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Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens

Phenotypic bacitracin resistance has been reported in Clostridium perfringens. However, the genes responsible for the resistance have not yet been characterized. Ninety-nine C. perfringens isolates recovered from broilers and turkeys were tested for phenotypic bacitracin resistance. Bacitracin MIC(9...

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Autores principales: Charlebois, Audrey, Jalbert, Louis-Alexandre, Harel, Josée, Masson, Luke, Archambault, Marie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435297/
https://www.ncbi.nlm.nih.gov/pubmed/22970221
http://dx.doi.org/10.1371/journal.pone.0044449
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author Charlebois, Audrey
Jalbert, Louis-Alexandre
Harel, Josée
Masson, Luke
Archambault, Marie
author_facet Charlebois, Audrey
Jalbert, Louis-Alexandre
Harel, Josée
Masson, Luke
Archambault, Marie
author_sort Charlebois, Audrey
collection PubMed
description Phenotypic bacitracin resistance has been reported in Clostridium perfringens. However, the genes responsible for the resistance have not yet been characterized. Ninety-nine C. perfringens isolates recovered from broilers and turkeys were tested for phenotypic bacitracin resistance. Bacitracin MIC(90) (>256 µg/ml) was identical for both turkey and chicken isolates; whereas MIC(50) was higher in turkey isolates (6 µg/ml) than in chicken isolates (3 µg/ml). Twenty-four of the 99 isolates showed high-level bacitracin resistance (MIC breakpoint >256 µg/ml) and the genes encoding for this resistance were characterized in C. perfringens c1261_A strain using primer walking. Sequence analysis and percentages of amino acid identity revealed putative genes encoding for both an ABC transporter and an overproduced undecaprenol kinase in C. perfringens c1261_A strain. These two mechanisms were shown to be both encoded by the putative bcrABD operon under the control of a regulatory gene, bcrR. Efflux pump inhibitor thioridazine was shown to increase significantly the susceptibility of strain c1261_A to bacitracin. Upstream and downstream from the bcr cluster was an IS1216-like element, which may play a role in the dissemination of this resistance determinant. Pulsed-field gel electrophoresis with prior double digestion with I-CeuI/MluI enzymes followed by hybridization analyses revealed that the bacitracin resistance genes bcrABDR were located on the chromosome. Semi-quantitative RT-PCR demonstrated that this gene cluster is expressed under bacitracin stress. Microarray analysis revealed the presence of these genes in all bacitracin resistant strains. This study reports the discovery of genes encoding for a putative ABC transporter and an overproduced undecaprenol kinase associated with high-level bacitracin resistance in C. perfringens isolates from turkeys and broiler chickens.
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spelling pubmed-34352972012-09-11 Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens Charlebois, Audrey Jalbert, Louis-Alexandre Harel, Josée Masson, Luke Archambault, Marie PLoS One Research Article Phenotypic bacitracin resistance has been reported in Clostridium perfringens. However, the genes responsible for the resistance have not yet been characterized. Ninety-nine C. perfringens isolates recovered from broilers and turkeys were tested for phenotypic bacitracin resistance. Bacitracin MIC(90) (>256 µg/ml) was identical for both turkey and chicken isolates; whereas MIC(50) was higher in turkey isolates (6 µg/ml) than in chicken isolates (3 µg/ml). Twenty-four of the 99 isolates showed high-level bacitracin resistance (MIC breakpoint >256 µg/ml) and the genes encoding for this resistance were characterized in C. perfringens c1261_A strain using primer walking. Sequence analysis and percentages of amino acid identity revealed putative genes encoding for both an ABC transporter and an overproduced undecaprenol kinase in C. perfringens c1261_A strain. These two mechanisms were shown to be both encoded by the putative bcrABD operon under the control of a regulatory gene, bcrR. Efflux pump inhibitor thioridazine was shown to increase significantly the susceptibility of strain c1261_A to bacitracin. Upstream and downstream from the bcr cluster was an IS1216-like element, which may play a role in the dissemination of this resistance determinant. Pulsed-field gel electrophoresis with prior double digestion with I-CeuI/MluI enzymes followed by hybridization analyses revealed that the bacitracin resistance genes bcrABDR were located on the chromosome. Semi-quantitative RT-PCR demonstrated that this gene cluster is expressed under bacitracin stress. Microarray analysis revealed the presence of these genes in all bacitracin resistant strains. This study reports the discovery of genes encoding for a putative ABC transporter and an overproduced undecaprenol kinase associated with high-level bacitracin resistance in C. perfringens isolates from turkeys and broiler chickens. Public Library of Science 2012-09-06 /pmc/articles/PMC3435297/ /pubmed/22970221 http://dx.doi.org/10.1371/journal.pone.0044449 Text en © 2012 Charlebois et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Charlebois, Audrey
Jalbert, Louis-Alexandre
Harel, Josée
Masson, Luke
Archambault, Marie
Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title_full Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title_fullStr Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title_full_unstemmed Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title_short Characterization of Genes Encoding for Acquired Bacitracin Resistance in Clostridium perfringens
title_sort characterization of genes encoding for acquired bacitracin resistance in clostridium perfringens
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435297/
https://www.ncbi.nlm.nih.gov/pubmed/22970221
http://dx.doi.org/10.1371/journal.pone.0044449
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