"Four - in - one" platform based on multifunctional nanozyme for ultra - accurate detection and on - demand disinfection of Listeria monocytogenes.

The inability to integrate detection and disinfection hindered building a unified pathogen monitoring platform, risking secondary contamination. Herein, a novel "four - in - one" platform for monitoring foodborne Listeria monocytogenes (L. monocytogenes) was presented. The magnetic daptomycin - functionalized Fe3O4 (Dap/Fe3O4) could selectively bind to L. monocytogenes, enhancing detection accuracy. The separated bacteria were captured by aptamers - functionalized Fe - doped - silica nanoparticles (Apt/Fe@SiNPs) for tri - mode detection. Besides fluorescence, the Apt/Fe@SiNPs converted 3,3',5,5' - tetramethylbenzidine (TMB) to oxidized TMB (oxTMB) via peroxidase activity, allowing colorimetric and subsequent photothermal detection upon irradiation, as low as 2.06 CFU/mL. Magnetic - induced aggregation of Apt/Fe@SiNPs generated toxic hydroxyl radicals around L. monocytogenes, achieving ∼99.6% disinfection. Furthermore, the biofilm of L. monocytogenes was effectively inhibited by the action of hydroxyl radicals. The platform might offer a promising prospect to control L. monocytogenes in food industries.

Assessment of Viability of Listeria monocytogenes by Flow Cytometry.

In food industry, Listeria monocytogenes contamination can occur accidentally despite the quality control of raw materials and factory. Decontamination processes or inhibitory effects of ingredients/additives in food products are set up to ensure compliance with hygiene and microbiological criteria. These actions represent stresses for the pathogenic agent, causing fluctuations in its physiological states. Moreover, during these environmental stresses, Listeria monocytogenes can enter in a viable but nonculturable (VBNC) state which is not detected by plate counting but by flow cytometry. This technique coupled with cell staining by fluorescent dyes offers the possibility to assess different physiological states based on different cellular parameters: enzymatic activity, transmembrane integrity, membrane potential, and respiratory activity. In this chapter, we present a method to assess the viability of foodborne pathogens using a double-staining principle based on the assessment of membrane integrity and intracellular esterase activity.

Maple compounds prevent biofilm formation in Listeria monocytogenes via sortase inhibition.

The Pss exopolysaccharide (EPS) enhances the ability of the foodborne pathogen Listeria monocytogenes to colonize and persist on surfaces of fresh fruits and vegetables. Eradicating listeria within EPS-rich biofilms is challenging due to their increased tolerance to disinfectants, desiccation, and other stressors. Recently, we discovered that extracts of maple wood, including maple sap, are a potent source of antibiofilm agents. Maple lignans, such as nortrachelogenin-8'-O-ß-D-glucopyranoside and lariciresinol, were found to inhibit the formation of, and promote the dispersion of pre-formed L. monocytogenes EPS biofilms. However, the mechanism remained unknown. Here, we report that these lignans do not affect Pss EPS synthesis or degradation. Instead, they promote EPS detachment, likely by interfering with an unidentified lectin that keeps EPS attached to the cell surfaces. Furthermore, the maple lignans inhibit the activity of L. monocytogenes sortase A (SrtA) in vitro. SrtA is a transpeptidase that covalently anchors surface proteins, including the Pss-specific lectin, to the cell wall peptidoglycan. Consistent with this, deletion of the srtA gene results in Pss EPS detachment from listerial cells. We also identified several additional maple compounds, including epicatechin gallate, isoscopoletin, scopoletin, and abscisic acid, which inhibit L. monocytogenes SrtA activity in vitro and prevent biofilm formation. Molecular modelling indicates that, despite their structural diversity, these compounds preferentially bind to the SrtA active site. Since maple products are abundant and safe for consumption, our finding that they prevent biofilm formation in L. monocytogenes offers a viable source for protecting fresh produce from this foodborne pathogen.

Inhibition of Salmonella typhimurium and Listeria monocytogenes in coconut juice by graphene-doped photocatalyst rGO/TiO2.

This research used the photocatalyst rGO/TiO2 prepared by hydrothermal method to inhibit the growth of these microorganisms in water and coconut juice. In coconut juice, the initial count of Salmonella typhimurium decreased from 3 × 105 CFU /mL to 6.3 × 104 CFU /mL, and the initial count of L. monocytogenes was reduced from 3 × 105 CFU/mL to 1.2 × 105 CFU/mL. Moreover, the chemical structure characterization rGO/TiO2 showed that the doping of rGO formed a compact composite, enhanced the transfer of photogenerated electrons, and improved the photocatalytic efficiency of TiO2. The active substances ·OH and ·O2- produced by photocatalysis directly destroyed the integrity of bacteria cells, led to leakage of protein and DNA in the cells, and resulted in inactivation of the microorganisms, although Salmonella typhimurium and Listeria monocytogenes have different cell structures. These results would provide a good candidate photocatalyst to resist Salmonella typhimurium and Listeria monocytogenes and promote the development of photocatalysis applications.

Clinical and microbiological characteristics and follow-up of invasive Listeria monocytogenes infection among hospitalized patients: real-world experience of 16 years from Hungary.

PURPOSE: Invasive Listeria monocytogenes infection is rare, but can lead to life-threatening complications among high-risk patients. Our aim was to assess characteristics and follow-up of adults hospitalized with invasive L. monocytogenes infection. METHODS: A retrospective observational cohort study was conducted at a national referral center between 2004 and 2019. Patients with proven invasive listeriosis, defined by the European Centre for Disease Prevention and Control criteria, were included. Data collection and follow-up were performed using the hospital electronic system, up until the last documented visit. The primary outcome was in-hospital all-cause mortality, secondary outcomes included residual neurological symptoms, brain abscess occurrence, and requirement for intensive care unit (ICU) admission. RESULTS: Altogether, 63 cases were identified (57.1% male, median age 58.8 ± 21.7 years), and 28/63 developed a complicated disease course (44.4%). At diagnosis, 38/63 (60.3%) presented with sepsis, 54/63 (85.7%) had central nervous system involvement, while 9/63 (14.3%) presented with isolated bacteremia. Frequent clinical symptoms included fever (53/63, 84.1%), altered mental state (49/63, 77.8%), with immunocompromised conditions apparent in 56/63 (88.9%). L. monocytogenes was isolated from blood (37/54, 68.5%) and cerebrospinal fluid (48/55, 87.3%), showing in vitro full susceptibility to ampicillin and meropenem (100% each), gentamicin (86.0%) and trimethoprim/sulfamethoxazole (97.7%). In-hospital all-cause mortality was 17/63 (27.0%), and ICU admission was required in 28/63 (44.4%). At discharge, residual neurological deficits (11/46, 23.9%) and brain abscess formation (6/46, 13.0%) were common. CONCLUSION: Among hospitalized adult patients with comorbidities, invasive L. monocytogenes infections are associated with high mortality and neurological complications during follow-up.

Effect of Nitrite-Embedded Packaging Film on Growth of Listeria monocytogenes in Nitrite-free and Conventionally-cured Bologna Sausage.

Listeria monocytogenes is a pathogen frequently associated with ready-to-eat (RTE) meat and poultry products. Nitrite is a key antimicrobial additive that can offer some degree of protection against L. monocytogenes when included in meat product formulations. The objectives of this study were to determine the potential of nitrite-embedded film to affect the growth of L. monocytogenes following postthermal processing of conventionally-cured and nitrite-free bologna. Two bologna treatment formulations, a conventionally-cured control formulation (CON) and a nitrite-free formulation (UCC), were manufactured, packaged in conventional (CF) or nitrite-embedded (NEF) film, inoculated with 3.5 log CFU/cm2 of a cocktail of L. monocytogenes strains, and stored at 10 ± 1 °C. CON-NEF and UCC-NEF treatments had significantly slower (P < 0.05) growth of L. monocytogenes than CON-CF and UCC-CF, with populations in UCC-CF (which contained no nitrite) increasing by 3.4 logs after 10 d of storage in UCC-CF and 3.6 logs after 50 d in CON-CF (which had formulated nitrite only), while in the NEF-packaged samples, with or without formulated nitrite, they did not exceed the inoculum level until after day 40. Initial (day 0) residual nitrite was significantly greater (P < 0.05) in the control formulation. Packaging in NEF, however, resulted in an increase of 27-28 ppm by day 3, regardless of formulation, after which it decreased rapidly. Results suggest NEF can be used as a post-lethality antimicrobial intervention in food safety intervention strategies, in both cured and uncured processed meat products.

Sanitizer Resistance and Persistence of Listeria monocytogenes Isolates in Tree Fruit Packing Facilities.

The foodborne pathogen Listeria monocytogenes can persist in produce processing environments, which increases the risk for food contamination. Increased resistance to antimicrobials commonly used in cleaning and sanitizing procedures may contribute to L. monocytogenes' persistence in these environments. This study aimed to evaluate sanitizer resistance in L. monocytogenes isolates collected from three tree fruit packing facilities (F1, F2, and F3) during packing seasons 2020-2021 (Y1) and 2021-2022 (Y2), and to assess evidence of persistence based on the genomic similarity of isolates to historical isolates collected in previous years. L. monocytogenes isolates collected in 2020-2022 (n = 44) were tested for resistance to peroxyacetic acid (PAA) and a proprietary biofilm-removing agent using a broth microdilution assay. Further, L. monocytogenes isolates were whole genome sequenced and screened for the presence of antimicrobial resistance and virulence genes, as well as to assess the genomic similarity of isolates using the CFSAN SNP bioinformatic pipeline. Over half (57%) of the tested isolates had a PAA minimum inhibitory concentration (MIC) of 250 ppm, which was similar to the applied concentration of the PAA sanitizer in the three facilities (230 ppm). In contrast, 80% of tested isolates had a biofilm remover MIC of 0.13 ppm, which was substantially below the concentration applied in the facilities (137 ppm). Genomes of all tested isolates carried antimicrobial resistance (fosX, lin, mdrL, mprF, and norB) and virulence (inlA, inlB, plcA, plcB, prfA, hly, mpl, and iap) genes. L. monocytogenes isolates collected between 2020 and 2022 belonged to three distinct lineages, with 22 multilocus sequence types (MLSTs) belonging to 22 different clonal complexes. Genomic similarity analysis with historical isolates collected from the same facilities in 2016-2017 demonstrated a 5-year persistence of the genotypes ST 1003 and ST 554 in F2, which were no longer detected in 2022. Overall, our results highlight the need to re-evaluate sanitizer concentrations to effectively control persistent L. monocytogenes strains in tree fruit packing facilities.

Antibacterial effect and mechanism of theaflavin against Listeria monocytogenes and its application on apple skins.

Theaflavin 3,3'-digallate (TF3), a major polyphenolic component of black tea, exhibits antibacterial effects against many foodborne pathogens. However, the antibacterial mechanisms of TF3 against Listeria monocytogenes remain unclear. In this study, we investigated the effects of TF3 on viability, biofilm, and membrane function of L. monocytogenes by the conventional plating method, crystal violet staining, and microscopy using fluorescent dyes JC-1 and Laurdan, respectively. It was found that TF3 showed excellent antibacterial activity against L. monocytogenes with the minimum inhibitory concentration of 62.5 mg/L. The viable count determined on TSA decreased by 3 log after the treatment for 2 h with TF3 at 62.5 mg/L. The viable count determined on TSA containing 4% NaCl decreased by more than 4 log after the treatment for 30 min with TF3 at the same concentration, suggesting that TF3 gave damage on the cells, enhancing the antibacterial action of 4% NaCl, but the damage was recoverable in the absence of 4% NaCl. To explore the antibacterial mechanisms of TF3, the effects of TF3 on membrane potential and membrane fluidity were investigated. TF3 reduced both membrane potential and fluidity of L. monocytogenes at 62.5 mg/L, suggesting that TF3 damaged the structural integrity of the cell membrane. TF3 reduced biofilm mass of mature biofilm of L. monocytogenes. Moreover, THEAFLAVIN TF40, a commercially available Camellia sinensis leaf extract containing TF3, reduced viable count of L. monocytogenes by 2 log on apple skin. These results suggest the potential of theaflavins as a natural anti-Listeria disinfectant.

Analysis of a case report of meningitis caused by Listeria monocytogenes.

Background: Listeria monocytogenes is a Gram-positive bacterium transmitted to humans through contaminated food, water, and animal faeces, posing a public health risk. Listeria monocytogenes is difficult to isolate and is not sensitive to first-line treatment with broad-spectrum cephalosporins for bacterial meningitis. Listeria meningitis is rare but can progress rapidly and may be accompanied by serious complications (hydrocephalus, ventricular inflammation, cerebral palsy, and brain abscess) and a high mortality rate. Case presentation: It is a retrospective analysis of the clinical characteristics and treatment of a rare case of Listeria monocytogenes infection. Using laboratory indicators such as white blood cells (WBC), C-reactive protein (CRP), and procalcitonin (PCT), three detection methods (cerebrospinal fluid/blood culture), Targeted gene sequencing technology (tNGS), and Metagenomic next-generation sequencing technology (mNGS) combined with clinical manifestations of patients, analyze the use plan and prognosis of antibiotics in patients. The patient in this case initially had neurological symptoms such as fever, headache, unclear consciousness, and vomiting; laboratory indicators include elevated WBC, CRP, and PCT. Listeria monocytogenes was cultured in both the patient's cerebrospinal fluid and blood samples. After treatment with penicillin and meropenem, the patient recovered and was discharged without any sequelae. Conclusion: Due to the rarity of Listeria monocytogenes, there may be deficiencies and difficulties in clinical differential diagnosis, making it difficult to achieve targeted antibiotic treatment. Therefore, accurate identification of Listeria monocytogenes and relevant laboratory inflammation indicator testing, combined with traditional culture methods and NGS testing, through empirical coverage of Listeria monocytogenes, targeted antibiotic treatment ultimately impacts clinical outcomes significantly.

A novel antimicrobial peptide WBp-1 from wheat bran: Purification, characterization and antibacterial potential against Listeria monocytogenes.

This study introduces a novel antimicrobial peptide (AMP), WBp-1, isolated from wheat bran and purified via reversed-phase high-performance liquid chromatography. The amino acid sequence, determined as IITGASSGIGKAIAKHFI by LC-MS/MS, was composed predominantly of alkaline and hydrophobic residues. WBp-1 was predicted to be a stable, hydrophobic, cationic peptide with an α-helical structure. Moreover, it displayed significant antibacterial efficacy against Listeria monocytogenes, with a minimum inhibitory concentration of 150 µg/mL. Further mechanistic studies suggest that WBp-1 exerts its bactericidal activity by disrupting cell membrane integrity, impeding peptidoglycan synthesis by binding to penicillin-binding protein 4 via hydrogen bonding, increasing cell permeability, altering membrane potential and fluidity, and altering surface hydrophobicity. Interestingly, WBp-1 showed minimal hemolytic activity and cytotoxicity against LO2 cells, even at 16× MIC. These findings highlight the strong potential of WBp-1 as a novel antibacterial agent and food preservative against Listeria monocytogenes.

Outbreak of Listeriosis Likely Associated with Baker's Yeast Products, Switzerland, 2022-2024.

We traced back a nationwide outbreak of human listeriosis in Switzerland to a persisting production line contamination of a factory producing baker's yeast with Listeria monocytogenes serotype 1/2a sequence type 3141. We used whole-genome sequencing to match clinical isolates to isolates from product samples.

Modeling Behavior of Salmonella spp. and Listeria monocytogenes in Raw and Processed Vegetables.

Given the persistent occurrence of foodborne illnesses linked to both raw and processed vegetables, understanding microbial behavior in these foods under distribution conditions is crucial. This study aimed to develop predictive growth models for Salmonella spp. and Listeria monocytogenes in raw (mung bean sprouts, onion, and cabbage) and processed vegetables (shredded cabbage salad, cabbage and onion juices) at various temperatures, ranging from 4 to 36 °C. Growth models were constructed and validated using isolated strains of Salmonella spp. (S. Bareilly, S. Enteritidis, S. Typhimurium) and L. monocytogenes (serotypes 1/2a and 1/2b) from diverse food sources. The minimum growth temperatures for Salmonella varied among different vegetable matrices: 8 °C for mung bean sprouts, 9 °C for both onion and cabbage, and 10 °C for ready-to-eat (RTE) shredded cabbage salad. Both pathogens grew in cabbage juice at temperatures above 17 °C, while neither demonstrated growth in onion juice, even at 36 °C. Notably, Salmonella spp. exhibited faster growth than L. monocytogenes in all tested samples. At 8 °C, the lag time (LT) and specific growth rate (SGR) for Salmonella spp. in mung bean sprouts were approximately tenfold longer and threefold slower, respectively, compared to those at 10 °C. A decrease in refrigerator storage temperature by 1 or 2 degrees significantly prevented the growth of Salmonella in raw vegetables. These findings offer valuable insights into assessing the risk of foodborne illness associated with the consumption of raw and processed vegetables and inform management strategies in mitigating these risks.

Optimization of High-Pressure Processing for Microbial Inactivation in Pigmented Rice Grass Juice and Quality Impact Assessment during Refrigerated Storage.

Pigmented rice grass juice (RGJ) is a good source of bioactive compounds, but fresh juice has a relatively short shelf life of only 7 days at 4 °C. The objectives of this study were to determine the optimal growth stage of pigmented rice grass, investigate the optimal condition of high-pressure processing (HPP) for bacterial inactivation in inoculated RGJ using response surface methodology (RSM), and evaluate quality changes in uninoculated HPP-treated juice during storage at 4 °C compared with heat-treated (85 °C/10 min) and untreated samples. Results revealed that the optimal growth stage of rice grass was 9 days with the highest total anthocyanin content of 158.92 mg/L. The optimal condition of HPP was determined to be 612 MPa, 11 min, and 36 °C, and inactivated Escherichia coli K12 and Listeria innocua with 6.43 and 5.02 log reductions, respectively, meeting FDA regulations. The lethality of bacteria after HPP treatment can be explained by damage to the cell membrane and the leakage of intracellular constituents such as protein and nucleic acid. During 12 weeks of storage at 4 °C, total plate counts and yeast and mold counts in uninoculated HPP-treated juice were not detected. Moreover, HPP did not significantly change phytochemical properties (p < 0.05), caused a minor impact on physicochemical properties of RGJ, and maintained the durability of juice samples during storage. Analysis of the phytochemical profile revealed that HPP treatment could preserve most of the phenolic compounds in RGJ and especially increase the contents of protocatechuic acid, 4-hydroxybenzoic acid, syringic acid, transcinnamic acid, isorhamnetin-3-o-glucoside, quercetin, and cyanidin-3-glucoside (p < 0.05). Overall, HPP is a potential pasteurization technique for microbial inactivation and nutritional preservation for rice grass juice.

Microbiological Quality and Antibiotic Resistance of Relevant Bacteria from Horsemeat.

The aim of this work was to assess the microbiological safety and quality of horsemeat. A total of 19 fresh horsemeat samples were analysed. Mesophile counts were 4.89 ± 1.08 log CFU/g, and Enterobacteriaceae, Staphylococcus spp., and enterococci were only isolated from 36.84%, 21.05%, and 15.79% of the samples, respectively. Neither Staphylococcus aureus nor Escherichia coli were found in any sample. Listeria spp. and Listeria monocytogenes were detected in 31.58% and 21.05% of the samples, respectively. Campylobacter jejuni was not detected in any sample. The dominant bacteria were lactic acid bacteria. Seven different Staphylococcus spp. were identified, the most common being S. delphini, S. saprophyticus, and S. warneri. S. delphini showed resistance against mupirocin and cefoxitin. All the L. monocytogenes strains showed resistance against ampicillin, cefotaxime, and oxacillin. Multi-resistant Yersinia enterocolitica, Stenotrophomonas maltophilia, and Vagococcus. fluvialis strains were found, with resistance to 11, 7, and 8 antibiotics, respectively, causing significant concern. Therefore, specific actions should be taken to decrease the contamination of horsemeat.

A Novel C-Terminal Truncated Bacteriocin Found by Comparison between Leuconostoc mesenteroides 406 and 213M0 Isolated from Mongolian Traditional Fermented Milk, Airag.

Bacteriocins produced by lactic acid bacteria are known to be useful tools for food biopreservation and fermentation control. Leuconostoc mesenteroides subsp. mesenteroides 406 and 213M0 isolated from different samples of Mongolian traditional fermented milk, airag, had been reported to produce listericidal bacteriocin-like inhibitory substances with similar but slightly different properties. In this study, the antibacterial properties and the related gene sequences of both strains were compared, and then their bacteriocins were purified and identified. Strain 406 was superior to strain 213M0 in cell growth and antibacterial activity against many strains. However, the activity of 213M0 was stronger than that of 406 against a few strains. DNA sequencing revealed two and three plasmids in 406 and 213M0, respectively, and each one of them harbored an almost identical mesentericin Y105-B105 gene cluster. Removal of these plasmids resulted in a complete loss of activity, indicating that the antibacterial activity of both strains was generated by bacteriocins encoded on the plasmids. Mesentericins Y105 and B105 were purified from both cultures, and another novel bacteriocin, named mesentericin M, was identified from the 213M0 culture only. Its structural gene was coded on a 213M0 plasmid and, surprisingly, its C-terminal three amino acid residues were post-translationally cleaved. To our knowledge, this is the first report of a C-terminal truncated bacteriocin. In conclusion, the novel bacteriocin should be mainly responsible for the difference in antibacterial properties between the two strains.

Effects of Thermally-Assisted and High-Pressure Processing on Background Microbiota and the Listeria monocytogenes Load of a Minimally Processed Commodity.

The current study investigated the impact of treatments with elevated hydrostatic pressure (500 MPa) for inactivation of Listeria monocytogenes on smoked rainbow trout (Oncorhynchus mykiss) at high and low inoculation levels. The temperature values of the trials were set at 4.4 and 60.0 °C, adjusted with a circulating water bath connected to a stainless steel jacket surrounding the pressure processing chamber. Before pressure processing, the counts (selective counts of PALCAM, mean ± SD) of L. monocytogenes were 6.45 ± 0.1 log CFU/g and were reduced (p < 0.05) to 3.72 ± 0.3, and <1.48 ± 0.8 log CFU/g after 10 min of treatment at 4.4 and 60.0 °C, respectively. Treatments of low inoculation level samples were similarly efficacious and resulted in a reduction (p < 0.05) of the pathogen to 1.62 ± 0.3 and <0.82 ± 0.0 log CFU/g for treatments at 4.4 and 60.0 °C, respectively. At 4.4 °C, linear D-value and non-linear kmax1 were 8.68 and 0.50, and 5.81 and 2.41 for high-inoculation and low-inoculation samples, respectively. Application of hydrostatic pressure at 500 MPa at cold and elevated temperatures was efficacious for up to 5.03 log CFU/g reduction of L. monocytogenes, illustrating the potential for further adaptation of this technology.

Effect of the Bioprotective Properties of Lactic Acid Bacteria Strains on Quality and Safety of Feta Cheese Stored under Different Conditions.

Lately, the inclusion of additional lactic acid bacteria (LAB) strains to cheeses is becoming more popular since they can affect cheese's nutritional, technological, and sensory properties, as well as increase the product's safety. This work studied the effect of Lactiplantibacillus pentosus L33 and Lactiplantibacillus plantarum L125 free cells and supernatants on feta cheese quality and Listeria monocytogenes fate. In addition, rapid and non-invasive techniques such as Fourier transform infrared (FTIR) and multispectral imaging (MSI) analysis were used to classify the cheese samples based on their sensory attributes. Slices of feta cheese were contaminated with 3 log CFU/g of L. monocytogenes, and then the cheese slices were sprayed with (i) free cells of the two strains of the lactic acid bacteria (LAB) in co-culture (F, ~5 log CFU/g), (ii) supernatant of the LAB co-culture (S) and control (C, UHT milk) or wrapped with Na-alginate edible films containing the pellet (cells, FF) or the supernatant (SF) of the LAB strains. Subsequently, samples were stored in air, in brine, or in vacuum at 4 and 10 °C. During storage, microbiological counts, pH, and water activity (aw) were monitored while sensory assessment was conducted. Also, in every sampling point, spectral data were acquired by means of FTIR and MSI techniques. Results showed that the initial microbial population of Feta was ca. 7.6 log CFU/g and consisted of LAB (>7 log CFU/g) and yeast molds in lower levels, while no Enterobacteriaceae were detected. During aerobic, brine, and vacuum storage for both temperatures, pathogen population was slightly postponed for S and F samples and reached lower levels compared to the C ones. The yeast mold population was slightly delayed in brine and vacuum packaging. For aerobic storage at 4 °C, an elongation in the shelf life of F samples by 4 days was observed compared to C and S samples. At 10 °C, the shelf life of both F and S samples was extended by 13 days compared to C samples. FTIR and MSI analyses provided reliable estimations of feta quality using the PLS-DA method, with total accuracy (%) ranging from 65.26 to 84.31 and 60.43 to 89.12, respectively. In conclusion, the application of bioprotective LAB strains can result in the extension of feta's shelf life and provide a mild antimicrobial action against L. monocytogenes and spoilage microbiota. Furthermore, the findings of this study validate the effectiveness of FTIR and MSI techniques, in tandem with data analytics, for the rapid assessment of the quality of feta samples.

In Vitro Utilization of Prebiotics by Listeria monocytogenes.

Listeria monocytognes is an emerging pathogen responsible for the serious foodborne disease, listeriosis. The commensal gut microbiota is the first line of defense against pathogen internalization. The gut microbiome can be modified by prebiotic substrates, which are frequently added to food products and dietary supplements. Prebiotics should selectively support the growth of beneficial microbes and thus improve host health. Nevertheless, little is known about their effect on the growth of L. monocytogenes. The aim of this study was to evaluate the growth ability of four L. monocytogenes strains, representing the most common serotypes, on prebiotic oligosaccharides (beta-(1,3)-D-glucan, inulin, fructooligosaccharides, galactooligosaccharides, lactulose, raffinose, stachyose and 2'-fucosyllactose and a mixture of human milk oligosaccharides) as a sole carbon source. The results showed that only beta-(1,3)-D-glucan was metabolized by L. monocytogenes. These cell culture data suggest that beta-(1,3)-D-glucan may not be selectively utilized by healthy commensal bacteria, and its role in intestinal pathogen growth warrants further exploration in vivo.

Genetic Diversity, Virulence Factors and Antibiotic Resistance of Listeria monocytogenes from Food and Clinical Samples in Southern Poland.

Listeriosis is one of the most serious foodborne diseases under surveillance, with an overall mortality rate in the EU currently being high at 18.1%. Therefore, this study aims to investigate Listeria monocytogenes strains isolated from clinical and food samples for susceptibility to antimicrobials, presence of virulence factors, and genetic diversity. Species were identified using the MALDI-TOF, resistance to 11 antibiotics was determined according to EUCAST guidelines, and multiplex PCR was used for serotyping and detecting virulence genes. Strains were genotyped using the PFGE method. Clinical strains showed full sensitivity to all tested antibiotics. In total, 33.3% of strains from food products were found to be resistant to ciprofloxacin and 4.2% to tetracycline. Most of the tested isolates (79.2%) belonged to serotype 1/2a-3a, and the rest (20.8%) belonged to serotype 4ab-4b,4d-4e. Five virulence genes (prfA, hlyA, plcB, inlA, and lmo2672) were detected in all strains studied. The llsX gene was the least common, in 37.5% of clinical strains and 18.75% of strains isolated from food products. Among the analyzed strains, 13 strains displayed unique PFGE profiles. The other 11 strains belong to 3 clusters of pulsotypes: cluster 1 (2 strains), cluster 2 (6 strains), and cluster 3 (2 strains). The percentage of hospitalizations and deaths of Polish patients with listeriosis indicates the seriousness of this disease, especially in an aging society, while the molecular testing of clinical strains has been rarely performed, which makes it difficult to determine the source of infection.

Rli51 Attenuates Transcription of the Listeria Pathogenicity Island 1 Gene mpl and Functions as a Trans-Acting sRNA in Intracellular Bacteria.

Listeria pathogenicity island 1 (LIPI-1) is a genetic region containing a cluster of genes essential for virulence of the bacterial pathogen Listeria monocytogenes. Main virulence factors in LIPI-1 include long 5' untranslated regions (5'UTRs), among which is Rli51, a small RNA (sRNA) in the 5'UTR of the Zn-metalloprotease-coding mpl. So far, Rli51 function and molecular mechanisms have remained obscure. Here, we show that Rli51 exhibits a dual mechanism of regulation, functioning as a cis- and as a trans-acting sRNA. Under nutrient-rich conditions, rli51-mpl transcription is prematurely terminated, releasing a short 121-nucleotide-long sRNA. Rli51 is predicted to function as a transcription attenuator that can fold into either a terminator or a thermodynamically more stable antiterminator. We show that the sRNA Rli21/RliI binds to a single-stranded RNA loop in Rli51, which is essential to mediate premature transcription termination, suggesting that sRNA binding could stabilize the terminator fold. During intracellular infection, rli51 transcription is increased, which generates a higher abundance of the short Rli51 sRNA and allows for transcriptional read-through into mpl. Comparative intracellular bacterial transcriptomics in rli51-null mutants and the wild-type reference strain EGD-e suggests that Rli51 upregulates iron-scavenging proteins and downregulates virulence factors from LIPI-1. MS2 affinity purification confirmed that Rli51 binds transcripts of the heme-binding protein Lmo2186 and Lmo0937 in vivo. These results prove that Rli51 functions as a trans-acting sRNA in intracellular bacteria. Our research shows a growth condition-dependent mechanism of regulation for Rli51, preventing unintended mpl transcription in extracellular bacteria and regulating genes important for virulence in intracellular bacteria.