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Objective. To study the antibacterial activity of lysostaphin against staphylococci various species, as well as its effect on the viability of Vero cells. Materials and Methods. Lysostaphin was obtained by genetic engineering. Purification of the protein was carried out on SP-sepharose, the purity was determined by electrophoresis in PAGE by Lamley. The susceptibility to lysostaphin of 9 species 175 strains of staphylococci was studied. Identification was performed by MALDI-TOF MS, antibiotic susceptibility by EUCAST (v. 11.0). The MIC of lysostaphin was studied by the method of serial dilutions with concentrations between 0.015 and 512 mg/l. For 72 hours, the viability of Vero cells with lysostaphin at concentrations of 0.5-32.0 mg/l was determined by the MTT method with counting of living cells according to their growth curve. The results were analyzed by ANOVA followed by Dunnett's test. Results. A kinetic study of S. aureus growth in the presence of revealed an inhibitory effect of endopeptidase (MIC 0.06 mg/l). Lysostaphin was characterized by pronounced activity against clinical methicillinsensitive S. aureus. The MIC ranged between 0.03 and 0.5 mg/l and the MIC50/90 was 0.125/0.5 mg/l. For methicillin-resistant S. aureus MIC50/90 0.25/0.5 mg/l. The MIC50 for MRSE was 2 times higher than for MSSE – 1 mg/l. The maximum MIC value was determined against isolates of S. warneri and S. hominis – 64 mg/l, the lowest for S. saprophyticus – 0.5 mg/l. MIC50 of lysostaphin against MRSA was 4 times lower than that of vancomycin, MIC90 was 3 times lower. Differences in viable cells depending on the concentration of lysostaphin were not found. Conclusions. Significant activity of lysostaphin against staphylococci was revealed, which is several times higher than vancomycin against MRSA. Lysostaphin was also effective against methicillin-resistant S. aureus. The high anti-staphylococcal activity and cytocompatibility of lysostaphin are promising for its further study in the prevention and treatment of staphylococcal orthopedic infections.

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.

Objective. To evaluate the resistance rate and production of carbapenemases in Klebsiella pneumoniae with phenotypic resistance to carbapenems isolated from patients with orthopedic infection. Materials and Methods. The materials for the study were tissue samples, aspirates and removed orthopedic devices of patients with orthopedic infection at the Vreden Russian Research Institute of Traumatology and Orthopedics, between 2017 and 2019. K. pneumoniae strains were identified in Microlatest by iEMS ReaderMF. These strains were tested for susceptibility to 15 antimicrobial agents by disk diffusion methods, as described by the EUCAST. The carbapenemase genes were investigated by RT-PCR. Results. Of 858 isolated cultures, 6.8% were resistant to carbapenems. Molecular genetic analysis showed that 43.1% of the cultures had blaNDM gene and 24.1% blaOXA-48. All isolates of K. pneumoniae were characterized by resistance to cefotaxime, moxifloxacin and ciprofloxacin. OXA-48-strains were MDR in 50.0% of cases, XDR in 42.9%, PDR in 7.1%. Strains with NDM-carbapenemases were XDR in 68.0% and PDR in 32.0% of cases. The most effective antibiotic was fosfomycin. Thus, 66.7% of NDM-isolates demonstrated sensitivity to fosfomycin. One isolate was PDR with both NDM and OXA-48. Conclusions. Over the period of three years, carbapenemase-producing K. pneumoniae were isolated in the orthopedic hospital. These isolates were not only resistant to carbapenems, but also to a number of other antimicrobial agents. Isolates differed in resistance phenotypes depending on the presence of carbapenemases group, while strains with gene blaNDM were more resistant than those with blaOXA-48.