Vorobey E. S., Voronkova O. S., Vinnikov A. I.

MICROBIOLOGY

Scentific article

Removal of mature biofilms is the main task of therapy, because biofilms are the basement of the pathogenic process. The ability of active matrix to bind antibiotics or reduce the diffusion through the biofilm leads to incomplete removal of microorganisms, which attend to their survival and formation of chronic processes. An influence on staphylococcal biofilms can be done by destruction of extracellular polysaccharide matrix. This treatment, which may influence on the structure of the biofilm, is more effective than standard antibiotic therapy. It was established that the phage progeny spread radially through the biofilm, infecting neighboring cells and destroying the biofilm matrix. For this purposes phages producing the depolimerase, which may destroy biofilms matrix that is significantly depends on the structure of the polymer matrix. It was showed that the effect of the drug «Bacteriophage staphylococcal liquid» on 72-hour biofilms of investigated strains, sensitive to cefotaxime and azithromycin, characterized by decreasing of absorbance level of eluted dye on 19.1%, while the number of cells in the biofilm decreased in 253.0 times. More significant effect of staphylococcal bacteriophage was determined for biofilms of strains, resistant to cefotaxime and azithromycin. This effect characterized as a decrease of optical density on 33.9%, while the number of cells decreased in 393.6 times. An important step of our research was to study the biochemical composition of the polymer matrix of staphylococcal biofilms. Study the composition of biofilms matrix based on the specific destruction of it components. The percent of exopolysaccharides in the composition of biofilms matrix, formed by 72-hour, was determine by treatment of sodium periodate. It was found that after treatment the decreasing of optical density of eluted dye at 47.00 ± 2.87% for antibiotic sensitive strain and 59.02 ± 2.10% — for resistant took place. The presence of proteins was determine by using of proteinase K and trypsin. After treatment of the 72-hour biofilm by proteinase K absorbance of eluted dye, delayed by biofilm, decreased on 8.52 ± 1.72% compare to 72-hour control biofilm without enzyme adding for sensitive to antibiotic strain and on 3.52 ± 1.11% — for resistant strain. After trypsin adding decrease of absorbance of eluted dye was less — 12.16 ± 1.49% for sensitive to antibiotic strain and 7.20 ± 2.12% — for resistant. The content of the extracellular matrix DNA was determined with use of DNAase. The decrease of biofilms absorbance on 15.72 ± 2.55% compare to control biofilm for strain sensitive to antibiotic and 14.50 ± 1.33% — for resistant. This indicate that the main component of the biofilm matrix of studied strains are exopolysaccharides, and their number are on 12.02% higher in the antibioticresistant strain. The results demonstrate the effectiveness of the phages drugs use to control the growth of biofilms formed by Staphylococcus aureus strains, the matrix of that consisting of a large number of exopolysaccharide. For inhibition of the biofilmformation the study of the relation of matrix structure and sensitivity to antibiotics is necessary and the search of therapeutic drugs capable to inhibit the production of exopolysaccharides is main task of microbiology.

staphylococci, biofilm, matrix, exopolysaccharides, polysacchariddepolimerase

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«Bulletin of problems biology and medicine» Issue 1 part 1 (126), 2016 year, 249-253 pages, index UDK 579.61: 616-095