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Whole genome sequencing is an essential cornerstone of pathogen surveillance and outbreak detection. Established sequencing technologies are currently being challenged by Oxford Nanopore Technologies (ONT), which offers an accessible and cost-effective alternative enabling gap-free assemblies of chromosomes and plasmids. Limited accuracy has hindered its use for investigating pathogen transmission, but recent technology updates have brought significant improvements. To evaluate its readiness for outbreak detection, we selected 78 Listeria monocytogenes isolates from diverse lineages or known epidemiological clusters for sequencing with ONT's V14 Rapid Barcoding Kit and R10.4.1 flow cells. The most accurate of several tested workflows generated assemblies with a median of one error (SNP or indel) per assembly. For 66 isolates, the cgMLST profiles from ONT-only assemblies were identical to those generated from Illumina data. Eight assemblies were of lower quality, with more than 20 erroneous sites each, primarily caused by methylations at the GAAGAC motif (5'-GAAG 6mA C-3'/5'-GT 4mC TTC-3'). This led to inaccurate clustering, failing to group isolates from a persistence-associated clone that carried the responsible restriction-modification system. Out of 50 methylation motifs detected among the 78 isolates, only the GAAGAC motif was linked to substantially increased error rates. Our study shows that most L. monocytogenes genomes assembled from ONT-only data are suitable for high-resolution genotyping, but further improvements of chemistries or basecallers are required for reliable routine use in outbreak and food safety investigations., Competing Interests: The authors declare no conflict of interest.

Listeria monocytogenes is a major foodborne pathogen affecting developing, and developed countries. The analysis of food contact surfaces in food industries is key for better controlling this pathogen. The current study focused on the development, optimization, and evaluation of a rapid and simple method for the detection of L. monocytogenes on stainless steel surfaces, suitable for decentralized setups, taking advantage of Loop-mediated isothermal amplification (LAMP). This was accomplished using a general pre-enrichment broth (TSB), with a simple DNA extraction based on a chelating resin, and final isothermal amplification. Two different detection strategies were tested, real-time fluorescence and naked-eye colorimetric, which were evaluated after 5, 7, and 24 h of pre-enrichment. Regardless the detection chemistry selected, after 5-7 h of pre-enrichment, 10 3 -10 4  CFU/cm 2 were needed to obtain a positive result, while after 24 h, it was possible to detect concentrations below 10 CFU/cm 2 . Within each given time, all the performance parameters calculated, relative sensitivity, specificity, and accuracy, reached values higher than 80-90%; likewise, a Cohen's k of concordance with a culture-based approach higher than 0.8. Overall, the most sensitive assay can be performed in roughly 25 h. This time-to-result outperforms commercial kits with the added value of specifically detecting L. monocytogenes instead of Listeria spp. KEY POINTS: • Real-time fluorescence and naked-eye colorimetric, were compared for the novel assay. • An LOD50 of 3.4 CFU/cm 2 and 4.2 CFU/cm 2 was calculated for the two assays. • Three pre-enrichment times were compared providing 24 h better results., Competing Interests: Declarations Ethics approval This article does not contain any studies with human participants or animals performed by any of the authors. Conflict of interest The authors declare no conflict of interest., (© 2024. The Author(s).)

Among the severe foodborne illnesses, listeriosis resulting from the pathogen Listeria monocytogenes exhibits one of the highest fatality rates. This study investigated the application of near infrared hyperspectral imaging (NIR-HSI) for the classification of three L. monocytogenes serotypes namely serotype 4b, 1/2a and 1/2c. The bacteria were cultured on Brain Heart Infusion agar, and NIR hyperspectral images were captured in the spectral range 900-2500 nm. Different pre-processing methods were applied to the raw spectra and principal component analysis was used for data exploration. Classification was achieved with partial least squares discriminant analysis (PLS-DA). The PLS-DA results revealed classification accuracies exceeding 80 % for all the bacterial serotypes for both training and test set data. Based on validation data, sensitivity values for L. monocytogenes serotype 4b, 1/2a and 1/2c were 0.69, 0.80 and 0.98, respectively when using full wavelength data. The reduced wavelength model had sensitivity values of 0.65, 0.85 and 0.98 for serotype 4b, 1/2a and 1/2c, respectively. The most relevant bands for serotype discrimination were identified to be around 1490 nm and 1580-1690 nm based on both principal component loadings and variable importance in projection scores. The outcomes of this study demonstrate the feasibility of utilizing NIR-HSI for detecting and classifying L. monocytogenes serotypes on growth media., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: [Paul James Williams reports financial support was provided by National Research Foundation of South Africa. Rumbidzai T Matenda reports financial support and travel were provided by South Africa Department of Science and Innovation. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.]., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Listeria monocytogenes (LM) is a zoonotic pathogen that causes sporadic infectious listeriosis, which is a foodborne disease associated with consumption of contaminated food or feed. The internal organs of an African lion from a zoo in Shanghai were analysed to determine the cause of death. LM infection was suspected on the basis of the clinical symptoms and pathological changes and confirmed by polymerase chain reaction, whole genome sequencing and phylogenetic analysis. This is the first report of LM infection of an African lion in China., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

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

Background: There is a high demand in the food industry and in public health for rapid automated methods capable of high-volume sample processing., Objective: In an unpaired study, Roka Atlas® System performance was compared with Health Canada reference method MFHPB-30 for Listeria spp. (LSP Roka assay) and Listeria monocytogenes (LmG2 Roka Assay) detection on plastic (PL), sealed concrete (SC), and stainless-steel (SS) surfaces (45 samples each per candidate or reference method)., Methods: Seeking shorter enrichment time for the candidate method, R2 medium pre-enrichment for 14, 16, and 24 h at 35°C was combined with the Roka assay. Listeria welshimeri, L. innocua, and L. monocytogenes were employed to individually inoculate each of the three surfaces, with two competing microorganisms within the 10-100-fold higher concentration range., Results: False negative, false positive, sensitivity, and specificity were 0, 0, 100, and 100%, respectively, for the plastic, sealed concrete, and stainless-steel surfaces, regardless of inoculation level (high, low, and uninoculated) and enrichment time. Candidate method detected 10, 7, and 9 true positives, versus 10, 6, and 10 by the reference method in individually inoculated SS, PL, and SC, respectively., Conclusions: Probability of detection for all the three surfaces for the Roka Atlas System was comparable to the reference method in this unpaired study., Highlights: The Roka Atlas System detected targets after as little as 14 h enrichment. Surface type did not negatively affect assay sensitivity or specificity. The Roka Atlas System was comparable to the reference method., (© The Author(s) 2024. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Listeria monocytogenes (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 4×10 2 to 4×10 7 CFU/mL (R 2 =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., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Between 2017 and 2019, pulsed-field gel electrophoresis was replaced by whole genome sequencing (WGS) for identifying enteric disease clusters in Canada. The number and characteristics of all clusters of Listeria monocytogenes , Salmonella , Shiga toxin-producing Escherichia coli (STEC), and Shigella spp. between 2015 and 2021 were analyzed. Following the transition to WGS, an increase in the number of Salmonella , STEC, and Shigella clusters was noted, whereas the number of clusters of L. monocytogenes decreased. Unlike previous subtyping methods, WGS provided increased resolution to identify discrete clusters of Salmonella Enteritidis. This led to the identification of a number of outbreaks linked to frozen raw breaded chicken products and ultimately a change in food safety policy to reduce the number of illnesses associated with these products. Other pathogens did not experience a similar increase in the number of outbreaks detected. Although WGS did provide increased confidence in the genetic relatedness of cases and isolates, challenges remained in collecting epidemiological data to link these illnesses to a common source.

Farmstead dairy processing facilities may be particularly susceptible to Listeria spp. contamination due to the close physical proximity of their processing environments (PE) to associated dairy farm environments (FE). In this case study, we supported the implementation of interventions focused on improving (1) cleaning and sanitation efficacy, (2) hygienic zoning, and (3) sanitary equipment and facility design and maintenance in a farmstead dairy processing facility, and evaluated their effect on Listeria spp. detection in the farmstead's PE over 1 yr. Detection of Listeria spp. in the farmstead's PE was numerically reduced from 50% to 7.5% after 1 yr of intervention implementation, suggesting that these interventions were effective at improving Listeria spp. control. In addition, environmental samples were also collected from the farmstead's FE to evaluate the risk of the FE as a potential source of Listeria spp. in the PE. Overall, detection of Listeria spp. was higher in samples collected from the FE (75%, 27/36) compared with samples collected from the PE (24%, 29/120). Whole-genome sequencing performed on select isolates collected from the PE and FE supported the identification of 6 clusters (range of 3 to 15 isolates per cluster) that showed ≤ 50 high-quality single nucleotide polymorphism differences. Of these 6 clusters, 3 (i.e., clusters 2, 4, and 5) contained isolates that were collected from both the PE and FE, suggesting that transmission between these 2 environments was likely. Moreover, all cluster 2 isolates represented a clonal complex of L. monocytogenes commonly associated with dairy farm environmental reservoirs (i.e., CC666), which may support that the farmstead's FE represented an upstream source of the cluster 2 isolates that were found in the PE. Overall, our data underscore that although the FE can represent a potential upstream source of Listeria spp. contamination in a farmstead dairy processing facility, implementation of targeted interventions can help effectively minimize Listeria spp. contamination in the PE., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Neurolisteriosis, a common disease of small ruminants, is most often caused by Listeria monocytogenes . Here we describe 25 cases of caprine neurolisteriosis diagnosed in our laboratory over a 5-y period and compare our fluorescent antibody test (FAT) results with immunohistochemistry (IHC) and polymerase chain reaction (PCR) testing for diagnostic confirmation. Neurohistologic changes consistent with neurolisteriosis affected the pons in all cases, extending rostrally to the mesencephalon in 6 cases, caudally to the medulla oblongata in 6 cases, and/or dorsally to the cerebellum in 4 cases. Acute inflammatory changes were observed in 17 cases, and included neuroparenchymal microabscesses, neuronal necrosis and neuronophagia, axonal swelling, microgliosis and astrogliosis, and perivascular neutrophils with macrophages, lymphocytes, and plasma cells that occasionally extended to the leptomeninges. Subacute-to-chronic changes (8 cases) consisted of neuroparenchymal and perivascular clusters of macrophages with rare neutrophils, lymphocytes, and plasma cells admixed with glial nodules. Bacterial bacilli were observed within neutrophils or macrophages in H&E-stained tissue sections in 4 cases. Gram stain highlighted gram-positive bacilli in 13 cases. Neurolisteriosis was confirmed by FAT in 2 cases, by IHC in 19 cases, and by PCR in 20 cases., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Listeria monocytogenes is a ubiquitous bacterium and opportunistic pathogen for immunocompromised patients. Peritoneal dialysis-related L. monocytogenes peritonitis is a rare clinical presentation, with a total of 23 cases reported to date and an overall mortality rate of 17.3%. The current report describes a case of peritoneal dialysis-related L. monocytogenes peritonitis in a woman in her 60s on long-term immunosuppressive therapies for a prior renal transplant. We treated this patient successfully with intraperitoneal ampicillin (125 mg/L in dialysate), in addition to intravenous ampicillin (1 g every 12 hours) for 10 days and subsequent oral amoxicillin 500 mg every 12 hours to complete a total 21-day course of therapy. This treatment regimen was informed by a literature review of pre-existing case studies, which demonstrated considerable variability in recommended antimicrobial dosing and frequency. A summary of this literature review is included in the current case report., Competing Interests: Competing interests: None declared., (© BMJ Publishing Group Limited 2024. No commercial re-use. See rights and permissions. Published by BMJ.)

Listeria monocytogenes, a zoonotic foodborne pathogen, presents a significant threat to global public health. Therefore, rapid and sensitive detection methods are crucial in mitigating the spread of L. monocytogenes induced diseases. This study introduced a loop-mediated isothermal amplification (LAMP) lyophilized powder detection reagent specifically designed for identifying Listeria monocytogenes. The reagent is user-friendly, quick, and can be easily transported and stored at room temperature. It exhibits no cross-reactivity with eight other types of bacteria and boasts a sensitivity of 10 1  CFU/mL. In a comparative study of 30 samples, the LAMP lyophilized powder detection reagent demonstrated higher sensitivity than the commercial Listeria monocytogenes qPCR detection kit. Additionally, the experimental time was reduced by approximately 30 min, making it highly suitable for rapid diagnosis. Preparation of lyophilized LAMP reagents may facilitate large-scale deployment, particularly in endemic areas or regions facing rapid outbreaks. This could greatly aid in controlling the transmission of pathogens, especially those transmitted through food., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

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., (© 2024. The Author(s).)

Aims: Plant-based alternatives to meat and dairy products have become increasingly popular in the UK. Despite a public perception that they have a relatively low microbiological risk, outbreaks of illness have been linked with these foods. This study aimed to assess the microbiological safety and quality of vegan alternatives to dairy and meat products available in England., Methods and Results: Samples were collected between September 2022 and March 2023 from retail, production, and catering premises, and tested for a range of bacterial pathogens and hygiene indicators using standard procedures. A total of 937 samples were tested, of which 92% were of a satisfactory microbiological quality, 3% were borderline, and 5% were unsatisfactory. Those interpreted as unsatisfactory were due to elevated counts of Enterobacteriaceae and Escherichia coli (indicators of poor hygiene) rather than pathogenic microorganisms. Listeria monocytogenes was present in five samples of tofu, all from the same producer (all at counts of <100 CFU g-1), while other Listeria species were detected at counts of <20 CFU g-1 in two burgers and two 'vegan chicken' products. The majority of samples did not have pH and water activity values that would significantly contribute to preventing microbial growth: 62.4% had pH > 5.0 and 82.4% had Aw > 0.94., Conclusions: The majority of vegan products examined were of a satisfactory quality, but results demonstrate that microbiological control must be maintained using appropriate processing and storage temperatures, and application of a safe length of shelf life., (© The Author(s) 2024. Published by Oxford University Press on behalf of Applied Microbiology International.)

Objectives: Using a sporadic case of listeriosis suspected to have been caused by consuming a pre-packaged cold-chain ready-to-eat (RTE) food in Beijing, China in 2021 as an exemplar, this study demonstrated the importance of thoroughly investigating the source of listeriosis up to the production point for mitigating infection risk during routine monitoring of Listeria in food facilities and national surveillance program using whole-genome sequencing (WGS)., Methods: Epidemiological, laboratory, traceback, and plant investigations were used to identify the source of infection., Results: WGS showed the isolate from the patient was genetically indistinguishable from that of the implicated food. During a plant investigation, L. monocytogenes was detected in 26% (9/35) of the environmental samples and one of two raw material samples, confirming the source., Conclusion: To our knowledge, this is the first investigation in China linking a case of L. monocytogenes infection to a suspected food and its production environment. This report highlights the risk of L. monocytogenes contamination of RTE food and demonstrates the role of food safety risk monitoring in identifying potential sources of infection. Reinforcing control programs in RTE processing plants, intensified surveillance of microorganisms in food products and targeted health education is required to mitigate the infection risk., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Listeria monocytogenes (Lm) is a pathogenic bacteria able to grow at refrigerated temperatures, widely distributed in the environment. This bacteria is susceptible to contaminate various food products of which refrigerated ready-to-eat foods (RTEF) may pose a risk for public health. In Europe, food business operators (FBOs) shall ensure that foodstuffs comply with the relevant microbiological criteria set out in the Regulation (EC) N°2073/2005. Food safety criteria for Lm are defined in RTEF throughout their shelf-life. FBOs should implement studies to demonstrate that the concentration of Lm does not exceed 100 CFU/g at the end of the shelf-life, taking into account foreseeable conditions of distributions, storage and use, including the use by consumers. However, this last part of the cold chain for food products is the most difficult to capture and control. For this purpose, the European Union Reference Laboratory for Lm (EURL Lm) launched an inquiry to its National Reference Laboratory network and reviewed the scientific literature from 2002 to 2020. The outcomes were integrated in the technical guidance document of the EURL Lm to assess shelf-life of RTEF which resulted in the recommendation to use 10 °C as the reference temperature to simulate the reasonably foreseen storage conditions in domestic refrigerators., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024. Published by Elsevier Ltd.)

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., (© 2024. The Author(s).)

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., (© 2024. The Author(s).)

Introduction: We describe the case of an immunocompromised patient in whom a severe infection with meningoencephalitis was detected. Listeria monocytogenes was the pathogen. Invasive listeriosis is a rare infectious disease that is notifiable in Switzerland and can be lethal. The most common route of contamination for the ubiquitous bacteria is the consumption of foods such as dairy products, raw vegetables or meat. Antibiotic treatment can lead to a complete cure., Competing Interests: Die Autoren haben keine Interessenkonflikte im Zusammenhang mit diesem Artikel deklariert, (© 2024 Aerzteverlag medinfo AG.)

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 T ambient 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., (© 2024 Institute of Food Technologists.)

Listeria monocytogenes , one of the main foodborne pathogens, is commonly found in milk and dairy products. This study aimed to estimate the presence of L. monocytogenes in milk and dairy product supply chains using a meta-analysis based on PubMed, Embase, Web of Science, and Scopus databases. A total of 173 studies were included in this meta-analysis. The pooled prevalence in the supply chain environment was 8.69% (95% confidence interval [CI]: 5.30%-12.78%), which was higher than that in dairy products (4.60%, 95% CI: 1.72%-8.60%) and milk products (2.93%, 95% CI: 2.14%-3.82%). Subgroup analysis showed that L. monocytogenes prevalence in raw milk (3.44%, 95% CI: 2.61%-4.28%) was significantly higher than in pasteurized milk (0.60%, 95% CI: 0.00%-2.06%). The highest prevalence of L. monocytogenes in milk and dairy products was observed in North America (5.27%, 95% CI: 2.19%-8.35%) and South America (13.54%, 95% CI: 3.71%-23.37%). In addition, studies using culture and molecular methods (5.17%, 95% CI: 2.29%-8.06%) had higher prevalence than other detection methods. Serogroup 1/2a and 3a (45.34%, 95% CI: 28.74%-62.37%), serogroup 1/2b and 3b (14.23%, 95% CI: 6.05%-24.24%), and serogroup 4b/4e (13.71%, 95% CI: 6.18%-22.83%) were dominant in these studies. The results of this study provide a better understanding of the prevalence of L. monocytogenes in milk and dairy product supply chains and suggest a potential foodborne pathogen burden.

Multilocus variable number tandem repeat analysis (MLVA) is a molecular subtyping technique that remains useful for those without the resources to access whole genome sequencing for the tracking and tracing of bacterial contaminants. Unlike techniques such as multilocus sequence typing (MLST) and pulsed-field gel electrophoresis, MLVA did not emerge as a standardized subtyping method for Listeria monocytogenes , and as a result, there is no reference database of virulent or food-associated MLVA subtypes as there is for MLST-based clonal complexes (CCs). Having previously shown the close congruence of a 5-loci MLVA scheme with MLST, a predictive model was created using the XGBoost machine learning (ML) technique, which enabled the prediction of CCs from MLVA patterns with ∼85% (±4%) accuracy. As well as validating the model on existing data, a straightforward update protocol was simulated for if and when previously unseen subtypes might arise. This article illustrates how ML techniques can be applied with elementary coding skills to add value to previous-generation molecular subtyping data in-built food processing environments.

Background: Rapid and sensitive detection of foodborne pathogens in food plays a crucial role in controlling outbreaks of foodborne diseases, of which Listeria monocytogenes and Salmonella typhimurium are representative and notable pathogens. Thus, it's of great importance to achieve the effective detection of these pathogens. However, the most common detection methods (culture-based technique, Polymerase Chain Reaction and immunological methods) have disadvantages that cannot be ignored, such as time-consuming, laborious, complex sample preparation process, and the possibility of cross-reaction. Hence, it is essential to develop a facile detection method for the pathogens with high sensitivity and specificity to avoid the above-mentioned disadvantages., Results: We report a label-free visual platform for the simultaneous capture and detection of Listeria monocytogenes and Salmonella typhimurium. For the first time, we have prepared polydimethylsiloxane-Chromotrope 2R membrane which serves as the substrate for bacterial capture and enrichment through the formation of specific recognition sites. The positively charged Pt-covalent organic framework combines with the pathogens through surface charge interaction, thereby the label-free sandwich platform is formed. Remarkable peroxidase activity of Pt-covalent organic framework converts the conversion of bacterial quantity into amplified color signal by catalyzing 3,3',5,5'-Tetramethylbenzidine to oxidized 3,3',5,5'-Tetramethylbenzidine. The platform demonstrates the capability to identify two representative food-borne pathogens within a time frame of 100 min, exhibiting high sensitivity and excellent specificity without the interference from non-target bacteria. The limit of detection of the visual platform toward Listeria monocytogenes and Salmonella typhimurium was 1.61 CFU mL -1 and 1.31 CFU mL -1 , respectively. And the limit of quantification toward Listeria monocytogenes and Salmonella typhimurium was 4.94 CFU mL -1 and 2.47 CFU mL -1 , respectively. The relative standard derivations of the visual platform for both bacteria were lower than 4.9 %. Furthermore, our proposed platform has obtained reliable and satisfactory results on analyzing diverse food samples., Significance: This research expands the application of a label-free platform combined with unlabeled nanocomponents in the rapid isolation and detection of diverse of food-borne pathogens. The platform possesses the advantages of simple operation and real-time monitoring, without complicated sample pretreatment process. The whole detection process can realize the simultaneous monitoring of Listeria monocytogenes and Salmonella typhimurium within 100 min. Furthermore, it is also of reference significance for the detection of other common pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Listeria monocytogenes in beef receives less attention compared to other pathogens such as Salmonella and Escherichia coli. To address this gap, we conducted a literature review focusing on the presence of L. monocytogenes in beef. This review encompasses the pathogenic mechanisms, routes of contamination, prevalence rates, and the laws and regulations employed in various countries. Our findings reveal a prevalence of L. monocytogenes in beef and beef products ranging from 2.5% to 59.4%. Notably, serotype 4b was most frequently isolated in cases of beef contamination during food processing, with the skinning and evisceration stages identified as critical points of contamination., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

For foodborne pathogens pose a serious threat to public health, a magnetic separation strategy and a nanozyme-based biosensor are proposed for biosensing of Listeria monocytogenes (L. monocytogenes). In this work, doripenem is selected as a recognized molecule for the modification of magnetic beads to capture L.monocytogenes in food and environmental samples. Furthermore, the POD-like MXene-Hemin-Au is constructed from perspective of charge transfer engineering which provides a vivid example to rational design of nanozymes. Finally, the captured L.monocytogenes is labeled with MXene-Hemin-Au@mAb, forming the sandwich complexes for quantitative determination. The current signals that generated by the complexes exhibit a good linear relationship with a limit of detection of 2.3 × 10 1  CFU/mL. The biosensor shows a satisfactory applicability in real samples with recoveries of 91.19% to 102.98%. Overall, the biosensor with integrated magnetic separation strategy presents a potential approach for high sensitivity biosensing of foodborne pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Listeria monocytogenes is a foodborne pathogen of significant concern for the food industry due to its remarkable ability to persist through safety control efforts, posing a subsequent health threat to consumers. Understanding the microbial communities coexisting with L. monocytogenes in food processing environments provides insights into its persistence mechanisms. We investigated the microbial communities on non-food contact surfaces in a facility producing ready-to-eat foods, known to harbour a ST121 L. monocytogenes strain over multiple years. A 10-week sampling period was coordinated with the company and public health authorities. Metagenomic analysis revealed a stable microbial composition dominated by Pseudomonas fluorescens. While highly related populations were present in high-care production zones, distinctive taxa characteristic of specific areas were observed (e.g., Sphingomonas aerolata). Although Listeria spp. were not detected in metagenomes, they were detected in cultured samples, suggesting low relative abundance in factory settings. The findings suggest that a stable resident microbiota, with distinct adaptations to different areas within the factory, was selected for by their collective ability to survive control efforts in this environment. Listeria spp. was a member of this microbial community, albeit at low abundance, and may likewise benefit from the mutualism of the overall microbial community., Competing Interests: Declaration of competing Interest none., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

To prevent foodborne illness, adequate cleaning and disinfection (C&D) is essential to remove pathogenic bacteria from the slaughter environment. The aim of this study was to determine the presence of Campylobacter spp., Listeria monocytogenes, and extended-spectrum beta-lactamase-producing Escherichia coli (ESBL E. coli) before and after C&D in slaughterhouses. Samples from food- and non-food contact surfaces taken before and after C&D in one red meat and one poultry slaughterhouse were analyzed for the target bacteria. Whole-genome sequencing and antimicrobial susceptibility testing were performed. In total, 484 samples were analyzed. Campylobacter spp. were isolated from 13.0% to 15.5% of samples before C&D in the red meat and poultry slaughterhouse, respectively. Listeria monocytogenes was isolated before C&D in 12.5% and 5.2% of samples in the red meat and poultry slaughterhouse, respectively. It was noted that C. jejuni was detected on multiple surfaces and that L. monocytogenes showed potential persistence in one slaughterhouse. After C&D, L. monocytogenes was found in one sample. ESBL E. coli was not detected either before or after C&D. These findings show the possibility to remove pathogenic bacteria from slaughter and meat processing facilities, but also indicate that deficiencies in slaughter hygiene pose a risk of cross-contamination of meat., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Listeria monocytogenes is a gram-positive bacterium that causes listeriosis in humans. This contaminates the ready-to-eat food products and compromises their safety. Thus, detecting its presence in food samples with high sensitivity and reliability is necessary. Herein, we propose a label-free electrochemical immunosensor based on a mussel-inspired polydopamine-modified zinc molybdate/MXene (PDA@ZnMoO 4 /MXene) composite for effective and rapid detection of L. monocytogenes in food products. Spectrophotometry approaches were employed to examine the resulting composites. Voltammetry and impedimetry techniques were used to confirm the step-by-step assembly of the immunosensor and its sensitive detection of L. monocytogenes in various food products, such as milk and smoked seafood. The results demonstrated the practicality of the constructed immunosensor, with an appreciable linearity of 10-10 7  CFU/ml and a reasonably low detection limit (LOD, 12 CFU/ml). Moreover, the immunosensor exhibited excellent selectivity for microbial cocktails and acceptable repeatability, reproducibility, and storage stability. Thus, we believe that the proposed sensitive, reliable, and label-free immunosensor based on the PDA surface modification technique for detecting L. monocytogenes can be extended to monitor various food-borne pathogens to ensure food safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Metabolomics is the study of low molecular weight biochemical molecules (typically <1500 Da) in a defined biological organism or system. In case of food systems, the term "food metabolomics" is often used. Food metabolomics has been widely explored and applied in various fields including food analysis, food intake, food traceability, and food safety. Food safety applications focusing on the identification of pathogen-specific biomarkers have been promising. This chapter describes a nontargeted metabolite profiling workflow using gas chromatography coupled with mass spectrometry (GC-MS) for characterizing three globally important foodborne pathogens, Escherichia coli O157:H7, Listeria monocytogenes, and Salmonella enterica, from selective enrichment liquid culture media. The workflow involves a detailed description of food spiking experiments followed by procedures for the extraction of polar metabolites from media, the analysis of the extracts using GC-MS, and finally chemometric data analysis using univariate and multivariate statistical tools to identify potential pathogen-specific biomarkers., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Foodborne pathogens remain a serious health issue in developed and developing countries. Safeness of food products has been assured for years with culture-based microbiological methods; however, these present several limitations such as turnaround time and extensive hands-on work, which have been typically address taking advantage of DNA-based methods such as real-time PCR (qPCR). These, and other similar techniques, are targeted assays, meaning that they are directed for the specific detection of one specific microbe. Even though reliable, this approach suffers from an important limitation that unless specific assays are design for every single pathogen potentially present, foods may be considered erroneously safe. To address this problem, next-generation sequencing (NGS) can be used as this is a nontargeted method; thus it has the capacity to detect every potential threat present. In this chapter, a protocol for the simultaneous detection and preliminary serotyping of Salmonella enterica serovar Enteritidis, Salmonella enterica serovar Typhimurium, Listeria monocytogenes, and Escherichia coli O157:H7 is described., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Foodborne pathogens continue to be a major health concern worldwide. Culture-dependent methodologies are still considered the gold standard to perform pathogen detection and quantification. These methods present several drawbacks, such as being time-consuming and labor intensive. The implementation of real-time PCR has allowed to overcome these limitations, and even reduce the cost associated with the analyses, due to the possibility of simultaneously and accurately detecting several pathogens in one single assay, with results comparable to those obtained by classical approaches. In this chapter, a protocol for the simultaneous detection of two of the most important foodborne pathogens, Salmonella spp. and Listeria monocytogenes, is described., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

A novel sensitive and reusable electrochemical biosensor for Listeria monocytegenes DNA has been constructed based on the recognition of water-soluble hydroxylated fullerene (fullerol) to single- and double-stranded DNA. First, the fullerol was electrodeposited on glassy carbon electrode (GCE), acting as a matrix for non-covalent adsorption of single-stranded probe DNA. Upon hybridization with the target DNA, the double helix structure was formed and desorbed from the electrode surface, driving synchronous regeneration of the biosensing interfaces. The biosensor showed a probe DNA loading density of 144 pmol∙cm -2 with the hybridization efficiency of 72.2%. The biosensor is applicable for the analysis of target DNA in actual milk samples with recoveries between 101.0% and 104.0%. This sensing platform provides a simple method for the construction of sensitive and reusable biosensor to monitor Listeria monocytogenes-related food pollution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

In recent years, nanozymes have been widely used in the field of biosensing and food safety testing due to their advantages of low cost, high stability, easy modification and adjustable catalytic activity. However, how to reduce the signal interference generated by reducing substances, macromolecules and colored substances in the food matrix in nanozymes-based colorimetric sensing is still a major challenge. In this paper, using Listeria monocytogenes as a model analyte, sodium sulfonyl methacrylate (SBMA) polymers were modified onto cotton swabs by photothermal polymerization and combined with Listeria monocytogenes-specific aptamer (Apt1) to prepare swabs that can specifically capture and isolate Listeria monocytogenes from complex matrices (SBMA/Apt1 cotton swab). In addition, in combination with the inhibitory effect of the aptamer (Apt2) on the oxidase activity of Mn 3 O 4 NPs, a colorimetric biosensor based on nanozymes that can quantitatively, sensitively, and specifically identify Listeria monocytogenes in food products was constructed. The results showed that the colorimetric signal of the method was linear with the concentration of Listeria monocytogenes in the range of 2.83-2.83 × 10 5  CFU/mL, and the limit of detection was 2.64 CFU/mL, which can be used for the detection of Listeria monocytogenes in complex environments and food samples., Competing Interests: Declaration of competing interest The authors declare that they have no known conflict of interest or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Semiconductor metal oxide gas sensors have been proven to be capable of detecting Listeria monocytogenes , one kind of foodborne bacteria, through monitoring the characteristic gaseous metabolic product 3-hydroxy-2-butanone. However, the detection still faces challenges because the sensors need to work at high temperatures and output limited gas sensing performance. The present study focuses on the design of single-atom Au-functionalized mesoporous SnO 2 nanospheres for the sensitive detection of ppb-level 3-hydroxy-2-butanone at low temperatures (50 °C). The fabricated sensors exhibit high sensitivity (291.5 ppm -1 ), excellent selectivity, short response time (10 s), and ultralow detection limit (10 ppb). The gas sensors exhibit exceptional efficacy in distinguishing L. monocytogenes from other bacterial strains (e.g., Escherichia coli ). Additionally, wireless detection of 3-hydroxy-2-butanone vapor is successfully achieved through microelectromechanical systems sensors, enabling real-time monitoring of the biomarker 3-hydroxy-2-butanone. The superior sensing performance is ascribed to the mesoporous framework with accessible active Au-O-Sn sites in the uniform sensing layer consisting of single-atom Au-modified mesoporous SnO 2 nanospheres, and such a feature facilitates the gas diffusion, adsorption, and catalytic conversion of 3-hydroxy-2-butanone molecules in the sensing layer, resulting in excellent sensing signal output at relatively low temperature that is favorable for developing low-energy-consumption gas sensors.

Background: Listeria monocytogenes brain abscess is a rare phenomenon that is common in immunocompromised patients. Streptococcus equinus brain abscess has never been reported in the literature to our knowledge. In this case report, we describe a case of brain abscess secondary to Listeria monocytogenes and Streptococcus equinus in an immunocompetent patient with transient low CD4 count., Case Presentation: A 27-year-old white, male patient, previously healthy, nonalcoholic, and occasional smoker, presented to the emergency department for confusion and headache. The patient was found to have a left parietal abscess, which was drained and the fluid was sent for culture. Culture grew Listeria monocytogenes and Streptococcus equinus. The patient was treated with intravenous ampicillin followed by oral amoxicillin for a total of 6 weeks. The CD4 count was low initially. However, after the resolution of the infection, the CD4 count came back within normal range. Another brain magnetic resonance imaging was done that showed a significantly decreased hyperintensity within the left parietal subcortical white matter at the site of surgery with significantly decreased enhancement and almost total resolution of the previous abscess., Conclusion: Transient low CD4 count is a rare phenomenon that exposes patients to unusual and atypical infections. Since low CD4 count is transient, patients treated promptly recover from their illness. Our patient developed a Listeria monocytogenes and Streptococcus equinus brain abscess, which is considered rare and has not been previously described in the literature to our knowledge., (© 2024. The Author(s).)

Pre-cut fresh fruits and vegetables are highly appealing to consumers for their convenience, however, as they are highly susceptible to microbial contamination in processing, the potential risks of foodborne illnesses to public health are not negligible. This study aimed to assess the prevalence, antibiotic susceptibility and molecular characteristics of major foodborne pathogens (Listeria monocytogenes, Escherichia coli, Staphylococcus aureus and Salmonella) isolated from fresh-cut fruits and vegetables in Beijing, China. 86 stains were isolated from 326 samples, with S. aureus being the highest prevalence (15.38 %), followed by E. coli (9.23 %) and L. monocytogenes (1.85 %), while no Salmonella was detected. The prevalence by type of food indicated that fruit trays and mixed vegetables were more susceptible to contamination by pathogens. 98 % of S. aureus were resistant to at least of one antibiotic, and showed a high resistance rate to benzylpenicillin (90 %) and oxacillin (48 %). Among 25 E. coli isolates, 57.67 % of which exhibited multi-drug resistance, with common resist to trimethoprim/sulfamethoxazole (66.67 %) and ampicillin (63.33 %). A total of 9 sequence types (STs) and 8 spa types were identified in 35 S. aureus isolates, with ST398-t34 being the predominant type (42.86 %). Additionally, analysis of 25 E. coli isolates demonstrated significant heterogeneity, characterized by 22 serotypes and 18 STs. Genomic analysis revealed that 5 and 44 distinct antibiotic resistance genes (ARGs) in S. aureus and E. coli, respectively. Seven quinolone resistance-determining regions (QRDRs) mutations were identified in E. coli isolates, of which GyrA (S83L) was the most frequently detected. All the S. aureus and E. coli isolates harbored virulence genes. ARGs in S. aureus and E. coli showed a significant positive correlation with plasmids. Furthermore, one L. monocytogenes isolate, which was ST101 and serogroupIIc from watermelon sample, harbored virulence genes (inlA and inlB) and LIPI-1 pathogenic islands (prfA, plcA, hly and actA), which posed potential risks for consumer's health. This study focused on the potential microbial risk of fresh-cut fruits and vegetables associated with foodborne diseases, improving the scientific understanding towards risk assessment related to ready-to-eat foods., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

A novel colorimetric aptasensor assay based on the excellent magnetic responsiveness and oxidase-like activity of Fe 3 O 4 @MIL-100(Fe) was developed. Fe 3 O 4 @MIL-100(Fe) absorbed with aptamer and blocked by BSA served as capture probe for selective isolation and enrichment of Listeria monocytogenes one of the most common and dangerous foodborne pathogenic bacteria. The aptamer absorbed on Fe 3 O 4 @MIL-100(Fe) was further used as signal probe that specifically binds with target bacteria conjugation of capture probe for colorimetric detection of Listeria monocytogenes, taking advantages of its oxidase-like activity. The linear range of the detection of Listeria monocytogenes was from 10 2 to 10 7  CFU mL -1 , with the limit of detection as low as 14 CFU mL -1 . The approach also showed good feasibility for detection of Listeria monocytogenes in milk and meat samples. The spiked recoveries were in the range 81-114% with relative standard deviations ranging from 1.28 to 5.19%. Thus, this work provides an efficient, convenient, and practical tool for selective isolation and colorimetric detection of Listeria monocytogenes in food., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Rationale: Listeria monocytogenes (LM) is an important foodborne bacterium, and LM meningoencephalitis is rare in clinical practice, with poor prognosis in severe patients. It is prone to misdiagnosis in clinical practice. We first reported a case of severe LM meningoencephalitis with muscle lesions and evaluated the comprehensive condition., Patient Concerns: A 48-year-old man had a fever and was admitted to the neurology department due to dizziness, nausea, and vomiting for 20 days., Diagnoses: LM meningoencephalitis complicated with muscle lesions., Interventions: We used moxifloxacin 0.4 g, qd, meropenem 2 g, q8h, and dexamethasone 10 mg, qd to reduce exudation and adhesion. Then due to consideration of side effects, we increased the dose of ampicillin by 2 g, q4h, stopped using meropenem and moxifloxacin, and turned to maintenance treatment with dexamethasone and ampicillin. We comprehensively managed his vital signs and physical organ functions, we also controlled some comorbidities. During the hospitalization period thereafter, we used intravenous anti-infection treatment with moxifloxacin 0.4 g, qd, ampicillin 0.5 g, q4h., Outcomes: Half a year later, the reexamination showed only protein elevation in cerebrospinal fluid and hydrocephalus in MRI. Afterward, the symptoms did not recur again. The patient recovered well after discharge., Lessons: LM meningoencephalitis complicated with lower limb muscle lesions is clinically rare. This report focuses on relevant treatment plans, which provide value for the examination and comprehensive management of patients with LM infection in the future., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2024 the Author(s). Published by Wolters Kluwer Health, Inc.)

Background: Listeria monocytogenes is a bacterial pathogen known for causing listeriosis, a foodborne illness with a wide spectrum of clinical presentations ranging from mild gastroenteritis to severe invasive disease, particularly affecting immunocompromised individuals, pregnant women, newborns, and the elderly. Successful treatment of patients with recurring listeria episodes due to colonised foreign material is often challenging, typically requiring a combination of antimicrobial treatment and surgical removal., Case Presentation: Here, we present a particularly complex case of chronic invasive listeriosis with a total of six relapses. After extensive investigations, the patient's ICD device was identified as the focus of infection., Conclusion: The confirmation of relapses through cgMLST analysis highlights the persistence of Listeria monocytogenes and the potential for recurrence even after apparent resolution of symptoms in patients with foreign material. It emphasises the necessity for a comprehensive assessment to identify and mitigate the risk of relapses, thereby ensuring optimal management and outcomes., (© 2024. The Author(s).)

Objectives: Listeria monocytogenes (LM) is a health threat worldwide given its high mortality and the growing of high-risk susceptible populations., Methods: All hospitalizations with a diagnosis of LM in the National Registry of Hospital Discharges were examined in Spain from 2000 to 2021., Results: A total of 8152 hospital admissions with LM were identified. The mean age was 59.5 years and 48% were immunosuppressed (IS). The rate of LM hospitalizations increased from 5 per 1 million population in 2000 to 8.9 in 2021 (p < 0.001). A foodborne outbreak in Andalusia determined a sharp increase in admissions with LM during 2019. The COVID-19 pandemic and lockdowns were associated with a decrease in LM admissions. The overall in-hospital mortality was 16.7%. The number of deaths in patients hospitalized with LM rose from 7.8 per 100,000 deceased in 2000 to 18 in 2021 (p < 0.001). After adjustment, age >65 years (odds ratio [OR] = 2.16), sepsis (OR = 2.60), meningoencephalitis (OR = 1.72), endocarditis (OR = 2.0), neonatal listeriosis (OR = 2.10) and IS (OR = 2.09) were associated with mortality., Conclusions: The number of patients hospitalized with LM in Spain has increased significantly from 2000 to 2021. The increase in the rate of admissions and deaths was largely driven by the growing proportion of elderly and IS patients., Competing Interests: Declaration of competing interests None declared., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

A microbiological study was conducted on 41 insect product samples (29 raw frozen [21 domestic and 8 imported], 10 powdered, and 2 processed), which were commercially available in Japan. The total aerobic count for raw frozen insects was 5.61 log cfu/g (range: 2.52-8.40), whereas the powdered insect count was 2.89 log cfu/g (range: 1.00-4.57). The bacterial count was significantly higher in raw frozen insects ( p < 0.05). The coliform count for the raw frozen insects ranged from <1 to 6.90 log cfu/g, and that for the powdered insects ranged from <1 to 1.00 log cfu/g. The number of samples with values above the detection limit was significantly higher in raw frozen insects ( p < 0.05). The detection frequencies of aerobic spores (<1-4.63 log cfu/g), anaerobic spores (<0-4.40 log cfu/g), and Bacillus cereus (<1.7-3.83 log cfu/g) showed no sample type-related significant difference. Listeria spp. was isolated from four samples of raw frozen insects, one of which was Listeria monocytogenes . We did not detect any of the following: Salmonella spp., Shiga toxin-producing E. coli (STEC), Campylobacter jejuni/coli , or pathogenic Yersinia . We isolated insect products retailed in Japan harboring food poisoning bacteria, including L. monocytogenes and B. cereus . In particular, raw frozen products displayed high levels of hygienic indicator bacteria.

Controlling foodborne pathogens in buffalo milk is crucial for ensuring food safety. This study estimated the prevalence of nine target genes representing seven critical foodborne bacteria in milk and milk products, and identified factors associated with their presence in buffalo milk chain nodes in Bangladesh. One hundred and forty-three milk samples from bulk tank milk (n = 34), middlemen (n = 37), milk collection centers (n = 37), and milk product shops (n = 35) were collected and analyzed using RT-PCR. Escherichia (E.) coli, represented through yccT genes, was the most prevalent throughout the milk chain (81-97%). Chi-squared tests were performed to identify the potential risk factors associated with the presence of foodborne bacteria encoded for different genes. At the middleman level, the prevalence of E. coli was associated with the Mymensingh, Noakhali, and Bhola districts (P = 0.01). The prevalence of Listeria monocytogenes, represented through inlA genes, and Yersinia (Y.) enterocolitica, represented through yst genes, were the highest at the farm level (65-79%). The prevalence of both bacteria in bulk milk was associated with the Noakhali and Bhola districts (P < 0.05). The prevalence of Y. enterocolitica in bulk milk was also associated with late autumn and spring (P = 0.01) and was higher in buffalo-cow mixed milk than in pure buffalo milk at the milk collection center level (P < 0.01). The gene stx2 encoding for Shiga toxin-producing (STEC) E. coli was detected in 74% of the milk products. At the middleman level, the prevalence of STEC E. coli was associated with the use of cloths or tissues when drying milk containers (P = 0.01). Salmonella enterica, represented through the presence of invA gene, was most commonly detected (14%) at the milk collection center. The use of plastic milk containers was associated with a higher prevalence of Staphylococcus aureus, represented through htrA genes, at milk product shops (P < 0.05). These results suggest that raw milk consumers in Bangladesh are at risk if they purchase and consume unpasteurized milk., (© 2024. The Author(s).)

Objectives: Much has been written about the utility of genomic databases to public health. Within food safety these databases contain data from two types of isolates-those from patients (i.e., clinical) and those from non-clinical sources (e.g., a food manufacturing environment). A genetic match between isolates from these sources represents a signal of interest. We investigate the match rate within three large genomic databases (Listeria monocytogenes, Escherichia coli, and Salmonella) and the smaller Cronobacter database; the databases are part of the Pathogen Detection project at NCBI (National Center for Biotechnology Information)., Results: Currently, the match rate of clinical isolates to non-clinical isolates is 33% for L. monocytogenes, 46% for Salmonella, and 7% for E. coli. These match rates are associated with several database features including the diversity of the organism, the database size, and the proportion of non-clinical BioSamples. Modeling match rate via logistic regression showed relatively good performance. Our prediction model illustrates the importance of populating databases with non-clinical isolates to better identify a match for clinical samples. Such information should help public health officials prioritize surveillance strategies and show the critical need to populate fledgling databases (e.g., Cronobacter sakazakii)., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Background: Listeriosis is a global health threat to both animals and humans, especially in developing countries. This study was designed to isolate Listeria monocytogenes from faeces; environmental samples; and cow, sheep and goat milk, as well as human stool, to study its molecular characteristics and antibiotic sensitivity in the New Valley and Beheira Governorates, Egypt. The isolation and identification of L. monocytogenes were carried out using traditional culture and biochemical methods, followed by antibiography, genus confirmation of some isolates and detection and sequencing of InlB genes via PCR., Results: Out of 2097 examined samples, the prevalence of L. monocytogenes was 13.4% in animals; the prevalence was 9.2%, 2.4%, 25.4%, 4%, 42.4%, and 6.4% in cattle faeces, cattle milk, sheep faeces, sheep milk, goat faeces, and goat milk, respectively. However, the prevalence of L. monocytogenes was 8.3% in human samples. Both animal and human isolates showed 100% resistance to trimethoprim-sulfamethoxazole, and the isolates showed the highest sensitivity to flumequine (100%), amikacin (99.2%), gentamicin (97.6%), and levofloxacin (94.6%). Multidrug resistance (MDR) was detected in 86.9% of the tested isolates. The 16 S rRNA and inlB genes were detected in 100% of the randomly selected L. monocytogenes isolates. Phylogenetic analysis of three isolates based on the inlB gene showed 100% identity between faecal, milk and human stool isolates., Conclusions: Faeces and milk are major sources of listeriosis, and the high degree of genetic similarity between animal and human isolates suggests the possibility of zoonotic circulation. The high prevalence of MDR L. monocytogenes in both animal and human samples could negatively impact the success of prevention and treatments for animal and human diseases, thereby imposing serious risks to public health., (© 2024. The Author(s).)

Bacterial genotyping through whole-genome sequencing plays a crucial role in disease surveillance and outbreak investigations in public health laboratories. This study assessed the effectiveness of Oxford Nanopore Technologies (ONT) sequencing in the genotyping of Listeria monocytogenes and Salmonella enterica serovar Enteritidis. Our results indicated that ONT sequences, generated with the R10.4.1 flow cell and basecalled using the Dorado 0.5.0 Super Accurate 4.3 model, exhibited comparable accuracy to Illumina sequences, effectively discriminating among bacterial strains from outbreaks. These findings suggest that ONT sequencing has the potential to be a promising tool for rapid whole-genome sequencing of bacterial pathogens in public health laboratories for epidemiological investigations., Importance: This study unveils that Oxford Nanopore Technologies sequencing, by itself, holds the potential to serve as a whole-genome sequencing-based genotyping tool in public health laboratories, enabling routine subtyping of bacterial isolates for disease surveillance and outbreak investigations., Competing Interests: The authors declare no conflict of interest.

Human listeriosis is an infectious disease caused by Listeria monocytogenes. The invasive form of this disease leads to a high rate of hospitalizations and fatality. The main mode of transmission is through contaminated ready-to-eat foods such as dairy, vegetables and meat products. The knowledge of the diversity and population dynamics of isolates collected from human and food sources is essential for the detection of clusters and the identification of common sites of infection. The aim of this study was the molecular characterization of L. monocytogenes isolates in Argentina. We sequenced a total of 63 isolates, 35 from human and 28 from food sources, collected between 2018 and 2023. Our genomic study divided the isolates into two lineages, four serogroups, 17 sequence types and 15 clonal complexes (CCs). The hypervirulent clone CC1 (lineage I; serogroup IVb) predominated in human and food samples. The phylogenomic analysis showed a high and possible epidemiological relationship between isolates from human and/or food sources, suggesting the presence of transmission chains in our country. These findings highlight the need to strengthen genomic surveillance of L. monocytogenes in Argentina. The identification of geographic distribution and characteristics of predominant and emerging clones from human and food sources might help to focus action plans and public health policies better directed at the control and prevention of listeriosis., (Copyright © 2024 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.)

In this study, a molecular beacon (MB) was designed for colorimetric loop-mediated isothermal amplification (cLAMP). The length of complementary bases on the MB, guanine and cytosine content (GC content), and hybridization sites of complementary bases were investigated as key factors affecting the design of the MB. We designed MBs consisting of 10, 15, and 20 complementary bases located at both ends of the HRPzyme. In the case of the long dumbbell DNA structure amplified from the hlyA gene of Listeria monocytogenes, possessing a flat region (F1c-B1) of 61 base pairs (bp), an MB was designed to intercalate into the flat region between the F1c and B1 regions of the LAMP amplicons. In the case of the short dumbbell DNA structure amplified from the bcfD gene of Salmonella species possessing a flat region (F1c-B1) length of 6 bp, another MB was designed to intercalate into the LoopF or LoopB regions of the LAMP amplicons. The results revealed that the hybridization site of the MB on the LAMP amplicons was not crucial in designing the MB, but the GC content was an important factor. The highest hybridization efficiencies for LAMP amplicons were obtained from hlyA gene-specific and bcfD gene-specific MBs containing 20- and 15-base complementary sequences, respectively, which exhibited the highest GC content. Therefore, designing MBs with a high GC content is an effective solution to overcome the low hybridization efficiency of cLAMP assays. The results obtained can be used as primary data for designing MBs to improve cLAMP accessibility., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Pretransplant mortality rates in the US remain high and are connected to effective organ donation and utilization. Thus, there is a need to maximize the utilization of available donors. In some cases, this has been safely achieved using organs from donors with infectious complications. For example, several studies describe the use of organs from donors with bacterial meningitis due to pathogens such as Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenza, and Escherichia coli, with good outcomes. Listeria is an aerobic and facultatively anaerobic, nonspore-forming, Gram-positive rod that can affect the central nervous system, causing meningitis and meningoencephalitis. Due to its virulence, ability to cause intracellular infection, and lack of clinical data, people dying with listeria may not be evaluated for organ donation, may not have organs recovered, or may have their organs recovered but not transplanted. Herein, we describe the outcomes of 7 solid organ transplant recipients who received organs from 2 donors with Listeria monocytogenes central nervous system infection., (Copyright © 2024 American Society of Transplantation & American Society of Transplant Surgeons. Published by Elsevier Inc. All rights reserved.)

Listeriosis is a globally rare foodborne disease that causes fetal-placental infection, leading to adverse pregnancy outcome, while limited research among pregnant women is available in China. This study was therefore aimed at analyzing the incidence, clinical manifestations, and clinical outcome of listeriosis among pregnant women and its associated dietary behavior risk factors in prevention. A hospital-based case-control study had been conducted from January 2017 to December 2021. Clinical data, laboratory information, and questionnaires including dietary behaviors and personal hygiene were collected within 2 days after case diagnosis. There were 48 pregnant women, including 12 cases and 36 controls, with an average age of 31.19 ± 3.75 years. The incidence of admission-based listeriosis among pregnant women was 1.6058 per 10,000. The 12 strains were divided into 3 serotypes: 1/2a(83.33%), 1/2b(8.33%), and 4b(8.33%). Among the cases, 5 cases (41.67%) resulted in abortion, 3 cases (25%) induced preterm labor, and 4 cases (33.33%) had full-term deliveries after treatment. There were 7 live births in the case group, among which 6 were admitted to the neonatal intensive care unit (NICU), while 1 case had a healthy fetal outcome. All patients in the control group gave birth to live fetuses. Epidemiological investigation revealed that pregnant women dining at restaurants three or more times per week might increase the risk of having Listeria infection. There were no significant differences in dietary consumed behaviors, hand hygiene, and refrigerator usage behaviors between case and control groups. The study suggested that dining at restaurants might be associated with Listeria infection among pregnant women. Therefore, it is essential to enhance education on listeriosis serious consequences and promote healthy dietary and hygiene habits among pregnant women.

Competing Interests: I declare no competing interests.