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Characterization of genes encoding for acquired bacitracin resistance in Clostridium perfringens

Audrey Charlebois1, Louis-Alexandre Jalbert, Josée Harel

  • 1Department of pathology and microbiology, Faculty of Veterinary Medicine of the University of Montreal, Centre de Recherche en Infectiologie Porcine, Saint-Hyacinthe, Québec, Canada.

Plos One
|September 13, 2012
PubMed

Insights

Genes responsible for high-level bacitracin resistance in Clostridium perfringens were identified. The study found an ABC transporter and undecaprenol kinase, encoded by the bcrABD operon, confer resistance in poultry isolates.

Area of Science:

  • Microbiology
  • Genetics
  • Antimicrobial Resistance

Background:

  • Phenotypic bacitracin resistance is observed in Clostridium perfringens.
  • The genetic basis for this resistance in C. perfringens remains largely uncharacterized.
  • Bacitracin resistance is a concern in poultry production due to its use as a feed additive.

Purpose of the Study:

  • To characterize the genes responsible for high-level bacitracin resistance in Clostridium perfringens.
  • To investigate the mechanisms of bacitracin resistance in C. perfringens isolates from poultry.
  • To identify the genetic elements and regulatory mechanisms involved in bacitracin resistance.

Main Methods:

  • Phenotypic testing of 99 C. perfringens isolates for bacitracin resistance (MIC determination).
  • Genetic characterization using primer walking, sequence analysis, and amino acid identity comparisons.
  • Molecular techniques including pulsed-field gel electrophoresis, hybridization, semi-quantitative RT-PCR, and microarray analysis.
  • Assessment of efflux pump inhibitor (thioridazine) effects on bacitracin susceptibility.

Main Results:

  • High-level bacitracin resistance (MIC >256 µg/ml) was observed in 24 out of 99 C. perfringens isolates.
  • Putative genes encoding an ABC transporter (bcrA) and overproduced undecaprenol kinase (bcrB) were identified in strain c1261_A.
  • These genes, along with a regulatory gene (bcrR), form the bcrABD operon, located on the chromosome.
  • The bcrABD cluster is expressed under bacitracin stress and present in all resistant strains.
  • An IS1216-like element was found flanking the bcr cluster, suggesting a role in dissemination.

Conclusions:

  • The bcrABD operon, encoding an ABC transporter and undecaprenol kinase, is responsible for high-level bacitracin resistance in Clostridium perfringens.
  • This resistance mechanism is present in C. perfringens isolates from both turkeys and broiler chickens.
  • The findings provide insights into the genetic basis of bacitracin resistance in a significant poultry pathogen.

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