Adaptive resistance to biocides in Salmonella enterica and Escherichia coli O157 and cross-resistance to antimicrobial agents

Abstract

The mechanisms by which bacteria resist killing by antibiotics and biocides are still poorly defined, although repeated exposure to sublethal concentrations of antibacterial agents undoubtedly contributes to their development. This study aimed both to investigate the potential of Salmonella enterica and Escherichia coli O157 for adaptive resistance to commonly used biocides and to determine any cross-resistance to antibiotics. Strains were repeatedly passaged in media containing increasing concentrations of a biocide or antibiotic until adaptive resistance was obtained. A wide panel of antimicrobial agents was then screened by using the adapted strain to determine cross-resistance, if any. Adaptive resistance was readily achieved for both S. enterica and E. coli O157. Cross-resistance in adaptively resistant S. enterica varied with the serotype; Salmonella enterica serovar Enteritidis expressed cross-resistance to chloramphenicol, whereas Salmonella enterica serovar Typhimurium expressed cross-resistance to chlorhexidine. Benzalkonium chloride-resistant Salmonella enterica serovar Virchow showed elevated resistance to chlorhexidine; however, chlorhexidine-resistant Salmonella serovar Virchow did not demonstrate reciprocal cross-resistance to benzalkonium chloride, suggesting specific rather than generic resistance mechanisms. E. coli O157 strains acquired high levels of resistance to triclosan after only two sublethal exposures and, when adapted, repeatedly demonstrated decreased susceptibilities to various antimicrobial agents, including chloramphenicol, erythromycin, imipenem, tetracycline, and trimethoprim, as well as to a number of biocides. These observations raise concern over the indiscriminate and often inappropriate use of biocides, especially triclosan, in situations where they are unnecessary, whereby they may contribute to the development of microbial resistance mechanisms.

FIG. 1.

(a) Adaptation of Salmonella serovar Enteritidis, Salmonella serovar Typhimurium, Salmonella serovar Virchow, and E. coli O157 to ERY. (b) Adaptation of Salmonella serovar Enteritidis, Salmonella serovar Typhimurium, Salmonella serovar Virchow, and E. coli O157 to BKC. (c) Adaptation of Salmonella serovar Virchow and E. coli O157 to CHX. (d) Adaptation of E. coli O157 to TLN.

FIG. 2.

RAPD profiles obtained from preadapted E. coli O157 (lane 1) and from E. coli O157 strains postadapted to ERY (lane 2), BKC (lane 3), CHX (lane 4), and TLN (lane 5). Lane M, 1-kbp molecular weight marker.

FIG. 3.

Cross-resistance between TLN-resistant E. coli O157 strains and several antibacterial agents by Stoke's method. See Materials and Methods for antibiotic and biocide abbreviations and concentrations.