Adaptation of Pseudomonas aeruginosa clinical isolates to benzalkonium chloride retards its growth and enhances biofilm production

The increasing percentage of Pseudomonas aeruginosa strains that are resistant to multiple antibiotics is a global problem. The exposure of P. aeruginosa isolates to repeated sub lethal concentrations of biocides in hospitals and communities may be one of the causes leading to increased antibiotic resistance. Benzalkonium chloride (BAC) is widely used as disinfectant and preservative. This study investigated the effect of exposure of P. aeruginosa clinical isolates to sub lethal concentrations of BAC on their antibiotic resistance, growth process and biofilm formation. The collected 43 P. aeruginosa clinical isolates were daily subjected to increasing sub lethal concentrations of BAC. The effect of adaptation on antibiotic resistance, growth process, cell surface hydrophobicity and biofilm formation of P. aeruginosa isolates were examined. Interestingly, Most P. aeruginosa isolates adapted to BAC showed an increase in antibiotic resistance and 66% of the isolates showed retardation of growth, 63% showed increased cell surface hydrophobicity and 23.5% exhibited enhanced biofilm formation by crystal violet assay. To define whether the effect of BAC adaptation on biofilm production was manifested at the transcriptional level, quantitative RT-PCR was used. We found that 60% of the tested isolates showed overexpression of ndvB biofilm gene. More efforts are required to diminish the increasing use of BAC to avoid bacterial adaptation to this biocide with subsequent retardation of growth and enhanced biofilm formation which could lead to antibiotic resistance and treatment failure of infections caused by this opportunistic pathogen.