Extracts of Achillea millefolium L. inhibited biofilms and biofilm-related virulence factors of pathogenic bacteria isolated from wounds

Abstract

Biofilm is a surface-attached community of bacterial cells implicated in the pathogenesis of chronic infections and is highly resistant to antibiotics. New alternatives for controlling bacterial infections have been proposed focusing on the therapeutic properties of medicinal plants. Achillea millefollium (Yarrow) is a widespread plant species that is widely used in traditional medicine, especially for wound healing. Therefore, the purpose of this study was to examine the antibiofilm activity of A. millefolium ethanol, acetone, and ethyl acetate extracts on biofilms of Staphylococcus aureus, Proteus spp. and Pseudomonas aeruginosa strains originating from human wounds. Additionally, the effects of the tested extracts on auto-aggregation, cell surface hydrophobicity, and bacterial motility were evaluated. Phytochemical analysis included FT-IR spectroscopy and spectrophotometric quantification of phenolic compound contents was performed. In a test with crystal violet, the extracts strongly inhibited initial cell attachment and biofilm formation, but the effects on mature biofilms were weaker. The effects were dose- and strain-dependent, which was confirmed by fluorescence microscopy. The acetone extract showed the strongest antibiofilm activity. Biofilms of S. aureus S3 and S2 clinical strains were the most susceptible (inhibition of ≥76 % and ≥72 % at all tested concentrations, respectively). The highest concentration of total flavonoids was measured in the acetone extract (100.01 ± 3.13 mg RUE/g). Additionally, the extracts reduced bacterial auto-aggregation, swimming and swarming motility of some strains but did not disturb bacterial cell hydrophobicity. These results suggest that A. millefolium extracts have potential roles as new antibiofilm agents against human pathogenic bacteria.

Keywords: Antibiofilm activity; Auto-aggregation; Bacterial motility; Biofilm; Phytochemical analysis.