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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| 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. |
| doi_str_mv | 10.1371/journal.pone.0044449 |
| format | Article |
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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.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0044449</identifier><identifier>PMID: 22970221</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>ABC transporter ; Amino acid sequence ; Amino acids ; Analysis ; Animals ; Antibiotics ; Antimicrobial agents ; Bacillus licheniformis ; Bacillus subtilis ; Bacitracin ; Bacitracin - pharmacology ; Bacteria ; Bacterial proteins ; Biology ; Blotting, Southern ; Breakpoints ; Chickens ; Chromosomes ; Clostridium perfringens ; Clostridium perfringens - drug effects ; Clostridium perfringens - genetics ; Clusters ; Digestion ; DNA microarrays ; Drug Resistance, Microbial - genetics ; E coli ; Efflux ; Electrophoresis, Gel, Pulsed-Field ; Enterococcus faecalis ; Enzymes ; Escherichia coli ; Food ; Gel electrophoresis ; Gene mapping ; Genes ; Genes, Bacterial ; Genetic aspects ; Hybridization ; Hybridization analysis ; Kinases ; Levels ; Medicine ; Microbial Sensitivity Tests ; Minimum inhibitory concentration ; Molecular Sequence Data ; Mutation ; Nucleic Acid Hybridization ; Oligonucleotide Array Sequence Analysis ; Operons ; Pathology ; Phosphatase ; Plasmids ; Polymerase Chain Reaction ; Poultry ; Primers ; Pulsed-field gel electrophoresis ; Stress ; Thioridazine ; Turkeys ; Undecaprenol kinase ; Veterinary medicine ; Veterinary Science</subject><ispartof>PloS one, 2012-09, Vol.7 (9), p.e44449-e44449</ispartof><rights>COPYRIGHT 2012 Public Library of Science</rights><rights>Charlebois et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: https://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2012 Charlebois et al 2012 Charlebois et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c725t-8efc622f2143e6bf12c3e9aa8cf8e1fdf05399ab07ff6554601a075c794e4edc3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435297/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435297/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79342,79343</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22970221$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Charlebois, Audrey</creatorcontrib><creatorcontrib>Jalbert, Louis-Alexandre</creatorcontrib><creatorcontrib>Harel, Josée</creatorcontrib><creatorcontrib>Masson, Luke</creatorcontrib><creatorcontrib>Archambault, Marie</creatorcontrib><title>Characterization of genes encoding for acquired bacitracin resistance in Clostridium perfringens</title><title>PloS one</title><addtitle>PLoS One</addtitle><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.</description><subject>ABC transporter</subject><subject>Amino acid sequence</subject><subject>Amino acids</subject><subject>Analysis</subject><subject>Animals</subject><subject>Antibiotics</subject><subject>Antimicrobial agents</subject><subject>Bacillus licheniformis</subject><subject>Bacillus subtilis</subject><subject>Bacitracin</subject><subject>Bacitracin - pharmacology</subject><subject>Bacteria</subject><subject>Bacterial proteins</subject><subject>Biology</subject><subject>Blotting, Southern</subject><subject>Breakpoints</subject><subject>Chickens</subject><subject>Chromosomes</subject><subject>Clostridium perfringens</subject><subject>Clostridium perfringens - drug effects</subject><subject>Clostridium perfringens - genetics</subject><subject>Clusters</subject><subject>Digestion</subject><subject>DNA microarrays</subject><subject>Drug Resistance, Microbial - genetics</subject><subject>E coli</subject><subject>Efflux</subject><subject>Electrophoresis, Gel, Pulsed-Field</subject><subject>Enterococcus faecalis</subject><subject>Enzymes</subject><subject>Escherichia coli</subject><subject>Food</subject><subject>Gel electrophoresis</subject><subject>Gene mapping</subject><subject>Genes</subject><subject>Genes, Bacterial</subject><subject>Genetic aspects</subject><subject>Hybridization</subject><subject>Hybridization analysis</subject><subject>Kinases</subject><subject>Levels</subject><subject>Medicine</subject><subject>Microbial Sensitivity Tests</subject><subject>Minimum inhibitory concentration</subject><subject>Molecular Sequence Data</subject><subject>Mutation</subject><subject>Nucleic Acid Hybridization</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Operons</subject><subject>Pathology</subject><subject>Phosphatase</subject><subject>Plasmids</subject><subject>Polymerase Chain Reaction</subject><subject>Poultry</subject><subject>Primers</subject><subject>Pulsed-field gel electrophoresis</subject><subject>Stress</subject><subject>Thioridazine</subject><subject>Turkeys</subject><subject>Undecaprenol kinase</subject><subject>Veterinary medicine</subject><subject>Veterinary 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one</jtitle><addtitle>PLoS One</addtitle><date>2012-09-06</date><risdate>2012</risdate><volume>7</volume><issue>9</issue><spage>e44449</spage><epage>e44449</epage><pages>e44449-e44449</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>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.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>22970221</pmid><doi>10.1371/journal.pone.0044449</doi><tpages>e44449</tpages><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_plos_journals_1326544679 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | ABC transporter Amino acid sequence Amino acids Analysis Animals Antibiotics Antimicrobial agents Bacillus licheniformis Bacillus subtilis Bacitracin Bacitracin - pharmacology Bacteria Bacterial proteins Biology Blotting, Southern Breakpoints Chickens Chromosomes Clostridium perfringens Clostridium perfringens - drug effects Clostridium perfringens - genetics Clusters Digestion DNA microarrays Drug Resistance, Microbial - genetics E coli Efflux Electrophoresis, Gel, Pulsed-Field Enterococcus faecalis Enzymes Escherichia coli Food Gel electrophoresis Gene mapping Genes Genes, Bacterial Genetic aspects Hybridization Hybridization analysis Kinases Levels Medicine Microbial Sensitivity Tests Minimum inhibitory concentration Molecular Sequence Data Mutation Nucleic Acid Hybridization Oligonucleotide Array Sequence Analysis Operons Pathology Phosphatase Plasmids Polymerase Chain Reaction Poultry Primers Pulsed-field gel electrophoresis Stress Thioridazine Turkeys Undecaprenol kinase Veterinary medicine Veterinary Science |
| title | Characterization of genes encoding for acquired bacitracin resistance in Clostridium perfringens |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-09T07%3A09%3A57IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Characterization%20of%20genes%20encoding%20for%20acquired%20bacitracin%20resistance%20in%20Clostridium%20perfringens&rft.jtitle=PloS%20one&rft.au=Charlebois,%20Audrey&rft.date=2012-09-06&rft.volume=7&rft.issue=9&rft.spage=e44449&rft.epage=e44449&rft.pages=e44449-e44449&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0044449&rft_dat=%3Cgale_plos_%3EA543310377%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1326544679&rft_id=info:pmid/22970221&rft_galeid=A543310377&rft_doaj_id=oai_doaj_org_article_9a1b88f138964b6caa0b576df79d8038&rfr_iscdi=true |