Human Listeriosis Research Articles

Genomic epidemiology of hypervirulent Listeria monocytogenes CC619: Population structure, phylodynamics and virulence

Listeria monocytogenes is a ubiquitous foodborne pathogen causing human and animal listeriosis with high mortality. Neurological and maternal-neonatal listeriosis outbreaks in Europe and the United States were frequently associated with clonal complexes CC1, CC2 and CC6 harboring Listeria Pathogenicity Island-1 (LIPI-1), as well as CC4 carrying both LIPI-1 and LIPI-4. However, human listeriosis in China was predominantly linked to CC87 and CC619 from serotype 1/2b. To understand the genetic evolution and distribution patterns of CC619, we characterized the epidemic history, population structure, and transmission feature of CC619 strains through analysis of 49,421 L. monocytogenes genomes globally. We found that CC619 was uniquely distributed in China, and closely related with perinatal infection. As CC619 strains were being mainly isolated from livestock and poultry products, we hypothesized that pigs and live chicken were the reservoirs of CC619. Importantly, all CC619 strains not only harbored the intact LIPI-1 and LIPI-4, but these also carried LIPI-3 that could facilitate host colonization and invasion. The deficiency of LIPI-3 or LIPI-4 markedly decreased L. monocytogenes colonization capacity in a model of intragastric infection in the mouse. Altogether, our findings suggest that the hypervirulent CC619 harboring three pathogenicity islands LIPI-1, LIPI-3 and LIPI-4 is a putatively persistent population in various foods, environment, and human population, warranting the further research for deciphering its pathogenicity and strengthening epidemiological surveillance.

Why does Listeria monocytogenes survive in food and food-production environments?

Listeria monocytogenes is one of the most dangerous food-borne pathogens and is responsible for human listeriosis, a severe disease with a high mortality rate, especially among the elderly, pregnant women and newborns. Therefore, this bacterium has an important impact on food safety and public health. It is able to survive and even grow in a temperature range from -0.4°C to 45°C, a broad pH range from 4.6 to 9.5 and at a relatively low water activity (aW < 0.90), and tolerates salt content up to 20%. It is also resistant to ultraviolet light, biocides and heavy metals and forms biofilm structures on a variety of surfaces in food-production environments. These features make it difficult to remove and allow it to persist for a long time, increasing the risk of contamination of food-production facilities and ultimately of food. In the present review, the key mechanisms of the pathogen's survival and stress adaptation have been presented. This information may grant better understanding of bacterial adaptation to food environmental conditions.

Pulsed-field gel electrophoresis typing and molecular characterization of &lt;em&gt;Listeria monocytogenes&lt;/em&gt; isolates in raw milk samples from Polonnaruwa District, Sri Lanka

Listeria monocytogenes is an important food borne human pathogen associated with severe diseases in humans and animals. Human infection is associated with high mortality rates. Our previous studies done in 2012 on milk and milk products revealed 4b as the dominant serotype followed by 1/2a, 1/2b and 1/2c. Listeria monocytogenes is an important food borne human pathogen associated with severe diseases in humans and animals. Human infection is associated with high mortality rates. Our previous studies done in 2012 on milk and milk products revealed 4b as the dominant serotype followed by 1/2a, 1/2b and 1/2c. Therefore, this study was conducted to determine the sources of contamination and to identify the strain differences (pulsotypes) by pulsed-field gel electrophoresis (PFGE) typing and serotype profiles of L. monocytogenes isolates from farm collected raw milk samples from randomly identified locations in Grama Niladhari divisions of the Polonnaruwa district, Sri Lanka. Eighty isolates obtained from raw milk samples were confirmed as L. monocytogenes by Polymerase Chain Reaction, belonging to serotypes 4b, 1/2a, 1/2b and 1/2c. The strains were identified by PFGE. PFGE analysis digested with AscI and ApaI enzymes revealed different banding patterns. All the pulsotypes were found to be serotype 4b. The sub typing indicated the diversity of the Listeria species. The presence of serotypes 1/2a, 1⁄2b, 1/2c, and 4b in raw milk is a public health concern, as these serotypes are frequently associated with foodborne outbreaks and sporadic cases of human listeriosis. In our study, PFGE analysis allowed discrimination among isolates of the same serogroup. Further PFGE analysis showed heterogeneity among isolates recovered from both same sampling areas and different areas.

Genetic and Phenotypic Diversity of Listeria monocytogenes in Pig Slaughterhouses in Korea.

Listeria monocytogenes is a foodborne pathogen that has variable subtypes associated with human listeriosis and occurs in food and processing environments. This study was conducted to provide the genetic and phenotypic characterization of L. monocytogenes in pig carcasses and environments of slaughterhouses in Korea. A total of 22 L. monocytogenes were isolated from eight of 26 pig slaughterhouses between 2020 and 2022, and the most common serotype was 1/2c (40.9%), followed by serotypes 1/2b (31.8%) and 1/2a (27.3%). The isolates showed a significantly high prevalence of virulence genes located in Listeria pathogenicity island-1 (LIPI-1) and internalins (90.9-100%; p < 0.05). However, the prevalence rates of llsX, ptsA, and stress survival islet-1 (SSI-1) located in LIPI-3, LIPI-4, and SSI were only 9.1%, 22.7%, and 31.8%, respectively. In addition, among the epidemic clones (EC), ECI, ECII, ECIII, and ECV, only one isolate was represented as ECV. Isolates identified from the same slaughterhouses were divided into two or more pulsotypes, except for two slaughterhouses. Furthermore, the seven STs were classified into seven clonal complexes (CCs) (CC8, CC9, CC37, CC87, CC121, CC155, and CC288), and all CCs belonged to lineages I (31.8%) and II (68.1%). Interestingly, the isolates showed a high prevalence of oxacillin resistance (59.1%), and most isolates of the serotypes 1/2a and 1/2b exhibited oxacillin resistance, whereas only one of nine serotype 1/2c isolates exhibited oxacillin resistance. These results provide the genetic diversity of L. monocytogenes in pig carcasses and environments of slaughterhouses, and continuous monitoring will be helpful in predicting food safety risks.

Design of a novel affinity probe using the cell wall-binding domain of a Listeria monocytogenes autolysin for pathogen detection.

Rapid identification of foods and food-processing environments contaminated with Listeria monocytogenes is crucial for controlling human listeriosis, a leading cause of fatality related to foodborne illnesses. The L. monocytogenes autolysin IspC anchors to the bacterial surface non-covalently via its C-terminal cell wall-binding domain (CWBD), which contains conserved Gly-Trp (GW) dipeptides. This study explored the binding property of CWBDIspC to design an affinity probe for detecting L. monocytogenes. Immunogold electron microscopy analysis of a ∆ispC mutant and its wild type (WT) revealed that exogenously added CWBDIspC bound to surface ligands that were evenly distributed and remained mostly unoccupied in the wild type. CWBDIspC detected, via recognizing the cell wall component lipoteichoic acid, L. monocytogenes strains primarily belonging to the serotype 4 group, as well as Listeria innocua, and Listeria welshimeri but did not detect three other Listeria spp. (Listeria seeligeri, Listeria ivanovii, and Listeria grayi) nor any non-Listeria spp. tested. CWBDIspC, when tagged with the enhanced green fluorescent protein, was capable of detecting L. monocytogenes by fluorescence microscopy, a colony lift filter assay, and a microtube binding assay. CWBDIspC, when covalently attached to magnetic beads, showed the ability to separate L. monocytogenes cells with a maximum capture rate of 90 to nearly 100% from 1 × 103 to 1 × 105 target cells. This study demonstrated for the first time that the CWBD derived from a GW surface protein IspC is a novel antibody alternative for the rapid detection and isolation of L. monocytogenes despite binding to two other Listeria species.IMPORTANCEHuman listeriosis is caused by consuming foods contaminated with the bacterial pathogen Listeria monocytogenes, leading to the development of a severe and life-threatening foodborne illness. Detection of L. monocytogenes present in food and food processing environments is crucial for preventing Listeria infection. The L. monocytogenes peptidoglycan hydrolase IspC anchors non-covalently to the bacterial surface through its C-terminal cell wall-binding domain (CWBD), CWBDIspC. This study explored the surface binding property of CWBDIspC to design, construct, characterize, and assess an affinity molecular probe for detecting L. monocytogenes. CWBDIspC recognized a cell wall ligand lipoteichoic acid that remains evenly displayed and mostly unoccupied on the bacterial surface for interaction with the exogenously added CWBDIspC. CWBDIspC, when fused to the enhanced green fluorescent protein reporter or covalently conjugated onto magnetic beads, exhibited the functionality as an antibody alternative for rapid detection and efficient separation of the pathogen.

Environmental persistence of Listeria monocytogenes and its implications in dairy processing plants.

Listeriosis, an invasive illness with a fatality rate between 20% and 30%, is caused by the ubiquitous bacterium Listeria monocytogenes. Human listeriosis has long been associated with foods. This is because the ubiquitous nature of the bacteria renders it a common food contaminant, posing a significant risk to the food processingsector. Although several sophisticated stress coping mechanisms have been identified as significant contributing factors toward the pathogen's persistence, a complete understanding of the mechanisms underlying persistence across various strains remains limited. Moreover, aside from genetic aspects that promote the ability to cope with stress, various environmental factors that exist in food manufacturing plants could also contribute to the persistence of thepathogen. The objective of this review is to provide insight into the challenges faced by the dairy industry because of the pathogens' environmental persistence. Additionally, it also aims to emphasize the diverse adaptation and response mechanisms utilized by L. monocytogenes in food manufacturing plants to evade environmental stressors. The persistence of L. monocytogenes in the food processing environment poses a serious threat to food safety and public health. The emergence of areas with high levels of L. monocytogenes contamination could facilitate Listeria transmission through aerosols, potentially leading to the recontamination of food, particularly from floors and drains, when sanitation is implemented alongside product manufacturing. Hence, to produce safe dairy products and reduce the frequency of outbreaks of listeriosis, it is crucial to understand the factors that contribute to the persistence of this pathogen and to implement efficient control strategies.

Viability and Virulence of Listeria monocytogenes in Poultry.

The prevalence of Listeria monocytogenes in 30 samples of poultry was determined using culture-dependent (isolation on OCLA and confirmation by conventional polymerase chain reaction -PCR-, OCLA&PCR) and culture-independent (real-time polymerase chain reaction, q-PCR) methods. L. monocytogenes was detected in 15 samples (50.0%) by OCLA&PCR and in 20 (66.7%) by q-PCR. The concentrations (log10 cfu/g) of L. monocytogenes (q-PCR) ranged from 2.40 to 5.22 (total cells) and from <2.15 to 3.93 (viable cells). The two methods, q-PCR using a viability marker (v-PCR) and OCLA&PCR (gold standard), were compared for their capacity to detect viable cells of L. monocytogenes, with the potential to cause human disease. The values for sensitivity, specificity and efficiency of the v-PCR were 100%, 66.7% and 83.3%, respectively. The agreement between the two methods (kappa coefficient) was 0.67. The presence of nine virulence genes (hlyA, actA, inlB, inlA, inlC, inlJ, prfA, plcA and iap) was studied in 45 L. monocytogenes isolates (three from each positive sample) using PCR. All the strains harbored between six and nine virulence genes. Fifteen isolates (33.3% of the total) did not show the potential to form biofilm on a polystyrene surface, as determined by a crystal violet assay. The remaining strains were classified as weak (23 isolates, 51.1% of the total), moderate (one isolate, 2.2%) or strong (six isolates, 13.3%) biofilm producers. The strains were tested for susceptibility to a panel of 15 antibiotics. An average of 5.11 ± 1.30 resistances per isolate was observed. When the values for resistance and for reduced susceptibility were taken jointly, this figure rose to 6.91 ± 1.59. There was a prevalence of resistance or reduced susceptibility of more than 50.0% for oxacillin, cefoxitin, cefotaxime, cefepime ciprofloxacin, enrofloxacin and nitrofurantoin. For the remaining antibiotics tested, the corresponding values ranged from 0.0% for chloramphenicol to 48.9% for rifampicin. The high prevalence and level of L. monocytogenes with numerous virulence factors in poultry underline how crucial it is to follow correct hygiene procedures during the processing of this foodstuff in order to reduce the risk of human listeriosis.

Antimicrobial resistance of Listeria monocytogenes serogroups IIa and IVb from food and food-production environments in Poland.

Listeria monocytogenes is an important foodborne pathogen responsible for human listeriosis, which is a disease with high hospitalisation and mortality rates. The bacteria are usually susceptible to most antibacterial substances, but resistance to some of them has been recently observed. The present study introduces the evidence on the emergence of antibiotic resistance among L. monocytogenes strains isolated from food and food-production environments in Poland. A total of 283 L. monocytogenes isolates classified into serogroups IIa and IVb which had been recovered from food and food production environments were tested with 17 antimicrobials. These included those that are recommended for treatment of severe listeriosis cases in humans. A multiplex PCR was used to identify serogroups, and a microbroth dilution method was applied for the determination of antibiotic resistance among the isolates tested. Only 34 (12.0%) strains were susceptible to all the antimicrobials used in the study. The remaining 249 (88.0%) strains displayed different instances of resistance to the antimicrobials tested, from insusceptibility to one (112 strains; 39.6%) to resistance to four antibacterial substances (6 strains; 2.1%). Among them, there were 38 strains (13.4%) with multiresistance patterns. Polish food and its processing environments may be a potential source of antimicrobial-resistant L. monocytogenes, which may pose a potential health risk to consumers in the country.

The Isolation, Genetic Analysis and Biofilm Characteristics of Listeria spp. from the Marine Environment in China.

Listeria monocytogenes is an important pathogen that can cause listeriosis. Despite the growing recognition of Listeria spp. as a foodborne and environmental pathogen, the understanding of its prevalence and characteristics of Listeria spp. in the marine environment remains unknown. In this study, we first investigated the genetic and phenotypic characteristics of Listeria species isolated in a coastal city in China. The findings revealed that the sequence type 87 (ST87) L. monocytogenes, a prevalent clinical and seafood strain in China, dominates in recreational beach sands and possesses a notable biofilm-forming capacity in seawater. The presence of ST87 L. monocytogenes in coastal environments indicates the potential health risks for both recreational activities and seafood consumption. Moreover, the ST121 isolates from sand had a versatile plasmid encoding multifunctional genes, including uvrX for UV resistance, gbuC for salt resistance, and npx for oxidative resistance and multiple transposases, which potentially aid in survival under natural environments. Black-headed gulls potentially facilitate the spread of L. monocytogenes, with similar ST35 strains found in gulls and beach sand. As a reservoir of microbes from marine environments and human/animal excrement, coastal sand would play an important role in the spread of L. monocytogenes and is an environmental risk for human listeriosis.

A Novel Two-Component Bacteriocin, Acidicin P, and Its Key Residues for Inhibiting Listeria monocytogenes by Targeting the Cell Membrane.

Listeria monocytogenes is an important pathogen which easily contaminates food and causes fatal systemic infections in human. Bacteriocins have received much attention regarding their natural methods of controlling health-related pathogens. Here, we investigated and characterized a novel two-component bacteriocin named acidicin P from Pediococcus acidilactici LAC5-17. Acidicin P showed obvious antimicrobial activity to L. monocytogenes. Through a sequence similarity network analysis for two-component bacteriocin precursors mined in the RefSeq database, acidicin P was observed to belong to an unusual group of two-component bacteriocins. Acidicin P contains two peptides designated Adpα and Adpβ which are assessed to interact with each other and form a helical dimer structure which can be inserted into the lipid bilayer of target cell membrane. We demonstrate that A5, N7, and G9 in the A5xxxG9 motif of Adpα and S16, R19, and G20 in the S16xxxG20 motif of Adpβ played crucial roles in stabilizing the helix-helix interaction of Adpα and Adpβ and were essential for the antilisterial activity of acidicin P by site-directed mutagenesis. A positive residue, R14, in Adpα and a negative residue, D12, in Adpβ are also important for acidicin P to fight against L. monocytogenes. These key residues are supposed to form hydrogen bonding, which is crucial for the interaction of Adpα and Adpβ. Furthermore, acidicin P induces severe permeabilization and depolarization of the cytoplasmic membrane and causes dramatic changes in L. monocytogenes cell morphology and ultrastructure. Acidicin P has the potential to be applied to inhibit L. monocytogenes efficiently both in the food industry and medical treatments. IMPORTANCE L. monocytogenes can cause widespread food contamination and severe human listeriosis, which amount to a large proportion of the public health and economic burdens. Today, L. monocytogenes is usually treated with chemical compounds in the food industry or antibiotics for human listeriosis. Natural and safe antilisterial agents are urgently required. Bacteriocins are natural antimicrobial peptides that have comparable narrow antimicrobial spectra and are attractive potentials for precision therapy for pathogen infection. In this work, we discover a novel two-component bacteriocin designated acidicin P, which shows obvious antilisterial activity. We also identify the key residues in both peptides of acidicin P and demonstrate that acidicin P is inserted into the target cell membrane and disrupts the cell envelop to inhibit the growth of L. monocytogenes. We believe that acidicin P is a promising lead for further development as an antilisterial drug.

Listeria monocytogenes an Emerging Pathogen: a Comprehensive Overview on Listeriosis, Virulence Determinants, Detection, and Anti-Listerial Interventions.

Listeria monocytogenes, the third most deleterious zoonotic pathogen, is a major causative agent of animal and human listeriosis, an infection related to the consumption of contaminated food products. Even though, this pathogen has been responsible for the outbreaks of foodborne infections in the early 1980s, the major outbreaks have been reported during the past two decades. Listeriosis infection in the host is a rare but life-threatening disease with major public health and economic implications. Extensive reports on listeriosis outbreaks are associated with milk and milk products, meat and meat products, and fresh produce. This bacterium can adapt to any environmental and stress conditions, making it a prime causative agent for major foodborne diseases. The pathogen could survive an antibiotic treatment and persist in the host cell, thereby escaping the standard diagnostic practices. The current review strives to provide concise information on the epidemiology, serotypes, and pathogenesis of the L. monocytogenes to decipher the knowledge on the endurance of the pathogen inside the host and food products as a vehicle for Listeria contaminations. In addition, various detection methods for Listeria species from food samples and frontline regimens of L. monocytogenes treatment have also been discussed.

Draft Genome Sequences of Heavy Metal-Resistant Listeria monocytogenes Strains of Sequence Type 14 (Clonal Complex 14) from Human Listeriosis Cases in Sweden.

Listeria monocytogenes of clonal complex 14 (CC14) is a potentially hypervirulent clone of serotype 1/2a but remains poorly characterized. We report the genome sequences of five sequence type 14 (ST14) (CC14) strains from human listeriosis cases in Sweden, which harbor a chromosomal heavy metal resistance island that is generally uncommon in serotype 1/2a.

Whole-genome sequencing of Listeria monocytogenes isolated from the first listeriosis foodborne outbreak in South Korea.

Listeria monocytogenes is a foodborne pathogen that causes listeriosis in humans with severe symptoms. In South Korea, listeriosis had only been reported sporadically among hospitalized patients until the first foodborne outbreak occurred in 2018. In this study, a L. monocytogenes strain responsible for this outbreak (FSCNU0110) was characterized via whole genome sequencing and compared with publicly available L. monocytogenes genomes of the same clonal complex (CC). Strain FSCNU0110 belonged to multilocus sequence typing (MLST)-based sequence type 224 and CC224, and core genome MLST-based sublineage 6,178. The strain harbored tetracycline resistance gene tetM, four other antibiotic resistance genes, and 64 virulence genes, including Listeria pathogenicity island 1 (LIPI-1) and LIPI-3. Interestingly, llsX in LIPI-3 exhibited a characteristic SNP (deletion of A in position 4, resulting in a premature stop codon) that was missing among all CC224 strains isolated overseas but was conserved among those from South Korea. In addition, the tetM gene was also detected only in a subset of CC224 strains from South Korea. These findings will provide an essential basis for assessing the characteristics of CC224 strains in South Korea that have shown a potential to cause listeriosis outbreaks.

Serotyping, RAPD PCR, Antibiotic Susceptibility Profiling of Listeria spp. Isolated from Raw Chicken Meat

The present study was aimed at finding the prevalence, serotyping, RAPD and Setyping of Listeria monocytogenes in raw chicken meat purchased from different retail outlets and local butcher shops across the Nagpur city. Cultural examination of 100 raw poultry meat samples, revealed the prevalence of Listeria spp. (35 %). L. grayi was predominant (15 %) among the isolates followed by L. monocytogenes (12 %), L. welshimeri (5 %), L. innocua (2 %) and L. seeligeri (1%) respectively. Among the majority of strains of L. monocytogenes, virulence marker genes hlyA and iap, all together or in combination were detected. Serotyping PCR study revealed that all L. monocytogenes isolates to be serotype 4b, which is the major serotype involved in the human listeriosis. RAPD PCR analysis of L. monocytogenes isolates revealed two types of banding patterns. All 35 strains of Listeria showed in-vitro susceptibility to antibiotics like vancomycin (93.88 %) ceftriaxone + Tazobactum (92.71%) and moderate sensitivity to enrofloxacin (38.78 %), ceftriaxone (34.90%) and ampicillin (34.69 %). Whereas, all the isolates were least sensitive to erythromycine (6.49%) and sulphasomidine (4.08 %). All the Listeriae isolates were refractory to penicillin. The present findings indicate the presence of multiple drug resistance among L. monocytogenes and other Listeria spp. isolated from raw poultry meat samples. Potentially pathogenic L. monocytogenes isolates recovered from raw poultry meat samples also signifies the zoonotic potential of listeriosis in the local area of Nagpur. High prevalence of L. monocytogenes and other Listeria spp. in raw poultry meat and their sensitivity to the antibiotics tested, clearly indicates that vancomycin and ceftriaxone +tazobactam should be used as a drug of choice for treatment of listeriosis. Keywords: Serotyping PCR, RAPD, Antibiotic Sensitivity test, ceftriaxone, sulphasomidine, tazobactam

Draft Genome Sequences of Closely Related Listeria monocytogenes Lineage III Strains Isolated from a Food Processing Environment and a Case of Human Listeriosis.

Listeria monocytogenes lineage III is genetically highly diverse, and closely related lineage III strains from food facilities and human listeriosis have not been reported. Here, we report the genome sequences of three closely related lineage III strains from Hawaii, namely, one isolated from a human case and two isolated from a produce storage facility.

Natural Antimicrobials for Listeria monocytogenes in Ready-to-Eat Meats: Current Challenges and Future Prospects

Listeria monocytogenes, an intra-cellular, Gram-positive, pathogenic bacterium, is one of the leading agents of foodborne illnesses. The morbidity of human listeriosis is low, but it has a high mortality rate of approximately 20% to 30%. L. monocytogenes is a psychotropic organism, making it a significant threat to ready-to-eat (RTE) meat product food safety. Listeria contamination is associated with the food processing environment or post-cooking cross-contamination events. The potential use of antimicrobials in packaging can reduce foodborne disease risk and spoilage. Novel antimicrobials can be advantageous for limiting Listeria and improving the shelf life of RTE meat. This review will discuss the Listeria occurrence in RTE meat products and potential natural antimicrobial additives for controlling Listeria.

High multidrug resistance of Listeria monocytogenes and association with water sources in sheep and goat dairy flocks in Jordan

Listeria monocytogenes is a significant pathogen that causes listeriosis in humans and small ruminants. This study aimed to estimate the prevalence, antimicrobial resistance and risk factors of L. monocytogenes in small dairy ruminants in Jordan. A total of 948 milk samples were collected from 155 sheep and goat flocks across Jordan. L. monocytogenes was isolated from the samples, confirmed and tested for 13 clinically important antimicrobials. Data were also collected on the husbandry practices to identify risk factors for the presence of L. monocytogenes. The results showed that the flock-level prevalence of L. monocytogenes was 20.0% (95% CI; 14.46%−26.99%) and the prevalence in the individual milk samples was 6.43% (95% CI; 4.92%−8.36%). The univariable (UOR=2.65, p = 0.021) and multivariable (AOR=2.49, p = 0.028) analyses showed that using water from municipality pipelines as a water source in the flock reduced L. monocytogenes prevalence. All L. monocytogenes isolates were resistant to at least one antimicrobial. High percentages of the isolates were resistant to ampicillin (83.6%), streptomycin (79.3%), kanamycin (75.0%), quinupristin/dalfopristin (63.8%) and clindamycin (61.2%). About 83.6% of the isolates (94.2% and 75% of the sheep and goat isolates) exhibited multidrug resistance (resistance to ≥3 antimicrobial classes). In addition, the isolates exhibited fifty unique antimicrobial resistance profiles. Thus, it’s recommended to restrict the misuse of clinically important antimicrobials and to chlorinate and monitor the water sources in sheep and goat flocks.

Whole-Genome Sequencing-Based Characterization of Clinical Listeria monocytogenes Isolates in China, 2013-2019.

Invasive listeriosis is a rare but serious foodborne disease that causes maternal-neonatal, central nervous system, and bloodstream infections. The aim of this study was to assess the whole-genome sequencing (WGS)-based genetic diversity of clinical Listeria monocytogenes isolates over a 7-year period and prove the effect of WGS application in food vehicle investigation. A total of 360 isolates were recovered during 2013 and 2019 through the national listeriosis special surveillance program. Two hundred twenty-six isolates (62.8%) were associated with pregnancy. All isolates belonged to lineage I (214 isolates) or lineage II (146 isolates), with 4 serogroups (46.9% IIb, 39.7% IIa, 12.5% IVb, and 0.8% IIc). All isolates were in 25 clonal complexes (CCs) and 3 singletons, with CC87, CC8, and CC5 being the most common causes of human listeriosis. All clinical isolates were positive for Listeria pathogenicity island 1 (LIPI-1), LIPI-3 was present in 21.4% of isolates and LIPI-4 was detected in 29.2% of isolates. LIPI-4-positive isolates, including CC87, sequence type (ST)619, ST382, CC4, and CC2, have been shown to confer hypervirulence. Fifteen isolates harbored at least one antimicrobial encoding gene, including tet (M), mef (A), msr (D), and dfr (G). The sublineage designations were consistent with CC designations, and 215 distinct cgMLST types (CTs) were classified, the most abundant being CT58 and CT750. In summary, there is a high level of genetic diversity among the clinical isolates. WGS has strengthened listeriosis surveillance and will be implemented for other foodborne bacteria in the National Molecular Tracing Network for Foodborne Disease.

Contamination of beef and beef products by Listeria spp. and molecular characterization of L. monocytogenes in Mpumalanga, South Africa

AbstractThis study determined the prevalence, risk factors, and molecular characteristics of Listeria species detected in beef and beef products sampled in Mpumalanga province, South Africa. Four hundred beef and beef products were collected from 30 retail outlets in three districts (Bronkhorstspruit, Emalahleni, and Middelburg) within the province. Standard bacteriological and polymerase chain reaction (PCR) assays were used in the study. The overall prevalence of L. monocytogenes and other Listeria spp. in the samples was 8.3% (33/400) and 30% (120/400) (p &lt; .05), respectively. For the five variables investigated, statistically significant effects were evident only for the region (p &lt; .001) and type of product (p &lt; .0001) for L. monocytogenes, the type of outlet (p = .011) and the type of product (p &lt; .0001) for Listeria spp. Of the 20 types of beef and beef products tested, 15 (75%) and 17 (85%) were contaminated by L. monocytogenes and Listeria spp., respectively (p = .429). Among the four categories of products tested, the prevalence of L. monocytogenes was 7.3% (8/109), 10.6% (11/104), 7.5% (8/106), and 7.4% (6/81) for raw beef, ready‐to‐eat (RTE) products, milled beef, and offal &amp; organs, respectively (p &gt; .799). Among the 33 L. monocytogenes isolates, PCR genoserogroup IIa (42.4%, 1/2a‐3a) was most frequently detected. All (100%) of the isolates carried one or more of the eight virulence‐associated genes assessed, with genes inlC and inlJ detected in all the isolates. The overall prevalence of L. monocytogenes (8.3%) and the high frequency of virulent serogroups of L. monocytogenes commonly associated with human listeriosis pose a food safety risk to consumers of beef and beef‐based products contaminated by L. monocytogenes.

Listeria monocytogenes Isolates from Meat Products and Processing Environment in Poland Are Sensitive to Commonly Used Antibiotics, with Rare Cases of Reduced Sensitivity to Ciprofloxacin.

Antibiotic resistance is a global health problem, causing not only an increased mortality rate of bacterial infections but also economic losses due to, among other reasons, the need for longer hospital stays. Listeria monocytogenes is one of the foodborne pathogens with the ability to induce a serious illness called listeriosis, with approximately 20-30% fatal outcomes. The treatment regimen of listeriosis in humans includes the administration of antibiotics (in most cases, ampicillin or trimethoprim with sulfamethoxazole in case of allergies to β-lactams), so the resistance of this pathogen to antibiotics can potentially lead to increased mortality. The antibiotic sensitivity status of n = 153 L. monocytogenes isolates originating from meat food samples (raw and processed) and meat-processing environment (both contacting and non-contacting with food) collected between October 2020 and November 2021 in Poland was examined in this study. Susceptibility to antibiotics was determined using the disc diffusion method on Mueller-Hinton agar plates. All collected samples were susceptible to 9 antibiotics: ampicillin (10 µg), chloramphenicol (30 µg), erythromycin (15 µg), gentamicin (10 µg), penicillin (10 IU), streptomycin (10 µg), sulfamethoxazole/trimethoprim (1.25/23.75 µg), tetracycline (30 µg) and vancomycin (30 µg). Some of the isolates (n = 10; 6.5%) showed reduced susceptibility to ciprofloxacin (5 µg), which was classified as an intermediate response. All these ten isolates were collected from surfaces contacting with food in food-processing facilities.