Infection with Listeria monocytogenes meningoencephalitis:A case report

BACKGROUND Listeria meningitis is an infectious disease of the central nervous system caused by Listeria monocytogenes.This bacterium is widely present in the natural environment and can be transmitted through channels such as food and water.Patients usually show symptoms such as fever,headache,and neck stiffness.In severe cases,coma,convulsions,or even death may occur.Traditional diagnostic methods,such as cerebrospinal fluid(CSF)culture and serological tests,have certain limitations.Although CSF culture is the“gold standard”for diagnosis,it is time-consuming and has a relatively low positivity rate.Serological detection may also result in false positive or false negative results.The emergence of metagenomic sequencing(mNGS)technology has led to a significant break-through in diagnosing Listeria meningitis,allowing quick and accurate detection of various pathogens in samples.CASE SUMMARY Here,we present the case of a previously healthy 64-year-old woman diagnosed with Listeria meningitis using mNGS.She was successfully treated with intravenous ampicillin and meropenem,without any complications.CONCLUSION Listeria meningitis must be considered,especially in patients who fail to show improvement with first-line antibiotic treatments.mNGS significantly reduces the diagnosis time,supporting timely treatment of patients.

Meningoencephalitis due to Listeria monocytogenes in a Young Immunocompetent Patient: Case Report with Literature Review

Meningoencephalitis secondary to Listeria monocytogenes (L. monocytogenes) mainly affects newborns, the elderly and immunocompromised people;there are extremely rare cases in which said infection occurs in immunocompetent individuals. We present the case of a young adult patient without immunocompromise, who developed meningoencephalitis due to L. monocytogenes;This case is exceptional, since it occurred in an individual outside the classic age group, in addition to not having risk factors, which is why it should be considered an atypical causal agent.

Transcriptomics reveals substance biosynthesis and transport on membranes of Listeria monocytogenes affected by antimicrobial lipopeptide brevilaterin B

Listeria monocytogenes is a worrisome food-borne pathogen threatening global food safety.Our previous study proved that lipopeptide brevilaterin B showed efficient antibacterial activity against L.monocytogenes by interacting with the cell membrane.This research further explored the antibacterial mechanism of brevilaterin B against L.monocytogenes at the sub-minimum inhibition concentration via transcriptomic analysis.Brevilaterin B induced growth inhibition rather than direct membrane lysis in L.monocytogenes at the minimum inhibitory concentration.Transcriptomic analysis showed 1779 difference expressed genes,including 895 up-regulated and 884 down-regulated genes.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis indicated that brevilaterin B influenced multiple pathways of L.monocytogenes,including peptidoglycan biosynthesis,membrane transport(ATP-binding cassette transports,ion transport),cellular metabolism(amino acid and lipid metabolism),ATP synthesis,and activation of the stress response(quorum sensing and bacterial chemotaxis).In conclusion,brevilaterin B affects gene expression related to biosynthesis,transport and stress response pathways on the membrane of L.monocytogenes.The present work provides the first transcriptomic assessment of the antibacterial mechanism of lipopeptide brevilaterin B at the gene level.

PCR-based methodologies for detection and characterization of Listeria monocytogenes and Listeria ivanovii in foods and environmental sources

Listeria monocytogenes is an important foodborne pathogen responsible for listeriosis,a fatal disease.It is widely distributed in various foods and environmental sources.In this review,we focused on addressing PCR-based technologies,including conventional PCR,qPCR and droplet digital PCR(ddPCR).Specifically,we described(a)conventional PCR and mono-,duplex-and multiplex-qPCR methodologies;(b)development and applications of gene HlyA-,Iap-,PrfA–and SsrA-based conventional and qPCR assays as well as PCR assays targeting newly identified gene targets for specific detection of L.monocytogenes;differentiation of viable from dead L.monocytogenes by qPCR in conjugation with propidium monoazide pretreatment;PCR-based serotype identification of L.monocytogenes isolates;PCR-based detection of L.ivanovii,infecting ruminants,differentiation of L.monocytogenes from other Listeria species;and sigB-gene based PCR identification of Listeria spp;(c)applications of ddPCR in detection of L.monocytogenes;and(d)application of qPCR assays in detection and subtyping of L.monocytogenes in milk and dairy products;meats,meat products and meat-processing environment;and seafood,seafood products and processing environment.Our goal was to provide a relatively comprehensive overview of PCR-based methodologies available in detection,characterization and subtyping of various strains of L.monocytogenes in foods and environmental sources.

Prevalence and methodologies for detection,characterization and subtyping of Listeria monocytogenes and L.ivanovii in foods and environmental sources

Listeria monocytogenes,one of the most important foodborne pathogens,can cause listeriosis,a lethal disease for humans.L.ivanovii,which is closely related to L.monocytogenes,is also widely distributed in nature and infects mainly warm-blooded ruminants,causing economic loss.Thus,there are high priority needs for methodologies for rapid,specific,cost-effective and accurate detection,characterization and subtyping of L.monocytogenes and L.ivanovii in foods and environmental sources.In this review,we(A)described L.monocytogenes and L.ivanovii,world-wide incidence of listeriosis,and prevalence of various L.monocytogenes strains in food and environmental sources;(B)comprehensively reviewed different types of traditional and newly developed methodologies,including culture-based,antigen/antibody-based,LOOP-mediated isothermal amplification,matrix-assisted laser desorption ionization-time of flight-mass spectrometry,DNA microarray,and genomic sequencing for detection and characterization of L.monocytogenes in foods and environmental sources;(C)comprehensively summarized different subtyping methodologies,including pulsed-field gel electrophoresis,multi-locus sequence typing,ribotyping,and phage-typing,and whole genomic sequencing etc.for subtyping of L.monocytogenes strains from food and environmental sources;and(D)described the applications of these methodologies in detection and subtyping of L.monocytogenes in foods and food processing facilities.

In vitro Selection of DNA Aptamers and Fluorescence-Based Recognition for Rapid Detection Listeria monocytogenes

Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides. Systematic evolution of ligands by exponential enrichment （SELEX） was used to select and PCR amplify DNA sequences （aptamers） capable of binding to and detecting Listeria monocytogenes, one of the major food-borne pathogens. A simplified affinity separation approach was employed, in which L. monocytogenes in exponential （log） phase of growth was used as the separation target. A fluorescently-labeled aptamer assay scheme was devised for detecting L. monoeytogenes. This report described a novel approach to the detection of L. monocytogenes using DNA aptamers. Aptamers were developed by nine rounds of SELEX. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. One of aptamers named e01 with the highest affinity was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. This study has proved the principle that the whole-cell SELEX could be a promising technique to design aptamer-based molecular probes for dectection of pathogenic microorganisms without tedious isolation and purification of complex markers or targets.

Antimicrobial Resistance,Virulence Profile,and Molecular Characterization of Listeria monocytogenes Isolated from Ready-to-eat Food in China,2013-2014

We aimed to investigate the potential pathogenic profile and antibiotic resistance of Listeria monocytogenes isolated from ready-to-eat food in China.Antimicrobial resistance was determined by broth microdilution following the Clinical and Laboratory Standards Institute protocol Molecular serotyping,virulence,and resistance genes were identified using PCR.Multi-locus

Detection of Listeria monocytogenes in Dairy Food by Loop-mediated Isothermal Amplification(LAMP)

[Objective] This study aimed to detect the reliability of LAMP method for detecting Listeria monocytogenes in dairy food. [Method] Based on the sequence of hlyA gene encoding listeriolysin O in Listeria monocytogenes, the LAMP method was established for detecting Listeria monocytogenes in dairy food. [Result] The es- tablished LAMP rapid detection method has .high specificity and sensitivity, which are equivalent to those of real-time fluorescent quantitative PCR. The detection re- sults of Listeria monocytogenes in dairy food by established LAMP method were completely consistent with those by bacterial isolation method. [Conclusion] The de- tection results of Listeria monocytogenes by LAMP method can be directly identified by naked eye, so the established LAMP method was suitable for the detection of Listeria monocytogenes in dairy food in emergency situations.

Combination Use of PFGE and Drug-resistant Analysis in the Epidemiological Investigation of Listeria Monocytogenes

Listeria monocytogenes is the pathogen of listeriosis and it causes severe infections like septicemia, encephalitis, and meningitis, especially in immunocompromised individuals, newborns, and pregnant women. Its wide distribution in the environment and ability to survive or even grow under adverse conditions has made L. monocytogenes an important public health concern and in food industry.

Brainstem abscesses caused by Listeria monocytogenes:A case report

BACKGROUNDIntracranial Listeria infections are common in newborns and immunocompromisedindividuals, but brainstem abscesses are rare.CASE SUMMARYWe report a rare case of brainstem abscesses caused by Listeria monocytogenes in apreviously healthy adult patient. The patient’s magnetic resonance imagingexamination showed multiple brain abscesses, and his second cerebrospinal fluidculture test indicated the presence of Listeria monocytogenes. Despite earlyempirical therapy, the patient’s condition progressively deteriorated. Because thepatient's abscesses were located in the brainstem and multiple lobes, surgery wasnot possible. The patient died 40 d after admission.CONCLUSIONThis case highlights the importance of rational clinical use of drugs to avoidpotentially serious infectious complications.

Inhibitory Effect of Live-Attenuated Listeria Monocytogenes-based Vaccines Expressing MIA Gene on Malignant Melanoma

Listeria monocytogenes(LM),a Gram-positive facultative intracellular bacterium,can be used as an effective exogenous antigen expression vector in tumor-target therapy.But for successful clinical application,it is necessary to construct attenuated LM stain that is safe yet retains the potency of LM based on the full virulent pathogen.In this study,attenuated LM and recombinants of LM expressing melanoma inhibitory activity(MIA) were constructed successfully.The median lethal dose(LD 50) and invasion efficiency of attenuated LM strains were detected.The recombinants were utilized for immunotherapy of animal model of B16F10 melanoma.The level of MIA mRNA expression in tumor tissue was detected by using real-time polymerase chain reaction(PCR) with specific sequence,meanwhile the anti-tumor immune response was assayed by flow cytometric analysis and enzyme-linked immunosorbent spot(ELISPOT) assay.The results showed the toxicity and invasiveness of attenuated LM were decreased as compared with LM,and attenuated LM expressing MIA,especially the double-genes attenuated LM recombinant,could significantly induce anti-tumor immune response and inhibit tumor growth.This study implicates attenuated LM may be a safer and more effective vector for immunotherapy of melanoma.

Application Progress of Recombinant Attenuated Listeria monocytogenes in Tumor Immunotherapy

Much progress of application of bacterial vaccine in treatment and prevention of tumor was acquired,which showed broad prospect in clinical study of animals and humans. Listeria monocytogenes( L. monocytogenes) was considered much important by virtue of its special characteristic of biology and immunology.L. monocytogenes was ingested by professional or part-time phagocytes,survived and proliferated in the phagocytes under synergism of toxic factor secreted by itself,meanwhile,the cellular and humoral immune response was induced. Antigenic gene of specific tumor was loaded in the attenuated L. monocytogenes,which can enhance immune response of host cells. Effective cell targeted to enter tumor tissue and acted on tumor cells to induce apoptosis of tumor cells. Tumor degenerated not easy to reappear. Therefore,recombinant attenuated L. monocytogenes was a safe and effective anti-cancer vaccine vector. Now the work of researchers mainly focuses on solving practical problem in clinical application. Biological characteristics of L. monocytogenes,feasibility and superiority of L. monocytogenes as targeted vaccine vector,problem and prospect of L. monocytogenes in clinical application of anti-tumor were reviewed in this paper.

Bacteria on Meat Abattoir Process Surfaces after Sanitation:Characterisation of Survival Properties of Listeria monocytogenes and the Commensal Bacterial Flora

Contamination of food with spoilage bacteria and pathogens from food processing environment remains a challenge for the food industry. Bacteria able to persist in such environments over time must survive several hygienic hurdles. The aim of this study was to identify bacteria surviving practical disinfection and compare their survival abilities with representative isolates of the pathogen Listeria monocytogenes. Bacteria isolated from processing surfaces after cleaning and disinfection in a meat abattoir were identified. Selected isolates of the most frequently isolated bacterial genera along with eight meat associated L. monocytogenes were further characterized with regard to biofilm formation abilities at 12℃ and 20℃, tolerance to desiccation (stainless steel at 70% RH at 12℃) and bactericidal effects of recommended in-use-concentrations of four commercial disinfectants on stainless steel surface. The most dominating bacterial genera based on counts on non-selective agar were Aerococcus, Acinetobacter, Pseudomonas, Serratia and Staphylococcus. Isolates of Citrobacter. Enterobacter and Serratia dominated on agar plates selective for Enterobacteriaceae. In general, Gram negative bacteria formed more biofilm than Gram positives, especially at 12℃ with the best biofilm formers being Acinetobacter, Citrobacter and Pseudomonas. Listeria monocytogenes were poor biofilm formers. Gram positives survived better air drying than Gram negatives. Strains of L. monocytogenes were more sensitive to desiccation than the other Gram positives;Aerococcus, Kocuria and Staphylococcus. Two disinfectants containing peracetic acid and a disinfectant containing alkylaminoacetate had limited or no antibacterial effect against bacteria dried on stainless steel. A quaternary ammonium compound-based disinfectant provided >2 log reductions of Aerococcus, Acinetobacter and Listeria. Only 0.5 log reductions were obtained against Staphylococcus and no bactericidal effect against Serratia. In this study the dominating flora in a meat abattoir was isolated and identified. Several of these bacteria were better biofilm formers and more resistant to desiccation and disinfection than L. monocytogenes. The disinfectants tested had limited bactericidal activity against surface associated bacteria.

Listeria monocytogenes bacteremia in a centenarian and pathogen traceability:A case report

BACKGROUND Early diagnosis and appropriate antibiotic treatment are important to survival of Listeria monocytogenes(L.monocytogenes)bacteremia.Penicillin tends to be the most commonly used antibiotic.However,there are limited data on antibiotic use in elderly patients with serious complications.We describe the clinical presentation,antibiotic therapy,and traceability of L.monocytogenes in a centenarian with a history of eating frozen food.CASE SUMMARY A 102-year-old man suffered from high fever with chill after hematochezia.Tentative diagnoses were lower gastrointestinal hemorrhage and localized peritonitis.Meropenem and ornidazole were the empirical therapy.The patient did not respond and developed multiple system dysfunction even after teicoplanin was added to the therapy.L.monocytogenes was identified from blood cultures on day 5 of admission.The patient had a history of consuming frozen dumplings.Meropenem/ornidazole/teicoplanin were replaced with meropenem/linezolid.The patient gradually became afebrile.He received meropenem/linezolid for 10 d,and piperacillin/tazobactam was applied as step-down treatment for 2 wk with good clinical results.There was no sign of relapse during follow-up after discharge.L.monocytogenes isolates from the patient and frozen dumplings belonged to different serotypes and sequence types(STs):1/2b and ST5 from the patient and 1/2c and ST9 from the dumplings.CONCLUSION More awareness of listeriosis should be raised.Linezolid might be an option for listeriosis in elderly people with serious complications.

Inhibition Effect of Herbal Preservatives on Listeria monocytogenes on Chilled Pork

This study investigated the growth situation of Listeria monocytogenes on chilled pork and the effect of herbal preservatives on this pathogen. The inhibitions of herbal preservatives were identified. The minimum inhibitory concentrations （MIC） of cinnamon and clove were all 0.79 mg.mL^-1, while the rosemary was 1.58 mg.mL^-1. And the composite herbal preservatives were got through orthogonal experiment. The optimum proportion was as following on agar medium： 1.16 mg.mL^-1 cinnamon＋2.38 mg. mL^-1 rosemary＋3.17 mg. mLl clove （herb combination number 5）, while on chilled pork, the strong inhibition of L. monocytogenes was showed, which demonstrated that the surface application of herb combination resulted in an effective delay of L. monocytogenes growth.

Altered Host Resistance to Listeria monocytogenes In Mice Exposed to 1-Chloroacetophenone (CN) Vapours

Short term repeated exposure of 1-chloroacetophenone (CN) vapours at a concentration of 0.153 mg per litre for 15 minutes daily on 10 consecuitve days in Swiss albino male mice resulted in increased mortality to Listeria monocytogenes. Significantly elevated bacterial growth was observed in the spleen and liver of the CN exposed animals. The increased bacterial count in these organs was evident within 4-6 days post challenge as compared to vehicle exposed infected and unexposed infected animals. Increased susceptibility to infection has been considered to be the function of immune alteration due to cumulative short term effects ofCN vapour inhalation. This may be attributed to immunotoxic effects of CN on Tcells mediated macrophage functions.

Extraintestinal infection of Listeria monocytogenes and susceptibility to spontaneous abortion during pregnancy:A systematic review and meta-analysis

BACKGROUND Listeria monocytogenes(L.monocytogenes)is one of the most important zoonotic bacteria that is transmitted to humans through infected animal products and is the cause of human listeriosis.Pregnant women and immunocompromised patients are more susceptible to the bacterium than healthy people.Recent studies have reported extensive evidence on the role of L.monocytogenes infection and the risk of spontaneous abortion.AIM To evaluate the possible connection with L.monocytogenes in the risk of spontaneous abortion in pregnancy.METHODS We conducted a systematic literature review using several databases to search the relevant case-control studies on the association between L.monocytogenes infection and spontaneous abortion.Finally,the impact of infection with L.monocytogenes and risk of spontaneous abortion was assessed via odds ratio at corresponding 95%confidence intervals.RESULTS In the present study,we evaluated the data of 4059 pregnant women who had a spontaneous abortion,and interestingly their colonization rate of L.monocytogenes was about 20.5%.CONCLUSION Therefore,based on statistical analysis,we found that there is a significant relationship between the infection with L.monocytogenes and spontaneous abortion.

Biofilm Determination of Listeria monocytogenes That Isolated from Different Sources

The study was conducted for the detection of Listeria monocytogenes bacteria from different sources （CSF and blood） obtained from patients in Pediatric hospital and from food sources like （yogurt, raw vegetables and raw milk, sausage）. Ten isolates were isolated from 150 specimens one of them from CSF and one isolate isolated from blood samples the others isolated from food specimens 6 isolates isolated from sausage and 2 from raw vegetables. Isolates were identified traditionally involved culture methods based on selective enrichment and plating followed by the characterization of Listeria spp. based on colony morphology, sugar fermentation and haemolytic properties, identification by Api listeria was done. Determine the isolates that produce biofilm by tissue culture plate method. The highest biofilm forming strains ofListeria monocytogenes isolates appear in No. （D10, El, E5 and E7） OD reading each of them is （0.13, 0.09, 0.11 and 0.19） respectively, the lowest or poor biofilm forming strains appear in No. （D11, D12, E2, E3, E4 and E6） that optical density （OD） reading are （0.04, 0.03, 0.05, 0.04, 0.05 and 0.03） respectively by comparing with control prepared in well （A12） that stained by crystal violate without putting any isolates and the OD reading is （0.003）. Confirmation by PCR was done only for four isolates that produce biofilm （D10 and El） that obtained from CSF and blood sample and for （E5 and E7） that obtained from food samples.

Phenotypic and Genotypic Characterisation of Antibiotic Resistance in Escherichia coli, Klebsiella spp., and Listeria monocytogenes Isolates from Raw Meat Sold in Nairobi

Worldwide, the increase in antimicrobial resistance (AMR) is a public health concern. Food-borne associated antibiotic-resistant pathogens can contaminate raw meat during slaughter, transportation, and at sale points. A cross-sectional study was conducted from March 2021 to December 2021 to determine antimicrobial susceptibility patterns and characterize the molecular basis of resistance in E. coli, Klebsiella spp., and L. monocytogenes contaminating raw meat collected from retail outlets in Nairobi. Isolation and identification of the strains were done using the standard culture methods and PCR. Antimicrobial susceptibilities of the recovered strains were determined using disk diffusion while the presence of antibiotic resistance gene determinants;blaTEM, blaCTX-M, blaOXA, sul, and qnrS was done using PCR. Of 270 samples collected, 163 (60%) Escherichia coli, 19 (7%) Klebsiella spp., and L. monocytogenes 3 (1.1%) were recovered. Among Escherichia coli, high antibiotic resistance was found to Erythromycin 161 (98%) and ampicillin 88 (54%) while low resistance was found against imipenem 2 (1%). Similarly, high resistance was found among Klebsiella spp. to Erythromycin 19 (100%) and ampicillin 12 (63%) low resistance to ceftazidime 1 (5%), cefotaxime 1 (5%), aztreonam 1 (5%), and chloramphenicol 1 (5%). One isolate among the three Listeria monocytogenes strains isolated was resistant to Trimethoprim-sulfamethoxazole. No resistance was exhibited to gentamycin by all Klebsiella spp. The prevalence of multidrug-resistant (resistance to three or more classes of antibiotics) isolates was 95/182 (52.2%). The common resistance pattern observed was Erythromycin, ampicillin, tetracycline, and trimethoprim-sulfamethoxazole with a prevalence of 19 (20%). ESBL was confirmed in isolates that harbored: blaTEM (65%), blaCTX-M (44%), blaOXA (33%) while sul and qnrS were detected in 46.7% and 13.6% respectively. Circulation of antibiotic-resistant and MDR isolates found in this study could play a role in the dissemination of AMR among food-borne bacteria and suggest potential food safety and public health risk. Therefore, enhanced surveillance for antibiotic-resistant organisms in raw meat for early detection of emerging resistant bacteria species in the food chain is recommended.

Detection of Listeria monocytogenes in Foods and Characterization by PFGE

The aims of the present study were to investigate the prevalence of Listeria monocytogenes in 1042 foods collected from different market to characterize the isolates by phenotypical and molecular methods. In particular, L. monocytogenes obtained from different types of foods such as RTE (kimbap), fish (smoked salmon and seasoned-dried slice fish) and meat (cut raw beef and pork) from 2009 to 2011, were used. Twelve samples (2.1%) were positive for L. monocytogenes. Detection rate of L. monocytogenes varied significantly by food type and ranged from 1.1% to 5.2%. Meat is the highest prevalence for L. monocytogenes (5.2%) followed by RTE (1.8%) and Fish (1.1%). Twelve isolates were also serotyped by the agglutination method. The most common serotypes detected in the 12 strains tested were 1/4b (75.0%), followed by 1/2a (16.7%), and 1/2b (8.3%). For this study, we used serotyping and detected 6 different virulence-associated genes (inlA, inlB, plcA, plcB, hlyA, and actA) and 16s rRNA using multiplex-PCR. PFGE was performed to determine genetic characterization of L. monocytogenes strains to define the genetic diversity.