Bifidogenic properties of cell-free extracts derived from probiotic strains of Bifidobacterium bifidum and Lactobacillus reuteri

Isayenko

Voyda

et al. 2019

Regul. Mech. Biosyst.

Self Cite

The development of new effective preparations for the correction of microecological disorders based on probiotic derivatives requires a comprehensive study of the biological activity of the latter. We studied the proliferative activity and biofilm formation by clinical isolates: Escherichia coli and Pseudomonas aeruginosa under the influence of cell-free extracts containing structural components and metabolites of the Bifidobacterium bifidum and Lactobacillus reuteri probiotic strains. Cell-free extracts were obtained from disintegrates and cultures of probiotics. Disintegrates were prepared by cyclic freezing-thawing of probiotic cell suspensions. The cultures were obtained by cultivating probiotic microorganisms in their own disintegrates. The obtained disintegrates and cultures were filtered. The proliferative activity of the test cultures was studied using the spectrophotometric microtiter plate method after an hour-long exposure in undiluted cell-free extracts and subsequent cultivation in a nutrient medium containing 30%vol of the studied extracts at 37 °C for 24 hours. The biofilm formation of the test cultures was studied with 30% vol content of cell-free extracts in the cultivation medium using the spectrophotometric microtiter plate method. All the studied extracts exerted a similar effect on the proliferative activity and biofilm formation by E. coli and P. aeruginosa. Exposure of the test cultures in all undiluted extracts during an hour led to a significant decrease in the optical density of the test samples: optical density of the test wells ranged from 36.5% to 49.8% of the control wells. The test cultures that were exposed to the extracts: filtrate of L. reuteri disintegrate (L), filtrate of В. bifidum disintegrate (B) and filtrate of В. bifidum culture, grown in В. bifidum disintegrate (MB) after dilution and subsequent cultivation over the next 24 hours completely restored the ability to proliferate. The proliferative activity of the test cultures that were exposed to the extracts: filtrate of L. reuteri culture, grown in L. reuteri disintegrate (ML) and filtrate of L. reuteri culture, grown in L. reuteri disintegrate supplemented with 0.8 M glycerol and 0.4 M glucose (MLG), was significantly inhibited after dilution and subsequent cultivation. The inhibition indices calculated for the ML extract were: 25.9% (E. coli) and 53.0% (P. aeruginosa). Inhibition indices calculated for the MLG extract were: 62.0% (E. coli) and 96.9% (P. aeruginosa). MLG extract had more pronounced inhibitory effect on the proliferation of the test cultures than ML extract. All the studied extracts exerted significant inhibitory effect on the biofilm formation of the test cultures. Analysis of the results of the study shows that cell-free extracts of L. reuteri culture grown in its disintegrate without supplementation or supplemented with glycerol and glucose have the highest antimicrobial activity and can be used as metabiotics to prevent overgrowth of potentially pathogenic bacteria, as well as inoculation and proliferation of pathogenic gram-negative bacteria in the gastrointestinal tract. They can be used alone or in combination with cellular probiotics to enhance their probiotic action. This study encourages further careful investigation of the biochemical composition of cell-free extracts and clarifying the mechanism of their action.

Section: Introductionmentioning

confidence: 99%

Influence of cell-free extracts of Bifidobacterium bifidum and Lactobacillus reuteri on proliferation and biofilm formation by Escherichia coli and Pseudomonas aeruginosa

Isayenko

Voyda

et al. 2019

Regul. Mech. Biosyst.

Self Cite

“…Застосування біологічно активних дериватів у складі безклітинних пробіотичних продуктів покликане вирішити проблему небажаних ефектів і недостатньої ефективності клітинних пробіотиків [4,15]. Нашими попередніми дослідженнями встановлено, що безклітинні екстракти, отримані шляхом дезінтеграції пробіотичних бактерій термоциклюванням з наступним культивуванням у власних дезінтегратах, здатні стимулювати проліферацію і біоплівкоутворення пробіотичних бактерій та пригнічувати ріст потенційних патогенів [7,8,9].…”

Section: вступunclassified

Adhesive Properties of Bacteria Under the Influence of Bifidobacterium Bifidum 1 and Lactobacillus Reuteri DSM 17938i Cell-Free Extracts

Act. Probl. of the Modern Med.

Kolpak

Pogorila

et al. 2020

Increasing in the adhesive potential of probiotic bacteria on the one hand and weakening the adhesive ability of the pathobionts on the other is a pathogenetically based approach in the fight against intestinal infectious diseases and intestinal dysbiosis. At present postbiotic products are considered as one of the most promising means of influencing the intestinal microbiota. The aim of the work was to study the effect of cell-free extracts containing derivatives of Bifidobacterium bifidum 1 and Lactobacillus reuteri DSM 17938 on the adhesive properties of probiotic (L. reuteri DSM 17938 and B. bifidum 1) and opportunistic bacteria (Staphylococcus aureus ATCC 25923 and Escherichia coli ATCC 25922). Four cell-free extracts were investigated: L – obtained from the L. reuteri disintegrate; ML – obtained from the L. reuteri culture, cultivated in its own disintegrate; B – obtained from the B. bifidum disintegrate; MB – obtained from the B. bifidum culture, cultivated in its own disintegrate. The effect of cell-free extracts on the adhesive properties of microorganisms was studied by using Brilis method. ML, MB, and B extracts were found to be able to significantly increase the adhesive potential of B. bifidum and L. reuteri. Under their influence adhesion index calculated for bifidobacteria increased by 41.1 % (ML), 41.5 % (MB) and 50.6 % (B), and the microorganism adhesion index calculated for lactobacilli increased by 30.77 % (ML), 44.45 % (MB) and 44.97 % (B). Both studied probiotic bacteria moved up from the category of «moderately adhesive» into the category of «strongly adhesive» with microorganism adhesion index > 4.0. Lactobacilli also became «strongly adhesive» under the influence of L extract, while microorganism adhesion index increased by 37.9 %. Among the studied extracts, only L extract caused a significant increase in the adhesion index of the S. aureus test culture. The other extracts caused minor changes in the adhesive potential of the test cultures. Under their influence, there was a tendency to increase or decrease the adhesive ability of bacteria. Under the influence of MB extract, the S. aureus test culture passed from the «strongly adhesive» category to the «moderately adhesive» category, and the E. coli test culture transferred from the «moderately adhesive» category to the «weakly adhesive» category. The data obtained are of great importance when developing new metabiotics based on the B. bifidum 1 and L reuteri DSM 17938 probiotic strains derivatives.

“…Для налагодження промислового виробництва постбіотичних засобів необхідними є науково і економічно обґрунтований спосіб отримання пробіотичних похідних та достатня доказова база їх біологічної активності, лікувальної ефективності та безпеки. Нами були розроблені оригінальні способи одержання дериватів пробіотичних штамів Bifidobacterium bifidum 1 і Lactobacillus reuteri DSM 17938 та в експериментах in vitro отримані дані, що демонструють високу інгібіторну щодо патобіонтів, стимуляторну щодо пробіотичних бактерій та імуномодуляторну активність дериват-вмісних безклітинних екстрактів (БКЕ) [12,13,14,15]. Зважаючи на те, що активність in vitro не можна ототожнювати з активністю in vivo, перед нами постало завдання дослідити ефективність застосування БКЕ при інфекційному процесі та дисбіозі у лабораторних тварин.…”

unclassified

Application Efficiency of Bifidobacterium Bifidum 1 and Lactobacillus Reuteri DSM 17938 Cell-Free Extracts in Vivo

RS Global - Science Review

Marina

Polianska

et al. 2020

Self Cite

Insufficient efficiency and safety of cellular probiotics encourages the search for new effective means of correction of microecological disorders. Most of the beneficial effects of probiotics are due to the biological activity of their structural components and metabolites. Recently, great hope is pinned on postbiotic products as a means of restoring the balance of intestinal microbial populations. The data obtained in this experimental study demonstrate the ability of cell-free extracts from Bifidobacterium bifidum 1 and Lactobacillus reuteri DSM 17938 cultures, cultivated in their own disintegrates supplemented with ascorbic acid, to provide anti-infection protection and correct microecological disturbances at modeling an infectious process against a background of antibiotic-induced dysbiosis in mice. The beneficial effects of cell-free extracts showed up in the acceleration of the pathogen elimination and an increase in the number of representatives of the positive intestinal microbiota. The results of the study justify the need for further clinical trials to determine the therapeutic efficacy of cell-free extracts when included in the protocols of dysbiosis treatment.