, it has also been shown that TRM may not maintain their identity and form circulating memory T cells after in vivo restimulation. Therefore, we set out to determine how TRM respond to antigenic activation in culture. Using Listeria monocytogenes and LCMV infection models, we found that TRM from the intraepithelial compartment of the small intestine expand in vitro after antigenic stimulation and subsequent resting in homeostatic cytokines. A large fraction of the expanded TRM retained their phenotype, including the expression of key TRM markers CD69 and CD103 (ITGAE). The optimal culture of TRM required low O2 pressure to maintain the expression of these and other TRM‐associated molecules. Expanded TRM retained their effector capacity to produce cytokines after restimulation, but did not acquire a highly glycolytic profile indicative of effector T cells. The proteomic analysis confirmed TRM profile retention, including expression of TRM‐related transcription factors, tissue retention factors, adhesion molecules, and enzymes involved in fatty acid metabolism. Collectively, our data indicate that limiting oxygen conditions supports in vitro expansion of TRM cells that maintain their TRM phenotype, at least in part, suggesting an opportunity for therapeutic strategies that require in vitro expansion of TRM. [ABSTRACT FROM AUTHOR]

The aim of using minimal food processing technologies is to preserve the raw material or to achieve a special technological goal with the least possible impact. When several technologies are used together, the intensity of each treatment can be reduced according to Leistner's hurdle principle. Does the order of application of the treatments result in a detectable difference? This research focuses on the effect of the combination of the sous-vide technology and the high hydrostatic pressure (HHP) treatment. The effect of the pressure level (300 and 600 MPa) and the influence of the treatment order was investigated on pork (longissimus thoracis and lumborum muscles (LTL)). Physico-chemical and microbiological measurements were carried out on day 0 and after 21-day storage at 2 °C and 8 °C. Significant differences were found for both the order of treatment and pressure level in weight loss (p < 0.001), CIELab color parameters a* and b* (p < 0.001), and denaturation enthalpy (p < 0.01). The texture (p < 0.001) and lipid oxidation TBARS values (p < 0.05) were only influenced by the pressure level. In the challenge test, the initial count of 105 CFU/g Listeria monocytogenes dropped below detection limit in all cases. Total aerobic and anaerobic viable numbers were below/near the detection limit in all combined-treated samples on day 0 and showed only slight or more notable growth after 21-day storage at 2 °C and 8 °C, respectively. An additional 300 MPa pressure treatment can increase the safety of sous-vide cooked pork samples while having only a minor effect on physicochemical properties. The 600 MPa pressure treatment results in a stable, albeit not shelf-stable product, but it also affects a considerable number of quality parameters such as color, texture, weight loss, and TBARS values. [ABSTRACT FROM AUTHOR]