Sensitivity of biofilms formed by Staphylococcus aureus bacteria isolated from patients with chronic osteomyelitis to antiseptics and disinfectants

Materials and methods Biofilms of S. aureus bacteria were grown in 96-well plates and on coverslips for 24 hours. Twenty-four hour old biofilms were treated with disinfectants according to the developed protocol. The effect of antiseptics and disinfectants on monospecies biofilms of S. aureus was assessed visually and quantitatively on glass coverslips using an Axio Lab. A1 microscope (Carl Zeiss, Germany) and an ELx808 photometer (BioTek, USA) at a wavelength of 630 nm. All experiments were repeated four times.

Results Disks impregnated with 3 % Н₂О₂ or 6 % H₂O₂ skin antiseptics had a bactericidal effect against all clinical isolates studied. S. aureus strains formed biofilms with varying intensity on the surface of coverslips and in 96-well plates. Impregnation with 3 % or 6 % hydrogen peroxide decreased the intensity of MSSA biofilm formation by 1.2 times compared to control values and of MRSA by 1.5 times. The application of a skin antiseptic to 24-hour old biofilms did not lead to a significant decrease in the level of biofilm formation. Biofilms in the groups differed in structure and morphology.

Discussion The varying activity of disinfectants against monospecies biofilms depends on their type and concentration. Low concentrations of H₂O₂ disrupt cell membranes, oxidize DNA, and destabilize enzymes and proteins. Sublethal concentrations of H₂O₂ can lead to the emergence of cells with the small colony variant (SCV) phenotype due to increased mutation rates and subsequent replication, which will facilitate the survival of the pathogen in body tissues.

Conclusion Hydrogen peroxide at a concentration of 3 % and 6 % suppresses the activity of biofilm formation by clinical strains of S. aureus and MRSA, but does not completely remove them from the abiotic surface, which indicates the bacteriostatic nature of the action of the disinfectant. The skin antiseptic is not effective against the biofilm form of bacteria, but can have a bactericidal effect on single adhered cells around which the exopolysaccharide matrix has not formed yet.

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