Evidence for an efflux pump in multidrug-resistant Campylobacter jejuni.
Mechanisms of drug resistance in Campylobacter jejuni were investigated. Mutant strains 34PEFr, which was resistant to pefloxacin (128-fold increase in the MIC), and 34CTXr, which was resistant to cefotaxime (32-fold increase in the MIC) and which was derived from the susceptible parent 34s, were obtained by serial passages on pefloxacin and cefotaxime gradient plates, respectively. Both mutants showed cross-resistance to erythromycin, chloramphenicol, tetracycline, beta-lactams, and quinolones. While the quinolone resistance of strain PEFr could be explained by a mutation at codon 86 of the gyrA gene, the multidrug resistance phenotype of both strains was further investigated. Accumulation of pefloxacin, ciprofloxacin, and minocycline was measured by fluorometry and was found to be lower in the mutant strains than in the parent strain. Preincubation of the cells with carbonyl cyanide m-chlorophenylhydrazone, however, completely abolished this difference. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of outer membrane preparations from both mutant strains showed overexpression of two proteins of 55 and 39 kDa which were absent from the outer membranes of the wild-type strain. These results indicate that in C. jejuni 34PEFr and 34CTXr, multidrug resistance is associated with an efflux system with a broad specificity.
Charvalos
E
E
Department of Bacteriology, Parasitology, Zoonoses and Geographical Medicine, School of Health Sciences, University of Crete, Herakleion, Greece.
Tselentis
Y
Y
Hamzehpour
M M
MM
Köhler
T
T
Pechere
J C
JC
D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D000890:Anti-Infective Agents / Q000378:metabolism / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D002439:Cefotaxime / Q000378:metabolism / Q000494:pharmacology; D002511:Cephalosporins / Q000378:metabolism / Q000494:pharmacology; D018432:Drug Resistance, Multiple / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D015366:Pefloxacin / Q000378:metabolism / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D001618:beta-Lactamases / Q000378:metabolism
D000900:Anti-Bacterial Agents; D000890:Anti-Infective Agents; D001425:Bacterial Outer Membrane Proteins; D002511:Cephalosporins; D015366:Pefloxacin; D001618:beta-Lactamases; D002439:Cefotaxime