Effects of bacteriophages on biofilms formed by Staphylococcus aureus isolated from patients with orthopedic infection

Abstract

Objective. To study effects of bacteriophages on biofilm formation and formed biofilm by S. aureus isolated from patients with orthopedic infection. Materials and Methods. A total of 50 clinical strains of S. aureus were tested. Species identification was performed by MALDI-TOF MS, antibiotic susceptibility – in accordance with the EUCAST v21. Isolates susceptibility to bacteriophages «Sextafag» (Microgen, Russia) was determined by MPA medium. The antibacterial activity of phages against S. aureus ATCC 29213 and S. aureus ATCC 43300 was evaluated by growth kinetic curves. Biofilms of bacteriophage-sensitive S. aureus strains were formed according to the protocol described by O’Toole. Isolates were divided into categories in accordance with the Stepanovic criteria. The effects of bacteriophages on the formation of S. aureus biofilm were studied by co-incubation of phages and bacteria followed by calculation of the percentage inhibition relative to the control without the introduction of the phages. The effect of phages on 24-hour biofilms formed by staphylococci was also evaluated in comparison with the control. Results. Out of 50 clinical S. aureus strains studied, 43 isolates (86%) were susceptible to phages, including 22 MSSA and 21 MRSA. All phage-susceptible cultures were characterized by biofilm-forming ability of varying degree: 28% – weak biofilm producer, 35% – moderate, 37% – strong. Inhibition of biofilm formation was determined in all tested MRSA strains, while in 73% of isolates the index of biofilm formation inhibition was more than 80%, which exceeded this indicator for MSSA by 2.5 times. In turn, the destruction of the formed biofilm under the action of the bacteriophage was 72% for all S. aureus. In 57% of MSSA strains, the decrease in biofilm biomass in comparison with the control was more than 80%, while this indicator was 2 times higher than for MRSA. Conclusions. The results demonstrated a high in vitro efficacy of bacteriophages against biofilm formation in S. aureus.

Keywords

• staphylococcus aureus
• biofilms
• bacteriophage
• orthopedic infection