Estimation of combined ex vivo effect of thermal ablation and vancomycin on the growth of Staphylococcus aureus culture

Aim. To study the ex vivo effect of high temperature exposure (55–56 °C) combined with vancomycin on culture behavior of pathogenic Staphylococcus aureus (S. aureus).

Materials and methods. Liquid cultures of methicillin-resistant S. aureus (MRSA) strain 43300 were heated ex vivo at 55–56 °C for 0–60 min, either with or without vancomycin (20 μg/ml), followed by incubation at 37 °C up to 120 min. A control suspension (100 or 250 microbial cells per 1 ml of isotonic saline) was maintained at 37 °C. Then, cultures were seeded on solid agar medium, and colony-forming units (CFU) were calculated using computer morphometry after 48h growth. Each experimental subgroup (growth control, thermal ablation, antibiotic, and thermal ablation + antibiotic) included at least three replicates.

Results. A semi-lethal heat exposure time (LD50) of 12.25 min was determined for a liquid microbial culture at 100 cells/ml. When the density was increased to 250 cells/ml, 30 min thermal ablation (55–56 °C) was insufficient for MRSA growth suppression. Vancomycin (20 μg/ml) alone did not affect CFU output. However, combined heat and antibiotic treatment resulted in 28% bacteriostatic effect (p < 0.001) on agar medium.

Conclusion. The study revealed a bacteriostatic effect of combined use of high-temperature exposure with vancomycin, which were ineffective when used separately. The obtained results have practical significance for reconstructive surgery of bone tissue, but require additional studies to clarify the mechanisms of the discovered phenomenon.

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