Antimicrobial Activity of Bifidobacteria Derivatives After Storage in a Frozen State

Knysh, O. V.; Isaenko, О. Yu.; Babych, E M; Kompaniets, A. M.; Pakhomov, Oleksandr; Polyanska, Valentyna P.; Zachepylo, Svitlana V.; Данілова, І. С.

doi:10.15407/cryo28.03.237

Cited by 6 publications

(2 citation statements)

References 7 publications

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“…Probiotic strains B. bifidum 1 (from dietary supplement "Bifidumbacterin-Biopharma", JSC "Biopharma", Ukraine) and L. reuteri DSM 17938 (from dietary supplement "BioGaia", BioGaia AB, Sweden) were used for obtaining of cell-free extracts (BbCFE and LrCFE). The procedure for obtaining CFEs was described in detail earlier (Knysh et al, 2018). Briefly, bifidobacteria and lactobacilli were cultured for 72 hours in their own disintegrates, obtained by tenfold thermal cycling.…”

Section: Methodsmentioning

confidence: 99%

In vitro immunomodulatory effect of Bifidobacterium bifidum and Lactobacillus reuteri cell free extracts

Knysh

Pogorila

Voyda

2020

Regul. Mech. Biosyst.

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Recent studies have shown that alterations of the immune response in the gastrointestinal mucosa are key components of the mechanism of the probiotic action of beneficial bacteria. Most of the beneficial effects of probiotics are due to the action of their structural components and metabolites. Macrophages are first-line defense cells of the immune system, which not only participate in the detection, phagocytosis and destruction of harmful microorganisms, but also determine the nature of the subsequent immune response by presenting antigens to T-cells and initiating inflammation by releasing cytokines. We researched the effect of two types of cell-free extracts (CFEs) containing probiotic derivatives (structural components and metabolites of bacteria) Bifidobacterium bifidum 1 (BbCFE) and Lactobacillus reuteri DSM 17938 (LrCFE) on the activity of mouse peritoneal macrophages and on the ability of peripheral human blood mononuclear cells to produce cytokines. CFEs were obtained by culturing probiotics in their own disintegrates and then removing cells and cell debris by centrifugation and filtration. Peritoneal macrophages were isolated from mice. Some of them were infected in vitro by Salmonella thyphimurium. Uninfected and infected macrophages were incubated in culture medium containing (30% vol) or not containing CFEs at 37 °С in a microaerobic atmosphere (5% СО2) for 18 hours. After incubation, peritoneal macrophages were lysed. The obtained suspensions were centrifuged and supernatants were carefully collected. Macrophages activity was assessed by the nitrites level, superoxide dismutase (SOD), lactate dehydrogenase (LDH) activity and antiinflammatory cytokines levels in supernatants using spectrophotometric method. Peripheral mononuclear cells were isolated from the blood of healthy volunteers. The ability of peripheral mononuclear blood cells to produce antiinflammatory cytokines was evaluated after cell stimulation with lipopolysaccharide (LPS) and incubation with or without CFEs. Cytokine levels in supernatants were determined using enzyme-linked immunosorbent assay (ELISA). After infection with S. thyphimurium in macrophages, nitrite levels increased 5.5-fold, SOD activity 4.8-fold, and LDH 2-fold. Both studied CFEs exerted a similar effect on the macrophages’ activity. Addition of BbCFE to the incubation medium of infected macrophages resulted in a 4-fold decrease in nitrite levels, and the addition of LrCFE was accompanied by a decrease in nitrite levels to levels in intact cells. Under the influence of both CFEs, the activity of SOD and LDH was significantly reduced and did not differ significantly from the activity of these enzymes in intact cells. BbCFE and LrCFE did not have a significant effect on nitrite levels, SOD and LDH activity in intact macrophages. Under the influence of BbCFE, there was a 2-fold decrease in the production of TNF, a 2-fold increase in IL10 production, and a 30% increase in IL6 production by mononuclear cells. LrCFE caused a decrease in TNF production by 26.7% and IL6 by 36%, and IL10 by 1.9 times. Thus, the studied CFEs normalized the nitrite levels in peritoneal macrophages infected with S. thyphymurium and infection-induced activation of SOD and LDH enzymes. This demonstrates their ability to modulate oxidative processes in macrophages. In addition, under the influence of the investigated CFEs, there was a decrease in the production of pro-inflammatory cytokines (TNFα and IL-6) and increased production of anti-inflammatory cytokine (IL-10) by human peripheral mononuclear cells. The results of the study indicate the ability of CFEs by influencing the functions of innate immunity cells to restrict the inflammatory response and oxidative stress. Based on this, CFEs can be considered as promising agents for the treatment of inflammatory diseases.

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Section: Methodsmentioning

confidence: 99%

In vitro immunomodulatory effect of Bifidobacterium bifidum and Lactobacillus reuteri cell free extracts

Knysh

Pogorila

Voyda

2020

Regul. Mech. Biosyst.

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“…Among antimicrobial, immunomodulatory, anti-inflammatory and antiviral drugs with native bifidobacteria cells at present such metabolic drugs design involves the development of medicines containing biologically active metabolic products and separate structural components of probiotic cells [16], as even cells fragments are considered useful in terms of their positive effect on the body.…”

Section: Introductionmentioning

confidence: 99%

Use of Highly Dispersed Silica in Biotechnology of Complex Probiotic Product Based on Bifidobacteria

Danylenko

Marynchenko

Bortnyk

et al. 2022

Innov Biosyst Bioeng

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Background. The probiotics immobilization technology is one of the most effective ways for controlled and continuous delivery of viable cells into the intestine. It is well known that multifaceted physiological roles of bifidobacteria are to normalize and stabilize the microbiocenosis, to form intestine colonization resistance, to synthesis amino acids, proteins and vitamins, to maintain non-specific resistance of the organism and so all. Such a wide range of positive effects on the macroorganism allows us to consider bifidobacteria as a basis for functional immobilized healthcare products development. Objective. Taxonomic position determination of the Bifidobacterium longum strain selected for immobilization, study of the viability of this bifidobacteria strain in a complex probionic product based on highly dispersed silica in simulated gastrointestinal tract's conditions and after freeze-drying. Methods. The production strain Bifidobacterium longum IMV B-7165 from the Institute of Food Resources of the National Academy of Agrarian Sciences of Ukraine collection of industrial strains has been used in the study. It was isolated from the healthy human infant's gastrointestinal tract. Commonly used bioinformatics, microbiological, biotechnological and statistical methods have been applied. Results. The best alignments for the sequence of bifidobacteria isolate "4202" 16S rRNA (it was previously deposited as Bifidobacterium longum IMV B-7165) and classic dendrograms based on these results were performed. According to the results of microscopic studies of samples of microorganisms with highly dispersed silica products ("Enterosgel", "Sillard P" and "Toxin.Net") it was found that the immobilization of the Streptococcus thermophilus and bifidobacteria cultures did not differ fundamentally. To study the immobilization effect on the bifidobacteria preservation and properties the following carriers were used: "Enterosgel", "Toxin.NET" and "Sillard P". The survival of immobilized bifidobacteria was further studied in simulated gastrointestinal conditions: immobilized cells are better protected from acid and bile, although with increasing acidity, survival decreases in both control and immobilized cells. Conclusions. The taxonomic position of a bifidobacterial isolate from the healthy human infants used in immobilization studies was clarificated (Bifidobacterium animalis subsp lactis). Under the simulated conditions of the upper gastrointestinal tract in the case of acid and bile impact, the best survival was demonstrated by immobilized cultures of bifidobacteria together with the Enterosgel sorbent (a content of 10% by weight of the culture). The survival of immobilized preparations after freeze-drying was slightly reduced in the case of immobilization on the "Enterosgel" and "Toxin.NET" samples of enterosorbents (a content from 15% to 25% by weight of the culture). The best results were observed in the case of immobilization of bifidobacteria with 5% content of the "Toxin.NET" enterosorbent (enterosgel + inulin).

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Impact of Biologically Active Complexes of L. rhamnosus GG and S. boulardii After Storage Under Low Temperatures on Biofilm Forming Ability in Corynebacteria

Isayenko¹,

Knysh²,

Popov³

et al. 2021

Probl Cryobiol Cryomed

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This paper describes the temperature regimens and shelf life of biologically active complexes (structural components and metabolites) of Lactobacillus rhamnosus GG and Saccharomyces boulardii, which have antimicrobial activity and reduce biofi lm formation in pathogenic agents of Corynebacterium spp. Preservation of biological activity of complexes after 6-month storage (observation period) at (−23 ± 1)°C and for 60 days (observation period) under hypothermia at (4 ± 1)°C has been demonstrated. The degree of inhibition of biofi lm formation in pathogenic corynebacteria depended on sensitivity of the Corynebacterium spp. test-culture strain to products of microbial origin. A mixture of lactobacillus and saccharomycete metabolites displayed weak biofi lm formation by toxicogenic strains of corynebacteria (p < 0.05). Our fi ndings testifi ed to a possible use of biologically active substances, stored at 4 and −23°C in designing the L. rhamnosus GG and S. boulardii complexes and in technological processes of their production to prevent the persistence of the diphtheria pathogens.

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Antimicrobial Activity of Bifidobacteria Derivatives After Storage in a Frozen State

Cited by 6 publications

References 7 publications

In vitro immunomodulatory effect of Bifidobacterium bifidum and Lactobacillus reuteri cell free extracts

In vitro immunomodulatory effect of Bifidobacterium bifidum and Lactobacillus reuteri cell free extracts

Use of Highly Dispersed Silica in Biotechnology of Complex Probiotic Product Based on Bifidobacteria

Impact of Biologically Active Complexes of L. rhamnosus GG and S. boulardii After Storage Under Low Temperatures on Biofilm Forming Ability in Corynebacteria

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