Low Potential Virulence Associated with Mutations in the inlA and prfA Genes in Listeria monocytogenes Isolated from Raw Retail Poultry Meat

2013 ◽  
Vol 76 (1) ◽  
pp. 129-132 ◽  
Author(s):  
VICTORIA LÓPEZ ◽  
JAIME NAVAS ◽  
JOAQUÍN V. MARTÍNEZ-SUÁREZ
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Pulsed Field Gel Electrophoresis ◽  
Poultry Meat ◽  
Pathogenic Potential ◽  
Molecular Serotyping ◽  
Potential Source ◽  
Raw Foods ◽  
Over Time

Packaged raw foods can represent a potential source of Listeria monocytogenes contamination when opened at home, and listeriosis is associated with the consumption of undercooked raw foods. The aim of this study was to characterize a group of L. monocytogenes strains isolated from 56 packages of raw chicken meat from a single brand in order to determine the diversity of the strains that dominate in a particular food over time, as well as their pathogenic potential. Forty (71%) samples were found to be positive for L. monocytogenes, and three isolates per sample were subjected to PCR molecular serotyping. Subtyping of 45 isolates from different manufacturing dates (n = 40) or different molecular serotype within the same sample (n = 5) identified 11 different L. monocytogenes subtypes as defined by pulsed-field gel electrophoresis and sequencing of virulence genes actA and inlA. Two of the subtypes accounted for 51% of the isolates. About 40% of isolates (three subtypes) were found to potentially present attenuated virulence because of the presence of mutations in the prfA and inlA genes.

scholarly journals Recurrent and Sporadic Listeria monocytogenes Contamination in Alheiras Represents Considerable Diversity, Including Virulence-Attenuated Isolates

2007 ◽  
Vol 73 (12) ◽  
pp. 3887-3895 ◽  
Author(s):  
M. T. S. Fel�cio ◽  
T. Hogg ◽  
P. Gibbs ◽  
P. Teixeira ◽  
M. Wiedmann
Keyword(s):  
Listeria Monocytogenes ◽  
Control Strategies ◽  
Tetracycline Resistance ◽  
The United States ◽  
Pulsed Field Gel Electrophoresis ◽  
Pathogenic Potential ◽  
Molecular Serotyping ◽  
Production And Distribution ◽  
Invasion Assays

ABSTRACT Microbiological characterization of alheiras, traditional smoked meat sausages produced in northern Portugal, had previously shown that more than 60% of the lots analyzed were contaminated with Listeria monocytogenes at levels higher than 100 CFU/g. In order to better understand L. monocytogenes contamination patterns in alheiras, we characterized 128 L. monocytogenes isolates from alheiras using a variety of subtyping techniques (i.e., molecular serotyping; arsenic, cadmium, and tetracycline resistance typing; and pulsed-field gel electrophoresis [PFGE]). Subtyping of isolates from products collected on two separate dates provided evidence for the persistence of specific L. monocytogenes PFGE types in the production and distribution chains of alheiras from four different processors. A subset of 21 isolates was further characterized using ribotyping and Caco-2 cell invasion assays to evaluate the pathogenic potential of L. monocytogenes present in alheiras. Caco-2 invasion assays revealed seven isolates with invasion efficiencies that were less than 20% of that of the control strain 10403S. All seven isolates had premature stop codons in inlA that represented three distinct mutations, which had previously been observed in isolates from the United States or France. Our findings indicate the need for a comprehensive approach to control L. monocytogenes in alheiras, including strategies to reduce persistence. The presence of considerable diversity in invasion phenotypes among L. monocytogenes strains present in alheiras, including the presence of subtypes likely to be virulence attenuated, may provide an opportunity to initially focus control strategies on the subtypes most likely to cause human disease.

Listeria monocytogenes From Farm to Fork in a Brazilian Pork Production Chain

2020 ◽  
Vol 83 (3) ◽  
pp. 485-490 ◽  
Author(s):  
DANILO A. L. SILVA ◽  
CLARISSE V. BOTELHO ◽  
BRUNA T. F. MARTINS ◽  
RAFAELA M. TAVARES ◽  
ANDERSON C. CAMARGO ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Pulsed Field Gel Electrophoresis ◽  
Control Measures ◽  
Production Chain ◽  
Pork Production ◽  
Pulsed Field ◽  
Genetic Profiles ◽  
Different Sources

ABSTRACT Listeria monocytogenes contamination was assessed in different steps of a pork production chain. Ten lots of pigs were sampled at termination barns, at slaughter (after bleeding, after buckling, after evisceration, and after final washing), at processing (knives, deboning tables, and employees' hands), and of end products (ribs, shoulder, ham, and sausage). All samples (n = 670) were subjected to L. monocytogenes detection, and the obtained isolates (n = 18, identified as Listeria spp.) were characterized by their biochemical characteristics, serogroups, virulence genes, pulsed-field gel electrophoresis profiles, antibiotic resistances (ampicillin, penicillin, gentamicin, and sulfamethoxazole-trimethoprim), and adhesion abilities. The results revealed the low occurrence of Listeria spp. in the evaluated pork production chain. However, four tested sausage samples (40%) were positive for Listeria spp., with L. monocytogenes identified in two (20%) of these samples. Ten isolates were identified as L. monocytogenes (eight from serogroup 1/2a or 3a and two from serogroup 4b, 4d, or 4e): all isolates were also positive for the virulence-related genes hlyA, iap, plcA, actA, inlA, inlB, inlC, and inlJ and susceptible to the tested antibiotics. One sausage sample was contaminated by both serogroups 1/2a or 3a and 4b, 4d, or 4e. Isolates from serogroup 1/2a or 3a obtained during visits 5 and 6 presented distinct genetic profiles by pulsed-field gel electrophoresis, indicating that contamination may come from different sources. The adhesion potential exhibited by Listeria spp. isolates (n = 18) ranged from weak (serogroup 4b, 4d, or 4e) to moderate (L. innocua and L. monocytogenes serogroup 1/2a or 3a). Despite the low occurrence of L. monocytogenes, pathogenic serogroups were detected in sausages, demanding control measures by the industry. HIGHLIGHTS

scholarly journals Characterization of epidemic clones of Listeria monocytogenes serotype 4b isolated from humans and meat products in Brazil

2015 ◽  
Vol 9 (09) ◽  
pp. 962-969 ◽  
Author(s):  
André Victor Barbosa ◽  
Aloysio de Mello Figueiredo Cerqueira ◽  
Leonardo Alves Rusak ◽  
Cristhiane Moura Falavina Dos Reis ◽  
Nilma Cintra Leal ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Foodborne Pathogen ◽  
Meat Products ◽  
Human Case ◽  
Pulsed Field Gel Electrophoresis ◽  
Clinical Samples ◽  
Pulsed Field ◽  
Serotype 4B

Introduction: Listeria monocytogenes is an important foodborne pathogen and the 4b serotype is responsible for many cases of human listeriosis reported in Brazil. Several listeriosis outbreaks worldwide have involved a small number of well-defined clonal groups, designated as epidemic clones (ECs). Methodology: We studied 71 strains of serotype 4b, including 25 isolates from human cases of listeriosis and 46 from meat-based foods, collected in Brazil between 1977 and 2010. The presence of ECs (I and II) markers and virulence genes (inlA, inlB, ilnC, inlJ and actA) were evaluated by PCR assay. The genetic relationship of ECs-positive strains was assessed by pulsed field gel electrophoresis. Results: ECI and ECII markers were found both in human and food strains, with 19.7% positive for the ECI marker and 40.8% for ECII. Most strains (97.2%) were positive for the virulence genes that were studied. Nevertheless, the actA gene amplicons showed two distinct sizes, with all ECI positive strains exhibiting a 105bp deletion. Pulsed field gel electrophoresis (PFGE) analysis allowed the recognition of highly related strains, particularly from two outbreaks of neonatal listeriosis in São Paulo State occurred in 1992 and 1997, both ECII-positive; and two ECI strains from a human case (1982) and from bovine meat (2009). Conclusions: The presence of ECs among clinical samples and beef isolates of serotype 4b from some regions of Brazil highlights the need for rigorous control of production procedures. Furthermore, the association of ECII with two nosocomial outbreaks suggests its ability to spread in these settings.

Multiplex PCR Assay Simplifies Serotyping and Sequence Typing of Listeria monocytogenes Associated with Human Outbreaks

2005 ◽  
Vol 68 (9) ◽  
pp. 1907-1910 ◽  
Author(s):  
WEI ZHANG ◽  
STEPHEN J. KNABEL
Keyword(s):  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Rapid Identification ◽  
Pulsed Field Gel Electrophoresis ◽  
Pcr Assay ◽  
Pulsed Field ◽  
Typing Scheme ◽  
Multiplex Pcr Assay

Listeria monocytogenes serotypes 1/2a and 4b are responsible for the majority of cases of human listeriosis worldwide. In this study, a multiplex PCR assay was developed to allow rapid identification and easily interpretable differentiation of serotypes 1/2a and 4b from other serotypes of L. monocytogenes by simultaneously targeting two virulence genes (inlB and inlC) and two serotype-specific genes (ORF2372 and lmo0171). A subsequent gel extraction and sequence typing analysis of the highly polymorphic intragenic regions in inlB and inlC simplified a previously developed multi–virulence-locus sequence typing scheme and provided discriminatory power for subtyping L. monocytogenes similar to pulsed-field gel electrophoresis analysis.

Low Occurrence of Listeria monocytogenes on Bovine Hides and Carcasses in Minas Gerais State, Brazil: Molecular Characterization and Antimicrobial Resistance

2014 ◽  
Vol 77 (7) ◽  
pp. 1148-1152 ◽  
Author(s):  
ANDERSON CARLOS CAMARGO ◽  
ANDREA LAFISCA ◽  
MARCUS VINÍCIUS COUTINHO COSSI ◽  
FREDERICO GERMANO PISCITELLI ALVARENGA LANNA ◽  
MARIANE REZENDE DIAS ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Antimicrobial Resistance ◽  
Multiplex Pcr ◽  
Gel Electrophoresis ◽  
Virulence Genes ◽  
Minas Gerais ◽  
Pulsed Field Gel Electrophoresis ◽  
International Organization ◽  
Minas Gerais State ◽  
Positive Results

Listeria monocytogenes occurrence was assessed in three slaughterhouses located in Minas Gerais state, Brazil, by analysis of 209 bovine carcasses. Four sponge samples were obtained from each carcass in different steps (A, from hide, before bleeding; B, after hide removal; C, after evisceration; and D, after end washing), resulting in a total of 836 samples. The samples were tested for the presence of L. monocytogenes according to the International Organization for Standardization 11290-1, and positive results were recorded in steps A (1 of 209) and D (1 of 209) from slaughterhouse 03. L. monocytogenes isolates (n = 5) were identified by multiplex PCR as belonging to serogroup IIc (representing serotypes 1/2c or 3c) and presented identical pulsed-field gel electrophoresis profiles; in addition, the isolates harbored the virulence genes inlA, inlB, inlC, inlJ, plcA, hlyA, actA, and iap and were sensitive to ampicillin, vancomycin, gentamicin, erythromycin, tetracycline, rifampin, chloramphenicol, trimethoprim, and sulfamethoxazole. The obtained data indicated a low occurrence of L. monocytogenes on bovine hides and carcasses from slaughterhouses located in Minas Gerais state, Brazil, and the presence of distinct virulence makers and susceptibility to a variety of antimicrobials by the obtained isolates.

scholarly journals Caracterização molecular de Listeria monocytogenes oriundas de cortes cárneos bovinos e de abatedouros frigoríficos de bovinos localizados no Distrito Federal, Brasil

2016 ◽  
Vol 36 (10) ◽  
pp. 957-964 ◽  
Author(s):  
Joana M. Palma ◽  
◽  
Rodrigo C. Lisboa ◽  
Dália P. Rodrigues ◽  
André F.M. Santos ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Distrito Federal

RESUMO: Este trabalho teve como objetivo realizar a detecção de cepas de Listeria monocytogenes de cortes cárneos bovinos bem como no ambiente de abatedouros frigoríficos localizados no Distrito Federal, promover a sorotipificação pela reação em cadeia da polimerase (PCR), realizar antibiograma e submeter às cepas à eletroforese de campo pulsado (Pulsed-field gel electrophoresis - PFGE). Foram analisados um total de 125 cortes cárneos bovinos, 45 amostras de swabs de carcaças e 43 amostras de swabs em que foram detectados 13 cepas de Listeria monocytogenes, sendo 11 em cortes cárneos bovinos e 2 swabs de ambiente em um abatedouro frigorifico. Não foram isoladas cepas de swabs de carcaça. Dentre as 13 cepas de Listeria monocytogenes foram encontradas seis cepas do sorotipo 4b, cinco do sorotipo 1/2c e duas cepas do sorotipo 1/2a. Dentre as 11 cepas de L. monocytogenes encontradas em cortes cárneos bovino, uma (9,1%) cepa apresentou resistência a eritromicina, outra (9,1%) cepa a gentamicina e outra a ciprofloxacina (9,1%) e todas as cepas (100%) apresentaram resistência ao Ác. Nalidíxico. Das duas (2) cepas oriundas de ralos de abatedouro frigorífico, todas (100%) apresentaram resistência ao Ác. Nalidíxico e a sulfonamidas. A análise por eletroforese de campo pulsante (PFGE) demonstrou 13 diferentes pulsotipos, em que foram agrupados em 3 diferentes grupos clonais, que coincidentemente se correlacionavam com os 3 diferentes sorotipos encontrados sugerindo uma ampla disseminação desses perfis no Distrito Federal.

Isolation and Pulsed-Field Gel Electrophoresis Typing of Listeria monocytogenes from Modified Atmosphere Packaged Fresh-Cut Vegetables Collected in Ireland

2006 ◽  
Vol 69 (10) ◽  
pp. 2524-2528 ◽  
Author(s):  
GILLIAN A. FRANCIS ◽  
DAVID O'BEIRNE
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Fruits And Vegetables ◽  
Pulsed Field Gel Electrophoresis ◽  
Leafy Vegetables ◽  
Modified Atmosphere ◽  
Pulsed Field ◽  
Fruit Samples ◽  
Fresh Cut ◽  
Pfge Profiles

The incidence of Listeria monocytogenes in modified atmosphere packaged fresh-cut fruits and vegetables from chill cabinets of a supermarket in Ireland was investigated over a 2-year period. Overall, 9.58% of fresh-cut produce was contaminated with Listeria spp. Various species of Listeria were isolated from samples, including L. monocytogenes, L. seeligeri, L. innocua, L. welshimeri, and L. ivanovii. No fruit samples contained detectable L. monocytogenes. Overall, a total of 21 L. monocytogenes isolates (2.9% of samples) were recovered from a range of products, including dry coleslaw mix (80% shredded cabbage and 20% shredded carrot), bean sprouts, and leafy vegetables such iceberg, romaine, and radicchio lettuce and mixed salad leaves (curly endive, escarole, and radicchio leaves). Dry coleslaw mix appeared to have the highest incidence of Listeria contamination (20%) compared with other products. Listeria contamination was more frequent (P < 0.05) during the summer and autumn months than during the winter and spring months. The 21 L. monocytogenes isolates were subsequently subtyped by genomic macrorestriction techniques using ApaI with pulsed-field gel electrophoresis (PFGE). PFGE of digested DNA produced bands of 79 to 518 kb. Four PFGE profiles were identified, and approximately 50% of the isolates were associated with profile 1. This study indicates that fresh-cut vegetables packaged under a modified atmosphere can support growth of numerous species of Listeria, including L. monocytogenes.

scholarly journals Sources of Listeria monocytogenes Contamination in a Cold-Smoked Rainbow Trout Processing Plant Detected by Pulsed-Field Gel Electrophoresis Typing

1999 ◽  
Vol 65 (1) ◽  
pp. 150-155 ◽  
Author(s):  
Tiina Autio ◽  
Sebastian Hielm ◽  
Maria Miettinen ◽  
Anna-Maija Sjöberg ◽  
Kaarina Aarnisalo ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Pulsed Field Gel Electrophoresis ◽  
Hot Water ◽  
Processing Plant ◽  
Pulsed Field ◽  
Eradication Program ◽  
Fish Samples ◽  
Raw Fish

ABSTRACT Sites of Listeria monocytogenes contamination in a cold-smoked rainbow trout (Oncorhynchus mykiss) processing plant were detected by sampling the production line, environment, and fish at different production stages. Two lots were monitored. The frequency of raw fish samples containing L. monocytogenes was low. During processing, the frequency of fish contaminated with L. monocytogenes clearly rose after brining, and the most contaminated sites of the processing plant were the brining and postbrining areas. A total of 303 isolates from the raw fish, product, and the environment were characterized by pulsed-field gel electrophoresis (PFGE). PFGE yielded nine pulsotypes, which formed four clusters. The predominating L. monocytogenespulsotypes of the final product were associated with brining and slicing, whereas contaminants of raw fish were not detected in the final product. Air-mediated contamination in the plant could not be proved. In accordance with these results, an L. monocytogenes eradication program was planned. The use of hot steam, hot air, and hot water seemed to be useful in eliminatingL. monocytogenes. None of the control samples taken in the 5 months after the eradication program was implemented containedL. monocytogenes.

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