A Study and Modeling of Bifidobacterium and Bacillus Coculture Continuous Fermentation under Distal Intestine Simulated Conditions.

The diversity and the stability of the microbial community are associated with microecological interactions between its members. Antagonism is one type of interaction, which particularly determines the benefits that probiotics bring to host health by suppressing opportunistic pathogens and microbial contaminants in food. Mathematical models allow for quantitatively predicting intrapopulation relationships. The aim of this study was to create predictive models for bacterial contamination outcomes depending on the probiotic antagonism and prebiotic concentration. This should allow an improvement in the screening of synbiotic composition for preventing gut microbial infections. The functional model (fermentation) was based on a three-stage continuous system, and the distal colon section (N2, pH 6.8, flow rate 0.04 h-1) was simulated. The strains Bifidobacterium adolescentis ATCC 15703 and Bacillus cereus ATCC 9634 were chosen as the model probiotic and pathogen. Oligofructose Orafti P95 (OF) was used as the prebiotic at concentrations of 2, 5, 7, 10, 12, and 15 g/L of the medium. In the first stage, the system was inoculated with Bifidobacterium, and a dynamic equilibrium (Bifidobacterium count, lactic, and acetic acids) was achieved. Then, the system was contaminated with a 3-day Bacillus suspension (spores). The microbial count, as well as the concentration of acids and residual carbohydrates, was measured. A Bacillus monoculture was studied as a control. The stationary count of Bacillus in monoculture was markedly higher. An increase (up to 8 h) in the lag phase was observed for higher prebiotic concentrations. The specific growth rate in the exponential phase varied at different OF concentrations. Thus, the OF concentration influenced two key events of bacterial infection, which together determine when the maximal pathogen count will be reached. The mathematical models were developed, and their accuracies were acceptable for Bifidobacterium (relative errors ranging from 1.00% to 2.58%) and Bacillus (relative errors ranging from 0.74% to 2.78%) count prediction.

The Combination of In Vitro Assessment of Stress Tolerance Ability, Autoaggregation, and Vitamin B-Producing Ability for New Probiotic Strain Introduction.

The health benefits of probiotics are beyond doubt. The positive effects of lactobacilli and bifidobacteria on the function of many body systems have been repeatedly proven by various studies. To completely realize the potential of probiotic microorganisms, the strains should be tested by the greatest combination of characteristics that contribute to the wellness of the host. In this work, for the first time, a combined assessment of the probiotic properties and vitamin B-producing potential of various species and strains of bifidobacteria and lactobacilli was carried out. The presence of an additional advantage, such as vitamin-producing ability, can prevent vitamin deficiency both at the level of the consumption of fermented foods, when the enrichment will occur naturally on the spot, and during colonization by these intestinal strains, when synthesis will occur in vivo. To select potential probiotics, the stress tolerance ability of 16 lactic acid bacteria and bifidobacteria strains to low pH values, bile, and proteolytic enzymes, as well as their ability to autoaggregate, were studied under conditions of modeling the gastrointestinal tract in vitro. The ability of the strains to extracellularly accumulate water-soluble B vitamins was evaluated by capillary electrophoresis. Among the tested strains of bifidobacteria, B. adolescentis VKPM AC-1662 is of interest; it was characterized by the greatest stress tolerance ability and the ability to autoaggregate, in addition to the extracellular synthesis of riboflavin and pyridoxine. Among lactic acid bacteria, L. sakei VKPM B-8936 demonstrated the greatest tolerance to low pH, L. plantarum VKPM B-11007 to duodenal conditions, L. acidophilus VKPM B-2213 to pepsin, and L. salivarius VKPM B-2214 to pancreatin. The highest percentage of autoaggregation was observed in L. salivarius VKPM B-2214, which also accumulated the largest amount of pantothenic acid, but it was sensitive to stress conditions. The obtained results could be used to create new products enriched with probiotics and B vitamins.

A Study on the Synbiotic Composition of Bifidobacterium bifidum and Fructans from Arctium lappa Roots and Helianthus tuberosus Tubers against Staphylococcus aureus.

A number of mechanisms have been proposed explaining probiotics and prebiotics benefit human health, in particular, probiotics have a suppression effect on pathogen growth that can be enhanced with the introduction of prebiotics. In vitro models enhanced with computational biology can be useful for selecting a composition with prebiotics from new plant sources with the greatest synergism. Water extracts from burdock root and Jerusalem artichoke tubers were purified by ultrafiltration and activated charcoal and concentrated on a rotary evaporator. Fructans were precipitated with various concentrations of ethanol. Bifidobacterium bifidum 8 VKPM AC-2136 and Staphylococcus aureus ATCC 43300 strains were applied to estimate the synbiotic effect. The growth of bifidobacteria and staphylococci in monocultures and cocultures in broths with glucose, commercial prebiotics, as well as isolated fructans were studied. The minimum inhibitory concentrations (MICs) of lactic and acetic acids for the Staphylococcus strain were determined. A quantitative model joining the formation of organic acids by probiotics as antagonism factors and the MICs of pathogens (as the measure of their inhibition) was tested in cocultures and showed a high predictive value (R2 ≥ 0.86). The synbiotic factor obtained from the model was calculated based on the experimental data and obtained constants. Fructans precipitated with 20% ethanol and Bifidobacterium bifidum have the greater synergism against Staphylococcus.

A quantitative model of Bacillus cereus ATCC 9634 growth inhibition by bifidobacteria for synbiotic effect evaluation.

The present study is dedicated to the development of novel criteria for assessing the synbiotic effect of prebiotic and probiotic composition against a specific pathogen. These criteria were obtained from the quantitative model of Bifidobacterium adolescentis ATCC 15703 and Bacillus cereus ATCC 9634 (as a model food contaminant) competition in co-culture fermentation. The model is based on the hypothesis that probiotics can reduce the specific growth rate of non-probiotics by producing short-chain fatty acids. To define the relationship between the specific growth rate of non-probiotics and short-chain fatty acid yields, the inhibition constants were determined separately for each inhibitor produced by bifidobacteria (lactic, acetic and propionic acids) in a pure culture of bacilli. Two different equations based on the minimum inhibitor concentration (MIC) and inhibition constant (Ki) were used to connect the specific growth rate and concentrations of inhibitors. The yields of the inhibitors mentioned above were obtained from co-culture experiments. The experimental values and the values predicted by the model of Bacillus count did not differ significantly (R2 not less than 0.83) in the competition experiments. Therefore, the general criterion of the synbiotic effect was derived from the model and presents the coefficient of non-probiotic specific growth rate reduction as a result of probiotic growth and inhibitor formation in the final co-culture fermentation. This criterion has been examined for different commercial prebiotics coupled with the Bifidobacterium adolescentis strain. The synergistic combination of inulin GR with bifidobacteria had the best effect against Bacillus cereus ATCC 9634.