Pathogen Listeria Monocytogenes Research Articles

Interspecies Interaction of Bacteria Listeria monocytogenes, Yersinia pseudotuberculosis, and Marine Saprotrophs during Long-Term Culturing in Biofilms.

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.

EⅡB Mutation Reduces the Pathogenicity of Listeria monocytogenes by Negatively Regulating Biofilm Formation Ability, Infective Capacity, and Virulence Gene Expression.

To explore the role of the membrane permease ⅡB (EⅡB) gene of Listeria pathogenicity island 4 (LIPI-4) in the virulence of Listeria monocytogenes, both an EⅡB deletion strain (∆EⅡB) and a complemented strain were constructed. In vitro experiments demonstrated that EⅡB deletion affected the biofilm formation ability of the wild-type strain (Lm928). Moreover, this deletion decreased the intracellular proliferation abilities of L. monocytogenes. Mice infected with ∆EⅡB survived longer and experienced less weight loss on days 1, 2, and 3 post-infection. The bacterial load in the liver tissue of ∆EⅡB-infected mice was significantly reduced, and a considerable decrease in the blood levels of inflammatory cytokines IL-β, IL-6, IL-10, and TNF-α were observed. Following EⅡB deletion, 65% (13/20) of genes were downregulated, 25% (5/20) were upregulated, and 10% (2/20) showed no change. These findings suggest that EⅡB deletion may reduce both the in vivo and in vitro virulence levels as well as the biofilm formation ability of Lm928 by downregulating the transcription levels of genes associated with virulence and biofilm formation. These findings provide a foundation for further examining the pathogenic mechanisms of LIPI-4 and EⅡB in L. monocytogenes.

Reaction kinetics and mechanism of UVC-LED265–285nm and UVC-LED265–285nm/hydrogen peroxide toward foodborne pathogenic Listeria monocytogenes cells and its chromosomally encoded virulent hly genes

To investigate the principle-based inactivation kinetics and mechanisms of the foodborne pathogen Listeria monocytogenes (Lm), along with its representative virulence gene (hly) encoded in both intracellular and extracellular genomic DNA, we applied disinfection treatments using UVC-LED265–285nm and UVC-LED265–285nm combined with hydrogen peroxide (10 mg/L). These treatments were conducted under controlled model reaction matrices (phosphate-buffered solutions at pH 6.2 and 7.9) and/or actual agricultural water, depending on filtration pre-treatment. Comprehensive analytical methods, such as plate count, flow cytometry, and quantitative real-time PCR, were adopted for the qualitative/quantitative determination of bacterial or gene inactivation parameters. According to the experimentally determined fluence-based first-order rate constants (kUVC-LED and kUVC-LED/H2O2), the k value of Lm viability and hly genes was 1.14–2.19 cm2/mJ and 2.18 × 10−2–2.42 × 10−1 cm2/mJ, respectively. Damage to the longer amplicon or the extracellular form of the hly gene was higher than that to the shorter amplicon or the intracellular form by a factor of 2∼10 fold. Increasing pH in reaction matrices were marginally affected to the rates of inactivation kinetics. Dose-dependent inactivation levels tested in agriculture water matrices were comparable with the kinetics from the PB solutions of both UVC-LED265–285nm and UVC-LED265–285nm/hydrogen peroxide owing to OH radical scavenging by the water matrix components. These results will be valuable for optimizing UVC-LED and UVC-LED-based advanced oxidation process disinfection systems, specifically tailored to the unique challenges of bacterial or gene inactivation in the (waste)water at food processing and agricultural sectors.

A Snapshot, Using a Multi-Omic Approach, of the Metabolic Cross-Talk and the Dynamics of the Resident Microbiota in Ripening Cheese Inoculated with Listeria innocua.

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

Is the persistence of Listeria monocytogenes in food processing facilities and its resistance to pathogen intervention linked to its phylogeny?

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.IMPORTANCEListeria monocytogenes substantially contributes to the mortality of foodborne disease in developed countries. The virulence of strains of four lineages of L. monocytogenes differs, indicating that risks associated with the presence of L. monocytogenes are lineage specific. Our study extends the current knowledge by documentation that the lineage-level phylogeny of L. monocytogenes plays a role in the source of isolation, in the persistence in food processing facilities, and in the resistance to pathogen intervention technologies. In short, the control of risks associated with the presence of L. monocytogenes in food is also lineage specific. Understanding the route of contamination L. monocytogenes is an important factor to consider when designing improved control measures.

In Vitro and In Silico Characterization of N-Formylated Two-Peptide Bacteriocin from Enterococcus faecalis CAUM157 with Anti-Listeria Activity.

Enterococcus faecalis CAUM157 (KACC 81148BP), a Gram-positive bacteria isolated from raw cow's milk, was studied for its bacteriocin production. The antimicrobial activity of CAUM157 was attributed to a two-peptide class IIb bacteriocin with potent activity against food-borne pathogen Listeria monocytogenes and periodontal disease-causing pathogens (Prevotella intermedia KCTC 15693T and Fusobacterium nucleatum KCTC 2488T). M157 bacteriocins exhibit high temperature and pH stability and resist hydrolytic enzyme degradation and detergent denaturation, potentially due to their structural conformation. Based on amino acid sequence, M157A and M157B were predicted to be 5.176kDa and 5.182kDa in size, respectively. However, purified bacteriocins and chemically synthesized N-formylated M157 peptides both showed 5.204kDa (M157A) and 5.209kDa (M157B) molecular mass, confirming the formylation of the N-terminal methionine of both peptides produced by strain CAUM157. Furthermore, the strain demonstrated favorable growth and fermentation with minimal bacteriocin production when cultured in whey-based media, whereas a 1.0% tryptone or soytone supplementation resulted in higher bacteriocin production. Although Ent. faecalis CAUM157 innately harbors genes for virulence factors and antimicrobial resistance (e.g., tetracycline and erythromycin), its bacteriocin production is valuable in circumventing the need for live microorganisms, particularly in food applications for pathogen control.

Designing a Multiepitope Vaccine against the Foodborne Pathogenic Bacteria Listeria monocytogenes Using Subtractive Immunoinformatics Approaches.

Listeria monocytogenes, a Gram-positive bacterium, is a prominent foodborne pathogen that causes listeriosis and poses substantial health hazards worldwide. The continuing risk of listeriosis outbreaks underlies the importance of designing an effective prevention strategy and developing a robust immune response by reverse vaccinology approaches. This study aimed to provide a critical approach for developing a potent multiepitope vaccine against this foodborne disease. A chimeric peptide construct containing 5 B-cell epitopes, 16 major histocompatibility complex I (MHC-I) epitopes, and 18 MHC-II epitopes were used to create a subunit vaccination against L. monocytogenes. The vaccine safety was evaluated by several online methods, and molecular docking was performed using ClusPro to determine the binding affinity. Immune simulation was performed using the C-ImmSimm server to demonstrate the immune response. The results validated the antigenicity, non-allergenicity, and nontoxicity of the chimeric peptide construct, confirming its suitability as a subunit vaccine. Molecular docking showed a good score of 1276.5 and molecular dynamics simulations confirmed the construct's efficacy, demonstrating its promise as a good candidate for listeriosis prophylaxis. The population coverage was as high as 91.04% with a good immune response, indicating good antigen presentation with dendritic cells and production of memory cells. The findings of this study highlight the potential of the designed chimeric peptide construct as an effective subunit vaccine against Listeria, paving the way for future advances in preventive methods and vaccine design.

Functionalized nanowires for miRNA-mediated therapeutic programming of naïve T cells.

Cellular programming of naïve T cells can improve the efficacy of adoptive T-cell therapy. However, the current ex vivo engineering of T cells requires the pre-activation of T cells, which causes them to lose their naïve state. In this study, cationic-polymer-functionalized nanowires were used to pre-program the fate of primary naïve CD8+ T cells to achieve a therapeutic response in vivo. This was done by delivering single or multiple microRNAs to primary naïve mouse and human CD8+ T cells without pre-activation. The use of nanowires further allowed for the delivery of large, whole lentiviral particles with potential for long-term integration. The combination of deletion and overexpression of miR-29 and miR-130 impacted the ex vivo T-cell differentiation fate from the naïve state. The programming of CD8+ T cells using nanowire-delivered co-delivery of microRNAs resulted in the modulation of T-cell fitness by altering the T-cell proliferation, phenotypic and transcriptional regulation, and secretion of effector molecules. Moreover, the in vivo adoptive transfer of murine CD8+ T cells programmed through the nanowire-mediated dual delivery of microRNAs provided enhanced immune protection against different types of intracellular pathogen (influenza and Listeria monocytogenes). In vivo analyses demonstrated that the simultaneous alteration of miR-29 and miR-130 levels in naïve CD8+ T cells reduces the persistence of canonical memory T cells whereas increases the population of short-lived effector T cells. Nanowires could potentially be used to modulate CD8+ T-cell differentiation and achieve a therapeutic response in vivo without the need for pre-activation.

Peculiarities of milk microflora of Saanen goats in the conditions of the Steppe zone of Ukraine

The quality of goat milk can be affected by mesophilic aerobic and facultatively anaerobic microorganisms (MAFAnM), intestinal bacteria, psychrotrophic, some pathogenic microorganisms, fungi, etc. Traditional microbiological methods, which are used to determine the safety of milk, do not provide an opportunity to obtain an instant result, require the preparation of nutrient media and glassware, which takes up work time and is energy-consuming. We have carried out studies of goat milk by classical (using conventional and selective nutrient media) and alternative modern (using test plates, which are widely used to control microbiological contamination in the food industry) methods. As a result of the conducted research, it was established that the bacterial contamination of goat milk was within the permissible limits. There were no significant differences in the results of using the test plates. According to the classical method, the average indicators of the number of mesophilic aerobic and facultatively anaerobic microorganisms were 4.97 ± 0.14 lg CFU/mL, according to the alternative method – 4.86 ± 0.13 lg CFU/mL. Escherichia coli was isolated in four samples (with an average concentration of microorganisms of 1.86 ± 0.62 lg CFU/mL), in the same samples Enterobacter spp. in the amount of 1.77 ± 0.61 lg CFU/mL. In three samples, the presence of Proteus spp. with average concentration values of 2.19 ± 0.15 lg CFU/mL. Using an alternative method, contamination of milk with Escherichia coli (1.53 ± 0.33 lg CFU/mL) was detected in five samples, coliform bacteria were isolated in four samples (1.49 ± 0.50 lg CFU/mL). Staphylococcus aureus was not detected by any of the methods, but other representatives of the genus Staphylococcus were detected in all samples by both classical and alternative methods (2.48 ± 0.05 and 3.01 ± 0.07 lg CFU/mL). Using the classic method, it was established that two samples were positive for Enterococcus spp. (2.46 ± 0.08 lg CFU/mL), Bacillus spp. were isolated in six samples (1.70 ± 0.09 lg CFU/mL), in three – Clostridium spp. (1.66 ± 0.06 lg CFU/mL), in all six samples psychrotrophic bacteria (2.09 ± 0.26 lg CFU/mL) and yeast (2.41 ± 0.24 lg CFU/mL), four of the samples contained single colonies of mold fungi of the genus Aspergillus. Pathogens Listeria monocytogenes and Salmonella spp. not found. It will be promising to study the dependence of the milk microbiome on environmental factors, as well as the influence of the milk microbiota on the course of technological processes, quality and safety indicators during the production of goat cheese or other dairy products.

Rapid and visual detection of Listeria monocytogenes based on polymerase spiral reaction in fresh-cut fruit

Widespread foodborne pathogen Listeria monocytogenes exhibits a pronounced fatality rate among individuals with compromised immune systems, the elderly, and pregnant women. A simple and user-friendly approach is required to identify L. monocytogenes. A distinctive thermostatic nucleic acid amplification technology called polymerase spiral reaction (PSR) has been extensively employed in the identification of foodborne pathogens. In the study, a straightforward and intuitive approach for identifying L. monocytogenes in fresh-cut fruit was established using PSR in conjunction with hydroxy naphthol blue (HNB). Following optimization, the system's primary constitutes-betaine, dNTP, and MgSO4 were found to be 0.5 M, 1.0 mM, and 6 mM, respectively. Six strains of non- L. monocytogenes were used to test the assay's specificity. The PSR assay performed at 65 °C for 50 min demonstrated the capability to detect L. monocytogenes at level as low as 1 × 10−4 ng/μL DNA per tube and 5.1 × 101 CFU/g of freshly cut fruit. Notably, the sensitivity of this approach surpassed that of PCR by10-fold (1 × 10−3 ng/μL). In order to detect L. monocytogenes, the PSR assay was designed to be rapid, accurate, effective, and apparatus free. This innovative assay has provided researchers a fresh perspective on identification harmful microorganisms.

Surface topography and chemistry of food contact substances, and microbial nutrition affect pathogen persistence and symbiosis in cocktail Listeria monocytogenes biofilms

The foodborne pathogen Listeria monocytogenes (L. monocytogenes) can form very persistent biofilms on food contact substances, either in monospecies, or in conjunction with background microflora. Although some studies have investigated how bacterial symbiosis facilitates L. monocytogenes survival in complex cocktail biofilms, very little is known about how this core symbiotic relationship is affected by environmental factors. Here, we investigate how L. monocytogenes persistence is affected by symbiosis with Escherichia coli O157:H7, Pseudomonas fluorescens, and Ralstonia insidiosa, and how these relationships are impacted by environmental factors, including food contact surface topography and chemistry, and microbial nutrition. We find that symbiosis can significantly enhance L. monocytogenes persistence in four-species cocktail biofilms (5.71 ± 0.03 Log CFU/cm2) by up to 1.9 Log CFU/cm2 compared to monospecies biofilms (3.81 ± 0.08 Log CFU/cm2). We also report that surface topography and chemistry, microbial nutrition, and symbiont species significantly impact the symbiotic relationships involving L. monocytogenes, revealing the adaptability of bacterial symbiosis to changing environmental conditions and the complex nature of cocktail pathogen biofilms. Antagonistic and synergistic interactions involving pathogens in cocktail biofilms, and factors affecting those interactions, are elucidated and can establish a foundation to study symbiosis and its role in mitigating pathogen persistence in food systems, and helping identify unanticipated increased food safety risks of pathogens due to enhanced persistence.

Genetic Identification and Technological Potential of Indigenous Lactic Acid Bacteria Isolated from Alheira, a Traditional Portuguese Sausage.

Alheira is a naturally fermented meat sausage traditionally made in the Portuguese region of Trás-os-Montes. Lactic acid bacteria (LAB) are the dominant microorganisms in alheira and can endow it with various technological properties. This study aimed (1) to characterize technological features and in vitro antimicrobial activity of LAB isolated from alheira, and (2) to reveal associations between such phenotypic characteristics and the isolates species identified through amplification and sequencing of the 16S ribosomal gene. Sixty-two LAB isolates were identified and Enterococcus (E.) faecium corresponded to 32.3% of isolates, followed by Leuconostoc (L.) mesenteroides (19.4%) and Latilactobacillus (Lb.) sakei (17.7%), aligning with previous research on traditional Portuguese fermented meat sausages. The phenotypic analysis of LAB isolates indicated diverse acidification capacities, proteolytic activities, and inhibitory effects against foodborne pathogens Listeria (L.) monocytogenes, Salmonella (S.) Typhimurium and Staphylococcus (S.) aureus. Overall, lactobacilli displayed high inhibition activity against the pathogens S. aureus, L. monocytogenes, and S. Typhimurium. Although the mechanisms for the inhibition of pathogen growth need to be further elucidated, these findings enhance our understanding of LAB diversity and functionality in alheira sausages, contributing to product safety and quality.

Alternative Additives for Organic and Natural Ready-to-Eat Meats to Control Spoilage and Maintain Shelf Life: Current Perspectives in the United States

Food additives are employed in the food industry to enhance the color, smell, and taste of foods, increase nutritional value, boost processing efficiency, and extend shelf life. Consumers are beginning to prioritize food ingredients that they perceive as supporting a healthy lifestyle, emphasizing ingredients they deem acceptable as alternative or “clean-label” ingredients. Ready-to-eat (RTE) meat products can be contaminated with pathogens and spoilage microorganisms after the cooking step, contributing to food spoilage losses and increasing the risk to consumers for foodborne illnesses. More recently, consumers have advocated for no artificial additives or preservatives, which has led to a search for antimicrobials that meet these demands but do not lessen the safety or quality of RTE meats. Lactates and diacetates are used almost universally to extend the shelf life of RTE meats by reducing spoilage organisms and preventing the outgrowth of the foodborne pathogen Listeria monocytogenes. These antimicrobials applied to RTE meats tend to be broad-spectrum in their activities, thus affecting overall microbial ecology. It is to the food processing industry’s advantage to target spoilage organisms and pathogens specifically.

Selection of a Probiotic for Its Potential for Developing a Synbiotic Peach and Grape Juice.

Due to recent interest in the potential of probiotics as health promoters and the impact of health and environmental concerns on eating habits, non-dairy probiotic food products are required. This study aimed to evaluate the viability of different probiotic microorganisms in peach and grape juice (PGJ) with or without the prebiotic inulin and their antimicrobial activity against the foodborne pathogen Listeria monocytogenes and the juice spoilage microorganism Saccharomyces cerevisiae. Firstly, the viability of seven probiotic strains was studied in PGJ with an initial concentration of 107 CFU/mL for 21 days at 4 °C and for 3 days at 37 °C. In parallel, the physicochemical effect, the antimicrobial effect and the lactic acid production in PGJ were evaluated. Secondly, the probiotic with the best viability results was selected to study its antimicrobial effect against L. monocytogenes and S. cerevisiae, as well as ethanol and acetaldehyde production by the latter. L. casei showed the highest viability and grew in both refrigerated and fermentation conditions (1 log), produced the greatest lactic acid (5.12 g/L) and demonstrated in vitro anti-Listeria activity. Although the addition of the prebiotic did not improve the viability, lactic acid production or anti-Listeria activity of the probiotics, under the conditions studied, the prebiotic potential of inulin, support the design of a synbiotic juice. Finally, although none of the probiotic, fermentation products, or postbiotics showed any antimicrobial activity against L. monocytogenes or S. cerevisiae, the addition of L. casei to the PGJ significantly reduced the production of S. cerevisiae metabolite ethanol (29%) and acetaldehyde (50%). L. casei might be a suitable probiotic to deliver a safe and functional PGJ, although further research should be carried out to determine the effect of the probiotic and fermentation on the nutritional profile of PGJ.

Unveiling the persistent threat: recent insights into Listeria monocytogenes adaptation, biofilm formation, and pathogenicity in foodborne infections.

Listeriosis is a severe disease caused by the foodborne pathogen Listeria monocytogenes, posing a significant risk to vulnerable populations such as the elderly, pregnant women, and newborns. While relatively uncommon, it has a high global mortality rate of 20-30%. Recent research indicates that smaller outbreaks of the more severe, invasive form of the disease occur more frequently than previously thought, despite the overall stable infection rates of L. monocytogenes over the past 10years. The ability of L. monocytogenes to form biofilm structures on various surfaces in food production environments contributes to its persistence and challenges in eradication, potentially leading to contamination of food and food production facilities. To address these concerns, this review focuses on recent developments in epidemiology, risk evaluations, and molecular mechanisms of L. monocytogenes survival in adverse conditions and environmental adaptation. Additionally, it covers new insights into strain variability, pathogenicity, mutations, and host vulnerability, emphasizing the important events framework that elucidates the biochemical pathways from ingestion to infection. Understanding the adaptation approaches of L. monocytogenes to environmental stress factors is crucial for the development of effective and affordable pathogen control techniques in the food industry, ensuring the safety of food production.

Inactivation of lmo0946 (sif) induces the SOS response and MGEs mobilization and silences the general stress response and virulence program in Listeria monocytogenes.

Bacteria have evolved numerous regulatory pathways to survive in changing environments. The SOS response is an inducible DNA damage repair system that plays an indispensable role in bacterial adaptation and pathogenesis. Here we report a discovery of the previously uncharacterized protein Lmo0946 as an SOS response interfering factor (Sif) in the human pathogen Listeria monocytogenes. Functional genetic studies demonstrated that sif is indispensable for normal growth of L. monocytogenes in stress-free as well as multi-stress conditions, and sif contributes to susceptibility to β-lactam antibiotics, biofilm formation and virulence. Absence of Sif promoted the SOS response and elevated expression of mobilome genes accompanied by mobilization of the A118 prophage and ICELm-1 mobile genetic elements (MGEs). These changes were found to be associated with decreased expression of general stress response genes from the σB regulon as well as virulence genes, including the PrfA regulon. Together, this study uncovers an unexpected role of a previously uncharacterized factor, Sif, as an inhibitor of the SOS response in L. monocytogenes.

Schisandrin A enhances pathogens resistance by targeting a conserved p38 MAPK pathway

Schizandrin A (SA), also known as deoxyschizandrin, is one of the most biologically active lignans isolated from the traditional Chinese medicine Fructus schisandrae chinensis. Schisandrin A has proven benefits for anti-cancer, anti-inflammation, hepatoprotection, anti-oxidation, neuroprotection, anti-diabetes. But the influence of Schisandrin A to the innate immune response and its molecular mechanisms remain obscure. In this study, we found that Schisandrin A increased resistance to not only the Gram-negative pathogens Pseudomonas aeruginosa and Salmonella enterica but also the Gram-positive pathogen Listeria monocytogenes. Meanwhile, Schisandrin A protected the animals from the infection by enhancing the tolerance to the pathogens infection rather than by reducing the bacterial burden. Through the screening of the conserved immune pathways in Caenorhabditis elegans, we found that Schisandrin A enhanced innate immunity via p38 MAPK pathway. Furthermore, Schisandrin A increased the expression of antibacterial peptide genes, such as K08D8.5, lys-2, F35E12.5, T24B8.5, and C32H11.12 by activation PMK-1/p38 MAPK. Importantly, Schisandrin A-treated mice also enhanced resistance to P. aeruginosa PA14 infection and significantly increased the levels of active PMK-1. Thus, promoted PMK-1/p38 MAPK-mediated innate immunity by Schisandrin A is conserved from worms to mammals. Our work provides a conserved mechanism by which Schisandrin A enhances innate immune response and boosts its therapeutic application in the treatment of infectious diseases.

Investigating the presence and antibiotic susceptibilities of Escherichia coli O157 and Listeria monocytogenes in ruminant feces and feed in Balıkesir province

The aim of this study is to determine the presence, virulence factors, and antibiotic susceptibilities of E. coli O157 and L. monocytogenes in ruminant feces and feed. This study was carried out for the first time in Balıkesir province. Feces, pellet feed, and silage samples were analyzed simultaneously for E. coli O157 and L. monocytogenes using feces of one gram and feed of twenty-five grams according to ISO 16654:2001/Amd 1:2017 and ISO 11290-1, respectively. 38 (38%) E. coli O157 strains were isolated and identified from a total of 100 ruminant feces. A total of 3 (3%) E. coli O157:H7 strains were detected by PCR from one hundred ruminant fecal samples. In the study, resistance to antibiotics increased, especially in E. coli O157 isolates. In this study, enterohaemolysin was the predominant virulence factor among the E. coli isolates, and it was thought that it was important for pathogenesis. The Sxt1 gene was higher than the Stx2 gene. A total of 24 L. monocytogenes strains were isolated from a total of 100 ruminant fecal samples and 50 silage samples. Three of these strains were isolated from silage samples taken from the farms, where L. monocytogenes was isolated from sheep feces. As a result, poor quality silage could be an important source of infection for listeriosis in Balıkesir province. Epidemiologically, poor quality silage was thought to be one of the sources of listeriosis. It was thought that ruminant feces played an important role as a reservoir in the spread and transmission of E. coli O157. The antibiotic resistance status of E. coli O157 and L. monocytogenes isolates should be monitored with epidemiological studies.

Thymbra capitata L. hydrolate as a washing solution for controlling Listeria monocytogenes and spoilage microorganisms on rocket salad

Ready-to-eat (RTE) vegetables, including fresh-cut ones, are exposed to the risk of the foodborne pathogen Listeria monocytogenes. Nowadays, fresh-cut products are widely consumed because of their convenience. Nevertheless, effective washing treatments are required for their preparation, to maintain freshness and microbiological safety. Currently, natural substances are increasingly demanded as alternatives to traditional synthetic washing agents. Therefore, this study investigates the application of Thymbra capitata hydrolate as washing solution for rocket salad (Eruca sativa) stored at 4 °C for 48 h, to evaluate a potential application both during the production process of fresh-cut vegetables, and also at the domestic level. The hydrolate applied at a concentration of 500 μL/mL for 5 min allowed to reduce L. monocytogenes load of about 1 log CFU/g at each time of analysis, and to determine a significant reduction of microbial groups normally found on rocket salad, particularly total mesophilic and psychrotrophic counts. The treatment showed no negative effects on pH, aw, and colour of the rocket salad. At a sensory level, the differences among control and treated samples were minimally perceived only on the first day of analysis but disappeared over time. Therefore, this alternative washing treatment revealed to counteract L. monocytogenes growth and to help maintaining the quality of rocket salad. Hence, it could be optimized for a concrete application, also considering the enhancement of the antilisterial activity, the sustainability and low cost of the hydrolate, which represents a by-product from the production of essential oils.

Automated centrifugal microfluidic system for the preparation of adaptor-ligated sequencing libraries.

The intensive workload associated with the preparation of high-quality DNA libraries remains a key obstacle toward widespread deployment of sequencing technologies in remote and resource-limited areas. We describe the development of single-use microfluidic devices driven by an advanced pneumatic centrifugal microfluidic platform, the PowerBlade, to automate the preparation of Illumina-compatible libraries based on adaptor ligation methodology. The developed on-chip workflow includes enzymatic DNA fragmentation coupled to end-repair, adaptor ligation, first DNA cleanup, PCR amplification, and second DNA cleanup. This complex workflow was successfully integrated into simple thermoplastic microfluidic devices that are amenable to mass production with injection molding. The system was validated by preparing, on chip, libraries from a mixture of genomic DNA extracted from three common foodborne pathogens (Listeria monocytogenes, Escherichia coli and Salmonella enterica serovar Typhimurium) and comparing them with libraries made via a manual procedure. The two types of libraries were found to exhibit similar quality control metrics (including genome coverage, assembly, and relative abundances) and led to nearly uniform coverage independent of GC content. This microfluidic technology offers a time-saving and cost-effective alternative to manual procedures and robotic-based automation, making it suitable for deployment in remote environments where technical expertise and resources might be scarce. Specifically, it facilitates field practices that involve mid- to low-throughput sequencing, such as tasks related to foodborne pathogen detection, characterization, and microbial profiling.