title: Comparative transcription analysis and toxin production of two fluoroquinolone-resistant mutants of Clostridium perfringens.
abstract: BACKGROUND
  Fluoroquinolone use has been listed as a risk factor for the emergence of virulent clinical strains of some bacteria. The aim of our study was to evaluate the effect of fluoroquinolone (gatifloxacin) resistance selection on differential gene expression, including the toxin genes involved in virulence, in two fluoroquinolone-resistant strains of Clostridium perfringens by comparison with their wild-type isogenic strains.
  
  
  RESULTS
  DNA microarray analyses were used to compare the gene transcription of two wild types, NCTR and ATCC 13124, with their gatifloxacin-resistant mutants, NCTRR and 13124R. Transcription of a variety of genes involved in bacterial metabolism was either higher or lower in the mutants than in the wild types. Some genes, including genes for toxins and regulatory genes, were upregulated in NCTRR and downregulated in 13124R. Transcription analysis by quantitative real-time PCR (qRT-PCR) confirmed the altered expression of many of the genes that were affected differently in the fluoroquinolone-resistant mutants and wild types. The levels of gene expression and enzyme production for the toxins phospholipase C, perfringolysin O, collagenase and clostripain had decreased in 13124R and increased in NCTRR in comparison with the wild types. After centrifugation, the cytotoxicity of the supernatants of NCTRR and 13224R cultures for mouse peritoneal macrophages confirmed the increased cytotoxicity of NCTRR and the decreased cytotoxicity of 13124R in comparison with the respective wild types. Fluoroquinolone resistance selection also affected cell shape and colony morphology in both strains.
  
  
  CONCLUSIONS
  Our results indicate that gatifloxacin resistance selection was associated with altered gene expression in two C. perfringens strains and that the effect was strain-specific. This study clearly demonstrates that bacterial exposure to fluoroquinolones may affect virulence (toxin production) in addition to drug resistance.
authors: Division of Microbiology, National Center for Toxicological Research, US Food and Drug Administration, Jefferson, AR, USA.
authors: Sunny
authors: S
authors: Park
authors: Miseon
authors: M
authors: Park
authors: Fatemeh
authors: F
authors: Rafii
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001427:Bacterial Toxins / Q000378:metabolism*; D003016:Clostridium perfringens / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D024841:Fluoroquinolones / Q000494:pharmacology*; D046228:Microarray Analysis; D060888:Real-Time Polymerase Chain Reaction; D012641:Selection, Genetic; D059467:Transcriptome
chemical_list: D000900:Anti-Bacterial Agents; D001427:Bacterial Toxins; D024841:Fluoroquinolones