Phenotypic and Genotypic Characterization of Food Animal Isolates ofSalmonellawith Reduced Sensitivity to Ciprofloxacin

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Abstract
Reports of nontyphoidal Salmonella enterica subsp. enterica showing reduced sensitivity to ciprofloxacin (RSC) have increased rapidly during the past decade. Infection in humans with Salmonella possessing RSC may compromise the effectiveness of ciprofloxacin therapy. Nineteen among 4,357 Salmonella strains isolated from food animals in Canada from 1998 to 1999 showed RSC; 17 were from turkeys and 2 from chickens. All were resistant to nalidixic acid and sulfisoxazole and possessed RSC at a level of 0.125-0.5 microg/ml. PCR-RFLP of the gyrA quinolone resistance-determining region (QRDR) with Hinfl revealed that S. Bredeney and S. Heidelberg isolates possessed a mutation in this region. Single-strand conformational polymorphism (SSCP) analysis showed that S. Schwarzengrund and S. Senftenberg isolates also possessed a point mutation in the QRDR. DNA sequencing confirmed the findings and showed that all isolates possessed a base substitution in the gyrA QRDR. Sequencing revealed no mutations in the gyrB and silent wobble mutations in the parC QRDR. Reserpine, a known efflux pump inhibitor, did not effect the MICs for ciprofloxacin, nalidixic acid, and tetracycline. The mar operon could be induced in all isolates at 37 degrees C and in 18 of 19 at 30 degrees C; induction resulted in a two- to four-fold increase in the MIC of ciprofloxacin. In 14 of the 19 isolates, the mutation rate was two-fold or higher than in a ciprofloxacin sensitive S. Bredeney and S. Typhimurium LT2 control strain. Examination of clonal relatedness using pulsed-field gel electrophoresis (PFGE) and plasmid profiles indicated that some degree of clonal dispersion may have occurred, but the majority of isolates may have arisen from de novo mutations.

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