Influence of sub-inhibitory concentrations of antiseptics on biofilm formation in S. epidermidis

Braz J Microbiol. 2025 Sep;56(3):1823-1833. doi: 10.1007/s42770-025-01702-0. Epub 2025 May 28.

Abstract

Clinical isolates of Staphylococcus epidermidis were evaluated for biofilm production/chemical nature, and for the effect of sub-inhibitory concentrations (sub-MICs) of ethanol and chlorhexidine on biofilm production. Moreover, some of the biofilm-positive samples were studied for the effect of sub-MICs of these antiseptics in the expression of icaA, icaR, sigB and sarA genes. Sub-MICs of ethanol (2 and/or 4%) led to an increase in biofilm production in almost all PIA-dependent and all PIA-independent strains tested, but did not induce biofilm production in primarily non- producers. Unlike ethanol, chlorhexidine sub-MICs determined reduction in biofilm production in biofilm-positive strains. In PIA-dependent strains, ethanol (1%) caused an increase in the relative expression of icaA and reduced expression of icaR, in addition to increased expression of global regulators (sarA and sigB), while the PIA-independent strain showed reduction in the expression of these global regulators. Unlike ethanol, chlorhexidine (0.5 µg/mL) determined increased expression of icaR and reduction of icaA in PIA-dependent strains, besides a reduction in the expression of sarA and sigB in the PIA-independent strain. The opposite effects of ethanol and chlorhexidine on the expression of icaA and icaR in PIA-dependent strains corroborated the phenotypic results obtained for biofilm production in isolates exposed to these antiseptics. The biofilm-inducing or -inhibiting effects of ethanol and chlorhexidine are of practical importance due to their widespread use as antiseptics in healthcare settings.

Keywords: Ica operon; Staphylococcus epidermidis; Biofilm; Chlorhexidine sub-MICs; Ethanol sub-MICs; PIA-dependent and PIA-independent biofilm.

MeSH terms

  • Anti-Infective Agents, Local* / pharmacology
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms* / drug effects
  • Biofilms* / growth & development
  • Chlorhexidine / pharmacology
  • Ethanol* / pharmacology
  • Gene Expression Regulation, Bacterial / drug effects
  • Humans
  • Microbial Sensitivity Tests
  • Staphylococcal Infections / microbiology
  • Staphylococcus epidermidis* / drug effects
  • Staphylococcus epidermidis* / genetics
  • Staphylococcus epidermidis* / isolation & purification
  • Staphylococcus epidermidis* / physiology

Substances

  • Anti-Infective Agents, Local
  • Chlorhexidine
  • Ethanol
  • Bacterial Proteins