Listeria Monocytogenes Isolates Research Articles

Genomic characteristics of listeria monocytogenes causing invasive listeriosis in Japan

We reviewed 18 listeriosis cases in Japan and performed molecular analysis of causative Listeria monocytogenes (LM) isolates. Strains genetically related to those from other countries caused various types of listeriosis, including vascular listeriosis in immunocompetent elderly people. Our results highlight the importance of integrated clinical and genomic analysis of LM.

Transmission of Listeriosis in a Neonatal Intensive Care Unit Supported by Whole-Genome Sequencing

Background: Listeriosis is a rare but serious infectious disease caused by Listeria monocytogenes (LM) and predominantly transmitted through contaminated food. Moreover, 15% of listeriosis cases in the United States are pregnancy associated; nosocomial neonatal transmission in hospitals is extremely rare. In July 2018, the California Department of Public Health (CDPH) was notified of 4 patients, a mother–neonate pair and twin neonates, with listeriosis at the same hospital. The CDPH and San Diego County Health and Human Services Agency initiated an investigation to determine transmission and prevent additional infections. Methods: We reviewed medical records of the neonates and their mothers, interviewed the mothers with a detailed food exposure questionnaire, interviewed healthcare personnel (HCP), and performed an infection control assessment of the neonatal intensive care unit (NICU). CDPH performed whole-genome sequencing (WGS) on LM isolates that were then analyzed by whole-genome multilocus sequence typing (wgMLST) by the Centers for Diseases Control and Prevention (CDC) to assess relatedness in PulseNet, a public health laboratory database. The CDC also performed testing for LM on formalin-fixed placentas from the mother of the twins. Results: During a 1-week period, 4 patients with LM were identified at the hospital. A mother was admitted at 31 weeks gestation with acute abdominal and back pain that progressed with precipitous vaginal delivery and postpartum sepsis. Her neonate was resuscitated, transported to the NICU, underwent a sepsis evaluation, received antibiotics, and was transferred to another hospital within 6 hours. Maternal blood, placenta, and neonatal blood cultures grew LM. Twin neonates, born to an asymptomatic mother and present in the NICU during the index neonate’s stay, developed acute infection 4 and 6 days after the index neonate’s transfer; blood cultures confirmed LM. The LM isolates from the 4 patients were indistinguishable by wgMLST and were not related to other PulseNet isolates. LM was not detected in the twin placentas. There were no common food exposures between the mothers. At least 1 common HCP cared for all 3 neonates. Infection control lapses included lack of proper hand hygiene during the index neonate’s resuscitation and potentially after cleaning and disinfection of the neonate’s incubator. Conclusions: This report provides supportive evidence that nosocomial transmission of LM can occur during a brief NICU stay due to lapses in infection control practices. Strict adherence to standard precautions in the delivery room and NICU is imperative to prevent cross transmission.Disclosures: NoneFunding: None

Antibacterial activity of plant extracts against Listeria monocytogenes isolated from ready-to-eat salads

Abstract Ready-to-eat salads are becoming more and more popular. However, due to their ingredients, they represent a suitable growth environment for different microbes. In the prevention of foodborne diseases, hygienic food preparation and appropriate storage conditions are very important. During this study, ten different ready-to-eat salads were analysed for the presence of Listeria monocytogenes. Five different selective agar mediums were used for the enumeration and isolation of Listeria monocytogenes. The isolated bacterial strains were subjected to morphological and biochemical confirmation tests. The antibacterial effects of five different freshly squeezed vegetable juices (carrots, celery, beets, horseradish, and onions) and of five essential oils (dill, thyme, oregano, lemongrass, and sage) were determined against Listeria monocytogenes, Listeria innocua, and L. monocytogenes strains isolated from ready-to-eat salads. Based on the results obtained from fresh vegetable juices, carrot juice exerted the highest antibacterial effect, while the others showed no or slight inhibitory effect (horseradish, beets, onions) against Listeria species. Among the essential oils, thyme, lemongrass, and oregano showed the strongest antibacterial effect against the studied Listeria species.

Serotype Assignment by Sero-agglutination, ELISA, and PCR.

For assessing isolates of Listeria monocytogenes, serotype designation is the first subtyping method used. Methodologies used to assign serotype are currently evolving and will eventually be replaced with whole genome sequencing. Traditionally, serotyping has been done with agglutination reactions; however, alternative methods utilizing enzyme linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) are common. Described here are the three non-genomic methods and the advantages and disadvantages of each.

Population Structure of Listeria monocytogenes in Emilia-Romagna (Italy) and Implications on Whole Genome Sequencing Surveillance of Listeriosis.

The population structure of human isolates of Listeria monocytogenes in Emilia-Romagna, Italy, from 2012 to 2018 was investigated with the aim of evaluating the presence of genomic clusters indicative of possible outbreaks, the proportion of cluster-associated vs. sporadic isolates and different methods and metrics of genomic analysis for use in routine surveillance. In the 2012–2018 period the notification rate of confirmed invasive cases in Emilia-Romagna was 0.91 per 100,000 population per year, more than twice the average rate of EU countries. Out of the total 283 cases, 268 (about 95%) isolates were typed through whole genome sequencing (WGS) for cluster detection with methods based on core-genome multi-locus sequence typing and single nucleotide polymorphisms. Between 66 and 72% of listeriosis cases belonged to genomic clusters which included up to 27 cases and lasted up to 5 years. This proportion of cluster-associated cases is higher than previously estimated in other European studies. Rarefaction analysis, performed by reducing both the number of consecutive years of surveillance considered and the proportion of isolates included in the analysis, suggested that the observed high proportion of cluster-associated cases can be ascribed to the long surveillance duration (7 years) and the high notification and typing rates of this study. Our findings show that a long temporal perspective and high surveillance intensity, intended as both exhaustiveness of the system to report cases and high WGS-typing rate, are critical for sensitive detection of possible outbreaks within a WGS-based surveillance of listeriosis. Furthermore, the power and complexity of WGS interpretation emerged from the integration of genomic and epidemiological information in the investigation of few past outbreaks within the study, indicating that the use of multiple approaches, including the analysis of the accessory genome, is needed to accurately elucidate the population dynamics of Listeria monocytogenes.

RAPD PCR and actA based molecular typing of L. monocytogenes isolated from human, food and domestic animals in northwest of Iran

Twenty-one Listeria monocytogenes isolates originated from food, animal, and clinical sources were evaluated by RAPD-PCR typing using four random primers (OPM-01, HLWL-74, HLWL-82, and HLWL85), both individually and in combination. The actA gene-typing and PCR-based serotyping were also used for the characterization of the isolates. Data were analyzed using the related software, and clusters were displayed in dendrogram. Among four primers, OPM-01 showed the highest discriminatory power. The dendrogram for human, food, and livestock isolates showed four different clusters (A-D) and two singletons. Using RAPD-PCR, lineages I, II, and the serogroups were discriminated. In actA gene-typing, 95% of isolates (all except one) belonged to actA type I. Results of PCR-serotyping and RAPD-PCR showed that animal isolates differed from food and clinical isolates. RAPD-PCR showed higher discriminatory power compared to other typing methods used in this study as isolates belonging to the same serotypes in PCR-based serotyping indicated different RAPD profiles.

The Composition of Microbial Communities in Six Streams, and Its Association With Environmental Conditions, and Foodborne Pathogen Isolation.

Surface water used for produce production is a potential source of pre-harvest contamination with foodborne pathogens. Decisions on how to mitigate food safety risks associated with pre-harvest water use currently rely on generic Escherichia coli-based water quality tests, although multiple studies have suggested that E. coli levels are not a suitable indicator of the food safety risks under all relevant environmental conditions. Hence, improved understanding of spatiotemporal variability in surface water microbiota composition is needed to facilitate identification of alternative or supplementary indicators that co-occur with pathogens. To this end, we aimed to characterize the composition of bacterial and fungal communities in the sediment and water fractions of 68 agricultural water samples collected from six New York streams. We investigated potential associations between the composition of microbial communities, environmental factors and Salmonella and/or Listeria monocytogenes isolation. We found significantly different composition of fungal and bacterial communities among sampled streams and among water fractions of collected samples. This indicates that geography and the amount of sediment in a collected water sample may affect its microbial composition, which was further supported by identified associations between the flow rate, turbidity, pH and conductivity, and microbial community composition. Lastly, we identified specific microbial families that were weakly associated with the presence of Salmonella or Listeria monocytogenes, however, further studies on samples from additional streams are needed to assess whether identified families may be used as indicators of pathogen presence.

Laboratory-based surveillance of invasive listeriosis in Northern Italy over a fourteen-year period: epidemiological and clinical results.

SummaryIntroductionInvasive listeriosis is a rare foodborne disease with a significant impact on public health worldwide, because of the severity of its clinical manifestations and high fatality rate. In this study, we provide a snapshot of epidemiology of listeriosis in Lombardy Region, Northern Italy, reviewing enhanced surveillance data collected over fourteen years, after the implementation of a voluntary laboratory-based surveillance system for the referral of clinical isolates of Listeria monocytogenes to a regional reference laboratory, since 2005.MethodsInvasive listeriosis cases data from 2005 to 2018 were extracted from the regional laboratory-based surveillance system database and compared with the regional mandatory notification disease system data.ResultsOver the fourteen-year period under study, 533 Listeria monocytogenes isolates were detected by the laboratory surveillance system, 55 of which from pregnancy-related cases. The median age of non-pregnancy-associated patients was 71 years, with 64.6% of cases observed in the elderly. Cases with underlying medical risk conditions accounted for 92.1%, and the fatality rate was 26.2%. By integrating data from the mandatory notification system and the laboratory-based surveillance system, a total of 935 cases were recorded. The collection of data through the laboratory surveillance system allowed to increase the surveillance sensitivity by 18%.ConclusionsOur results documented the growing epidemiological relevance of listeriosis through the analysis of two information sources, the regional mandatory notification system and the regional laboratory-based surveillance system. The data we obtained were consistent with the literature, except for pregnancy-related cases, which are often underdiagnosed. This study highlighted the importance of laboratory-based surveillance system, which led to a significant increase in the sensitivity of the mandatory notification system.

Listeria monocytogenes infections: Analysis of 41 patients

ObjectiveWe describe and analyze Listeria-related demographics and clinical features to determine the predisposing conditions for severe infections. MethodsWe performed a retrospective study using positive isolation of Listeria monocytogenes from blood, cerebrospinal fluid, and other organic fluids. Electronic health records were used to determine the epidemiological and clinical features of infections caused by L. monocytogenes. Mortality and sepsis were considered dependent variables in the statistical analyses. ResultsWe included 41 patients in an observation period of 15 years (2003–2018), with an annual incidence rate of 1.3 cases per 100,000 population. Three main population profiles were identified: newborns, pregnant women, and other adults (17.1%, 12.2%, and 82.9%, respectively). Neuroinvasive infection was present in 17 patients (41.5%). In both univariate and multivariate analyses, neurological infections, whether meningoencephalitis, rhombencephalitis, or brain abscesses, were the main risk factors for severe forms of Listeria-related infections (odds ratio 1.8, 95% CI 1.52–2.14, p=0.01). Malignancies, whether solid tumors or hematological neoplasms, immunosuppression, and chronic diseases were not related to either mortality or severe clinical syndromes. ConclusionInfections caused by L. monocytogenes were uncommon but could cause severe sepsis and mortality, especially in susceptible populations. Our study focused on neurological involvement and severe invasive forms of listeriosis. Neuroinvasive forms were the most important risk factors for severe illness but not for mortality.

Characterisation of Listeria monocytogenes isolates from cattle using a bovine caruncular epithelial cell model

Listeria monocytogenes is an important foodborne pathogen in human and veterinary health, causing significant morbidity and mortality including abortion. It has a particular tropism for the gravid uterus, however, the route of infection in reproductive tissues of ruminants (i.e. placentome), is much less clear. In this study, we aimed to investigate a bovine caruncular epithelial cell (BCEC) line as a model for L. monocytogenes infection of the bovine reproductive tract. The BCEC infection model was used to assess the ability of 14 different L. monocytogenes isolates to infect these cells. Lysozyme sensitivity and bacterial survival in 580 μg lysozyme/ml correlated with attenuated ability to proliferate in BCEC (p = 0.004 and p = 0.02, respectively). Four isolates were significantly attenuated compared to the control strain 10403S. One of these strains (AR008) showed evidence of compromised cell wall leading to increased sensitivity to ß-lactam antibiotics, and another (7644) had compromised cell membrane integrity leading to increased sensitivity to cationic peptides. Whole genome sequencing followed by Multi Locus Sequence Type analysis identified that five invasive isolates had the same sequence type, ST59, despite originating from three different clinical conditions. Virulence gene analysis showed that the attenuated isolate LM4 was lacking two virulence genes (uhpT, virR) known to be involved in intracellular growth and virulence.In conclusion, the BCEC model was able to differentiate between the infective potential of different isolates. Moreover, resistance to lysozyme correlated with the ability to invade and replicate within BCEC, suggesting co-selection for surviving challenging environments as the abomasum.

Large Nationwide Outbreak of Invasive Listeriosis Associated with Blood Sausage, Germany, 2018-2019.

Invasive listeriosis is a severe foodborne infection in humans and is difficult to control. Listeriosis incidence is increasing worldwide, but some countries have implemented molecular surveillance programs to improve recognition and management of listeriosis outbreaks. In Germany, routine whole-genome sequencing, core genome multilocus sequence typing, and single nucleotide polymorphism calling are used for subtyping of Listeria monocytogenes isolates from listeriosis cases and suspected foods. During 2018–2019, an unusually large cluster of L. monocytogenes isolates was identified, including 134 highly clonal, benzalkonium-resistant sequence type 6 isolates collected from 112 notified listeriosis cases. The outbreak was one of the largest reported in Europe during the past 25 years. Epidemiologic investigations identified blood sausage contaminated with L. monocytogenes highly related to clinical isolates; withdrawal of the product from the market ended the outbreak. We describe how epidemiologic investigations and complementary molecular typing of food isolates helped identify the outbreak vehicle.

A Study to Evaluate the Need for Comminution in the Preparation of Food Materials for Testing of Listeria monocytogenes, Staphylococcus Species, and Generic Escherichia coli.

Comminution reduces the sampling error arising from distributional heterogeneity of the target contaminant/target analyte in the material, facilitating the selection of a more representative test portion. A laboratory sampling method incorporating comminution prior to selection of the test portion (Sampling Method B) was compared to current sampling methods that used no comminution step (Sampling Method A). This required the development of an efficient process for comminution of food samples prior to removal of the test portion for the detection and isolation of Listeria monocytogenes and the enumeration of Staphylococcus species and Escherichia coli. From December 2016 to December 2017, 2742 tests were conducted on 778 unique food samples. For all food samples, a test portion (TPA) was first removed using Sampling Method A, and then the remainder of the material was comminuted and a second test portion (TPB) was removed using Sampling Method B and tested alongside the first portion. Across all food matrices and microbial targets, 17 additional targets were detected using only Sampling Method B, and positive detections of target analytes increased by 77% using Sampling Method B from the test portions taken using Sampling Method A. Utilizing a sample preparation method that includes a comminution step resulted in an increased number of pathogen detections. The introduction of a comminution step in the preparation of food samples for detection of three common microbial contaminants resulted in an increase in the rate of detection of natural contaminates in a variety of ready to eat foods. An efficient aseptic process for commutation that can be adapted to a wide range of laboratory settings was identified.

Genetic diversity and profiles of genes associated with virulence and stress resistance among isolates from the 2010-2013 interagency Listeria monocytogenesmarket basket survey.

Whole genome sequencing (WGS) was performed on 201 Listeria monocytogenes isolates recovered from 102 of 27,389 refrigerated ready-to-eat (RTE) food samples purchased at retail in U.S. FoodNet sites as part of the 2010–2013 interagency L. monocytogenes Market Basket Survey (Lm MBS). Core genome multi-locus sequence typing (cgMLST) and in-silico analyses were conducted, and these data were analyzed with metadata for isolates from five food groups: produce, seafood, dairy, meat, and combination foods. Six of 201 isolates, from 3 samples, were subsequently confirmed as L. welshimeri. Three samples contained one isolate per sample; mmong the 96 samples that contained two isolates per sample, 3 samples each contained two different strains and 93 samples each contained duplicate isolates. After 93 duplicate isolates were removed, the remaining 102 isolates were delineated into 29 clonal complexes (CCs) or singletons based on their sequence type. The five most prevalent CCs were CC155, CC1, CC5, CC87, and CC321. The Shannon’s diversity index for clones per food group ranged from 1.49 for dairy to 2.32 for produce isolates, which were not significantly different in pairwise comparisons. The most common molecular serogroup as determined by in-silico analysis was IIa (45.6%), followed by IIb (27.2%), IVb (20.4%), and IIc (4.9%). The proportions of isolates within lineages I, II, and III were 48.0%, 50.0% and 2.0%, respectively. Full-length inlA was present in 89.3% of isolates. Listeria pathogenicity island 3 (LIPI-3) and LIPI-4 were found in 51% and 30.6% of lineage I isolates, respectively. Stress survival islet 1 (SSI-1) was present in 34.7% of lineage I isolates, 80.4% of lineage II isolates and the 2 lineage III isolates; SSI-2 was present only in the CC121 isolate. Plasmids were found in 48% of isolates, including 24.5% of lineage I isolates and 72.5% of lineage II isolates. Among the plasmid-carrying isolates, 100% contained at least one cadmium resistance cassette and 89.8% contained bcrABC, involved in quaternary ammonium compound tolerance. Multiple clusters of isolates from different food samples were identified by cgMLST which, along with available metadata, could aid in the investigation of possible cross-contamination and persistence events.

Genomics-Metabolomics Profiling Disclosed Marine Vibrio spartinae 3.6 as a Producer of a New Branched Side Chain Prodigiosin.

A wide range of prescreening tests for antimicrobial activity of 59 bacterial isolates from sediments of Ria Formosa Lagoon (Algarve, Portugal) disclosed Vibrio spartinae 3.6 as the most active antibacterial producing strain. This bacterial strain, which has not previously been submitted for chemical profiling, was subjected to de novo whole genome sequencing, which aided in the discovery and elucidation of a prodigiosin biosynthetic gene cluster that was predicted by the bioinformatic tool KEGG BlastKoala. Comparative genomics led to the identification of a new membrane di-iron oxygenase-like enzyme, annotated as Vspart_02107, which is likely to be involved in the biosynthesis of cycloprodigiosin and analogues. The combined genomics-metabolomics profiling of the strain led to the isolation and identification of one new branched-chain prodigiosin (5) and to the detection of two new cyclic forms. Furthermore, the evaluation of the minimum inhibitory concentrations disclosed the major prodigiosin as very effective against multi-drug-resistant pathogens including Stenotrophomonas maltophilia, a clinical isolate of Listeria monocytogenes, as well as some human pathogens reported by the World Health Organization as prioritized targets.

Phylogenetic Analysis and Antibiotics Resistance of Listeria Monocytogenes Contaminating Chicken Meat in Surabaya, Indonesia.

The objective of this study was to identify the phylogenetic analysis and antibiotic resistance of Listeria monocytogenes contaminating chicken meat in Surabaya. 60 chicken meat samples were collected from supermarkets, mobile vendors, and traditional markets in Surabaya. A selective medium is used for isolation and identification of Listeria monocytogenes by chopping 25 grams of the chicken meat and to put it into the sterilized Erlenmeyer flasks. Some methods were used for the identification procedures, such as biochemical and morphological tests, antibiotic resistance test, PCR, and sequencing; also a phylogenetic analysis was conducted by a neighbor-joining analysis using Genetix Mac ver 8.0 with hlyA genes of Listeria monocytogenes recorded in GenBank, such as Lineage I (KC808543), Lineage II (AY229462, AY229346, AY229499, and AY229404), Lineage III (KJ504139, HQ686043, KJ504116, and DQ988349), and Lineage IV (EU840690, EF030606). The result shows that the prevalence of L. monocytogenes in Surabaya contaminating the chicken meat samples from the supermarkets was 10% (2/20), from the mobile vendors was 0/20 (0%), and from the traditional markets was 5% (1/20). It was seen from the band at 456 bp fragment. Furthermore, three isolates found in Surabaya were included in the new lineages which were resistant to old-generation antibiotics such as sulfamethonazole-trimetophrim (SXT) and amoxyllin sulbactam (MAS), but they were still sensitive to new-generation antibiotics such as cefotaxime (CTX) and meropenem (MEM).

Outbreak of Listeriosis in South Africa Associated with Processed Meat

BackgroundAn outbreak of listeriosis was identified in South Africa in 2017. The source was unknown.MethodsWe conducted epidemiologic, trace-back, and environmental investigations and used whole-genome sequencing to type Listeria monocytogenes isolates. A case was defined as laboratory-confirmed L. monocytogenes infection during the period from June 11, 2017, to April 7, 2018.ResultsA total of 937 cases were identified, of which 465 (50%) were associated with pregnancy; 406 of the pregnancy-associated cases (87%) occurred in neonates. Of the 937 cases, 229 (24%) occurred in patients 15 to 49 years of age (excluding those who were pregnant). Among the patients in whom human immunodeficiency virus (HIV) status was known, 38% of those with pregnancy-associated cases (77 of 204) and 46% of the remaining patients (97 of 211) were infected with HIV. Among 728 patients with a known outcome, 193 (27%) died. Clinical isolates from 609 patients were sequenced, and 567 (93%) were identified as sequence type 6 (ST6). In a case–control analysis, patients with ST6 infections were more likely to have eaten polony (a ready-to-eat processed meat) than those with non-ST6 infections (odds ratio, 8.55; 95% confidence interval, 1.66 to 43.35). Polony and environmental samples also yielded ST6 isolates, which, together with the isolates from the patients, belonged to the same core-genome multilocus sequence typing cluster with no more than 4 allelic differences; these findings showed that polony produced at a single facility was the outbreak source. A recall of ready-to-eat processed meat products from this facility was associated with a rapid decline in the incidence of L. monocytogenes ST6 infections.ConclusionsThis investigation showed that in a middle-income country with a high prevalence of HIV infection, L. monocytogenes caused disproportionate illness among pregnant girls and women and HIV-infected persons. Whole-genome sequencing facilitated the detection of the outbreak and guided the trace-back investigations that led to the identification of the source.

Control of Growth and Persistence of Listeria monocytogenes and β-Lactam-Resistant Escherichia coli by Thymol in Food Processing Settings

The main objective of this study was to evaluate the efficacy of thymol in controlling environmental contamination in food processing facilities. The effect of thymol was tested as an agent to prevent planktonic and bacterial biofilm growth of twenty-five Listeria monocytogenes isolates from a variety of foods and five Escherichia coli isolates from a farm. The E. coli isolates were positive for extended spectrum β-lactamase (ESBL) genes. All isolates and reference strains were susceptible to thymol at Minimum inhibitory concentration (MIC) values ranging from 250 to 800 μg/mL. An interesting activity of interference with biofilm formation of L. monocytogenes and E. coli was found for thymol at sub-MIC concentrations of 200, 100, 75, and 50 μg/mL. Anti-biofilm activity ranging from 59.71% to 66.90% against pre-formed 24-h-old L. monocytogenes biofilms at concentrations of 500 or 800 µg/mL, corresponding to 2× MIC, was determined against free-living forms of six isolates chosen as the best or moderate biofilm producers among the tested strains. The property of thymol to attack L. monocytogenes biofilm formation was also observed at a concentration of 100 µg/mL, corresponding to 1/4 MIC, by using a stainless-steel model to simulate the surfaces in food industries. This study gives information on the use of thymol in food processing setting.

Pathogenicity, haematology and pathology of Listeria monocytogenes in rat

Listeria monocytogenes is a gram positive bacterium of public health concern with low morbidity but high mortality rate. Listeriosis in human is usually caused by the consumption of infected animal and food. It is noteworthy that an infected animal such as rat could be potential carrier and vector. Therefore, this study examined the clinic pathological changes associated with Listeria monocytogenes infection in rat. Six (6) Winstar laboratory rats divided into 2 groups. Group 1 was challenged with 0.1-mL saline water as control while the second group was challenged intraperitoneally with 0.1 mL molecularly characterised Listeria monocytogenes (1 × 104 CFU/mL) isolates. The rats were observed for clinical signs and mortality. Haematology, serum and enzymes indices were examined using standard procedures. Then, the liver and kidney of the rats from the two groups were aseptically examined for histopathological changes. The result shows that 33.3% of the rats challenged with Listeria monocytogenes died within 24 h (1 day) while 100% mortality were recorded within 48 h (2 days). There were significant decreases (P < 0.05) in the values of haematological parameters and biochemical of the rats than the control group (P < 0.05) while a significant increase in hepatic and renal blood enzymes of infected rats was observed. The rat exposed to Listeria infection revealed moderate glomerular atrophy in the kidney, and zonal degeneration of hepatocytes was observed in the liver. The study indicates that Listeria monocytogenes is a pathogenic organism to rats which could cause huge mortality within 3 days post infection. The infection also causes derangement of haematological, biochemical and tissue pathological changes in organs especially liver.

Isolation and Identification of Listeria monocytogenes from Raw Vegetables and Meat Sold in Quetta, Pakistan

Isolation and Identification of Listeria monocytogenes from Raw Vegetables and Meat Sold in Quetta, Pakistan

Population structure-guided profiling of antibiotic resistance patterns in clinical Listeria monocytogenes isolates from Germany identifies pbpB3 alleles associated with low levels of cephalosporin resistance

ABSTRACT Numbers of listeriosis illnesses have been increasing in Germany and the European Union during the last decade. In addition, reports on the occurrence of antibiotic resistance in Listeria monocytogenes in clinical and environmental isolates are accumulating. The susceptibility towards 14 antibiotics was tested in a selection of clinical L. monocytogenes isolates to get a more precise picture of the development and manifestation of antibiotic resistance in the L. monocytogenes population. Based on the population structure determined by core genome multi locus sequence typing (cgMLST) 544 out of 1220 sequenced strains collected in Germany between 2009 and 2019 were selected to cover the phylogenetic diversity observed in the clinical L. monocytogenes population. All isolates tested were susceptible towards ampicillin, penicillin and co-trimoxazole – the most relevant antibiotics in the treatment of listeriosis. Resistance to daptomycin and ciprofloxacin was observed in 493 (91%) and in 71 (13%) of 544 isolates, respectively. While all tested strains showed resistance towards ceftriaxone, their resistance levels varied widely between 4 mg/L and >128 mg/L. An allelic variation of the penicillin binding protein gene pbpB3 was identified as the cause of this difference in ceftriaxone resistance levels. This study is the first population structure-guided analysis of antimicrobial resistance in recent clinical isolates and confirms the importance of penicillin binding protein B3 (PBP B3) for the high level of intrinsic cephalosporin resistance of L. monocytogenes on a population-wide scale.