Priming with biocides: a pathway to antibiotic resistance?

Aims

To investigate the priming effects of sub-inhibitory concentrations of biocides on antibiotic resistance in bacteria.

Methods and results

Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus were exposed to sub-inhibitory concentrations of biocides via a gradient plate method. Minimum inhibitory concentration (MIC) and antibiotic susceptibility were determined, and efflux pump inhibitors (thioridazine and chlorpromazine) were used to investigate antibiotic resistance mechanism(s).

Escherichia coli displayed a 2-fold increase in MIC (32 to 64 mg l-1) to H2O2 which was stable after 15 passages, but lost after 6 weeks, and P. aeruginosa displayed a 2-fold increase in MIC (64 to 128 mg l-1) to BZK which was also stable for 15 passages. There were no other tolerances observed to biocides in E. coli, P. aeruginosa or S. aureus, however stable cross-resistance to antibiotics was observed in the absence of a stable increased tolerance to biocides. Six-fold increases in MIC to cephalothin and four-fold to ceftriaxone and ampicillin were observed in hydrogen peroxide primed E. coli. Chlorhexidine primed S. aureus showed a four-fold increase in MIC to oxacillin, and glutaraldehyde-primed P. aeruginosa showed four-fold (sulphatriad) and eight-fold (ciprofloxacin) increases in MIC. Thioridazine increased the susceptibility of E. coli to cephalothin and cefoxitin by four and two-fold respectively, and both thioridazine and chlorpromazine increased the susceptibility S. aureus to oxacillin by eight and four-fold respectively.

Conclusions

These findings demonstrate that sub-inhibitory concentrations of biocides can prime bacteria to become resistant to antibiotics even in the absence of stable biocide tolerance and suggests activation of efflux mechanisms may be a contributory factor.

Significance and impact of the study

This study demonstrates the effects of low-level exposure of biocides (priming) on antibiotic resistance even in the absence of obvious increased biocidal tolerance.