Реферат: На сьогодні єдина ефективна технологія тривалого зберігання метаболіт-вмісних продуктів мікробного походження остаточно не розроблена. Інформація щодо переваг та доцільності застосування того чи того методу зберігання таких продуктів у наукових літературних джерелах відсутня. Визначення оптимальних умов дозволить уникати втрати біологічної активності метаболітів під час зберігання як на етапах наукових досліджень, так і на виробництві. У роботі отримували метаболіти шляхом відділення від продуцентів за допомогою центрифугування та фільтрації після культивування пробіотиків у рідкому поживному середовищі. Досліджено протимікробну активність фільтратів культуральних рідин пробіотиків із різними посівними дозами одразу після отримання та зберігання протягом 60 діб у рідкому стані за температури (4 ± 1)°С, у замороженому стані при (-23 ± 1)°С та в ліофілізованому стані за гіпотермічних умов ((4 ± 1)°С). Встановлено, що зберігання фільтратів бульйонних культур пробіотиків у вищезазначених умовах не призводить до суттєвого зниження їхньої протимікробної активності. Отримані результати можуть бути використані під час створення комерційних препаратів метабіотиків.Ключові слова: пробіотики, Lactobacillus rhamnosus GG, Saccharomyces boulardii, продукти метаболізму, протимікробна активність, заморожування, ліофілізація.Реферат: На сегодняшний день единственная эффективная технология длительного хранения метаболит-содержащих продуктов микробного происхождения окончательно не разработана. Информация о преимуществах и целесообразности применения того или иного метода хранения таких продуктов в научных литературных источниках отсутствует. Определение оптимальных условий хранения позволит избежать потери биологической активности метаболитов при хранении как на этапах научных исследований, так и на производстве. В работе получали метаболиты путем отделения от продуцентов с помощью центрифугирования и фильтрации после культивирования пробиотиков в жидкой питательной среде. Исследовали противомикробную активность фильтратов культуральных жидкостей пробиотиков с разными посевными дозами сразу после получения и хранения в течение 60 суток в жидком состоянии при температуре (4 ± 1)°С, в замороженном состоянии при (-23 ± 1)°С и в лиофилизированном состоянии в условиях гипотермии ((4 ± 1)°С). Установлено, что хранение фильтратов бульонных культур пробиотиков в вышеупомянутых условиях не приводит к существенному снижению или потере проти-вомикробной активности продуктов их метаболизма. Полученные результаты могут быть использованы при создании коммерческих препаратов метабиотиков.Ключевые слова: пробиотики, Lactobacillus rhamnosus GG, Saccharomyces boulardii, продукты метаболизма, противо-микробная активность, замораживание, лиофилизация. Abstract:Nowadays there is no standard efficient technology for long-term storage of metabolite-containing products of microbial origin as well as no data about the advantages and expediency of using one or another method of storage for these products. Determining the optimal storage condition...

The aim of the work – to produce metabolic complexes with significant antibacterial properties using a new proprietary method and to substantiate the prospects of their use for designing of antimicrobial polyfunctional drugs. Materials and methods. Metabolic complexes of Lactobacillus rhamnosus GG, Lactobacillus plantarum, Saccharomyces boulardii, Enterococcus faecium were obtained by culturing the producers in ultrasonic disintegrates of other probiotic microorganisms. Sensitivity of antibiotic-resistant strains of Escherichia coli PR and Staphylococcus haemolyticus PR was determined by qualitative method. The suspension of test-cultures (optical density of 1.0 units on the McFarland scale) after incubation with metabolites (2, 24 and 48 hours at 37 °C) was inoculated into Mueller–Hinton agar. The absence of growth was indicative of the metabolic complexes antibacterial activity against the microorganism. Results. Cultivation of S. boulardii in the S. boulardii / L. rhamnosus / L. plantarum / E. faecium disintegrates, L. plantarum – in the L. rhamnosus / E. faecium disintegrates and L. rhamnosus – in the L. plantarum disintegrates was accompanied by an increase in the biomass of isolated microorganisms (P ≤ 0.03) and production of metabolites. Along with a similar increase in S. boulardii cells in the S. boulardii / L. rhamnosus / L. plantarum / E. faecium disintegrates, the metabolic products of lactobacterial and enterococcal disintegrates exhibited more active inhibitory effects against E. coli PR and S. haemolyticus PR. The increased antibacterial activity indicates the advantage of the new method. Another improvement is the extended spectrum of metabolic complexes owing to producer cultivation in the other probiotic disintegrates to obtain original biologically active substances with high antibacterial properties against pathogens. Among the strength of the method is that all the stages are unified into a single process avoiding multiphase procedure for the separate preparation of one probiotic microorganism disintegrate and the metabolic products of another. Conclusions. Disintegrates as a nutrient medium can be used not only for their own producers, but also for other strains/species and various probiotic microorganisms (fungi and bacteria). The increase in antibacterial activity of metabolic complexes has been found using the new method of production. The prospects of antimicrobial polyfunctional drugs designing on this basis have been proved.

The paper presents the results of the study on proliferation and biofilm formation by Staphylocoсcus aureus and Staphylocoсcus epidermidis under the influence of cell-free extracts obtained by the author's method and containing derivatives of probiotic strains Вifidobacterium bifidum and Lactobacillus reuteri.The aim of this work was to study the ability of cell-free extracts containing derivatives of probiotics Bifidobacterium bifidum and Lactobacillus reuteri to influence proliferation and biofilm formation by staphylococci in vitro, to evaluate the prospects of their use for the correction of microecological disorders and adjuvant therapy of staphylococcal infection.Materials and methods. Cell-free extracts were obtained from commercial strains B. bifidum and L. reuteri by the authors' method. Reference strain of S. aureus AТСС 25923 and clinical isolate of S. epidermidis were used as a test cultures. The investigation of the proliferation and biofilm formation by staphylococci was carried out by spectrophotometric method using a microtiter-plate reader "Lisa Scan EM" (Erba Lachema s.r.o., Czech Republic).Results. It has been established that the effect of cell-free extract on proliferation and biofilm formation depends on the type of extract and on the species of staphylococcus. Among the five studied extracts, only one significantly inhibits the proliferation and biofilm formation of both staphylococci species. It is the cell-free extract, obtained from L. reuteri culture, grown in its own disintegrate supplemented with glycerol and glucose. The proliferative activity of S. aureus is sensitive to the L. reuteri derivatives while the proliferative activity of S. epidermidis is sensitive to the B. bifidum derivatives. The filtrates of disintegrates have stimulatory effect, while the filtrates of cultures have inhibitory effect on the staphylococcal proliferation. The biofilm formation by S. aureus is significantly inhibited by B. bifidum derivatives and is stimulated by L. reuteri derivatives. The biofilm formation by S. epidermidis is stimulated by derivatives of bifidobacteria and does not change in the presence of derivatives of lactobacteria in the growth medium. Conclusions.Obtained results indicate a high bioregulatory potential of cell-free extracts of probiotic origin and the possibility of drugs development for microecological disorders correction on their basis. They also confirm that the method of obtaining probiotic derivatives with bacteriotropic activity through precursor-directed biosynthesis is promising. Cell-free extract, obtained from L. reuteri culture, grown in its own disintegrate supplemented with glycerol and glucose, exhibits pronounced anti-staphylococcal activity in vitro. After confirming efficacy in vivo, it can be recommended for the adjuvant therapy of staphylococcal infections.

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The aim of the work -to determine the sensitivity of reference strains to structural-metabolic complexes of both Lactobacillus rhamnosus GG alone and in combination with Saccharomyces boulardii to justify the possibility of developing antimicrobial drugs with polyfunctional activity. Materials and methods.Proprietary structural-metabolic complexes of lactobacteria and lactobacteria with saccharomycetes were obtained without the use of culture media. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by the serial dilution micromethod in a culture medium in a 96-well plate. To estimate the MIC, the optical density of the samples was spectrophotometrically measured using a Lisa ScanTM EM analyzer (Erba Mannheim, Czech Republic), and broth was plated on a solid culture medium for the MBC estimation. The concentrations of the test substances ranged from 1.10 to 0.02 mg/ml by the total protein. The test cultures were reference strains of S. aureus ATCC 25923, E. coli ATCC 25922, P. aeroginosa ATCC 27853.Results. The antimicrobial effect was found to be in direct proportion to the exposure time, concentration and activity of structural-metabolic complexes of lactobacteria and lactobacteria with saccharomycetes. The MBC of lactobacteria filtrates for S. aureus ATCC 25923, E. coli ATCC 25922, P. aeruginosa ATCC 27853 was 0.27 mg/ml by protein, and lactobacteria and saccharomycetes -0.21 mg/ml by protein. The structural-metabolitic complex of Lactobacillus with a concentration of 0.14 mg/ml by protein also presented bactericidal effect on the culture of P. aeruginosa ATCC 27853. The lowest tested concentrations of the studied Lactobacillus rhamnosus GG (0.03 mg/ml by protein) filtrates and combinations with Saccharomyces boulardii (0.02 mg/ml by protein) caused a decrease in the optical density of the reference strains of S.