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Listeria monocytogenes is a bacterial pathogen that can cause life-threatening central nervous system (CNS) infections. While mechanisms by which L. monocytogenes and other pathogens traffic to the brain have been studied, a quantitative understanding of the underlying dynamics of colonization and replication within the brain is still lacking. In this study, we used barcoded L. monocytogenes to quantify the bottlenecks and dissemination patterns that lead to cerebral infection. Following intravenous (IV) inoculation, multiple independent invasion events seeded all parts of the CNS from the blood, however, only one clone usually became dominant in the brain. Sequential IV inoculations and intracranial inoculations suggested that clones that had a temporal advantage (i.e., seeded the CNS first), rather than a spatial advantage (i.e., invaded a particular brain region), were the main drivers of clonal dominance. In a foodborne model of cerebral infection with immunocompromised mice, rare invasion events instead led to a highly infected yet monoclonal CNS. This restrictive bottleneck likely arose from pathogen transit into the blood, rather than directly from the blood to the brain. Collectively, our findings provide a detailed quantitative understanding of the L. monocytogenes population dynamics that lead to CNS infection and a framework for studying the dynamics of other cerebral infections.

Listeria monocytogenes (Lm) is a Gram-positive facultative intracellular pathogen that leads a biphasic lifecycle, transitioning its metabolism and selectively inducing virulence genes when it encounters mammalian hosts. Virulence gene expression is controlled by the master virulence regulator PrfA, which is allosterically activated by the host- and bacterially derived glutathione (GSH). The amino acid cysteine is the rate-limiting substrate for GSH synthesis in bacteria and is essential for bacterial growth. Unlike many bacteria, Lm is auxotrophic for cysteine and must import exogenous cysteine for growth and virulence. GSH is enriched in the host cytoplasm, and previous work suggests that Lm utilizes exogenous GSH for PrfA activation. Despite these observations, the import mechanism(s) for GSH remains elusive. Analysis of known GSH importers predicted a homologous importer in Lm comprised of the Ctp ABC transporter and the OppDF ATPases of the Opp oligopeptide importer. Here, we demonstrated that the Ctp complex is a high-affinity GSH/GSSG importer that is required for Lm growth at physiologically relevant concentrations. Furthermore, we demonstrated that OppDF is required for GSH/GSSG import in an Opp-independent manner. These data support a model where Ctp and OppDF form a unique complex for GSH/GSSG import that supports growth and pathogenesis. In addition, we show that Lm utilizes the inorganic sulfur sources thiosulfate and H2S for growth in a CysK-dependent manner in the absence of other cysteine sources. These findings suggest a pathoadaptive role for partial cysteine auxotrophy in Lm, where locally high GSH/GSSG or inorganic sulfur concentrations may signal arrival to distinct host niches.

Listeriamonocytogenes (L. monocytogenes) is a prevalent food-borne pathogen that can cause listeriosis, which manifests as meningitis and other symptoms, potentially leading to fatal outcomes in severe cases. In this study, we developed an aptasensor utilizing carboxylated magnetic beads and Cas12a to detect L. monocytogenes. In the absence of L. monocytogenes, the aptamer maintains its spatial configuration, keeping the double-stranded DNA attached and preventing the release of a startup template and activation of Cas12a's trans-cleavage capability. Conversely, in the presence of L. monocytogenes, the aptamer undergoes a conformational change, releasing the double-stranded DNA to serve as a startup template, thereby activating the trans-cleavage capability of Cas12a. Consequently, as the concentration of L. monocytogenes increases, the observable brightness in a blue light gel cutter intensifies, leading to a rise in fluorescence intensity difference compared to the control. This Cas12a aptasensor demonstrates excellent sensitivity towards L. monocytogenes, with a lowest detection limit (LOD) of 57.15 CFU/mL and a linear range of 4×102 to 4×107 CFU/mL (R2=0.9858). Notably, the proposed Cas12a aptasensor exhibited outstanding selectivity and recovery in beef samples, and could be employed for precise monitoring. This Cas12a aptasensor not only provides a novel fluorescent and visual rapid detection method for L. monocytogenes but also offers simplicity, speed, and stability compared to previous detection methods. Furthermore, it is suitable for on-site detection of beef samples. [ABSTRACT FROM AUTHOR]

Summary: Pork cutlets are susceptible to cross‐contamination with foodborne pathogens during processing. Food irradiation effectively improves food hygiene by eliminating microbes. We investigated the application of X‐ray irradiation (0.5–10 kGy) on pathogen inactivation (D10 value), quality changes [total aerobic bacteria (TAB), yeast and mould (Y&M), total coliform (TC), colour, thiobarbituric acid reactive substance (TBARS), and pH], and the hydrocarbon profile of pork cutlets. D10 values of Escherichia coli O157:H7, Salmonella typhimurium, Listeria monocytogenes, and Staphylococcus aureus were 0.37, 0.62, 0.52, and 0.36 kGy, respectively. At least 3.10 kGy (5‐D10 values) is required to ensure the microbiological safety of pork cutlets. X‐ray irradiation dose‐dependently reduced all microbial counts, and up to 7 kGy had no adverse effects on colour, TBARS, and pH. Furthermore, 1,7‐hexadecadiene and 8‐heptadecene were potential markers for irradiated pork cutlets. X‐ray irradiation of up to 7 kGy is effective in improving hygiene levels while maintaining pork cutlet quality. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes (LM) is a Gram‐positive (G+) bacterium that secretes nanoscale membrane vesicles (MVs). LM MVs comprise various bacterial components and may have potential as an antigen or drug‐delivery vehicle; however, the low yield of the LM MVs limits related research. G+‐bacterial MVs germinate from the bacterial plasma membrane and must pass through a thick crosslinked peptidoglycan layer for release. Herein, we aimed to increase the release of MVs by reducing the degree of crosslinking of peptidoglycan. We knocked out two genes related to the longitudinal crosslinking of peptidoglycan, dal and dat, and supplemented the knocked‐out dal gene through plasmid expression to obtain a stably inherited recombinant strain LMΔdd::pCW633. The structure, particle size, and main protein components of MVs secreted by this recombinant strain were consistent with those secreted from the wild strain, but the yield of MVs was considerably increased (p < 0.05). Furthermore, Listeria ivanovii (LI) was found to secrete MVs that differed in the composition of the main proteins compared with those of LM MVs. The abovementioned method was also feasible for promoting the secretion of MVs from the attenuated LM strain and LI wild‐type and attenuated strains. Our study provides a new method to increase the secretion of MVs derived from Listeria that could be extended to other G+ bacteria. [ABSTRACT FROM AUTHOR]

Infection by bacteria leads to tissue damage and inflammation, which need to be tightly controlled by host mechanisms to avoid deleterious consequences. It is previously reported that TMEM16F, a calcium‐activated lipid scramblase expressed in various immune cell types including T cells and neutrophils, is critical for the control of infection by bacterium Listeria monocytogenes (Lm) in vivo. This function correlated with the capacity of TMEM16F to repair the plasma membrane (PM) damage induced in T cells in vitro, by the Lm toxin listeriolysin O (LLO). However, whether the protective effect of TMEM16F on Lm infection in vivo is mediated by an impact in T cells, or in other cell types, is not determined. Herein, the immune cell types and mechanisms implicated in the protective effect of TMEM16F against Lm in vivo are elucidated. Cellular protective effects of TMEM16F correlated with its capacity of lipid scrambling and augment PM fluidity. Using cell type‐specific TMEM16F‐deficient mice, the indication is obtained that TMEM16F expressed in liver Kupffer cells (KCs), but not in T cells or B cells, is key for protection against Listeria in vivo. In the absence of TMEM16F, Listeria induced PM rupture and fragmentation of KCs in vivo. KC death associated with greater liver damage, inflammatory changes, and dysregulated liver metabolism. Overall, the results uncovered that TMEM16F expressed in Kupffer cells is crucial to protect the host against Listeria infection. This influence is associated with the capacity of Kupffer cell‐expressed TMEM16F to prevent excessive inflammation and abnormal liver metabolism. [ABSTRACT FROM AUTHOR]

In recent years, the awareness of healthier lifestyles among consumers has driven to an increased interest in more natural, nutritious, and low-processed foods. Ferulic acid, one of the most abundant phenolic acids in plants, has demonstrated a wide spectrum of antimicrobial activities and a range of biomedical effects, including antioxidant, antiallergic, hepatoprotective, anticarcinogenic, anti-inflammatory, and antithrombotic, among others. The objective of this work was to study the antilisterial effect of ferulic acid (FA, 1500 mg/L) on fresh, ready-to-eat orange (FOJ) and apple (FAJ) juices and evaluate its effect on product quality. The results showed that FA reduced the Listeria monocytogenes population after 9 days of storage at 4 °C, but no effect on the indigenous microbiota was observed. The titratable acidity and color significantly changed. The antioxidant capacity and total phenolic content significantly increased with the addition of FA, being at least two times greater in fortified juices. FAJ and FOJ containing FA were scored lower (6.8 and 5.7 on a 9-point hedonic scale, respectively) than their respective controls. Overall, our results demonstrated that FA treatment could be a useful strategy to maintain the safety of fresh apple and orange juices and increase the antioxidant activity and phenolic content. The potential industrial applications and health benefits of the fortification of fruit juices with FA should be further explored. [ABSTRACT FROM AUTHOR]

Calcium (Ca2+) signaling is fundamental to cellular processes in both eukaryotic and prokaryotic organisms. While the mechanisms underlying eukaryotic Ca2+ transport are well documented, an understanding of prokaryotic transport remains nascent. LMCA1, a Ca2+ adenosine triphosphatase (ATPase) from Listeria monocytogenes, has emerged as a prototype for elucidating structure and dynamics in prokaryotic Ca2+ transport. Here, we used a multidisciplinary approach integrating kinetics, structure, and dynamics to unravel the intricacies of LMCA1 function. A cryo-electron microscopy (cryo-EM) structure of a Ca2+-bound E1 state showed ion coordination by Asp720, Asn716, and Glu292. Time-resolved x-ray solution scattering experiments identified phosphorylation as the rate-determining step. A cryo-EM E2P state structure exhibited remarkable similarities to a SERCA1a E2-P* state, which highlights the essential role of the unique P-A domain interface in enhancing dephosphorylation rates and reconciles earlier proposed mechanisms. Our study underscores the distinctiveness between eukaryotic and prokaryotic Ca2+ ATPase transport systems and positions LMCA1 as a promising drug target for developing antimicrobial strategies. [ABSTRACT FROM AUTHOR]

Background: Foodborne infections such as listeriosis caused by the bacterium Listeria monocytogenes represent a significant public health concern, particularly when outbreaks affect many individuals over prolonged time. Systematic collection of pathogen isolates from infected patients, whole genome sequencing (WGS) and phylogenetic analyses allow recognition and termination of outbreaks after source identification and risk profiling of abundant lineages. Methods: We here present a multi-dimensional analysis of > 1800 genome sequences from clinical L. monocytogenes isolates collected in Germany between 2018 and 2021. Different WGS-based subtyping methods were used to determine the population structure with its main phylogenetic sublineages as well as for identification of disease clusters. Clinical frequencies of materno-foetal and brain infections and in vitro infection experiments were used for risk profiling of the most abundant sublineages. These sublineages and large disease clusters were further characterised in terms of their genetic and epidemiological properties. Results: The collected isolates covered 62% of all notified cases and belonged to 188 infection clusters. Forty-two percent of these clusters were active for > 12 months, 60% generated cases cross-regionally, including 11 multinational clusters. Thirty-seven percent of the clusters were caused by sequence type (ST) ST6, ST8 and ST1 clones. ST1 was identified as hyper- and ST8, ST14, ST29 as well as ST155 as hypovirulent, while ST6 had average virulence potential. Inactivating mutations were found in several virulence and house-keeping genes, particularly in hypovirulent STs. Conclusions: Our work presents an in-depth analysis of the genomic characteristics of L. monocytogenes isolates that cause disease in Germany. It supports prioritisation of disease clusters for epidemiological investigations and reinforces the need to analyse the mechanisms underlying hyper- and hypovirulence. [ABSTRACT FROM AUTHOR]

Sprouts are popular due to their high nutritional content, including vitamins, minerals, antioxidants, and enzymes. However, the conditions favorable for sprouting, such as warm and humid environments, are also ideal for the growth of bacteria, including food‐borne pathogens. Here, we analyzed the growth and developed predictive models of locally isolated and commercial strains of Salmonella enterica and Listeria monocytogenes in alfalfa sprouts under various constant temperatures, ranging from 5°C to 25°C. Our findings indicated that these pathogens could grow at 5°C in sprouts, albeit with a low growth rate. A rapid increase in concentration occurred at temperatures of 10°C and above. The fitted models demonstrated high performance, with R2 values ranging from 0.964 to 0.997 and RMSE values ranging from 0.15 to 0.51, respectively. Based on the fitted values, bias factor (Af) values varied between 1.01 and 1.06, with all accuracy factor (Bf) values at 1.00. Acceptable prediction zone (APZ) values ranged from 81.8% to 100%. Validation of the models under dynamic temperature conditions for specific strains showed acceptable performance. This study enhances our understanding of S. enterica and L. monocytogenes growth in alfalfa sprouts. The findings of this study could be used to improve the risk assessment of these pathogens in alfalfa sprouts. [ABSTRACT FROM AUTHOR]

Bacteria adapt the biosynthesis of their envelopes to specific growth conditions and prevailing stress factors. Peptidoglycan (PG) is the major component of the cell wall in Gram‐positive bacteria, where PASTA kinases play a central role in PG biosynthesis regulation. Despite their importance for growth, cell division and antibiotic resistance, the mechanisms of PASTA kinase activation are not fully understood. ReoM, a recently discovered cytosolic phosphoprotein, is one of the main substrates of the PASTA kinase PrkA in the Gram‐positive human pathogen Listeria monocytogenes. Depending on its phosphorylation, ReoM controls proteolytic stability of MurA, the first enzyme in the PG biosynthesis pathway. The late cell division protein GpsB has been implicated in PASTA kinase signalling. Consistently, we show that L. monocytogenes prkA and gpsB mutants phenocopied each other. Analysis of in vivo ReoM phosphorylation confirmed GpsB as an activator of PrkA leading to the description of structural features in GpsB that are important for kinase activation. We further show that ReoM phosphorylation is growth phase‐dependent and that this kinetic is reliant on the protein phosphatase PrpC. ReoM phosphorylation was inhibited in mutants with defects in MurA degradation, leading to the discovery that MurA overexpression prevented ReoM phosphorylation. Overexpressed MurA must be able to bind its substrates and interact with ReoM to exert this effect, but the extracellular PASTA domains of PrkA or MurJ flippases were not required. Our results indicate that intracellular signals control ReoM phosphorylation and extend current models describing the mechanisms of PASTA kinase activation. [ABSTRACT FROM AUTHOR]

Infective endocarditis (IE) is a disease associated with significant morbidity and mortality. It is more commonly caused by Gram-positive cocci, but Gram-positive bacilli may seldom cause the disease. Listeria monocytogenes is an aerobic Gram-positive coccobacillus and a foodborne and opportunistic pathogen most commonly causing gastrointestinal infections, even though bacteremia, sepsis, meningitis, and fetal infections may also occur. Listeria IE has rarely been described, with most reports being case reports or case series. Thus, the characteristics of this disease remain largely unknown. This systematic review aimed to present all published Listeria IE studies and describe their characteristics. A search of PubMed, Scopus, and the Cochrane Library for studies providing information on epidemiology, clinical findings, treatment, and outcome of Listeria IE cases was performed. A total of 54 studies containing data from 62 patients were included. Among all patients, 64.5% were male; the median age was 69 years. Among all patients, 54.8% had a history of a prosthetic valve. The aortic valve was the most commonly affected, followed by the mitral. Fever, heart failure, and embolic phenomena were the most commonly encountered clinical findings. The only isolated species was L. monocytogenes. Antimicrobial resistance was relatively low for aminopenicillins and aminoglycosides, the most commonly used antimicrobials for treating L. monocytogenes IE. Surgery was performed in 27.4% of patients. Mortality was 37.1%. Patients who survived were more likely to have had a prosthetic valve, to have necessitated transesophageal echocardiography for the diagnosis, to have mitral valve IE, and to have had surgical management; however, no factor was identified in a multivariate logistic regression model as an independent factor for overall mortality. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Summary: Persistent bacteria are subset of cells capable of surviving bactericidal treatments longer. They originate in response to environmental stressors and show a longer lag phase compared to the rest of the population they are part of. Impact of persistent cells on food safety and role of common food preservation techniques in their formation are still unclear. This study aims to investigate whether osmotic stress induces formation of persistent cells in Listeria monocytogenes capable of resisting longer during cooking cycle of pork sausages. Lag‐phase duration at single‐cell level of a strain grown in 6%‐NaCl broth using image processing software was first investigated. Then, trend of the strain inactivation curve was evaluated in experimentally contaminated pork sausages cooked at 65 °C for 11 min. Results showed that salt triggers the formation of persistent cells characterised by a longer lag phase and increased heat resistance, posing a concern especially for ready‐to‐eat foods. [ABSTRACT FROM AUTHOR]

Biofilm formation on abiotic surfaces in the food industry is a major public health problem. Indeed, biofilms represent a constant source of pathogens, such as Listeria monocytogenes and bacteria of the genus Salmonella. Formation of multispecies biofilms by pathogens and spoilage microorganisms significantly contributes to the persistence of microorganisms in meat processing plant environments and thus poses a serious risk for product safety. In this study, 46 strains of microorganisms isolated from food products and various elements of food industry environment were tested for their ability to form biofilms at different temperatures. The analyzed pathogenic strains (Listeria monocytogenes and Salmonella sp.) and spoilage microorganisms (Pseudomonas sp.) exhibited adhesion to abiotic surfaces with subsequent formation of persistent biofilms. Low positive temperature was not a limiting factor in their ability to form biofilms. After 24 h of incubation at 4°C, Listeria and Salmonella sp. developed stable biofilms. It was found that these bacteria were capable of forming biofilms on various abiotic surfaces present in the meat industry (ceramic tiles, glass, and plastic). Analysis of synergistic interactions among members of the genera Listeria, Salmonella, and Pseudomonas in the formation of mixed biofilms at 4°C showed that combinations of a pathogenic microorganism and a Pseudomonas member formed biofilms significantly more efficiently than combinations of two pathogens. This indicates the importance of Pseudomonas species in synergistic interaction among microorganisms. [ABSTRACT FROM AUTHOR]

New nitrobenzyl-pyridyl ether ligands were synthesized by reacting nitrobenzyl bromide (2-, 3- and 4-nitro) and halogen-substituted hydroxy pyridine (2-chloro-3-hydroxy-pyridine, 2-bromo-3-hydroxy-pyridine) compounds in DMF. By interacting the obtained ligands (L1, L3, L4 and L6) with silver (I) nitrate, transition metal complexes were prepared (AgL1, AgL3, AgL4 and AgL6). The structures of the synthesized ligands and complexes were characterized using FTIR, HRMS, 1H-NMR and 13C-NMR spectroscopic techniques. In addition, fluorescence spectra of the ligands (L1-L6) were detected in the presence of different metal cations (Li+, Na+, Mg2+, Al3+, K+, Ca2+, Cr3+, Fe3+, Co2+, Ni2+, Cu2+, Zn2+ and Ag+). In addition, the thermal behavior of all compounds were examined by TGA-DTA methods and the data of silver complexes and ligands were compared. Within the scope of the study, the antibacterial activities of all compounds were investigated against 4 pathogens, 2 of which were plant-borne (Xanthomonas vesicotoria, Clavibacter michiganensis subsp. Michiganensis) and 2 were food-borne (Escherichia coli, Listeria monocytogenes). [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is a human opportunistic foodborne pathogen that produces life-threatening infections with a high mortality rate. The control of Listeria in the food production environment and effective clinical management of human listeriosis are challenging due to the emergence of antibiotic resistance. Hence we evaluate the in vitro anti-Listeria activity of two synthetic cruzioseptins reproducing their natural sequences CZS-9, and CZS-12, and one engineered sequence based on CZS-1, named [K4K15]CZS-1. The assessment of the in vitro potential of cruzioseptins, highlighted the promising antibacterial effect of [K4K15]CZS-1 in very low concentrations (0.91 μM) and its thermal stability at high-temperature conditions, is compatible with the food industry. Microscopic and metabolomic analyses suggest cruzioseptin induces anti- Listeria bioactivity through membrane disruption and changes in the intracellular metabolome. We also report that [K4K15]CZS-1 is not resistant to peptidases/proteases emphasizing a key advantage for their use as a food preservative. However, there is a need for further structural and functional optimisations for the potential clinical application as an antibiotic. In conclusion, [K4K15]CZS-1 stand out as membrane-active peptides with the ability to induce shifts in the bacteria metabolome and inspire the development of strategies for the prevention of L. monocytogenes emergence and dissemination. • Frog Antimicrobial Peptides (AMPs) are an alternative bioactive for controlling listeriosis. • The cationic net charge is important to the potency of anti-Listeria activity. • Cruzioseptins can induce intracellular changes in the biological pathways of bacteria. • Engineered cruzioseptin is thermostable and retains bioactivity. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is an important foodborne pathogen. It can adhere to food or food contact surface for a long time and form biofilm, which will lead to equipment damage, food deterioration, and even human diseases. As the main form of bacteria to survive, the mixed biofilms often exhibit higher resistance to disinfectants and antibiotics, including the mixed biofilms formed by L. monocytogenes and other bacteria. However, the structure and interspecific interaction of the mixed biofilms are very complex. It remains to be explored what role the mixed biofilm could play in the food industry. In this review, we summarized the formation and influence factors of the mixed biofilm developed by L. monocytogenes and other bacteria, as well as the interspecific interactions and the novel control measures in recent years. Moreover, the future control strategies are prospected, in order to provide theoretical basis and reference for the research of the mixed biofilms and the targeted control measures. [ABSTRACT FROM AUTHOR]

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Listeria monocytogenes is a Gram-positive bacterium that causes listeriosis, a severe foodborne illness that is particularly dangerous during pregnancy. It thrives in diverse environments, including refrigerated conditions and food production facilities, due to its adaptability to varying temperatures, pH levels, and salt concentrations. Its virulence stems from the ability to invade host cells, particularly macrophages and epithelial cells, and avoid, or at least postpone, immune detection by utilizing virulence factors such as internalins, listeriolysin O, and actin assembly-inducing protein. This intracellular motility and biofilm formation make LM a persistent pathogen in food safety and public health. Pregnant women are at a much higher risk of listeriosis, which can result in serious fetal complications such as miscarriage, stillbirth, and preterm labor due to LM's affinity for placental tissues. The vertical transmission of LM from mother to fetus can lead to neonatal listeriosis, which can result in sepsis and meningitis, with high mortality rates if not promptly treated. Early diagnosis and treatment with antibiotics, such as ampicillin or gentamicin, are crucial for maternal and neonatal outcomes. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes (Lm) is responsible for listeriosis, a serious foodborne disease, with high hospitalization and mortality rates worldwide. The main cause of listeriosis in humans is the consumption of ready-to-eat (RTE) foods; Commission Regulation (EC) No 2073/2005 establishes microbiological criteria for Lm in RTE foods. Raw fish products are widely consumed, e.g., in sushi and various seafood recipes (e.g., carpaccio, sashimi, maki, nigiri, tartare, etc.), but are not subjected to RTE food safety criteria. The aim of our study was to assess the growth potential of Lm in raw sea bass fillets obtained from a leading aquaculture company in Greece. In order to assess the growth of Lm in raw sea bass fillets, we applied the "challenge test", a scientific experiment designed to assess the growth of Lm within a specific food product under controlled conditions. According to our results, and taking into consideration the health risk for the listeriosis-vulnerable population, raw fish products utilized in the preparation of RTE foods, including sushi and an array of seafood dishes, should be incorporated in the Category of Safety Criteria of Regulation (EC) No 2073/2005 "Ready-to-eat food able to support the growth of Listeria monocytogenes". [ABSTRACT FROM AUTHOR]

Adding essential oils in an edible coating could be an alternative for the food industry to control foodborne pathogens. In 2014, EFSA published a report highlighting the risk associated with Salmonella spp. and Norovirus in fresh and frozen berries. This study aimed to evaluate the efficacy of an edible coating (RP-7) with carvacrol and citral on reducing the population of Salmonella enterica, Escherichia coli O157:H7, Listeria monocytogenes, and murine Norovirus (MNV-1) in frozen strawberries and blueberries. Before evaluating the efficacy, the best method for applying the coating on fruit was studied. The immersion method was selected, with an optimal drying time of 45 min. After this, the berries were frozen and stored for one, two, three, four, and eight weeks at −18 °C. In strawberries, all bacteria were reduced to below 0.7 log cfu/strawberry in the eighth week, and the MNV-1 infectivity showed a reduction of nearly 2 logarithmic units. In blueberries, S. enterica and E. coli O157:H7 were reduced to 0.8 log cfu/blueberries within a week, and MNV-1 achieved a reduction of 0.8 logarithmic units at the end of the assay. The application of RP-7 affected the studied microorganisms in frozen strawberries and blueberries. [ABSTRACT FROM AUTHOR]

The consumption of raw or smoked fish entails significant microbiological risks, including contamination by pathogens such as Listeria monocytogenes, which can cause severe foodborne illnesses. This study explores the preservative use of piscicolin CM22, a novel bacteriocin derived from the psychrotolerant strain Carnobacterium maltaromaticum CM22, in two types of edible coatings (EC): chitosan-based and fish gelatin-based. An initial in vitro characterization of the technological and antimicrobial properties of these ECs with and without bacteriocin was conducted. The efficacy of the edible coatings was subsequently evaluated through shelf life and challenge tests against L. monocytogenes in raw and smoked fish products. The results demonstrated significant antimicrobial activity, with the chitosan-based coating containing piscicolin CM22 being the most effective in reducing microbial counts and maintaining pH and color stability. Furthermore, in the challenge test studies, both ECs effectively controlled L. monocytogenes, showing significant reductions in bacterial counts compared to the controls in fresh tuna, salmon, and smoked salmon. The ECs containing piscicolin CM22 reduced Listeria counts by up to 4 log CFU/g in raw and smoked fish samples, with effective control in smoked salmon for up to 15 days at refrigeration temperature. While further research is required to fully assess their preservation potential, these findings strongly indicate that piscicolin CM22-functionalized edible coatings hold significant potential for improving the quality and safety of fish products. [ABSTRACT FROM AUTHOR]

Essential oils (EOs) have antimicrobial properties, but their low solubility in water and strong flavor pose challenges for direct incorporation into food, as they can negatively impact organoleptic properties. To overcome these issues, strategies such as oil-in-water (O/W) nanoemulsions have been developed to improve EO dispersion and protection while enhancing antimicrobial efficacy. The objective of this study was to create sodium alginate-pink pepper essential oil (PPEO) nanoemulsions using microfluidization. Various formulations were assessed for physicochemical, physical, and antimicrobial properties to evaluate their potential in food applications. The microfluidized emulsions and nanoemulsions had droplet sizes ranging from 160 to 443 nm, polydispersity index (PdI) ranging from 0.273 to 0.638, and zeta potential (ζ) ranging from −45.2 to 66.3 mV. The nanoemulsions exhibited Newtonian behavior and remarkable stability after 20 days of storage. Antimicrobial testing revealed effectiveness against Staphylococcus aureus and Listeria monocytogenes, with minimum inhibitory concentrations (MIC) of 200 µg/mL for both microorganisms and minimum bactericidal concentrations (MBC) of 800 µg/mL and 400 µg/mL, respectively, proving that encapsulation of PPEO in nanoemulsions significantly increased its antibacterial activity. These results present the possibility of using PPEO nanoemulsions as a more effective natural alternative to synthetic preservatives in food systems. [ABSTRACT FROM AUTHOR]

Fermented nut-based products, obtained after soaking and fermentation, are gaining increasing interest as animal food substitutes because of ethical, environmental and health reasons. In these products, Lactic Acid Bacteria (LAB) perform the fermentation, leading to matrix acidification and contributing to controlling spoilage and pathogenic microbiota. In this work, LAB strains isolated from an artisanal product and combined with a commercial strain were added as starter cultures during nut soaking to produce a cheese-like fermented plant-based product. Three different LAB consortia were used in challenge tests at laboratory scale against Listeria monocytogenes, Escherichia coli or Salmonella Enteritidis, inoculated in nuts at 5 log CFU/g, and monitored for pathogen survival and matrix acidification. The combination of Lactiplantibacillus plantarum 82 and Leuc. carnosum 4010 resulted in faster acidification (pH value < 4.4 after 18 h instead of 48 h) and the reduction of target pathogens; L. monocytogenes was already absent after seven days from production, and the counts of E. coli or S. Enteritidis were lower with respect to other samples. Thus, this microbial consortium was used for a pilot-scale production in which, beyond safety, the fermented plant-based product was also characterized for aroma profile and phenolic compounds, parameters that are known to be affected by LAB fermentation. The results showed an enhancement of the aroma profile, with an accumulation of molecules able to confer cheese-like notes (i.e., acetoin and diacetyl) and higher phenolic content, as well as the presence of compounds (i.e., phenyllactic acid and hydroxyphenyllactic acid) that could exert antimicrobial activity. This study allowed us to set up a guided fermentation for a cheese-like vegan product, guaranteeing safety and improving aromatic and functional features. [ABSTRACT FROM AUTHOR]

Salmonella typhimurium and Listeria monocytogenes are relevant foodborne bacterial pathogens which may cause serious intoxications and infectious diseases in humans. In this study, a sensitive immunochromatographic analysis (ICA) for the simultaneous detection of these two pathogens was developed. For this, test strips containing two test zones with specific monoclonal antibodies (MAb) against lipopolysaccharides of S. typhimurium and L. monocytogenes and one control zone with secondary antibodies were designed, and the double-assay conditions were optimized to ensure high analytical parameters. Prussian blue nanoparticles (PBNPs) were used as nanozyme labels and were conjugated with specific MAbs to perform a sandwich format of the ICA. Peroxidase-mimic properties of PBNPs allowed for the catalytic amplification of the colorimetric signal on test strips, enhancing the assay sensitivity. The limits of detection (LODs) of Salmonella and Listeria cells were 2 × 102 and 7 × 103 cells/mL, respectively. LODs were 100-fold less than those achieved due to the ICA based on the traditional gold label. The developed double ICA was approbated for the detection of bacteria in cow milk samples, which were processed by simple dilution by buffer before the assay. For S. typhimurium and L. monocytogenes, the recoveries from milk were 86.3 ± 9.8 and 118.2 ± 10.5% and correlated well with those estimated by the enzyme-linked immunosorbent assay as a reference method. The proposed approach was characterized by high specificity: no cross-reactivity with other bacteria strains was observed. The assay satisfies the requirements for rapid tests: a full cycle from sample acquisition to result assessment in less than half an hour. The developed ICA has a high application potential for the multiplex detection of other foodborne pathogens. [ABSTRACT FROM AUTHOR]

AbstractListeria monocytogenes (L. monocytogenes), a gram-positive foodborne pathogen that can easily cause listeriosis. It secretes extracellular polymers and forms biofilms that are highly resistant to disinfection methods, such as UV light and germicides, posing risks to food processing equipment and food quality. Dispersion of biofilm is the cycle of its formation in which the bacteria return to planktonic state and become susceptible to antimicrobials, the strategic manipulation of biofilm dispersion is thus heralded as a novel and promising approach for the effective control of biofilm-related infections. Compared to the traditional methods, it is more effective to start with the composition of biofilms, cut off the production of their constituent substances, and genetically reduce the probability of biofilm formation. Meanwhile, the dispersion of bacteria can be supplemented with exogenous substances, making long-term control possible. This paper provides a brief but comprehensive overview of the mechanisms of L. monocytogenes biofilms or cross-contamination and their resistance properties, and facilitates our understanding and control of the prevention and containment of L. monocytogenes biofilm contamination based on the biofilm's active and passive diffusion strategies. This work provides practical guidelines for the food industry to guard against the enduring threat to food safety due to L. monocytogenes biofilms. [ABSTRACT FROM AUTHOR]

Objective. Bloodstream infections (BSIs) caused by Listeria monocytogenes are linked to high mortality of the patients. Case‐specific details related to this disease and causative strains in different districts remain to be characterized. Methods. In this study, medical data of BSIs admitted to West China Hospital from October 2017 to March 2023 were retrieved from the hospital information system. The in vitro antimicrobial susceptibility testing and whole‐genome sequencing were performed for L. monocytogenes strains isolated from blood specimens. The genetic relationship of these strains with those in public databases was also analyzed. Result. The in‐hospital mortality of L. monocytogenes BSIs was 25.7% (9/35). The changes in consciousness and elevated serum C‐reactive protein (CRP) level were found to be the differential factors of L. monocytogenes BSIs (P < 0.05). All the 27 strains studied were susceptible to ampicillin, meropenem, and erythromycin. Only 22.2% of them were susceptible to trimethoprim‐sulfamethoxazole. The Listeria pathogenicity islands 1 (LIPI‐1), truncated LIPI‐2, and multiple virulence‐related genes outside the LIPIs were determined from these strains. Also, 12 sequence types (STs) and 12 clonal complexes (CCs) were identified and classified into clonal lineages I (9/27, 33.3%) and lineages II (18/27, 66.7%), demonstrating genetic differences with the strains in the database. ST451/CC11 (5/27, 18.5%) and ST8/CC8 (4/27, 14.8%) were the common genotypes. Conclusions. The consciousness change and elevated serum CRP level were found to be the differential factors of L. monocytogenes BSIs. Considering the high virulence of the strains, it is needed to pay more attention to the dissemination of the predominant genotype. [ABSTRACT FROM AUTHOR]

Listeria monocytogenes is ubiquitous in nature and persistent in food‐processing facilities, farms, retail stores, and home and restaurant kitchens. Current research suggests ready‐to‐eat (RTE) products (including RTE hummus and fresh produce) to be of increasing interest and concern. These foods are typically stored at refrigeration temperatures suited to the survival of L. monocytogenes and are consumed without further processing. Since L. monocytogenes is ubiquitous in agricultural environments, the cultivation of fresh produce predisposes it to contamination. The contamination of RTE foods originates either from raw ingredients or, more commonly, from cross‐contamination within food‐processing facilities. Research on the food‐processing environment has been recommended to reduce the incidence of L. monocytogenes in foods. The consumption of contaminated foods by immunocompromised individuals causes invasive listeriosis, with a 20% to 30% fatality rate despite treatment. The emergence of antibiotic‐resistant strains has reduced the effectiveness of modern medicine and may increase morbidity and mortality. Without epidemiological surveillance and identifying trends in disease determinants, no action can be taken to improve food safety and mitigate the risk of such outbreaks. [ABSTRACT FROM AUTHOR]

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-β-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. [ABSTRACT FROM AUTHOR]

Several medium chain fatty acids and different chemical forms of these acids were evaluated in vitro as treatments of aerobically-exposed corn silage against spoilage and pathogenic microbes and for effects on rumen fermentation. Treatments were control (no additive), myristate (MY), laurate (LA), monolaurin (MLA), methyl ester laurate (MELA), a blend of mono-, di- and triglycerides of laurate (BLA), and monocaprylate (MC). After 24 h of aerobic incubation (37°C), yeast and mold growth were not influenced (P > 0.05) by treatments, while the net growth of lactic acid bacteria was decreased, albeit slightly, compared to that by untreated controls (P < 0.01) by all treatments of the air-exposed corn silage. Compared with controls, wild-type enterococci were decreased (P < 0.01) in MLA, MELA, and BLA. Staphylococcus aureus was reduced (P < 0.01) with MLA, MELA, BLA, and MC. Total aerobes showed reductions (P < 0.01) with MLA, BLA, and MC. Listeria monocytogenes numbers were reduced (P < 0.01) with MELA. Anaerobic incubation (24 h; 39°C) of ruminal fluid (10mL) with 0.2 g air-exposed and MCFA-treated corn silage revealed higher hydrogen accumulations (P < 0.01) with MLA and MC over the control treatment. Methane was decreased (P < 0.01) solely by MLA. There was an increase (P < 0.01) of acetate with MELA and MC; of propionate with MELA or by BLA; and of butyrate with MLA, MELA, BLA, and MC. Total VFA, hexose fermented, and ammonia were increased (P < 0.01) with MELA, BLA, and MC. The acetate:propionate ratio was increased (P < 0.01) with MC. The results showed that treatment of air-exposed corn silage with esterified MCFA had no effect on yeasts and molds but prevented propagation or reduced populations of some unwanted and potentially desirable bacteria. Modest methane reduction was seen during in vitro incubation of rumen fluid suspensions with MLA-treated silage and ammonia accumulations were increased in esterified MCFA-treated silage. Little, if any, other detrimental effects on beneficial ruminal fermentation characteristics were observed. [ABSTRACT FROM AUTHOR]

Food contamination has emerged as a significant global health concern, posing substantial challenges to the food industry. Bacteria are the primary cause of foodborne diseases. Consequently, it is crucial to develop accurate and efficient sensing platforms to detect foodborne bacteria in food products. Among various detection methods, biosensors have emerged as a promising solution due to their portability, affordability, simplicity, selectivity, sensitivity, and rapidity. Electrospun nanofibers have gained increasing popularity in enhancing biosensor performance. These nanofibers possess a distinctive three-dimensional structure, providing a large surface area and ease of preparation. This review provides an overview of the electrospinning technique, nanofibers and nanofiber-based biosensors. It also explores their mechanisms and applications in the detection of foodborne bacteria such as Salmonella, Listeria monocytogenes (L. monocytogenes), Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Pseudomonas putida (P. putida). [ABSTRACT FROM AUTHOR]

Ozone is a molecule that has gained increasing interest in recent years by food industries for sanitization of food-grade surfaces. Compared to chemical sanitizers such as chlorine, hydrogen peroxide, or peracetic acid, ozone shows undeniable advantages, such as the absence of by-products that should affect human health or the possibility of generating it when needed. Therefore, the aim of this paper was the assessment of the resistance to ozonized water of two pathogenic bacteria (Listeria monocytogenes, Salmonella) and of three airborne food-spoiling fungi (Aspergillus brasiliensis, Hyphopichia burtonii, and Penicillium nordicum) inoculated on stainless steel tiles and treated in static conditions with 1 to 6 mg L−1 (pathogens) or 8.5 mg L−1 (filamentous fungi). Ozonized water gave different results based on the tested microorganisms: pathogenic bacteria proved markedly more sensible to ozone than filamentous fungi, even if great differences were observed at inter- and intra-specific levels for both categories of microorganisms. Nevertheless, the non-linear inactivation kinetics of the studied strains made the calculation of a punctual F-value difficult, so in industrial practice, adequate tailoring of the treatments to be applied, based on the registered extrinsic factors and the industrial bio-burden, would be appropriate. [ABSTRACT FROM AUTHOR]

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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. [ABSTRACT FROM AUTHOR]

L. monocytogenes is considered one of the most dangerous foodborne pathogens. This study aimed to determine the occurrence of L. monocytogenes in RTE meat products from southern Poland, including serogroups and antimicrobial susceptibility, and to assess the usefulness of MALDI-TOF MS as a tool for identifying L. monocytogenes. A total of 848 production batches of RTE meat products were analyzed for L. monocytogenes. All L. monocytogenes isolates were serotyped using the multiplex PCR method, tested for antimicrobial susceptibility using the disk diffusion method and identified using the MALDI-TOF MS method. L. monocytogenes was detected in 52/848 batches of RTE meat products (6.13%). The isolates belonged to four serogroups: 17/52 (33%) isolates to IVb; 15/52 (29%) isolates to IIa; 10/52 (19%) isolates to IIc and 10/52 (19%) isolates to IIb. All isolates (52/52) showed susceptibility to the tested antimicrobials. Using MALDI-TOF MS, 10/52 isolates (19.2%) were identified at the level of secure genus identification, probable species identification; 37/52 isolates (71.2%) were identified at the level of probable genus identification; 3/52 isolates (5.8%) were incorrectly identified as L. innocua; and 2/52 isolates (3.8%) were not identified. The occurrence of L. monocytogenes in RTE meat products was low. Almost half of the analyzed isolates were L. monocytogenes of serogroups, which are most often associated with listeriosis in humans in Poland. All isolates showed susceptibility to five commonly used antimicrobials for treating listeriosis. The use of MALDI-TOF MS as a tool for the identification of L. monocytogenes indicated its limitations related to the insufficient representation of the pathogen in the reference database. [ABSTRACT FROM AUTHOR]

Listeria are Gram-negative intracellular foodborne pathogens that can cause invasive infections with high mortality rates. In this work, the antibacterial activity of ten essential oils, infusion extracts, and decoction extracts of some medicinal plants was tested against Listeria monocytogenes and listeria ivanovii strains. The effects of different physical conditions including temperature, pH, sodium chloride, and some organic acids were studied. The results showed that the water extracts gave the maximum bacterial inhibition, while ethanolic extract was inactive against the tested Listeria spp. The antibiotic sensitivity of L. monocytogenes LMG10470 and L. ivanovii LMZ11352 was tested against five antibiotics including imipenem, levofloxacin, amikacin, ampicillin, and amoxicillin. Imipenem was the most effective antibiotic, resulting in inhibition zones of 40 mm and 31 mm for L. monocytogenes and L. ivanovii, respectively. When imipenem mixed with Syzygium aromaticum oil, Salvia officinalis oil, Pimpinella anisum infusion, and Mentha piperita infusion each, the water extract of Moringa oleifera leaves and seeds against LMG10470 and LMZ11352 resulted in broader antibacterial activity. The antimicrobial activity of both Pimpinella anisum and Mentha piperita plant extracts is related to a variety of bioactive compounds indicated by gas chromatography–mass spectrometry analysis of these two plant extracts. These two plant extracts seemed to contain many chemical compounds elucidated by gas chromatography–mass spectrometry (GC-MS) and infrared radiation spectra. These compounds could be classified into different chemical groups such as ethers, heterocyclic compounds, aromatic aldehydes, condensed heterocyclic compounds, ketones, alicyclic compounds, aromatics, esters, herbicides, saturated fatty acids, and unsaturated fatty acids. The use of these natural compounds seems to be a useful technological adjuvant for the control of Listeria spp. in foods. [ABSTRACT FROM AUTHOR]

The present study describes the genetic characterization of L. monocytogenes strains found in the Republic of Kosovo's food chain. From 2016 to 2022, 995 samples were collected. Overall, 648 samples were from ready-to-eat (RTE) food products, 281 from food products consumed cooked (FPCC), 60 from raw materials, and 6 from environmental samples. Overall, 11.76% (117 out of 995) of the samples were contaminated by L. monocytogenes, comprising 6.33% (41 out of 648) from RTE products, 14.95% (42 out of 281) from FPCC, 55.00% (33 out of 60) from raw materials, and 16.66% (1 out of 6) from environmental samples. All isolates were subjected to molecular serotyping and clonal complex (CC) identification by using real-time PCR, as well as multilocus sequence typing. All isolates were grouped into four molecular serotypes, IIa (34.19%), IIb (3.48%), IIc (32.48%), and IVb (29.91%), as well as Lineage I (33.33%) and Lineage II (66.66%). In total, 14 CCs were identified from 41 RTE isolates; however, CC29 (7), CC2 (6), and CC6 (6) were the most dominant. By contrast, CC9 was by far the most represented CC in both FPCC (21) and RM (14). Moreover, 30 isolates expressed CC1, CC2, CC4, or CC6, which are particularly associated with severe human infections. [ABSTRACT FROM AUTHOR]

Preclinical mouse models are critical for understanding the pathophysiological response to infections and developing treatment strategies for sepsis. In keeping with ethical values, researchers follow guidelines to minimize the suffering of the mice. Weight loss is a criteria used as a humane end point, but there is no official recommendation for a maximum weight loss leading to euthanasia. To evaluate whether the thresholds used in daily practice are optimal, we performed a comprehensive retrospective analysis of data generated over 10 years with > 2300 mice used in models of infection with Listeria monocytogenes, Streptococcus pneumoniae, Candida albicans and H1N1 influenza virus. Weight loss segregated mice that survived from those that did not. Statistical analyses revealed that lowering the weight loss thresholds used (none, 30% or 20%) would have increased mortality rates due to the sacrifice of mice that survived infections (p < 0.01–0.001). Power calculations showed high variability and reduction of power as weight loss thresholds approached 20% for S. pneumoniae and L. monocytogenes models. Hence, weight loss thresholds need to be adapted to each model of infection used in a laboratory. Overall, weight loss is a valuable predictor of mortality that contributes to the robustness of composite scores. To our knowledge, this is the most extensive study exploring the relationship between weight loss threshold and sepsis outcome. It underscores the importance of the infection-model-specific evaluation of weight loss for use in clinical scores defining humane endpoints to minimize mouse suffering without compromising statistical power and scientific objectives. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: The foodborne bacterium Listeria monocytogenes (Lm) causes a range of diseases, from mild gastroenteritis to invasive infections that have high fatality rate in vulnerable individuals. Understanding the population genomic structure of invasive Lm is critical to informing public health interventions and infection control policies that will be most effective especially in local and regional communities. Methods: We sequenced the whole draft genomes of 936 Lm isolates from human clinical samples obtained in a two-decade active surveillance program across 58 counties in New York State, USA. Samples came mostly from blood and cerebrospinal fluid. We characterized the phylogenetic relationships, population structure, antimicrobial resistance genes, virulence genes, and mobile genetic elements. Results: The population is genetically heterogenous, consisting of lineages I–IV, 89 clonal complexes, 200 sequence types, and six known serogroups. In addition to intrinsic antimicrobial resistance genes (fosX, lin, norB, and sul), other resistance genes tetM, tetS, ermG, msrD, and mefA were sparsely distributed in the population. Within each lineage, we identified clusters of isolates with ≤ 20 single nucleotide polymorphisms in the core genome alignment. These clusters may represent isolates that share a most recent common ancestor, e.g., they are derived from the same contamination source or demonstrate evidence of transmission or outbreak. We identified 38 epidemiologically linked clusters of isolates, confirming eight previously reported disease outbreaks and the discovery of cryptic outbreaks and undetected chains of transmission, even in the rarely reported Lm lineage III (ST3171). The presence of animal-associated lineages III and IV may suggest a possible spillover of animal-restricted strains to humans. Many transmissible clones persisted over several years and traversed distant sites across the state. Conclusions: Our findings revealed the bacterial determinants of invasive listeriosis, driven mainly by the diversity of locally circulating lineages, intrinsic and mobile antimicrobial resistance and virulence genes, and persistence across geographical and temporal scales. Our findings will inform public health efforts to reduce the burden of invasive listeriosis, including the design of food safety measures, source traceback, and outbreak detection. [ABSTRACT FROM AUTHOR]

The present study investigated the microbiological safety of the increasingly popular plant‐based milk alternatives. No (10/27) or only very low microbial counts (17/27) were detected in the tested products. These were mainly identified as spore formers via MALDI‐ToF‐MS. Three products contained Bacillus cereus group isolates, which were able to form considerable amounts of enterotoxins and exhibited cytotoxicity towards CaCo‐2 cells. Preliminary tests showed good growth of B. cereus, Listeria monocytogenes, and Salmonella enterica in all tested products (maximum bacterial counts: 5 × 1012 cfu/mL). These experiments also revealed strain‐, time‐, and temperature‐, but especially product‐specific enterotoxin production of B. cereus. In propagation and persistence tests according to DIN EN ISO 20976‐1:2019‐09, rapid bacterial proliferation was also detected in all products. B. cereus generally showed lower bacterial counts (106–107 cfu/mL) compared to L. monocytogenes and S. enterica (108–109 cfu/mL), but was detectable in a larger number of products over the test period of 6 weeks. pH values decreased (20/27) over time and visual and/or olfactory alterations (24/27) were observed. The present study provides information on the occurrence, growth and persistence of pathogenic bacteria in plant‐based drinking milk alternatives. It also points out that the accompanying changes in pH, odor, and appearance are not necessarily recognizable to the consumer. Practical Application: The present study contributes to the understanding of the microbial risk related to plant‐based drinking milk alternatives. It is crucial that the manufacturer ensures that particularly spore formers have been effectively eliminated from the products. Among them, especially toxin‐producing bacteria can pose a risk to the consumer, as these products promote proliferation and persistence of the bacteria. [ABSTRACT FROM AUTHOR]

Time‐temperature data for queso fresco (QF) cheese varieties stored in a residential refrigerator operating at 5°C and a predictive microbiology secondary model for Listeria monocytogenes in QF were used to estimate a refrigerator performance indicator (RPI) of microbial preservation. RPI values were used to assess how compressor technology (single [SS] and variable speed [VS]), ambient temperature (21.1°C [LT] and 32.2°C [HT]), and refrigerator load (22.5 kg regular load and 39 kg higher load) affected preservation performance. All deterministic and probabilistic RPI estimations slightly exceeded the desirable 1.0 value, i.e., the variable temperatures for the QF kept in the refrigerator were worse than keeping it constantly at the temperature recommended by food safety agencies for QF. Furthermore, the mean comparison of estimates of the time‐temperature equivalent indicator previously developed by French researchers showed similar behavior to those observed for RPI. Finally, statistical analysis showed that Tambient was the factor with the highest impact on refrigerator performance because of its impact on the sample temperature increase during door openings and when exposed to ambient temperature during product use. This highlights the need to reduce the time for product temperature recovery by improving the compressor operation logic. Also important are consumer behavior changes such as a reduction in product exposure to ambient temperature and in the door opening duration and frequency. Practical Application: This study demonstrated how a quantitative tool (RPI) can assess refrigerator preservation performance. Although the findings presented can be applied to any cold chain segment, the data used was collected for its weakest link, the domestic refrigerator. Surveys show that 77% of them operate above the recommended 4°C. The RPI methodology is ready for use by refrigerator designers to assess performance improvements possible by modifications of the compressor operation logic. Moreover, it can be integrated into smart‐hubs monitoring the frequency and duration of refrigerator door openings to inform consumers when their habits are compromising the preservation performance of the refrigerator. [ABSTRACT FROM AUTHOR]

The effects of juice pH, type of acidulant, and post-treatment refrigeration on the high-pressure processing (HPP) inactivation of Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes in acid beverages were evaluated. Inoculated apple, orange, and grape juices (at their original pH and adjusted to pH 4.00, 4.50, and 5.00) were treated at 550 MPa for 1 min at 5 °C. In addition, inoculated model solutions acidified to a pH of 5.00 with acetic, citric, malic, and tartaric acids were treated at 400 MPa for 1 min at 5 °C. The effect of refrigerated storage for 24 h after treatment on pathogen inactivation in both experiments was also assessed. A greater than 5-log reduction of the three pathogens inoculated was achieved in all juices immediately after HPP at the juices' original pH, and of L. monocytogenes under all experimental conditions. Refrigerated storage for 24 h after HPP treatment improved the inactivation of E. coli O157:H7, to >5-log reduction, at pH 4.00 in apple juice and of Salmonella in the three juices at pH 4.00. The type of acidulant did not significantly (p > 0.01) affect E. coli or Salmonella inactivation in acidified model solutions but a greater than 5-log reduction after HPP was only achieved for L. monocytogenes when acetic acid was used. The effectiveness of HPP for pathogen inactivation depended largely on product pH and the target pathogen of concern. [ABSTRACT FROM AUTHOR]

Polar habitats offer excellent sites to isolate unique bacterial strains due to their diverse physical, geochemical, and biological factors. We hypothesize that the unique environmental conditions of polar regions select for distinct strains of lactic acid bacteria (LAB) with novel biochemical properties. In this study, we characterized ten strains of psychrotrophic LAB isolated from hitherto poorly described sources—High Arctic and maritime Antarctic soils and soil-like materials, including ornithogenic soils, cryoconites, elephant seal colonies, and postglacial moraines. We evaluated the physiological and biochemical properties of the isolates. Based on 16S rRNA and housekeeping genes, the four LAB strains were assigned to three Carnobacterium species: C. alterfunditum, C. maltaromaticum, and C. jeotgali. The remaining strains may represent three new species of the Carnobacterium genus. All isolates were neutrophilic and halophilic psychrotrophs capable of fermenting various carbohydrates, organic acids, and alcohols. The identified metabolic properties of the isolated Carnobacterium strains suggest possible syntrophic interactions with other microorganisms in polar habitats. Some showed antimicrobial activity against food pathogens such as Listeria monocytogenes and human pathogens like Staphylococcus spp. Several isolates exhibited unique metabolic traits with potential biotechnological applications that could be more effectively exploited under less stringent technological conditions compared to thermophilic LAB strains, such as lower temperatures and reduced nutrient concentrations. Analysis of extrachromosomal genetic elements revealed 13 plasmids ranging from 4.5 to 79.5 kb in five isolates, featuring unique genetic structures and high levels of previously uncharacterized genes. This work is the first comprehensive study of the biochemical properties of both known and new Carnobacterium species and enhances our understanding of bacterial communities in harsh and highly selective polar soil ecosystems. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Using sous-vide technology in combination with essential oils offers the potential to extend the preservation of food items while preserving their original quality. This method aligns with the growing consumer demand for safer and healthier food production practices. This study aimed to assess the suitability of minimal processing of game meat and the effectiveness of vacuum packaging in combination with Piper nigrum essential oil (PNEO) treatment to preserve red deer meat samples inoculated with Listeria monocytogenes. Microbial analyses, including total viable count (TVC) for 48 h at 30 °C, coliform bacteria (CB) for 24 h at 37 °C, and L. monocytogenes count for 24 h at 37 °C, were conducted. The cooking temperature of the sous-vide was from 50 to 65 °C and the cooking time from 5 to 20 min. Additionally, the study monitored the representation of microorganism species identified through mass spectrometry. The microbiological quality of red deer meat processed using the sous-vide method was monitored over 14 days of storage at 4 °C. The results indicated that the TVC, CB, and L. monocytogenes counts decreased with the temperature and processing time of the sous-vide method. The lowest counts of individual microorganism groups were observed in samples treated with 1% PNEO. The analysis revealed that PNEO, in combination with the sous-vide method, effectively reduced L. monocytogenes counts and extended the shelf life of red deer meat. Kocuria salsicia, Pseudomonas taetrolens, and Pseudomonas fragi were the most frequently isolated microorganism species during the 14-day period of red deer meat storage prepared using the sous-vide method. [ABSTRACT FROM AUTHOR]

In the dairy industry, traditional heat treatments are known for their high water and energy consumption, and more economically and environmentally friendly solutions are being sought. Infrared (IR) technology offers advantages in energy efficiency and environmental sustainability; however, its effectiveness in milk processing, particularly in pathogen inactivation, remains relatively unexplored. In this study, homogenized raw milk was subjected to IR treatment, and its impact on Listeria monocytogenes, Salmonella spp., and Enterobacteriaceae was assessed. Results indicate that IR treatment effectively reduces the microbial load, achieving levels of inactivation comparable to conventional pasteurization methods (around 6 Log10 CFU/mL). Moreover, the treatment maintains milk pH levels, suggesting minimal alteration to its composition. Further research is needed to explore the full extent of IR treatment on milk sanitation efficacy, deeply exploring IR technology to fully assess its applicability and integration into dairy processing practices. Despite regulatory challenges, the Wir System Milk shows promise as a cost-effective and eco-friendly alternative for raw milk treatment. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. The use of essential oils (EO) has attracted interest in the food industry because of their wide range of beneficial properties. In this study, a new functional yogurt was developed using 2 EO, marjoram and geranium, at 3 different concentrations (0.2%, 0.4%, and 0.6% vol/vol). The physicochemical properties, including syneresis, viscosity, pH, and chemical composition; bioactivities, including antioxidant activity, anticancer and antibacterial effects, total phenolic content (TPC), and total flavonoid content (TFC); and sensory characteristics of the developed yogurt were evaluated. The findings indicated that the yogurts fortified with 0.6% marjoram or geranium exhibited higher viscosity and lower syneresis compared with other treatments. The yogurt supplemented with 0.6% marjoram displayed significant antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, Salmonella typhimurium , and Escherichia coli. In addition, the yogurt enriched with geranium and marjoram oils at a concentration of 0.6% had notably significant (P < 0.05) higher TFC levels compared with the control sample and other concentrations. In the same context, in terms of TPC, yogurt supplemented with 0.6% marjoram displayed significantly (P < 0.05) elevated levels in comparison to the other samples tested. Yogurt enriched with marjoram oil exhibited noteworthy antioxidant activity, followed by geranium oil, compared with the control samples. The yogurt supplemented with 0.6% marjoram demonstrated strong radical scavenging activity, and the yogurt fortified with 0.6% geranium showed higher anticancer activity against HepG2 human liver carcinoma cells and oxidative stress enzyme activities. Among the various concentrations of EO tested, the yogurts fortified with 0.6% marjoram or geranium EO exhibited the most favorable outcomes, followed by 0.4% marjoram or geranium. To summarize, geranium and marjoram EO can be used as a potential nutritious ingredient and as a natural preservative for milk and related products. [Display omitted] [ABSTRACT FROM AUTHOR]