Nosocomial Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus: Sensitivity to Chlorhexidine-Based Biocides and Prevalence of Efflux Pump Genes

Int J Mol Sci. 2025 Jan 3;26(1):355. doi: 10.3390/ijms26010355.

Abstract

The widespread use of disinfectants and antiseptics has led to the emergence of nosocomial pathogens that are less sensitive to these agents, which in combination with multidrug resistance (MDR) can pose a significant epidemiologic risk. We investigated the susceptibility of nosocomial Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus to a 0.05% chlorhexidine (CHX) solution and a biocidal S7 composite solution based on CHX (0.07%) and benzalkonium chloride (BAC, 0.055%). The prevalence of efflux pump genes associated with biocide resistance and their relationship to antibiotic resistance was also determined. Both biocides were more effective against Gram-positive S. aureus than Gram-negative bacteria. The most resistant strains were P. aeruginosa strains, which were mainly killed by 0.0016% CHX and by 0.0000084% (CHX)/0.0000066% (BAC) S7. The S7 bactericidal effect was observed on P. aeruginosa and S. aureus after 10 min, while the bactericidal effect of CHX was only observed after 30 min. qacEΔ1 and qacE efflux pump genes were prevalent among E. coli and K. pneumoniae, while mexB was more often detected in P. aeruginosa. norA, norB, mepA, mdeA, and sepA were prevalent in S. aureus. The observed prevalence of efflux pump genes highlights the potential problem whereby the sensitivity of bacteria to biocides could decline rapidly in the future.

Keywords: Escherichia coli; Klebsiella pneumoniae; Pseudomonas aeruginosa; Staphylococcus aureus; antimicrobial resistance; biocides; biofilm formation; chlorhexidine; efflux pumps; quaternary ammonium compounds.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Bacterial Proteins / genetics
  • Benzalkonium Compounds / pharmacology
  • Chlorhexidine* / pharmacology
  • Cross Infection / epidemiology
  • Cross Infection / microbiology
  • Disinfectants* / pharmacology
  • Drug Resistance, Bacterial / genetics
  • Escherichia coli* / drug effects
  • Escherichia coli* / genetics
  • Humans
  • Klebsiella pneumoniae* / drug effects
  • Klebsiella pneumoniae* / genetics
  • Membrane Transport Proteins / genetics
  • Microbial Sensitivity Tests*
  • Pseudomonas aeruginosa* / drug effects
  • Pseudomonas aeruginosa* / genetics
  • Staphylococcus aureus* / drug effects
  • Staphylococcus aureus* / genetics

Substances

  • Chlorhexidine
  • Disinfectants
  • Membrane Transport Proteins
  • Bacterial Proteins
  • Anti-Bacterial Agents
  • Benzalkonium Compounds