title: Spontaneous mutation frequency and emergence of ciprofloxacin resistance in Campylobacter jejuni and Campylobacter coli.
abstract: OBJECTIVE
  We analysed the contribution of spontaneous mutation frequency to the evolution of ciprofloxacin resistance and the diversity of mutations in the quinolone resistance-determining region (QRDR) of gyrA and in the intergenic region, cmeR-cmeA, of the CmeABC efflux pump in Campylobacter jejuni and Campylobacter coli.
  
  
  METHODS
  The mutation frequency was measured in 11 quinolone-susceptible C. jejuni and 5 C. coli strains, with and without ox bile. MICs and target-specific and efflux-associated mutations were studied for a number of colonies of each strain selected from plates containing 1 mg/L ciprofloxacin.
  
  
  RESULTS
  The spontaneous mutation frequency level among susceptible C. jejuni and C. coli strains ranged from hypomutable (approximately 4 x 10(-9)) to strongly mutable (approximately 7 x 10(-3)). Ox bile had no effect on mutation frequency. Pre-existing ampicillin and tetracycline resistance increased the MICs of ciprofloxacin for two strains from 0.032-0.125 to 0.5 mg/L. MICs of ciprofloxacin for the colonies selected from plates containing 1 mg/L ciprofloxacin varied from 1 to 16 mg/L, with the Thr-86-->Ile or Asp-90-->Asn mutation detected in the QRDR of gyrA. In 21.5% (14/65) of the selected colonies, no specific mutations existed. Two types of mutations in CmeR promoter-binding inverted sequences were identified both in the parent strains and in the colonies selected from ciprofloxacin plates.
  
  
  CONCLUSIONS
  The variation in mutation frequencies between most C. jejuni and C. coli strains was up to 700-fold. Mutation in the QRDR of gyrA or in the intergenic region was not associated with differences in spontaneous mutation frequencies. Previously acquired resistance to tetracycline and ampicillin predisposed strains to high-level ciprofloxacin resistance and to multiple antibiotic resistance.
authors.affiliation: Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland. marja-liisa.hanninen@helsinki.fi
authors.forename: Marja-Liisa
authors.initials: ML
authors.lastname: Hänninen
authors.forename: Minna
authors.initials: M
authors.lastname: Hannula
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017000:Campylobacter coli / Q000187:drug effects* / Q000235:genetics; D016123:Campylobacter jejuni / Q000187:drug effects* / Q000235:genetics; D002939:Ciprofloxacin / Q000494:pharmacology*; D027081:DNA Gyrase / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009154:Mutation
chemical_list: D000890:Anti-Infective Agents; D001426:Bacterial Proteins; D021901:DNA, Intergenic; D002939:Ciprofloxacin; D027081:DNA Gyrase