Keywords
Abstract
The widespread occurrence of resistant P. aeruginosa strains, the ability to form films, complex regulatory pathways of biofilm formation and the numerous virulence factors require the search for new compounds and the development of drugs capable of inhibiting biofilm formation.
The aim of the study was to investigate the genes expression that regulate P. aeruginosa biofilm formation after exposure to an adamantane-containing compound.
The antibiofilm activity of the adamantane derivative AM-166 against a clinical isolate of P. aeruginosa was studied at concentrations of 0.15 MIC, 0.5 MIC and 2.0 MIC by the method of adsorption of gentian violet on biofilm structures. Detection of the genes algL, pslA, pelA, algD, algR, algU, mucA in P. aeruginosa strain 449 was performed by classic PCR with subsequent gel electrophoresis. The effect of compound AM-166 on the transcriptional activity of genes was studied by the 2–ΔΔCt method using real-time PCR.
The results of the study indicate that compound AM-166 has a dose-dependent antibiofilm effect against P. aeruginosa 449. The biomass of the biofilm after exposure to the adamantane derivative at concentrations of 0.15 MIC, 0.5 MIC, 2.0 MIC decreased by 22.8 %, 49.4 %, 62.9 %, respectively. After exposure to the subinhibitory concentration of AM-166, an increase in the expression level of the biofilm formation genes pelA, pslA, algD, algR and a decrease in the expression of algU, algL, mucA (р < 0.05), which regulate the synthesis of alginate, Pel- and Psl-polysaccharide, was observed.
Thus, the adamantane derivative 4-(adamantyl-1)-1-(1-aminobutyl) benzol showed inhibitory activity against P. aeruginosa biofilms. Both an increase and a decrease in the expression of genes regulating the synthesis of matrix components was observed.