Effects of Formulation on Microbicide Potency and Mitigation of the Development of Bacterial Insusceptibility

Cowley, Nicola L.; Forbes, Sarah; Amézquita, Alejandro; McClure, Peter; Humphreys, Gavin J.; McBain, Andrew J.

Abstract

Risk assessments of the potential for microbicides to select for reduced bacterial susceptibility have been based largely on data generated through the exposure of bacteria to microbicides in aqueous solution. Since microbicides are normally formulated with multiple excipients, we have investigated the effect of formulation on antimicrobial activity and the induction of bacterial insusceptibility. We tested 8 species of bacteria (7 genera) before and after repeated exposure (14 passages), using a previously validated gradient plating system, for their susceptibilities to the microbicides benzalkonium chloride, benzisothiozolinone, chlorhexidine, didecyldimethyl ammonium chloride, DMDM-hydantoin, polyhexamethylene biguanide, thymol, and triclosan in aqueous solution (nonformulated) and in formulation with excipients often deployed in consumer products. Susceptibilities were also assessed following an additional 14 passages without microbicide to determine the stability of any susceptibility changes. MICs and minimum bactericidal concentrations (MBC) were on average 11-fold lower for formulated microbicides than for nonformulated microbicides. After exposure to the antimicrobial compounds, of 72 combinations of microbicide and bacterium there were 19 ≥4-fold (mean, 8-fold) increases in MIC for nonformulated and 8 ≥4-fold (mean, 2-fold) increases in MIC for formulated microbicides. Furthermore, there were 20 ≥4-fold increases in MBC (mean, 8-fold) for nonformulated and 10 ≥4-fold (mean, 2-fold) increases in MBC for formulated microbicides. Susceptibility decreases fully or partially reverted back to preexposure values for 49% of MICs and 72% of MBCs after further passage. In summary, formulated microbicides exhibited greater antibacterial potency than unformulated actives and susceptibility decreases after repeated exposure were lower in frequency and extent.

Citation

Cowley, N. L., Forbes, S., Amézquita, A., McClure, P., Humphreys, G. J., & McBain, A. J. (2015). Effects of Formulation on Microbicide Potency and Mitigation of the Development of Bacterial Insusceptibility. Applied and Environmental Microbiology, 81(20), 7330-7338. https://doi.org/10.1128/aem.01985-15

• Journal Article Type: Article
• Acceptance Date: May 8, 2015
• Publication Date: Oct 15, 2015
• Deposit Date: Mar 11, 2024
• Journal: Applied and Environmental Microbiology
• Print ISSN: 0099-2240
• Electronic ISSN: 1098-5336
• Publisher: American Society for Microbiology
• Peer Reviewed: Peer Reviewed
• Volume: 81
• Issue: 20
• Pages: 7330-7338
• DOI: https://doi.org/10.1128/aem.01985-15
• Keywords: Ecology; Applied Microbiology and Biotechnology; Food Science; Biotechnology