The effect of nanobiocomposites of carrageenan and galactomannan — perspective prebiotics on the formation of the extracellular polysaccharide complexes of Bacillus amyloliquefaciens subsp. plantarum UCM B-5139 and UCM B-5140 probiotic strains is studied. Analysis of data showed that bacterial strains had differences in the content of various sugar residues within their exopolysaccharide matrices. Both compounds, carrageenan and galactomannan, and their nanocomposites showed a potency to change the content of the extracellular matrix of bacilli probiotic cells. The effect of these compounds depended, presumably, on the natural structural properties of polysaccharides of these strains. However, the synthesis of extracellular polysaccharides by Bacillus probiotic strains was not bloc ked in the presence of the investigated nanocomposites. The obtained results indicate the possible dual (direct and indirect) action of polysaccharides and their nanocomposites in case of their use as the prebiotic components of synbiotic preparations: through the direct biologi cal action on the properties of the mammalian epithelial cells or gut microflora, and indirectly through the changes of properties of the extracellular polysaccharide matrix of probiotic strains.

The purpose of this study was to investigate the antimicrobial and immunomodulatory action of a probiotic composition of Bacillus subtilis and B. megatherium strains (UnicaUro, Sirion (Ukraine)) for experimental bacterial vaginitis. Methods. Experimental studies were conducted on female BALB/c mice; we used Staphylococcus aureus strain B-918 (ATCC 6538) to induce bacterial vaginitis. The strain was vaginally introduced into mice before treatment with probiotic bacteria. In the vagina of mice, aerobic and optionally anaerobic bacteria, including representatives of the genera Staphylococcus, Streptococcus, Lactobacillus, Bifidobacterium, Pseudomonas, coliform bacteria, and microscopic fungi were identified in different periods of observation using generally accepted microbiological methods. Serum antibody titer to S. aureus was determined by the bacterial agglutination reaction. The phagocytic activity and oxygen-dependent bactericidal activity of peritoneal exudate macrophages (PEM) were evaluated using generally accepted immunological methods. Results. The formation of bacterial vaginitis in the BALB/c mice line infected with S. aureus B-918 (ATCC 6538) was evidenced by the appearance of external clinical manifestations of the infectious and inflammatory process against the background of the increased number of aerobic and optionally anaerobic microorganisms, including representatives of the genus Staphylococcus and Streptococcus, microscopic fungi, and decreased number of lactobacilli in different observation periods. The probiotic introduction to mice with bacterial vaginitis led to a dynamic change in the vaginal microbiota: the number of aerobic and optionally anaerobic microorganisms decreased, primarily due to the normalization of the number of representatives of Staphylococcus genus accompanied by a decrease in the antibody titer to staphylococcus in the blood serum. The effective therapeutic action of the probiotic was confirmed by the gradual disappearance of the external clinical signs of the infectious-inflammatory process in the vagina against the background of the functional activity of PEM. Conclusions. The probiotic composition of B. subtilis and B. megatherium (UnicaUro, Sirion, Ukraine) is a promising antimicrobial formulation that may be used in the treatment of bacterial vaginitis; however, further studies are required to confirm its therapeutic, antimicrobial, and immunomodulatory efficacy.

Probiotics based on bacteria of the genus Bacillus with a multifactorial mechanism of action are considered as a possible alternative to antibiotics in the treatment of purulent wounds. The aim of the study was to determine the antimicrobial and therapeutic effect of the Arederma probiotic preparation containing probiotic strains of the genus Bacillus in an experimental model of a purulent wound in animals. The antimicrobial efficacy of the probiotic against test strains and clinical isolates of pathogenic and opportunistic microorganisms was studied using the method of delayed antagonism. Staphylococcus aureus ATCC 6538 and Streptococcus pyogenes K-7 were used to model a purulent wound. From the surface of the wounds, bacteria of Staphylococcus, Streptococcus, Enterobacteriaceae, Pseudomonas genera and Enterobacteriaceae family were sown on appropriate selective media for the cultivation and enumeration of different groups of microorganisms by generally accepted microbiological research methods. The formation of a purulent wound in rabbits caused by mechanical skin damage and subsequent double infection with Staphylococcus aureus ATCC 6538 and Streptococcus pyogenes K-7 strains was accompanied with a pronounced inflammatory process, necrosis, the formation of purulent exudate and general intoxication. Representatives of the genera Staphylococcus and Streptococcus, microscopic fungi and, to a lesser extent, members of the family Enterobacteriaceae and Pseudomonas were found on the surface of purulent wounds, which confirmed the development of the infectious-inflammatory process. Treatment of purulent wounds with a suspension of probiotic preparation once a day for 4 days led to their faster healing (gradual attenuation of the inflammatory process, reduction of edema and discharge, as well as their disappearance) compared with untreated purulent wounds (control). Representatives of the Staphylococcus and Streptococcus genera, as well as microscopic fungi, presented in purulent wounds treated with probiotic preparation in much smaller numbers than in the control, and bacteria of the Pseudomonas genus and the Enterobacteriaceae family were not detected at all. The effective antimicrobial effect of this probiotic preparation against opportunistic and pathogenic microorganisms was confirmed by in vitro studies. Therefore, the Arederma probiotic preparation showed an effective therapeutic and antimicrobial effect in the experimental model of a purulent wound in animals, so it can be recommended for further preclinical and clinical studies.

The application of biocontrol biopesticides based on plant growth–promoting rhizobacteria (PGPR), particularly members of the genus Bacillus, is considered a promising perspective to make agricultural practices sustainable and ecologically safe. Recent advances in genome sequencing by third-generation sequencing technologies, e.g., Pacific Biosciences’ Single Molecule Real-Time (PacBio SMRT) platform, have allowed researchers to gain deeper insights into the molecular and genetic mechanisms of PGPR activities, and to compare whole genome sequences and global patterns of epigenetic modifications. In the current work, this approach was used to sequence and compare four Bacillus strains that exhibited various PGPR activities including the strain UCMB5140, which is used in the commercial biopesticide Phytosubtil. Whole genome comparison and phylogenomic inference assigned the strain UCMB5140 to the species Bacillus velezensis. Strong biocontrol activities of this strain were confirmed in several bioassays. Several factors that affect the evolution of active PGPR B. velezensis strains were identified: (1) horizontal acquisition of novel non-ribosomal peptide synthetases (NRPS) and adhesion genes; (2) rearrangements of functional modules of NRPS genes leading to strain specific combinations of their encoded products; (3) gain and loss of methyltransferases that can cause global alterations in DNA methylation patterns, which eventually may affect gene expression and regulate transcription. Notably, we identified a horizontally transferred NRPS operon encoding an uncharacterized polypeptide antibiotic in B. velezensis UCMB5140. Other horizontally acquired genes comprised a possible adhesin and a methyltransferase, which may explain the strain-specific methylation pattern of the chromosomal DNA of UCMB5140. • Whole genome sequence of the active PGPR Bacillus velezensis UCMB5140. • Identification of genetic determinants responsible for PGPR activities. • Role of methyltransferases and epigenetic mechanisms in evolution of bacteria.

Охарактеризованы аэробные спорообразующие

The aim of this review was to present the results of more than ten-year study of gaupsin biopreparation created on the basis of two strains Pseudomonas chlororaphis subsp. aureofaciens UCM В-111 and UCM В-306 with antifungal, entomopathogenic and antiviral activities. Data about antibiotic substances produced by these strains — phenazine and phenylpyrrole derivatives — are presented. Entomocidal properties against the wide spectrum of insect pests have been found out in the strains-producers. Antiviral activity of gaupsin due to the production of thermostable exopolymers containing neutral monosaccharides has been shown using the tobacco mosaic virus as a model. Lipopolysaccharides of the strains В-111 and В-306 also appeared to be highly active antiviral agents. Structure of their O-specific polysaccharides has been established. The last one are structurally heterogenic, presented by linear tri-and tetrasaccharide repeated links and have specific structure that has not been described previously.

The aim of the work was to study the lytic activity of probiotic strains of B. amyloliquefaciens subsp. plantarum and the possibility of using amino acids for its stimulation. Methods. The research objects were B. amyloliquefaciens subsp. plantarum UCM B-5139, UCM B-5140 – the basis of therapeutic and prophylactic preparation endosporin. The amino acid (DL-) and dry hydrolyzate mixture of amino acids were used. The lytic enzyme activity of bacilli was determined by the turbidimetric method and expressed as the percentage decrease in the optical density of live cell suspension of the test cultures. Results. Stimulation of the growth and lytic activity of studied bacilli strains against Gram negative bacteria cells by the mixtures of amino acids was installed. However, the separate amino acids such as arginine, alanine, tryptophan, histidine and phenylalanine stimulated the growth of B. amyloliquefaciens subsp. plantarum and the synthesis of lytic enzymes were more effective. The degree of destruction of the E. coli cell walls increased more than 30–100% and S. aureus in 2.3–4 times. Conclusion. High lytic activity of bacilli characterizing their antimicrobial properties, provided spore-forming bacteria harmless for warm-blooded may be no less important than their antagonistic activity against pathogenic and conditionally pathogenic microorganisms in the selection of strains for development of probiotics.

Proteases from probiotic strains of the genus Bacillus, just like the antibiotics, bacteriocins and other hydrolytic enzymes, are one of the main factors that determine their biological activity. The aim of this work was to study the synthesis and biochemical properties of proteases from two strains Bacillus amyloliquefaciens subsp. pl antarum UCM B-5139 and UCM B-5140 that included in the probiotic Endosporin. The cultivation of strains was carried out in flasks under rotating for two days. The influence of physicochemical parameters of the reaction medium on proteolytic activity was studied on partially purified protease preparations. Lytic activity was determined by turbidimetric method. On the second day of cultivation B. amyloliquefaciens subsp. plantarum UCM В-5139 and UCM В-5140 synthesized the metaldependent peptidase and serine protease, respectively. The optimum conditions of their action were the following: temperature 37–40 C and pH 6.5–7.0. Isolated proteases are able to lyse the living cells of Staphylococcus aureus and Candida albicans. Thus we demonstrated that B. amyloliquefaciens subsp. plantarum UCM B-5140 and UCM B-5139, included in the probiotic veterinary preparation Endosporin, produced proteolytic enzymes that hydrolyze the native insoluble proteins (elastin, fibrin and collagen). These enzymes belong to the group of neutral metal-dependent and serine proteases. They are active under physiological conditions against gram-positive bacteria and yeasts. The application of these proteases in biotechnology is considered.

The immunomodulatory properties of perspective prebiotics--beta-galactose-containing--polysaccharides arabinogalactan, arabinogalactan with flavonoids, carrageenan, galaktomannan have been studied. It was established that oral administration of the studied beta-polysaccharides to experimental animals leads to a significant activation of immune cells and the synthesis of humoral factors in their organism which significantly increases the cytotoxic activity of blood serum. The most pronounced immunomodulatory effect exhibits arabinogalactan with flavonoids and carrageenan.

Вивчено імуномодулюючі властивості перспективних пребіотиків – галактозовмісних β-полісахаридів арабіногалактану, арабіногалактану з флавоноїдами, карагінану, галактоманану та їх гідролізованих форм. Встановлено, що пероральне введення експериментальним тваринам досліджуваних β-полісахаридів призводить до значної активації клітин імунної системи і синтезу гуморальних факторів у їх організмі, які значно підвищують цитотоксичну активність сироватки крові. Найбільш виражений імуномодулюючий ефект проявляють арабіногалактан з флавоноїдами і карагінан.

A wide variety of genetic data about organisms of interest has become available with the advancement to next generation sequencing (NGS). For many potential new users, to process this huge amount of genetic data released by NGS and to utilize this information to resolve practical questions remains a challenge. Genetic barcoding of microorganisms is the first obvious area where NGS has met the requirements of applied microbiology. In general, barcoding in microbiology is a comparative genome approach to differentiate between species or strains that are hard to distinguish by traditional methods. In this chapter, we introduce the conceptual background of bacterial barcoding and present several basic bioinformatics tools and approaches to provide solutions to NGS data handling. While working with a putative industrial strain or potentially hazardous pathogen, the following questions arise: (i) is this strain unique and if so, what makes it unique genetically or practically speaking?; (ii) how can it be detected in the environment?; (iii) are there any genetic markers for its extraordinary activity? The possibility of barcoding of whole bacterial communities is considered and both the benefits and limitations of the traditional 16S rRNA based barcoding and multi-locus sequence typing are discussed.

Is established that at intestinal dysbacteriosis in the organism of experimental animals develop reactions of inflammatory type (early and late), leading to an accumulation in the blood serum of humoral immunity factors, blocking reactions mediated by lymphocytes ormacrophages, while maintaining their cytotoxic activity. Introduction of animals of tne probiotic endosporin promotes restoration of quantitative and qualitative indicators of intestinal microbiocenosis and the normalization of the immune system.

Due to the widespread of the ecological disorders and diseases of the gastrointestinal tract of humans and animals there is an increased inte rest in probiotic products (probiotics, prebiotics and synbiotic), which showed its efficacy and safety in their treatment and prevention. Yeast Saccharomyces cerevisiae and cell culture of a human larynx cancer НЕр-2 were used to study cytoand DNA-damage effects of several poly-saccharides (arabinogalactan and nanobiocomposite of arabinogalactan with flavonoids, karraginan, galactomannan and their hydrolyzed derivatives), which are new perspective prebio tic substances. Cytotoxic potential of substances was determined from MTT-assay and DNA-damage effects were determined by morphological and structural features of cell nuclei. The synergic action of the polysaccharides with chemical triggers of cellular damages (hydrogen peroxide, acetic acid, nitrosomethylurea) was evaluated. The studied substances possessed no cytoand DNA-damage action on the yeast and mammalian cells. It was shown that polysaccharides were able to enhance the effects of hydrogen peroxide, acetic acid and nitrosomethylurea. Nature of the marked effects remains unclear, but it may be assumed that the studied prebiotic substances can be used to enhance the effects of some chemical compounds and appear promising for use in integrated biomedical preparations of varied spectrum.

Показано, що на фоні дисбактеріозу кишечника в організмі експериментальних тварин розвиваються реакції запального типу (ранні та пізні), що призводять до накопичення в сироватці крові гуморальних факторів імунітету, за рахунок яких блокуються реакції, опосередковані лімфоцитами або макрофагами при збереженні їх здатності до прояву цитотоксичної активності. Введення тваринам пробіотика ендоспорину сприяє відновленню кількісних і якісних показників мікробіоценозу кишечника та нормалізації стану імунної системи.

Due to the widespread of the ecological disorders and diseases of the gastrointestinal tract of humans and animals there is an increased interest in probiotic products (probiotics, prebiotics and synbiotic), which showed its efficacy and safety in their treatment and prevention. Yeast Saccharomyces cerevisiae and cell culture of a human larynx cancer НЕр-2 were used to study cyto- and DNA-damage effects of several polysaccharides (arabinogalactan and nanobiocomposite of arabinogalactan with flavonoids, karraginan, galactomannan and their hydrolyzed derivatives), which are new perspective prebiotic substances. Cytotoxic potential of substances was determined from MTT-assay and DNA-damage effects were determined by morphological and structural features of cell nuclei. The synergic action of the polysaccharides with chemical triggers of cellular damages (hydrogen peroxide, acetic acid, nitrosomethylurea) was evaluated. The studied substances possessed no cyto- and DNA-damage action on the yeast and mammalian cells. It was shown that polysaccharides were able to enhance the effects of hydrogen peroxide, acetic acid and nitrosomethylurea. Nature of the marked effects remains unclear, but it may be assumed that the studied prebiotic substances can be used to enhance the effects of some chemical compounds and appear promising for use in integrated biomedical preparations of varied spectrum.

Natural nanocomposites hemicellulose arabinogalactan and flavonoids isolated (from Siberian larch) and characterized. Additionally nitro- and sulfo-esters of arabinogalactan and its calcium salt are synthesized and. characterized. All of the derivatives of the beta-hemicellulose arabinogalactan. are water-soluble and are promising prebiotics on the example test-strain Bifidobacterium bifidum. (except for the nitrate esters of arabinogalactan).