Effect of triclosan or a phenolic farm disinfectant on the selection of antibiotic-resistant Salmonella enterica.
OBJECTIVE
To determine the effect of growth of five strains of Salmonella enterica and their isogenic multiply antibiotic-resistant (MAR) derivatives with a phenolic farm disinfectant or triclosan (biocides) upon the frequency of mutation to resistance to antibiotics or cyclohexane.


METHODS
Strains were grown in broth with or without the biocides and then spread on to agar containing ampicillin, ciprofloxacin or tetracycline each at 4x MIC or agar overlaid with cyclohexane. Incubation was for 24 and 48 h and the frequency of mutation to resistance was calculated for strains with and without prior growth with the biocides. MICs were determined and the presence of mutations in the acrR and marR regions was determined by sequencing and the presence of mutations in gyrA by light-cycler analysis, for a selection of the mutants that arose.


RESULTS
The mean frequency of mutation to antibiotic or cyclohexane resistance was increased approximately 10- to 100-fold by prior growth with the phenolic disinfectant or triclosan. The increases were statistically significant for all antibiotics and cyclohexane following exposure to the phenolic disinfectant (P </= 0.013), and for ampicillin and cyclohexane following exposure to triclosan (P </= 0.009). Mutants inhibited by >1 mg/L ciprofloxacin arose only from strains that were MAR. Reduced susceptibility to ciprofloxacin (at 4x MIC for parent strains) alone was associated with mutations in gyrA. MAR mutants did not contain mutations in the acrR or marR region.


CONCLUSIONS
These data renew fears that the use of biocides may lead to an increased selective pressure towards antibiotic resistance.
Department of Food and Environmental Safety, Veterinary Laboratories Agency (Weybridge), New Haw, Addlestone, Surrey, UK. l.randall@vla.defra.gsi.gov.uk
L P
LP
Randall
S W
SW
Cooles
L J V
LJ
Piddock
M J
MJ
Woodward
D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000891:Anti-Infective Agents, Local / Q000494:pharmacology*; D002622:Chemistry, Agricultural; D002939:Ciprofloxacin / Q000494:pharmacology; D003510:Cyclohexanes / Q000494:pharmacology; D027081:DNA Gyrase / Q000235:genetics; D017931:DNA Primers; D004202:Disinfectants / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D026901:Membrane Transport Proteins / Q000235:genetics; D009154:Mutation / Q000235:genetics / Q000502:physiology; D010636:Phenols / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019779:Salmonella enterica / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D014260:Triclosan / Q000494:pharmacology*
C477884:AcrR protein, Klebsiella pneumoniae; D000900:Anti-Bacterial Agents; D000891:Anti-Infective Agents, Local; D003510:Cyclohexanes; D017931:DNA Primers; D004202:Disinfectants; D026901:Membrane Transport Proteins; D010636:Phenols; D014260:Triclosan; D002939:Ciprofloxacin; D027081:DNA Gyrase