Comparative evaluation of disinfectant efficacy against biofilm-residing microorganisms

Abstract

Introduction. Bacteria in biofilms (BFs) have increased resistance to antibacterial agents, including disinfectants; however, the efficacy level varies depending on the chosen treatment. Therefore, evaluation of efficacy of main disinfectants against BF-residing microorganisms is of scientific and practical interest. The purpose of the study was to explore the effect of disinfectants from various chemical groups on gram-positive and gram-negative bacteria residing in BFs. Materials and methods. The effect of the following disinfectants has been evaluated: alkyldimethylbenzylammonium chloride (ADBAC), tertiary amine (TA), polyhexamethylene guanidine chloride (PHMG), hydrogen peroxide (HP), chloramine (CA), dichloroisocyanuric acid sodium salt (Na DCC), sodium hypochlorite (HC), ethyl alcohol (EA), glutaraldehyde (GA)) against Pseudomonas aeruginosa ATCC 15442 and Staphylococcus aureus ATCC 6538-P BFs. BFs were grown in 96-well plates at 37ºC for 24 hours and then exposed to biocide solutions. The efficacy of disinfectants was evaluated by the number of remaining viable cells and BF relative density. Results. The analyzed bacterial strains formed moderate BFs; the average number of viable cells in BFs was 6.51 ± 0.19 lg. The viable bacterial cell counts in BFs reduced by more than 4 lg when exposed to HP solutions at a concentration of 6%, Na DCC solution — 0.1% (by active chlorine), HC — 1% (by active chlorine), CA – 1% (by product), PHMG — 0.05%, TA — 1.0 %. The BF density decreased by more than 70%. ADBAC solutions at concentrations of 0.1–1.0%, TA — 0.05%, HP — 3%, Na DCC solution — 0.05% (by active chlorine) caused a 2-lg reduction in viable cell counts in BFs. The efficacy of chlorine-active compounds and HP increased when 0.5% sulfonol was added. GA (0.25–1.00%) and EA (40–70%) solutions were ineffective against BF microorganisms. Conclusion. A promising potential in combating microbial biofilms is demonstrated by disinfectants from the group of oxidizing agents (chlorine-active and oxygen-containing), TA and PHMG; using ADBAC as an individual compound is ineffective; aldehydes and alcohols are unable to destroy BFs and eliminate microorganisms in them.