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Virulence and Antibiotic Resistance Genes in Listeria monocytogenes Strains Isolated From Ready-to-Eat Foods in Chile.

ABSTRACT: Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS). The overall positivity of L. monocytogenes in RTE food samples was 3.1% and 14 strains were isolated. L. monocytogenes ST8, ST2763, ST1, ST3, ST5, ST7, ST9, ST14, ST193, and ST451 sequence types were identified by average nucleotide identity, ribosomal multilocus sequence typing (rMLST), and core genome MLST. Seven isolates had serotype 1/2a, five 1/2b, one 4b, and one 1/2c. Three strains exhibited in vitro resistance to ampicillin and 100% of the strains carried the fosX, lin, norB, mprF, tetA, and tetC resistance genes. In addition, the arsBC, bcrBC, and clpL genes were detected, which conferred resistance to stress and disinfectants. All strains harbored hlyA, prfA, and inlA genes almost thirty-two the showed the bsh, clpCEP, hly, hpt, iap/cwhA, inlA, inlB, ipeA, lspA, mpl, plcA, pclB, oat, pdgA, and prfA genes. One isolate exhibited a type 11 premature stop codon (PMSC) in the inlA gene and another isolate a new mutation (deletion of A in position 819). The Inc18(rep25), Inc18(rep26), and N1011A plasmids and MGEs were found in nine isolates. Ten isolates showed CAS-Type II-B systems; in addition, Anti-CRISPR AcrIIA1 and AcrIIA3 phage-associated systems were detected in three genomes. These virulence and antibiotic resistance traits in the strains isolated in the RTE foods indicate a potential public health risk for consumers.

SUBMITTER: Parra-Flores J

PROVIDER: S-EPMC8921925 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature

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Virulence and Antibiotic Resistance Genes in Listeria monocytogenes Strains Isolated From Ready-to-Eat Foods in Chile.

Parra-Flores Julio J Holý Ondrej O Bustamante Fernanda F Lepuschitz Sarah S Pietzka Ariane A Contreras-Fernández Alejandra A Castillo Claudia C Ovalle Catalina C Alarcón-Lavín María Paula MP Cruz-Córdova Ariadnna A Xicohtencatl-Cortes Juan J Mancilla-Rojano Jetsi J Troncoso Miriam M Figueroa Guillermo G Ruppitsch Werner W

Frontiers in microbiology 20220221

Listeria monocytogenes is causing listeriosis, a rare but severe foodborne infection. Listeriosis affects pregnant women, newborns, older adults, and immunocompromised individuals. Ready-to-eat (RTE) foods are the most common sources of transmission of the pathogen This study explored the virulence factors and antibiotic resistance in L. monocytogenes strains isolated from ready-to-eat (RTE) foods through in vitro and in silico testing by whole-genome sequencing (WGS) ...[more]

PMID: 35299835

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Project description:Listeria monocytogenes is a pathogen transmitted through food that can cause severe infections in high-risk groups such as pregnant women, elderly, young children and immunocompromised individuals. It is a ubiquitous bacterium that can survive in harsh conditions, such as dry environments, at low temperatures, in brine conditions and at low pH values. It also has the capacity to form biofilms, which makes it particularly successful even in colonizing surfaces within food processing plants. This study analyzed the presence of L. monocytogenes in ready-to-eat food (RTE) such as sausage, cheese, fresh salads, and other types of raw food. 850 samples of refrigerated and packaged food collected in 2008 and 2009 were analyzed. It was found that 25% of these samples were contaminated with L. monocytogenes strains. Serotyping and virulence genes detection by polymerase chain reaction (PCR) identified that strains belonging to serotype 4b, and containing one or more genes encoded by pathogenicity island (LIPI-1), were significantly associated with specific food types. Furthermore, using pulse field gel electrophoresis (PFGE), it was possible to associate isolates from cheese with strains from clinical cases of listeriosis outbreaks that occurred during the same time period within the same geographic regions. In addition, a strong correlation was observed between isolates from frozen seafood and from clinical strains obtained from sporadic cases of listeriosis. In agreement with reports described in other countries, our results shown that Chilean strains of L. monocytogenes from food products include the most virulent serotypes, encoding for the main virulence genes of the LIPI-1, and were clonally related to clinical isolates from sporadic cases and outbreaks of listeriosis. In conclusion, we show that Chilean isolates of L. monocytogenes from RTE and raw food products can cause disease in humans, representing a public health risk that justifies permanent surveillance.

Project description:Antimicrobial resistance is an existential threat to the health sector, with far-reaching consequences in managing microbial infections. In this study, one hundred and ninety-four Listeria monocytogenes isolates were profiled for susceptibility using disc diffusion techniques. Possible foodborne listeriosis risk associated with ready-to-eat (RTE) foods (RTEF) and the risk of empirical treatment (EMPT) of L. monocytogenes infections, using multiple antimicrobial resistance indices (MARI) and antimicrobial resistance indices (ARI), respectively, were investigated. Twelve European Committee on Antimicrobial Susceptibility Testing (EUCAST) prescribed/recommended antimicrobials (EPAS) for the treatment of listeriosis and ten non-prescribed antimicrobials (non-PAS)] were evaluated. Antimicrobial resistance > 50% against PAs including sulfamethoxazole (61.86%), trimethoprim (56.19%), amoxicillin (42.27%), penicillin (41.24%), and erythromycin (40.21%) was observed. Resistance > 50% against non-PAS, including oxytetracycline (60.89%), cefotetan (59.28%), ceftriaxone (53.09%), and streptomycin (40.21%) was also observed. About 55.67% and 65.46% of the isolates had MARI scores ranging from 0.25-0.92 and 0.30-0.70 for EPAs and non-PAs, respectively. There was a significant difference (p < 0.01) between the MARI scores of the isolates for EPAs and non-PAs (means of 0.27 ± 0.21 and 0.31 ± 0.14, respectively). MARI/ARI scores above the Krumperman permissible threshold (>0.2) suggested a high risk/level of antimicrobial-resistant L. monocytogenes. The MARI risks of the non-success of empirical treatment (EMPT) attributed to EPAs and non-PAs were generally high (55.67% and 65.463%, respectively) due to the antimicrobial resistance of the isolates. MARI-based estimated success and non-success of EMPT if EUCAST-prescribed antimicrobials were administered for the treatment of listeriosis were 44.329% and 55.67%, respectively. The EMPT if non-prescribed antimicrobials were administered for the treatment of listeriosis was 34.53% and 65.46%, respectively. This indicates a potentially high risk with PAs and non-PAs for the treatment of L. monocytogenes infection. Furthermore, ARI scores ≤ 0.2 for EPAs were observed in polony, potato chips, muffins, and assorted sandwiches, whereas ARI scores for non-PAs were >0.2 across all the RTE food types. The ARI-based estimate identified potential risks associated with some RTE foods, including fried fish, red Vienna sausage, Russian sausage, fruit salad, bread, meat pies, fried chicken, cupcakes, and vetkoek. This investigation identified a high risk of EMPT due to the presence of antimicrobial-resistant L. monocytogenes in RTE foods, which could result in severe health consequences.

Project description:Listeria monocytogenes is a foodborne pathogen with a high mortality rate in humans. This study aimed to identify the pathogenic potential of L. monocytogenes isolated from ready-to-eat (RTE) foods and pasteurized milk in China on the basis of its phenotypic and genotypic characteristics. Approximately 7.7% (44/570) samples tested positive for L. monocytogenes among 10.8% (39/360) RTE and 2.4% (5/210) pasteurized milk samples, of which 77.3% (34/44) had < 10 MPN/g, 18.2% (8/44) had 10-110 MPN/g, and 4.5% (2/44) had > 110 MPN/g. A total of 48 strains (43 from RTE foods and five from milk samples) of L. monocytogenes were isolated from 44 positive samples. PCR-serogroup analysis revealed that the most prevalent serogroup was II.2 (1/2b-3b-7), accounting for 52.1% (25/48) of the total, followed by serogroup I.1 (1/2a-3a) accounting for 33.3% (16/48), serogroup I.2 (1/2c-3c) accounting for 12.5% (6/48), and serogroup II.1 (4b-4d-4e) accounting for 2.1%. All isolates were grouped into 11 sequence types (STs) belonging to 10 clonal complexes (CCs) and one singleton (ST619) via multi-locus sequence typing. The most prevalent ST was ST87 (29.2%), followed by ST8 (22.9%), and ST9 (12.5%). Virulence genes determination showed that all isolates harbored eight virulence genes belonging to Listeria pathogenicity islands 1 (LIPI-1) (prfA, actA, hly, mpl, plcA, plcB, and iap) and inlB. Approximately 85.4% isolates carried full-length inlA, whereas seven isolates had premature stop codons in inlA, six of which belonged to ST9 and one to ST5. Furthermore, LLS (encoded by llsX gene, representing LIPI-3) displays bactericidal activity and modifies the host microbiota during infection. LIPI-4 enhances neural and placental tropisms of L. monocytogenes. Results showed that six (12.5%) isolates harbored the llsX gene, and they belonged to ST1/CC1, ST3/CC3, and ST619. Approximately 31.3% (15/48) isolates (belonging to ST87/CC87 and ST619) harbored ptsA (representing LIPI-4), indicating the potential risk of this pathogen. Antimicrobial susceptibility tests revealed that > 95% isolates were susceptible to 16 antimicrobials; however, 60.4 and 22.9% isolates were intermediately resistant to streptomycin and ciprofloxacin, respectively. The results show that several isolates harbor LIPI-3 and LIPI-4 genes, which may be a possible transmission route for Listeria infections in consumers.

Dataset Information

Virulence and Antibiotic Resistance Genes in Listeria monocytogenes Strains Isolated From Ready-to-Eat Foods in Chile.

Publications

Virulence and Antibiotic Resistance Genes in <i>Listeria monocytogenes</i> Strains Isolated From Ready-to-Eat Foods in Chile.

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