Monocytogenes Isolates Research Articles

High density genomic surveillance and risk profiling of clinical Listeria monocytogenes subtypes in Germany

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

Oxford Nanopore's 2024 sequencing technology for Listeria monocytogenes outbreak detection and source attribution: progress and clone-specific challenges.

Whole genome sequencing is an essential cornerstone of pathogen surveillance and outbreak detection. Established sequencing technologies are currently being challenged by Oxford Nanopore Technologies (ONT), which offers an accessible and cost-effective alternative enabling gap-free assemblies of chromosomes and plasmids. Limited accuracy has hindered its use for investigating pathogen transmission, but recent technology updates have brought significant improvements. To evaluate its readiness for outbreak detection, we selected 78 Listeria monocytogenes isolates from diverse lineages or known epidemiological clusters for sequencing with ONT's V14 Rapid Barcoding Kit and R10.4.1 flow cells. The most accurate of several tested workflows generated assemblies with a median of one error (SNP or indel) per assembly. For 66 isolates, the cgMLST profiles from ONT-only assemblies were identical to those generated from Illumina data. Eight assemblies were of lower quality, with more than 20 erroneous sites each, primarily caused by methylations at the GAAGAC motif (5'-GAAG6mAC-3'/5'-GT4mCTTC-3'). This led to inaccurate clustering, failing to group isolates from a persistence-associated clone that carried the responsible restriction-modification system. Out of 50 methylation motifs detected among the 78 isolates, only the GAAGAC motif was linked to substantially increased error rates. Our study shows that most L. monocytogenes genomes assembled from ONT-only data are suitable for high-resolution genotyping, but further improvements of chemistries or basecallers are required for reliable routine use in outbreak and food safety investigations.

Sanitizer Resistance and Persistence of Listeria monocytogenes Isolates in Tree Fruit Packing Facilities

The foodborne pathogen Listeria monocytogenes can persist in produce processing environments, which increases the risk for food contamination. Increased resistance to antimicrobials commonly used in cleaning and sanitizing procedures may contribute to L. monocytogenes’ persistence in these environments. This study aimed to evaluate sanitizer resistance in L. monocytogenes isolates collected from three tree fruit packing facilities (F1, F2, and F3) during packing seasons 2020–2021 (Y1) and 2021–2022 (Y2), and to assess evidence of persistence based on the genomic similarity of isolates to historical isolates collected in previous years. L. monocytogenes isolates collected in 2020–2022 (n = 44) were tested for resistance to peroxyacetic acid (PAA) and a proprietary biofilm−removing agent using a broth microdilution assay. Further, L. monocytogenes isolates were whole genome sequenced and screened for the presence of antimicrobial resistance and virulence genes, as well as to assess the genomic similarity of isolates using the CFSAN SNP bioinformatic pipeline. Over half (57%) of the tested isolates had a PAA minimum inhibitory concentration (MIC) of 250 ppm, which was similar to the applied concentration of the PAA sanitizer in the three facilities (230 ppm). In contrast, 80% of tested isolates had a biofilm remover MIC of 0.13 ppm, which was substantially below the concentration applied in the facilities (137 ppm). Genomes of all tested isolates carried antimicrobial resistance (fosX, lin, mdrL, mprF, and norB) and virulence (inlA, inlB, plcA, plcB, prfA, hly, mpl, and iap) genes. L. monocytogenes isolates collected between 2020 and 2022 belonged to three distinct lineages, with 22 multilocus sequence types (MLSTs) belonging to 22 different clonal complexes. Genomic similarity analysis with historical isolates collected from the same facilities in 2016–2017 demonstrated a 5-year persistence of the genotypes ST 1003 and ST 554 in F2, which were no longer detected in 2022. Overall, our results highlight the need to re-evaluate sanitizer concentrations to effectively control persistent L. monocytogenes strains in tree fruit packing facilities.

Molecular serogrouping and virulence of Listeria monocytogenes from sheep and goat milk in Jordan

Listeria monocytogenes is a serious pathogen with several serogroups that differ in virulence and distribution among countries. This study determined the serogroups and virulence of 116 L. monocytogenes from sheep milk (n = 52) and goat milk (n = 64) in Jordan by several multiplex PCR. The results showed that the L. monocytogenes isolates belong to five serogroups. Most isolates (37.9%) belonged to serogroup 4b (d, e), followed by serogroup 1/2a (3a) (31.9%), serogroup 1/2b (26.7%), and serogroup 4a/c (2.6%) and serogroup IV (serotypes 1/2c and 3c) (0.9%). All isolates carried the internalin (inlA, inlC, inlJ) and lmo 2672 genes. Additionally, 36.4% and 29.0 % of 4b and 1/2b isolates carried the Listeriolysin S gene (llsX). Therefore, L. monocytogenes in sheep and goat milk are potential public health hazards because they belong to the major listeriosis-causing serogroups (4b, 1/2a, and 1/2b), and carry major virulence genes. Small ruminants’ milk and cheeses should be considered high-risk foods for L. monocytogenes infection. Thus, safe production practices should be monitored and enforced in small dairy shops.

Some virulence genes and biofilm formation capabilities of Listeria monocytogenes isolates from different sources

In this study, it was aimed to determine the biofilm-forming abilities of both clinical and food-borne isolates of Listeria monocytogenes, to investigate the presence of nine different virulence genes, and to consider the current threat status of this agent. A total of 28 isolates, 21 from food and seven from clinical origin, were used in the study. To determine the biofilm formation abilities of isolates, two different methods namely “tube adherence” and “microplate” were used respectively. For the determination of nine different virulence genes of Listeria monocytogenes (inlA, inlC, inlJ, hylA, luxS, flaA, prfA, inlB, actA), the method of polymerase chain reaction (PCR) was used. As a result, all isolates were found to be able to form a biofilm to varying degrees by both tube and microplate methods. These two methods were similar in terms of their results. All nine different virulence gene regions were detected at various rates in the isolates. Whereas the genes directly related to biofilm formation for the isolates weren't detected, to form biofilm was observed. The virulence genes detected in clinical origin isolates were proportionally higher than in food-borne isolates (except for flaA and prfA gene regions). It was concluded that bacteria of Listeria monocytogenes continue to form biofilm and carry virulence genes regardless they are from food or clinical origin. Also, food-borne contaminations continue to be a severe threat to human health. So, to prevent listeriosis cases of both humans and animals should be taken required precautions and all cases should be considered carefully.

Detection of Phenotypic and Genotypic Antibiotic Resistance among Listeria monocytogenes Isolated from Different Food Samples in Yola

Study’s Novelty/Excerpt This study investigates the phenotypic and genotypic antibiotic resistance of Listeria monocytogenes isolated from food samples in Yola, revealing significant multi-drug resistance (MDR) in 72.72% of the isolates. The research highlights alarming resistance rates to commonly used antibiotics, including 100% resistance to ceftriaxone and the presence of resistance genes blaCTX-M, blaTEM, and sul1 in all MDR isolates. These findings underscore a potential public health crisis, indicating that future outbreaks of L. monocytogenes in the area may be difficult to control with the current antibiotic treatments. Full Abstract Listeria monocytogenes is an opportunistic bacterial pathogen implicated in several lethal illness outbreaks. Future outbreaks may be more complicated to manage because of the surfacing of antibiotic resistance among L. monocytogenes strains in food products. Accordingly, this study aimed to determine the phenotypic and genotypic antibiotic resistance of L. monocytogenes previously isolated from different food samples in Yola. Antibiotic resistance of L. monocytogenes was determined via disc diffusion and polymerase chain reaction (PCR). Exactly 72.72% of the isolates were multi-drug resistant (MDR) with high resistance to Beta-Lactams and sulfonamides. Percentage resistance to ceftriaxone, ampicillin, augmentin, cefuroxime, and sulfamethoxazole/trimethoprim were 100%, 72.7%, 64.0%, 64.0%, and 54.5% respectively. The MDR isolates were subjected to PCR, and antibiotic resistance genes blaCTX-M, 7(100%), blaTEM, 7(100%), and sul1 7(100%) were detected in (72.72%) i.e. all the MDR isolates. L. monocytogenes isolates from food samples in this study area exhibited phenotypic and genotypic resistance to multiple antibiotics. This implies that future outbreaks of L. monocytogenes in the study area may be complicated to manage using the commonly used antibiotics tested in this study, representing a major public health concern.

Antimicrobial Resistance Profiles of Listeria Species Recovered from Retail Outlets in Gauteng Province, South Africa

The study determined the antimicrobial resistance (AMR) profiles of Listeria spp. (L. monocytogenes, L. innocua, and L. welshimeri) recovered from beef and beef products sold at retail outlets in Gauteng Province, South Africa. A total of 112 isolates of Listeria spp., including L. monocytogenes (37), L. innocua (65), and L. welshimeri (10), were recovered from beef and beef products collected from 48 retail outlets. Listeria spp. was recovered by direct selective plating following selective enrichment, and PCR was used to confirm and characterize recovered isolates. The disc diffusion method determined the resistance to 16 antimicrobial agents. All 112 isolates of Listeria spp. exhibited resistance to one or more antibiotics (P < 0.05). The prevalence of AMR in Listeria isolates was high for nalidixic acid (99.1%) and cefotaxime (80.4%) but low for gentamycin (2.7%), sulfamethoxazole-trimethoprim (3.6%), azithromycin (5.4%), and doxycycline (6.3%). Overall, for the three species of Listeria, the prevalence of resistance varied significantly only for streptomycin (P = 0.016) and tetracycline (P = 0.034). Multidrug-resistant isolates were detected in 75.7% (28/37), 61.5% (40/65), and 80% (8/10) isolates of L. monocytogenes, L. innocua, and L. welshimeri, respectively. The prevalence of AMR was significantly affected by the location and size of retail outlets, type of beef and beef products, and serogroups of L. monocytogenes. The high prevalence of AMR, particularly among the L. monocytogenes isolates, poses potential therapeutic implications for human consumers of contaminated beef products. There is, therefore, a need to regulate and enforce the use of antimicrobial agents in humans and animals in South Africa.

The Relationship between Biofilm Phenotypes and Biofilm-Associated Genes in Food-Related Listeria monocytogenes Strains.

Listeria monocytogenes is an important pathogen responsible for listeriosis, a serious foodborne illness associated with high mortality rates. Therefore, L. monocytogenes is considered a challenge for the food industry due to the ability of some strains to persist in food-associated environments. Biofilm production is presumed to contribute to increased L. monocytogenes resistance and persistence. The aims of this study were to (1) assess the biofilm formation of L. monocytogenes isolates from a meat processing facility and sheep farm previously characterized and subjected to whole-genome sequencing and (2) perform a comparative genomic analysis to compare the biofilm formation and the presence of a known set of biofilm-associated genes and related resistance or persistence markers. Among the 37 L. monocytogenes isolates of 15 sequence types and four serogroups involved in this study, 14%, 62%, and 24% resulted in the formation of weak, moderate, and strong biofilm, respectively. Increased biofilm-forming ability was associated with the presence of the stress survival islet 1 (SSI-1), inlL, and the truncated inlA genes. Combining the phenotypic and genotypic data may contribute to understanding the relationships between biofilm-associated genes and L. monocytogenes biofilm-forming ability, enabling improvement in the control of this foodborne pathogen.

Prevalence and characterization of foodborne pathogens isolated from fresh-cut fruits and vegetables in Beijing, China

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

Antimicrobial Resistance of Listeria monocytogenes Strains Isolated in Food and Food-Processing Environments in Italy

Listeria monocytogenes, along with various other pathogenic bacteria, may show resistance against a broad spectrum of antibiotics. Evaluating the extent of resistance in harmful microorganisms like Listeria monocytogenes holds significant importance in crafting novel therapeutic strategies to mitigate or combat the rise of infections stemming from antibiotic-resistant bacteria. The present work aims to investigate the occurrence of antimicrobial resistance among Listeria monocytogenes strains in meat products (n = 173), seafood (n = 54), dairy products (n = 19), sauces (n = 2), confectionary products (n = 1), ready-to-eat rice dishes (n = 1), and food-processing environments (n = 19). A total of 269 Listeria monocytogenes strains belonging to eight different serovars were tested against 10 antimicrobials. In the classes of antibiotics, most of the strains were resistant antibiotics belonging to the family of β-lactams (92.94%). High proportions of L. monocytogenes isolates were resistant to oxacillin (88.48%), followed by fosfomycin (85.87%) and flumenique (78.44%). The lowest level of resistance was observed against gentamycin (1.49%). A total of 235 strains (n = 87.36%) showed a profile of multidrug resistance. In conclusion, a high occurrence of resistant and multidrug-resistant strains of Listeria monocytogenes was observed among the examined serotypes isolated from different food sources. This understanding enables the adoption of suitable measures to avert contamination and the spread of resistant bacteria via food.

Prevalence, Genotypic Characteristics, and Antibiotic Resistance of Listeria monocytogenes From Retail Foods in Huzhou, China

Listeria monocytogenes are considered to be the major foodborne pathogen worldwide. To understand the prevalence and potential risk of L. monocytogenes in retail foods, a total of 1243 retail foods in 12 food categories were sampled and screened for L. monocytogenes from 2020 to 2022 in Huzhou, China. A total of 46 out of 1234 samples were confirmed to be L. monocytogenes positive with a total rate of 3.7%. The contamination rate of seasoned raw meat (15.2%) was the highest, followed by raw poultry meat and raw livestock meat (9.9%) and salmon sashimi (9.5%). The L. monocytogenes isolates belonged to four serotypes, 1/2a,1/2b, 1/2c, and 4b, with the most prevalent serotype being 1/2a (47.9%). All isolates were grouped into 15 sequence types (STs) belonging to 14 clonal complexes (CCs) via multilocus sequence typing (MLST). The most prevalent ST was ST9/CC9 (23.9%), followed by ST3/CC3 (19.6%) and ST121/CC121 (17.4%). Notably, 11 STs were detected from ready-to-eat (RTE) foods, some of them have been verified to be strongly associated with clinical origin listeriosis cases, such as ST3, ST2, ST5, ST8, and ST87. Listeria pathogenicity islands 1 (LIPI-1) and LIPI-2 were detected in approximately all L. monocytogenes isolates, whereas the distribution of both LIPI-3 genes and LIPI-4 genes exhibited association with specific ST, with LIPI-3 in ST3 and ST288, and LIPI-4 in ST87. The strains carrying LIPI-3 and LIPI-4 virulence genes in this study were all isolated from RTE foods. Antimicrobial susceptibility tests showed that >90% of isolates were susceptible to PEN, AMP, ERY, CIP, SXT, VAN, CHL, and GEN, indicating the antibiotic treatment might be still efficient for most of the L. monocytogenes strains. However, for the three clinical first-line antibiotics (PEN, AMP, and GEN), we also observed three and four strains showing MIC values greater than the susceptibility standards for PEN and AMP, respectively, and one strain showing resistance to GEN.

High Prevalence of Virulence-Associated Genes and Length Polymorphism in actA and inlB Genes Identified in Listeria monocytogenes Isolates from Meat Products and Meat-Processing Environments in Poland.

Listeria monocytogenes is a human pathogen that has the ability to cause listeriosis, a disease with possible fatal outcomes. The typical route of infection is ingestion of the bacteria with contaminated food. In this study, 13 virulence-associated genes were examined with PCR in the genomes of 153 L. monocytogenes isolates collected from meat products and processing environments in Poland. All isolates possessed genes from LIPI-1-hly, actA, plcA, plcB and mpl-as well as four internalins: inlA, inlB, inlC, inlJ. Invasion-associated protein iap, as well as genes prfA and sigB, encoding regulatory proteins, were also detected in all isolates. Gene flaA, encoding flagellin, was detected in 113 (74%) isolates. This was the only gene that was not detected in all isolates, as its presence is serotype-dependent. Gene actA showed polymorphism with longer and shorter variants in PCR amplicons. Two isolates were characterized by truncated inlB genes, lacking 141 bp in their sequence, which was confirmed by gene sequencing. All isolates were positive in hemolysis assays, proving the synthesis of functional PrfA and Hly proteins. Four genotypes of L. monocytogenes based on actA polymorphism and two genotypes based on inlB polymorphism were distinguished within the isolates' collection.

Biofilm formation by Listeria monocytogenes from the meat processing industry environment and the use of different combinations of detergents, sanitizers, and UV-A radiation to control this microorganism in planktonic and sessile forms.

This study aimed to evaluate the ability of biofilm formation by L. monocytogenes from the meat processing industry environment, as well as the use of different combinations of detergents, sanitizers, and UV-A radiation in the control of this microorganism in the planktonic and sessile forms. Four L. monocytogenes isolates were evaluated and showed moderate ability to form biofilm, as well as carried genes related to biofilm production (agrB, agrD, prfA, actA, cheA, cheY, flaA, sigB), and genes related to tolerance to sanitizers (lde and qacH). The biofilm-forming isolates of L. monocytogenes were susceptible to quaternary ammonium compound (QAC) and peracetic acid (PA) in planktonic form, with minimum inhibitory concentrations of 125 and 75 ppm, respectively, for contact times of 10 and 5min. These concentrations are lower than those recommended by the manufacturers, which are at least 200 and 300 ppm for QAC and PA, respectively. Biofilms of L. monocytogenes formed from a pool of isolates on stainless steel and polyurethane coupons were subjected to 14 treatments involving acid and enzymatic detergents, QAC and PA sanitizers, and UV-A radiation at varying concentrations and contact times. All treatments reduced L. monocytogenes counts in the biofilm, indicating that the tested detergents, sanitizers, and UV-A radiation exhibited antimicrobial activity against biofilms on both surface types. Notably, the biofilm formed on polyurethane showed greater tolerance to the evaluated compounds than the biofilm on stainless steel, likely due to the material's surface facilitating faster microbial colonization and the development of a more complex structure, as observed by scanning electron microscopy. Listeria monocytogenes isolates from the meat processing industry carry genes associated with biofilm production and can form biofilms on both stainless steel and polyurethane surfaces, which may contribute to their persistence within meat processing lines. Despite carrying sanitizer tolerance genes, QAC and PA effectively controlled these microorganisms in their planktonic form. However, combinations of detergent (AC and ENZ) with sanitizers (QAC and PA) at minimum concentrations of 125 ppm and 300 ppm, respectively, were the most effective.

Whole Genome Sequence Analysis of Listeria monocytogenes Isolates Obtained from the Beef Production Chain in Gauteng Province, South Africa.

The study used whole-genome sequencing (WGS) and bioinformatics analysis for the genomic characterization of 60 isolates of Listeria monocytogenes obtained from the beef production chain (cattle farms, abattoirs, and retail outlets) in Gauteng province, South Africa. The sequence types (STs), clonal complexes (CCs), and the lineages of the isolates were determined using in silico multilocus sequence typing (MLST). We used BLAST-based analyses to identify virulence and antimicrobial genes, plasmids, proviruses/prophages, and the CRISPR-Cas system. The study investigated any association of the detected genes to the origin in the beef production chain of the L. monocytogenes isolates. Overall, in 60 isolates of Listeria monocytogenes, there were seven STs, six CCs, forty-four putative virulence factors, two resistance genes, one plasmid with AMR genes, and three with conjugative genes, one CRISPR gene, and all 60 isolates were positive for proviruses/prophages. Among the seven STs detected, ST204 (46.7%) and ST2 (21.7%) were the most prominent, with ST frequency varying significantly (p < 0.001). The predominant CC detected were CC2 (21.7%) and CC204 (46.7%) in lineages I and II, respectively. Of the 44 virulence factors detected, 26 (across Listeria Pathogenicity Islands, LIPIs) were present in all the isolates. The difference in the detection frequency varied significantly (p < 0.001). The two AMR genes (fosX and vga(G)) detected were present in all 60 (100%) isolates of L. monocytogenes. The only plasmid, NF033156, was present in three (5%) isolates. A CRISPR-Cas system was detected in six (10%), and all the isolates carried proviruses/prophages. The source and sample type significantly affected the frequencies of STs and virulence factors in the isolates of L. monocytogenes. The presence of fosX and vga(G) genes in all L. monocytogenes isolates obtained from the three industries of the beef production chain can potentially cause therapeutic implications. Our study, which characterized L. monocytogenes recovered from the three levels in the beef production chain, is the first time genomics was performed on this type of data set in the country, and this provides insights into the health implications of Listeria.

Antimicrobial susceptibility patterns of Listeria monocytogenes isolated from fresh produce in KwaZulu-Natal Province, South Africa

Fresh, ready-to-eat produce is frequently irrigated with untreated water, making it a leading cause of food-borne illness outbreaks worldwide. This study investigated the presence of Listeria monocytogenes in fresh produce that was grown using river water. Standard biochemical tests were used for the identification of L. monocytogenes isolated from river water used for irrigation, and from fresh produce including lettuce, spinach, and pumpkin. The inlA gene of L. monocytogenes was molecularly identified using PCR amplification. The susceptibility of L. monocytogenes isolates to antimicrobial agents was assessed using the Kirby-Bauer disc diffusion method. The presence of the amplified inlA gene (800 bp) indicated that all of the fresh produce and river water samples were contaminated with virulent L. monocytogenes. Lettuce and spinach exhibited higher quantities of L. monocytogenes, with lettuce recording 87 CFU/g and spinach recording 71 CFU/g. The L. monocytogenes isolates from spinach and lettuce sources showed significant resistance to colistin (56.2% and 53.3%, respectively) as well as ampicillin (68.8% and 53.3%, respectively). Moreover, lettuce (40%) and spinach (31%) exhibited a common resistance pattern of AMP-CHL-CT-KAN-PIP-ERY-TET, with a maximum MAR index value of 0.54. Our research demonstrates the transmission of multidrug-resistant L. monocytogenes from irrigation river water to fresh produce. Hence, the ingestion of ready-to-eat fresh produce carries the potential for human listeriosis, particularly among individuals with compromised immune systems.

Characteristics of Clinical Isolates of Listeria monocytogenes in Sichuan, China, in 2022 Based on Whole Genome Sequencing Analysis.

Listeria monocytogenes is a foodborne pathogen. In 2022, we collected 15 strains of L. monocytogenes isolated from patients in some foodborne disease sentinel monitoring hospitals in Sichuan Province. Through whole genome sequencing (WGS), we obtained the virulence genes carried by the strains, multi-locus sequence typing (MLST), core genome MLST (cgMLST), clonal complex (CC), and serum groups and constructed a phylogenetic tree and minimum spanning tree with nonhuman strains. An analysis shows that all 15 strains of L. monocytogenes carry virulence genes LIPI-1 and LIPI-2, whereas the carrying rates of LIPI-3 and LIPI-4 virulence genes are relatively low. The MLST typing results showed a total of 10 sequence types (ST), including 10 CCs, with ST7 being the dominant type. The cgMLST clearly distinguishes strains of different lineages and CC types. The serum group is divided into three types: IIa, IIb, and IVb, with IIa being the dominant serum group. An analysis of antibiotic genes showed that all 15 strains carried FosX, lin, mprF, and norB with high carrying rates. The minimum inhibitory concentration results indicated that all were susceptible to eight antibiotics (ampicillin, penicillin, tetracycline, meropenem, erythromycin, vancomycin, ciprofloxacin, and trimethoprim-sulfamethoxazole). The analysis of strains isolated from different sources of Listeria revealed varying degrees of diversity, and the contamination of meat and environment within the province is closely related to clinical cases. L. monocytogenes isolated from clinical cases in Sichuan Province carry multiple virulence and antibiotic genes, with high potential pathogenicity. It is necessary to further strengthen the monitoring and control of food and environment by L. monocytogenes within Sichuan Province.

Surveillance of stewed meat products in Shanghai, China: Importance of controlling processing environment contamination for Listeria monocytogenes

Human listeriosis is a foodborne disease caused by Listeria monocytogenes contamination. Most cases occur due to consumption of ready-to-eat (RTE) foods. Stewed meat in seasoning, a type of pre-packaged ready-to-eat (RTE) food, is a popular traditional product in China. We used whole genome sequencing to analyze core genome multilocus sequence typing (cgMLST), serogroups and multilocus typing sequence typing (ST) of 67 L. monocytogenes isolates. The L. monocytogenes serogroup IIb-ST5 was predominantly isolated from different sources and linked to stewed meat in seasoning of Brand A, B, and C with <7 allelic differences by cgMLST, suggesting that this kind of food may be a high-risk food causing sporadic cases of listeriosis. Furthermore, L. monocytogenes isolates from different batches of stewed meat in seasoning products processed in the same processing plant had <5 five allelic differences, confirming that contamination occurred during manufacture. Additionally, 16 L. monocytogenes isolates from Plant A were the same clone (0 allelic difference), which confirmed the strong ability of L. monocytogenes to survive in the processing environment. Thus, increased and frequent monitoring, as well as constant technical support for the control of L. monocytogenes in food processing plants is necessary.

Prevalence, Antimicrobial Resistance, and Characterization of Listeria Spp. Isolated from Various Sources in Ethiopia: A Comprehensive Review.

Listeriosis is an important foodborne zoonotic disease affecting humans and animals in Ethiopia. This review aims to synthesize the epidemiology, prevalence, distribution, and antimicrobial resistance of Listeria species in the country. The literature reveals a widespread occurrence of Listeria infection in humans, animals, and food products, with an average prevalence of 21.6% for Listeria species and 6.9% for L. monocytogenes. Three sequence types (STs) of L. monocytogenes (2, 145, and 18) and twelve STs of L. innocua (1489, 1619, 603, 537, 1010, 3186, 492, 3007, 1087, 474, 1008, and 637) were reported from milk and dairy products. Contamination rates ranged from 4.1% to 42.9% across livestock, dairy, slaughterhouses, and processing facilities, indicating faults in production practices. Sporadic human listeriosis outbreaks have occurred since 1967, causing meningitis, perinatal infections, and deaths, with recent studies showing L. monocytogenes isolation in up to 10.4% of febrile patients, confirming foodborne transmission. Non-pathogenic Listeria species were also common on farms and in facilities. Ovine listeriosis poses a threat to Ethiopia's sheep and goat industries, with over 40% seroprevalence in some herds. Comprehensive control measures across the food chain are needed to curb contamination and protect public health. Isolates from various foods show antibiotic resistance to first-line agents but susceptibility to others like gentamicin and cephalosporins. In conclusion, this review synthesizes evidence on Listeria distribution in Ethiopia's food system and disease burden, highlighting the need for improved food safety policies and awareness.

Investigation of the seasonal prevalence, phenotypic, and genotypic characteristics of Listeria monocytogenes in slaughterhouses in Burdur.

This study examined Listeria monocytogenes isolates from two slaughterhouses in Burdur province, southern Turkey, over four seasons for antibiotic resistance, serogroups, virulence genes, in vitro biofilm forming capacity, and genetic relatedness. Carcass (540) and environment-equipment surface (180) samples were collected from two slaughterhouses (S1, S2) for 1 year (4 samplings). Of the 89 (12.4%) positive isolates, 48 (53.9%) were from animal carcasses, and 41 (46.1%) from the environment-equipment surfaces. Autumn was the peak season for Listeria monocytogenes compared to summer and spring (P<0.05). In addition, the most common serotype between seasons was 1/2c. Except for plcA and luxS genes, all isolates (100%) harbored inlA, inlC, inlJ, hlyA, actA, iap, flaA genes. Listeria monocytogenes isolates were identified as belonging to IIc (1/2c-3c; 68.5%), IVb (4b-4d-4e; 29.2%), and IIa (1/2a-3a; 2.2%) in the screening using multiplex polymerase chain reaction-based serogrouping test. A total of 65 pulsotypes and 13 clusters with at least 80% homology were determined by using pulsed field gel electrophoresis on samples that had been digested with ApaI. Thirty-four (38.2%) of the isolates were not resistant to any of the 14 antibiotics tested. The antibiotic to which the isolates showed the most resistance was rifampicin (44.9%). Serotype 1/2c was the most resistant serotype to antibiotics. Despite having biofilm-associated genes (inlA, inlB, actA, flaA, and luxS), a minority (11%) of isolates formed weak biofilm. This study revealed seasonal changes prevalence of Listeria monocytogenes, particularly higher in autumn, posing a greater risk of meat contamination. Notably, Serotype 1/2c showed significant prevalence and antibiotic resistance. Indistinguishable isolates indicated cross-contamination, underscoring the importance of prioritized training for slaughterhouse personnel in sanitation and hygiene protocols.

Draft genome of three isolates of Listeria monocytogenes isolated from Stenella coeruleoalba in Italy.

Listeria monocytogenes is the etiological agent of the listeriosis. Here, we described three draft genome sequences of L. monocytogenes isolated in Italy from stranded individuals of the striped dolphin Stenella coeruleoalba. All the genomes have been molecular typed through the multilocus sequence typing to identify the phylogenetic lineage, clonal complex, sublineage, and serogroup.