Object of study: probiotic and clinical strains of Lactobacillus spp., reference strains and clinical isolates of opportunistic and pathogenic bacteria, metabolites of Lactobacillus spp., experimental samples of native surface antigens of S. aureus.
The purpose of the study: to theoretically and experimentally substantiate new approaches to increase the effectiveness of prevention and control of staphylococcal carriers by reducing the persistence of S. aureus on mucous membranes due to the antagonistic properties of Lactobacillus spp. and blocking the adhesive processes by native surface antigens of the pathogen. Methods of research: general clinical, microscopic, bacteriological, physicochemical, biochemical, immunological, immunobiological, molecular genetic, analytical and medical statistical. Equipment: pH meter millivoltmeter pH-410; microscopes: "Biolam", "Primo Star", MPI-5; analyzers: "Lisa Scan EM", "Elise microplate reader with PC software Utrao SM 600", "Stat-Fax 3000+", Sapphire-400, "AAA 339M"; spectrophotometers: SF-56, "Specord UV VIS", "Bruker Avance II 600"; equipment for gel chromatography, for electrophoresis, chromato-mass spectrometric system Agilent 6890N/5973 inert, amplifier "Tertsyk".
The dissertation is devoted to the solution of the actual scientific and practical problem - increase of efficiency of prevention and struggle against bacteriocarriers of S. aureus on the basis of development and application of immunobiological drugs of new generation with native surface microbial antigens and probiotics that prevent the persistence of the pathogen on the mucous membranes of the upper respiratory tract (URT). For the first time, a combined mucosal candidate vaccine for the treatment / prevention of staphylococcal bacteriocarriers has been proposed, which prevents the persistence of the pathogen on mucous membranes and at the same time normalizes the microbiocenosis of the upper respiratory mucosa and stimulates their anti-infective resistance. For the first time, chemically unaltered (native) S. aureus surface antigens have been obtained as possible platforms for the development of a new class of non-injectable immunobiological drugs. The mode of application of physical factors for obtaining surface antigens of bacteria with anticolonization properties is substantiated. The biological and molecular genetic properties of the original strain of Lactobacillus plantarum IIB B-7679 isolated from the intestines of bees were studied. The safety of this strain was established and its possibility of use as a probiotic was substantiated. The high antagonistic ability of L. plantarum strain IIB B-7679 against representatives of the genera Staphylococcus, Streptococcus, Corynebacterium and Escherichia has been proved. Useful biological properties of L. plantarum strain IIB B-7679 make it possible to classify it as probiotic and recommend it for the development of new domestic immunobiological drugs. The biochemical composition of cell-free supernatants (CF) of Lactobacillus spp., Obtained under different conditions of the gas composition of the atmosphere of cultivation of producers and cell-free antigenic complexes (CFA) of S. aureus, obtained by physical factors, their immunobiological properties were studied. According to the results of in vivo immunobiological screening for the first time established the biological safety of the original method of SF and CFA in tests to determine the general toxic and dermatonecrotic action and the possibility of long-term nasal use under strict dosing. Adequate laboratory models of chronic tonsillitis (CT) and chronic rhinitis (CR) of staphylococcal origin have been developed. It was first shown that the use of Lactobacillus spp. and their metabolites for treatment in animals with laboratory-modulated chronic infections of staphylococcal origin increases the anti-infective resistance of their mucous membranes by increasing the population level of lactobacilli and the content of lysozyme and sIgA in the secretions of URT. The possibility of creating a combined mucosal vaccine based on native surface bacterial antigens of S. aureus and probiotic strains of Lactobacillus spp. and / or their metabolic compounds are experimentally substantiated. It was determined that intranasal administration of the candidate vaccine restores the population level of lactobacilli, the functioning of the local immune system and promotes complete eradication of S. aureus from the nasal cavity of laboratory animals. The degree of implementation: 1 patent of Ukraine for an invention, 5 patents for a utility model, 4 industry innovations, 1 information letter, 2 methodical recommendations, 6 acts of implementation. Sphere (industry) of use: health care (medicine: prevention / treatment of staphylococcal carriers, correction of airway dysbiosis, treatment of infectious and inflammatory diseases).
