Bifidobacterium Bifidum Strain Research Articles

Bifidobacterium bifidum Ameliorates DSS-Induced Colitis in Mice by Regulating Microbial Metabolome and Targeting Gut Microbiota.

Inflammatory bowel disease (IBD) is a recurrent inflammatory condition affecting the gastrointestinal tract, and its clinical treatment remains suboptimal. Probiotics have shown effectiveness in alleviating dextran sulfate sodium salt (DSS)-induced colitis, exhibiting strain-specific anti-inflammatory properties. In this study, we compared the therapeutic effects of five strains of Bifidobacterium bifidum isolated from healthy adult feces on DSS-induced colitis in mice. Additionally, we investigated the underlying mechanisms by examining gut microbiota composition and microbial metabolome. Our findings highlighted the superior efficacy of B. bifidum M1-3 compared to other strains. It significantly improved colitis symptoms, mitigated gut barrier disruption, and reduced colonic inflammation in DSS-treated mice. Moreover, gut microbiota composition analysis revealed that B. bifidum M1-3 treatment increased the abundance and diversity of gut microbiota. Specifically, it significantly increased the abundance of Muribaculaceae, Lactobacillus, Bacteroides, and Enterorhabdus, while decreasing the abundance of Escherichia-Shigella. Furthermore, our nontargeted metabolomics analysis illustrated that B. bifidum M1-3 treatment had a regulatory effect on various metabolic pathways, including tyrosine metabolism, lysine degradation, and tryptophan metabolism. Importantly, we confirmed that the therapeutic efficiency of B. bifidum M1-3 was dependent on the gut microbiota. These results are conducive to the development of probiotic products for alleviating colitis.

Bifidobacterium bifidum inhibits the TNF-α induced increase in intestinal epithelial tight junction permeability via a TLR-2/TLR-6 and PPAR-γ dependent inhibition of NF-κB p50/p65

Background: Bifidobacterium bifidum (BB) is one of the most widely used probiotic bacterial species and BB has been shown to have a protective effect against intestinal inflammation by an enhancement of the intestinal epithelial tight junction (TJ) barrier. Previous studies from our laboratory showed that a specific BB strain, referred to as BB1, caused a marked enhancement of intestinal epithelial TJ barrier, while strain BB4 had no effect. TNF-α contributes to the development of intestinal inflammation in inflammatory conditions of the gut, in part by increasing intestinal permeability and antigenic penetration. TNF-α induced increase in intestinal epithelial TJ permeability is mediated by an NF-κB p50/p65 activation of myosin light chain kinase (MLCK) gene and MLCK-induced opening of the intestinal TJ barrier. Peroxisome proliferator-activated receptor gamma (PPAR-γ) is an important nuclear receptor known to have anti-inflammatory activity by interfering with NF-κB activation. However, the protective effect of BB1 against TNF-α induced increase in intestinal TJ permeability remain unclear. Aims: The major purpose of this study was to delineate the protective effect of BB1 against the TNF-α induced increase in intestinal TJ permeability and to elucidate the intracellular mechanism involved. Methods: Filter-grown Caco-2 monolayers ( in vitro) and recycling intestinal perfusion of live mice ( in vivo) were the model systems used to assess intestinal TJ permeability. Results: 1) TNF-α caused an increase Caco-2 TJ permeability, as assed by Caco-2 trans-epithelial resistance (TER) and trans-epithelial flux of paracellular marker, FITC-Dextran 10kD. 2) Addition of BB1 (1x108 CFU/ml) to the apical membrane compartment inhibited the TNF-α increase in Caco-2 epithelial TJ permeability, while strain BB4 had no effect. 3) BB1 but not BB4, caused an increase in Caco-2 TLR-2 expression on the apical membrane surface, and siRNA induced knockdown of TLR-2 prevent the BB1 inhibition of TNF-α induced increase in Caco-2 TJ permeability. 4) TNF-α caused a rapid phosphorylation and degradation of Caco-2 IKK-α, as well as and phosphorylation and nuclear translocation of NF-κB p50/p65, indicating NF-κB activation. BB1, but not BB4, inhibited the NF-κB activation. 5) TNF-α activation of NF-κB was associated with an increase in MLCK gene (increase in MLCK promoter activity and mRNA transcription) and kinase activity. BB1, but not BB4, inhibited the TNF-α induced increase in MLCK gene and kinase activity. 6) BB1 but not BB4, caused a rapid PPAR-γ activation (PPAR-γ phosphorylation and nuclear translocation), and siRNA induced knockdown of PPAR-γ inhibited the TNF-α induced activation of NF-κB and increase in Caco-2 TJ permeability. 7) Intraperitoneal injection of TNF-α (5 mg) also caused time-dependent increase in mouse small intestinal permeability (IP) to FITC-Dextran 10kD. TNF-α caused a degradation of IkB-α in intestinal tissue and nuclear translocation of NF-kB in mouse enterocyte. 8) BB1, but not BB4, selectively activated PPAR-γ in mouse enterocytes and inhibited the TNF-α-induced activation of NF-κB, resulting in a decrease in mouse IP. 9) BB1 inhibition of TNF-α activation of mouse enterocyte NF-κB and increase in IP was prevented in mice with intestinal epithelial cell-specific knockout of PPAR-γ Villin-cre and TLR-2 Villin-cre mice. Conclusion: BB1 protects against the TNF-α induced increase in intestinal TJ permeability in a strain-specific manner. BB1 protects against the TNF-α induced increase in intestinal TJ permeability via a novel intracellular mechanism involving BB1/TLR-2 pathway activation of PPAR-γ, and PPAR-γ mediated inhibition of NF-κB p50/p65 and MLCK gene in enterocyte. This research project was supported by the National Institute of Diabetes and Digestive and Kidney Diseases R01-DK-121073-01. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

A novel infant microbiome formula (SIM03) improved eczema severity and quality of life in preschool children

Altered gut microbiome composition has been reported in children with eczema and interventions that restore beneficial bacteria in the gut may improve eczema. This open-label pilot study aimed to investigate the efficacy of a novel infant microbiome formula (SIM03) in young children with eczema. Pre-school Chinese children aged 1–5 years old with eczema received SIM03 twice daily for three months. The novelty of SIM03 consists of both the use of a patented microencapsulation technology to protect the viability of unique Bifidobacterium bifidum and Bifidobacterium breve strains identified through big data analysis of large metagenomic datasets of young Chinese children. Paired stool samples at baseline and following SIM03 were analyzed by metagenomics sequencing. Generalized estimating equation was used to analyze changes in eczema severity, skin biophysical parameters, quality of life and stool microbiome. Twenty children aged 3.0 ± 1.6 years (10 with severe eczema) were recruited. Treatment compliance was ≥ 98%. SCORing Atopic Dermatitis score decreased significantly at two months (P = 0.008) and three months (P < 0.001), while quality of life improved significantly at 1, 2, and 3 months. The relative abundance of B. breve and microbial pathways on acetate and acetyl-CoA synthesis were enriched in stool samples at one month (P = 0.0014). Children who demonstrated increased B. bifidum after SIM03 showed improvement in sleep loss (P = 0.045). Relative abundance of B. breve correlated inversely with eczema extent (P = 0.023) and intensity (P = 0.019) only among patients with increased B. breve at Month 3. No serious adverse event was observed. In conclusion, SIM03 is well tolerated. This patented microbiome formula improves disease severity and quality of life in young eczematous children by enhancing the delivery of B. bifidum and B. breve in the gut. SIM03 is a potential treatment option for childhood eczema.

Scientific and practical aspects of the development of feed additives

Relevance. An actual approach to solving the problems of improving animal health and increasing productive indicators is the development of biologically active additives based on microbial consortiums of probiotic microorganisms that synthesize active metabolites that increase the selective benefits of the beneficial microflora of the gastrointestinal tract and its biological activity in the correction of microbiocenosis and metabolism.Methods. The object of research was 5%, 10%, 15%, 20%, 25% sugar beet molasses. When cultivating in molasses, probiotic microorganisms Clostridium thermocellulociticus, Ruminococcus olbus, Clostridium lochheadii and Bifidobacterium bifidum strain 1 were used. A daily culture of microorganisms was used for inoculation on a nutrient medium, which was standardized to 1 × 106 CFU/cm3.Results. It has been established that the number of probiotic microorganisms that are part of the culture liquid during the cultivation of Clostridium thermocellulociticus, Ruminococcus olbus, Clostridium lochheadii on the eighth day in a 15% molasses-based nutrient medium is 9.9 × 106 CFU/cm³. In studies during the cultivation of B. bifidum, a significant increase in the content of Histidine by 1.13 times, serine, leucine + isoleucine, methionine by 5.5–7.0 times, and other non-essential and conditionally essential amino acids was found, which indicates a significant activity of probiotic microorganisms in nutrient medium. A significant activity of the complex of microorganisms Clostridium thermocellulociticus, Ruminococcus olbus, Clostridium lochheadii was determined, an increase in amino acids was found: lysine and glycine by 3.39 times, leucine + isoleucine by 1.3 times, meteonine and valine by 2.36 times, proline by 7.5 times.

Production of xylo-oligosaccharides with degree of polymerization 3–5 from wheat straw xylan by a xylanase derived from rumen metagenome and utilization by probiotics

In this study, a new xylanase gene (RuXyn) was obtained from a rumen metagenome and heterologously expressed in Escherichia coli to produce xylo-oligosaccharides (XOS) as a prebiotic. RuXyn, which belongs to the glycosyl hydrolase 10 (GH10) family, exhibited maximum activity at pH 6.0 and 50 °C, with excellent temperature resistance at 30–50 °C. RuXyn showed preferential hydrolysis of xylan, especially rice straw xylan. RuXyn activity was significantly increased in the presence of 5 mM dithiothreitol, 0.5% Tween-20, and 0.5% Triton X-100 (31%, 37%, and 43%, respectively). Reaction time (10, 15, 30, and 45 min) and dosage (0.15, 0.31, and 0.62 U) of RuXyn affected XOS yield and composition, with a maximum yield at 0.62 U and 30 min. The primary degradation products of wheat straw xylan, resulting from RuXyn activity, were xylotetraose, xylotriose, and xylopentose, which accounted for 33.4%, 33.0%, and 26.4% of the total sugar yield, respectively. The prebiotic effects of XOS were evaluated by in vitro fermentation of XOS using known probiotic Bifidobacterium bifidum and Levilactobacillus brevis strains. Results showed that regardless of incubation time and XOS concentration, XOS stimulated the growth and xylanolytic enzyme secretion of the two probiotics compared to the controls. Supplementation with XOS significantly enhanced lactate production in B. bifidum and L. brevis. Thus, these results indicate that RuXyn is capable of hydrolyzing wheat xylan to produce XOS with a degree of polymerization 3 and 5 and to subsequently promote the growth of probiotics.

Characteristics of Iranian Probiotic UF White Cheese

Cheese is able to transport live probiotics due to its dry matter, fat, and higher pH than yogurt-like products. According to health organizations’ emphasis on promoting the consumption of healthy dairy products and consumers’ desire to eat healthier foods, in this study, the feasibility of producing probiotic UF cheese containing probiotic strains of Bifidobacterium bifidum sp. IR007-113 and Enterococcus faecium sp. IBRC-M 10836 as a single or combined study was investigated. The experimental cheeses used in this study include A: UF cheese as a control sample, B: probiotic UF cheese containing Bifidobacterium bifidum, C: probiotic UF cheese containing Enterococcus faecium, and D: probiotic UF cheese containing a combination of Bifidobacterium bifidum and Enterococcus faecium strains. Viability of probiotic organisms on the basis of log cfu/g and aroma compounds including (acetaldehyde, diacetyl, acetone, and acetic acid all in micrograms per gram) was assessed during the 60-day ripening period. The results showed that the probiotic bacterial population decreased during the 60-day storage period. However, at the end of the storage period, all experimental cheeses contained probiotic strains above 106 cfu/g. The acetaldehyde compound increased during the 60-day storage period, and diacetyl, acetoin, and acetic acid contents decreased during the 60-day storage period except for the control sample. A total of 12 free fatty acids were identified in Iranian probiotic UF white cheese, which had the highest concentration of palmitic acid among saturated fatty acids and oleic acid among unsaturated fatty acids. The sensory scores of flavor increased during the 60-day storage period. Texture sensory scores of all experimental cheeses decreased during the storage period. In terms of acceptability, all experimental cheeses showed an increasing trend. In general, the results showed that it is possible to produce UF probiotic cheese with minimal adverse effects on quality characteristics and sensory acceptability. Also, treatment D had relatively better characteristics compared to other cheese variants.

Bifidobacterium bifidum prevents the IL-1B induced increase in intestinal permeability by a novel mechanism: TLR-2 dependent activation of PPAR-gamma and inhibition of NF-kB signaling pathway

Background: Bifidobacterium bifidum (BB) is one of the most common probiotic bacteria strains that have been shown to have protective effects against intestinal inflammation, primarily by enhancing the intestinal epithelial tight junction (TJ) barrier function. IL-1β contributes to the development of intestinal inflammation in inflammatory bowel disease (IBD) in part by causing an increase in intestinal permeability. The IL-1β induced increase in intestinal epithelial TJ permeability is mediated by an NF-κB-dependent activation of myosin light chain kinase (MLCK) gene and disruption of the intestinal TJ barrier. The peroxisome proliferator-activated receptor gamma (PPAR-γ) is an important nuclear receptor in enterocytes which is known to have anti-inflammatory activity by interfering with NF-κB activation. However, the role of BB/PPAR-γ (and the possible mechanism involved) in protecting against IL-1β increase in intestinal permeability remain unclear. Aims: The major purpose of this study was to delineate the protective effect of BB against the IL-1β induced increase in intestinal TJ permeability and the intracellular mechanism involved. Methods: Filter-grown Caco-2 monolayers ( in vitro) and recycling intestinal perfusion of live mice ( in vivo) were used to assess intestinal permeability by using a paracellular marker (dextran-10kDa). Results: IL-1β caused a rapid activation of NF-κB and NF-κB and MLCK-dependent increase in intestinal epithelial TJ permeability in vitro and in vivo. A specific strain of BB (1x108 CFU/ml) referred to as BB1 strain inhibited the IL-1β increase in intestinal TJ permeability while strain BB4 had no effect. Other BB strains examined (5 total strains including BB4) did not inhibit the IL-1β increase in intestinal TJ permeability, suggesting that the BB enhancement of Caco-2 TJ barrier function was strain-specific. BB1, not BB4 enhancement of the TJ barrier was associated with an increase in TLR-2 expression, and PPAR-γ activation; BB1 also inhibited NF-kB activation and MLCK activity in Caco-2 monolayers and in mouse enterocyte. The inhibitory effect of BB on 1L-1β induced increase intestinal permeability, NF-κB activation, MLCK expression and activity was blocked by siRNA-induced knockdown of TLR-2 or PPAR-γ in Caco-2 monolayers. We also generated Villin-cre intestinal epithelial specific PPAR-γ knockout mice to study the involvement of enterocytes TLR-2/PPAR-γ on BB1 effect. BB1 did not inhibit the IL-1β-induced activation of NF-κB, increase in MLCK expression and increase in mouse intestinal permeability in TLR-2 or PPAR-γ intestinal deficient mice. Conclusion: These studies provide a novel insight into the BB the protective mechanism against the IL-1β-induced increase in intestinal TJ permeability in -vitro and in-vivo. Our data show that BB protects against the IL-1β induced increase in intestinal TJ permeability by a novel mechanism involving TLR-2/PPAR-g mediated inhibition of NF-kB and MLCK gene activatiation. NIH funding This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Analysis of the live weight dynamics of male turkeys when using eubiotics based on bifidobacteria of Bifidobacterium bifidum strain in the diet

In poultry farming, in order to increase the productivity and livability of poultry stock, antibiotics and biologically active compounds are used as feed additives, which, along with positive properties, cause pathological changes. On this basis, the aim of the research was to determine the effects of various doses of medications based on bifidobacteria on productivity, to find out whether the factor of belonging to a particular subgroup affects the average value of live weight in this subgroup and to conduct a pair comparison of live weight values in subgroups on the principle of “round-robin”. In the diet of turkeys of the first three experimental groups, medication No. 1 – an eubiotic, containing live bifidobacteria of Bifidobacterium bifidum strain was added; the turkeys of the rest three experimental groups received medication No. 2 – an eubiotic, containing live, lyophilized Bifidobacterium bifidum cells sorbed on activated carbon particles. The results of the variance analysis showed that at the age of 4 weeks, the factor of belonging of turkeys to a particular group had a significant impact on the value of the live weight of animals. Then, at the age of 8 weeks, the situation changed. In groups from II to VII, there was practically no significant difference in the average values of live weight. Only the control group I was singled out. At the age of 12 and 16 weeks, male turkeys of the II, III, V, VI groups, where eubiotics based on bifidobacteria were used, gradually began to stand out in a big way. This trend continued until the 24th week. It has been established that medications, which contain bifidobacteria, have a great potential to increase the productivity of poults due to the activation of metabolic processes in the body. The most pronounced effect has biopreparation No. 2.

Screening of Bifidobacteria with Probiotic Potential from Healthy Infant Feces by Using 2'-Fucosyllactose.

Using 2'-fucosyllactose (2'-FL) as the sole carbon source can be an efficient way to screen bifidobacteria with superior probiotic capabilities since 2'-FL is a key element in promoting the growth of intestinal bifidobacteria in newborns. This approach was used in this work to screen eight bifidobacteria strains, including one strain of Bifidobacterium longum subsp. infantis BI_Y46 and seven strains of Bifidobacterium bifidum (BB_Y10, BB_Y30, BB_Y39, BB_S40, BB_H4, BB_H5 and BB_H22). Studies on their probiotic properties showed that BI_Y46 had a unique morphology with pilus-like structure, a high resistance to bile salt stimulation and a potent inhibitory action on Escherichia coli ATCC 25922. Similarly, BB_H5 and BB_H22 produced more extracellular polysaccharides and had a higher protein content than other strains. In contrast, BB_Y22 displayed considerable auto-aggregation activity and a high resistance to bile salt stimulation. Interestingly, BB_Y39 with weak self-aggregation ability and acid resistance had very excellent bile salt tolerance, extracellular polysaccharides (EPS) production and bacteriostatic ability. In conclusion, 2'-FL was used as sole carbon source to identify eight bifidobacteria with excellent probiotic properties.

Towards the isolation of more robust next generation probiotics: The first aerotolerant Bifidobacterium bifidum strain

This work reports on the first described aerotolerant Bifidobacterium bifidum strain, Bifidobacterium bifidum IPLA60003, which has the ability to form colonies on the surface of agar plates under aerobic conditions, a weird phenotype that to our knowledge has never been observed in B. bifidum. The strain IPLA60003 was generated after random UV mutagenesis from an intestinal isolate. It incorporates 26 single nucleotide polymorphisms that activate the expression of native oxidative-defense mechanisms such as the alkyl hydroxyperoxide reductase, the glycolytic pathway and several genes coding for enzymes involved in redox reactions. In the present work, we discuss the molecular mechanisms underlying the aerotolerance phenotype of B. bifidum IPLA60003, which will open new strategies for the selection and inclusion of probiotic gut strains and next generation probiotics into functional foods.

Structural analysis and prebiotic activity of exopolysaccharide produced by probiotic strain Bifidobacterium bifidum EPS DA-LAIM.

The online version contains supplementary material available at 10.1007/s10068-022-01213-w.

Synergistic effects of probiotic Bifidobacterium isolated from chicken's intestine in combination with polyvinylpyrrolidone on reduction of aflatoxin B1.

Food contamination with aflatoxin is one of the most critical concerns of health professionals. One of the best ways to reduce aflatoxin content in food is probiotics. Therefore, this study was performed to isolate Bifidobacterium from the chick's intestine; evaluate its probiotic activities and its application with Polyvinylpyrrolidone (PVP) to reduce aflatoxin B1 (AFB1 ) in the medium were investigated. Samples were isolated from the chick's intestine, and Bifidobacterium was isolated and identified by biochemical and molecular methods. Next, the potential probiotic characterization was assessed. Afterwards, the effect of selected isolate and PVP on reducing AFB1 in the medium was studied using ELISA and HPLC. Biochemical and molecular evaluations indicated isolation of Bifidobacterium bifidum strain from chick's intestine. One of the B. bifidum strains was selected for the next steps, which showed potential probiotic characterization and the ability to reduce the concentration of AFB1 in the medium (50% reduction). When used in combination with PVP showed synergistic effects in reducing the concentration of AFB1 from the medium (up to 90%). In the conclusion, it was found that selected B. bifidum strains and PVP could have synergistic effects in reducing AFB1 toxin in a medium up to 90%.

Evaluation of treatment in acute gastroenteritis: A comparative study.

To compare the efficacy of only dietary recommendations, zinc, probiotics and combination therapies in children admitted with acute gastroenteritis. The comparative, prospective, double-blind, placebo-controlled study was conducted from October 2020 to April 2021 at the Paediatric Emergency Service after approval from the ethics review committee of Diyarbakir Gazi Yasargil Training and Research Hospital, Turkey, and comprised infants with a diagnosis of acute gastroenteritis who were divided into four groups. Only appropriate dietary recommendations were given to the control group 1, while group 2 was given a single probiotic containing bifidobacterium breve, bifidobacterium bifidum, bifidobacterium infantis and bifidobacterium longum strains. Group 3 was given zinc and group 4 was given probiotics and zinc. Demographic data of the patients, admission complaints, physical examination findings, dehydration degrees, and laboratory findings were recorded and analysed. Of the 132 subjects, 79 (59.8%) were males. The overall mean age was 27.5±3.6 months. There were 22 (16.7%) patients in group 1, 34 (25.8%) in group 2, 28 (21.2%) in group 3, and 48 (36.4%) in group 4. The mean duration time to diarrhoea termination was 84.5±10.7 hours (range: 79-89 hours) in group 1, 73.05±6.8 hours (range: 70.5-75.4 hours) in group 2, 80.1±10.3 hours (range: 76-84 hours) in group 3, and 43.5±9.6 hours (range: 46-48 hours) in group 4. Group 4 outcome was statistically significant (p<0.001). The efficiency of combined treatment with probiotics and zinc was found to be significantly better in the treatment of childhood acute gastroenteritis.

Antagonistic activity of synbiotics: Response surface modeling of various factors

Synbiotic compositions have a great potential for curing microbial intestinal infections. Novel targeted synbiotics are a promising field of the modern functional food industry. The present research assessed the effect of various fructan fractions, initial probiotic counts, and test strains on the antagonistic properties of synbiotics.&#x0D; The research involved powdered roots of Arctium lappa L. and strains of Bifidobacterium bifidum, Bacillus cereus, and Salmonella enterica. The experiment was based on the central composite rotatable design. A water extract of A. lappa roots was purified and concentrated. Fructan fractions were precipitated at various concentrations of ethanol, dried, and sub jected to carbon-13 nuclear magnetic resonance (13C-NMR) spectrometry. The bifidobacteria and the test strains were co-cultivated in the same medium that contained one of the fractions. Co-cultivation lasted during 10 h under the same conditions. The acid concentrations were determined by high-performance liquid chromatography to define the synbiotic factor.&#x0D; The obtained fructans were closer to commercial oligofructose in terms of the number and location of NMR peaks. However, they were between oligofructose and inulin in terms of signal intensity. The response surface analysis for bacilli showed that the minimal synbiotic factor value corresponded to the initial probiotic count of 7.69 log(CFU/mL) and the fructan fraction precipitated by 20% ethanol. The metabolites produced by the bacilli also affected their growth. The synbiotic factor response surface for the experiments with Salmonella transformed from parabolic to saddle shape as the initial test strain count increased. The minimal synbiotic factor value corresponded to the lowest precipitant concentration and the highest probiotic count. &#x0D; The research established a quantitative relationship between the fractional composition of fructans and the antagonistic activity of the synbiotic composition with bifidobacteria. It also revealed how the ratio of probiotic and pathogen counts affects the antagonism. The proposed approach can be extrapolated on other prebiotics and microbial strains in vivo.

Development and Quality Evaluation of Synbiotic Yoghurt Incorporated with Oat Flour and Bifidobacterium bifidum NCDC-255

Objectives: Development of synbiotic yoghurt using non-fat milk and by incorporating oat flour and probiotic strain of Bifidobacterium bifidum, evaluating the product for its physico-chemical, rheological and sensory characteristics and checking the viability of Bifidobacterium bifidum strain in final product during storage period. Methods: In the current study synbiotic yoghurt was prepared using skim milk and incorporating, probiotic strain of Bifidobacterium bifidum (NCDC 255) and oats as prebiotic at two different level (T1-1%) and (T2-2%). Plain yoghurt (C1) and probiotic yoghurt (C2) served as control. Physico-chemical properties of the synbiotic yoghurt and its comparison with control yoghurt were carried out during 1st, 3rd, 5th and 7th days of storage at 4oC. Quality parameters such as fat, total solids, acidity, pH, syneresis, water holding capacity, and probiotic count were assessed. Sensory evaluation was also carried out to check the acceptability of the product. Findings: The product T2 with two per cent oat flour was the best with right proportion of ingredients and lowest syneresis. There was a statistically significant (p<0.01) increase in titratable acidity of yoghurt samples during storage. The water holding capacity of samples reduced gradually during storage. Test samples (T1 and T2) showed significantly higher Bifidobacterium count when compared with control. Minimum therapeutic requirement of probiotic organism was maintained within the product even after seven days of storage, in order to transfer the probiotic effect. Overall acceptability and keeping quality was observed to be high in developed product. Novelty: Synbiotic products have opened up novel perspectives for dairy based functional foods because of their health-benefits and worldwide popularity. Study emphasises development of synbiotic yoghurt that can provide benefits of oats and Bifidobacterium bifidum. Keywords: Probiotic; Beta glucan; Synbiotic yoghurt; Bifidobacterium; Oats

Gene-trait matching analysis reveals putative genes involved in Bifidobacterium spp. biofilm formation

Biofilm formation by bacteria represents an adaptation strategy to the environment, and some special genes may lead to a strong biofilm phenotype. In this study, we attempted to find functional genes associated with bifidobacterial biofilm formation. Firstly, we evaluated the biofilm formation ability of bifidobacterial strains from six species, which showed that Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium adolescentis, and Bifidobacterium pseudocatenulatum had biofilm-forming and non-biofilm-forming strains, while all Bifidobacterium bifidum strains could form strong biofilms. Then 48 strains were selected for genome sequencing and comparative analysis. The gene-trait matching analysis revealed that B. bifidum biofilm formation phenotype may associate with their unique genes, involving in stress response, quorum sensing, two components, and peptide synthesis. B. pseudocatenulatum biofilm formation was positively correlated with the eps cluster (rfbX). While no genotype related to the biofilm phenotype was found in B. longum using this analysis, but all contain autoinducer-2 (AI-2) receptor genes. Moreover, luxS, rbsB, rfbX were selected for real-time qPCR analysis, suggesting that their expression are important to biofilm formation. These results indicated that strains carrying certain genes tend to form stronger biofilms than those formed by strains without these genes.

Bifidobacterium bifidum relieved DSS-induced colitis in mice potentially by activating the aryl hydrocarbon receptor

Inflammatory bowel disease (IBD) characterized by relapsed intestinal inflammation and barrier function disruption is still a great therapeutic challenge. This study aimed to screen probiotics that have the potential to help alleviate IBD and further elucidate their mechanism of action. Caco-2 cell differentiated monolayers and RAW264.7 cells stimulated by lipopolysaccharide (LPS) were used for probiotic screening in vitro, and then the efficacies of the obtained six bacterial strains were evaluated in mice with dextran sulfate sodium (DSS)-induced colitis. The results showed that all of the strains at varying degrees could increase the transepithelial electrical resistance (TEER) value, decrease the influx of FITC-dextran in Caco-2 cell monolayers and attenuate the production of nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in LPS-stimulated RAW264.7 cells. In vivo experiments indicated that Bifidobacterium bifidum FL-276.1 (FL-276.1) and Bifidobacterium bifidum FL-228.1 (FL-228.1) showed the best efficacies to ameliorate body weight loss, colon shortening, and intestinal barrier disruption. Accordingly, in FL-276.1 and FL-228.1 groups, the genes of zonula occludens-1 (ZO-1), claudin-4, occludin and mucin 2 (Muc2) in mouse colonic tissues were significantly upregulated, while TNF-α, IL-1β and IL-6 were downregulated. Further results showed that strains FL-276.1 and FL-228.1 could activate the aryl hydrocarbon receptor (AhR) in the intestine. Our study showed that the two Bifidobacterium bifidum strains, FL-276.1 and FL-228.1, ameliorated DSS-induced colitis by enhancing the intestinal barrier and anti-inflammation potentially via the AhR pathway.

The effect of Lactococcus lactis and Bifidobacterium bifidum probiotics cell free supernatants on the expression of HDC and TDC genes in Staphylococcus strains isolated from milk

The most important microorganisms contaminating milk, such as staphylococci, exert their influence by producing toxins and biogenic amines (BA). The purpose of this study was to evaluate the influence of cell-free supernatants (CFS) related to the probiotic strains of Lactococcus lactis and Bifidobacterium bifidum on the expression of BA-producing genes, histamine decarboxylase, and tyramine decarboxylase in staphylococci isolated from milk. Staphylococci isolates of cow raw milk were identified and isolated using standard methods. Samples containing these strains were analyzed by HPLC for BA production. Bacteria with target genes were treated with L. lactis and B. bifidum CFS, and the expression of target genes was measured by Real time PCR. Generally, 60 strains of Staphylococcus were isolated, and 90% of strains had HDC and TDC genes in genome. The levels of BAs were significantly higher on the second and third days after sampling compared with the first day (p <0.02). The analysis RT-PCR indicated that the under expression of target genes was statistically significant (P<0.05). This study showed that the use of probiotic bacteria can reduce the production of these amines and increase the quality of milk through reducing the expression of tyramine and histamine producing genes.

Integration of Transcriptome and Metabolome Reveals the Genes and Metabolites Involved in Bifidobacterium bifidum Biofilm Formation.

Bifidobacterium bifidum strains, an important component of probiotic foods, can form biofilms on abiotic surfaces, leading to increased self-resistance. However, little is known about the molecular mechanism of B. bifidum biofilm formation. A time series transcriptome sequencing and untargeted metabolomics analysis of both B. bifidum biofilm and planktonic cells was performed to identify key genes and metabolites involved in biofilm formation. Two hundred thirty-five nonredundant differentially expressed genes (DEGs) (including vanY, pstS, degP, groS, infC, groL, yajC, tadB and sigA) and 219 nonredundant differentially expressed metabolites (including L-threonine, L-cystine, L-tyrosine, ascorbic acid, niacinamide, butyric acid and sphinganine) were identified. Thirteen pathways were identified during the integration of both transcriptomics and metabolomics data, including ABC transporters; quorum sensing; two-component system; oxidative phosphorylation; cysteine and methionine metabolism; glutathione metabolism; glycine, serine and threonine metabolism; and valine, leucine and isoleucine biosynthesis. The DEGs that relate to the integration pathways included asd, atpB, degP, folC, ilvE, metC, pheA, pstS, pyrE, serB, ulaE, yajC and zwf. The differentially accumulated metabolites included L-cystine, L-serine, L-threonine, L-tyrosine, methylmalonate, monodehydroascorbate, nicotinamide, orthophosphate, spermine and tocopherol. These results indicate that quorum sensing, two-component system and amino acid metabolism are essential during B. bifidum biofilm formation.

USING Sargassum SEAWEEDS GROWN IN COASTAL AREAS OF SOUTH ASIA, AS COATING MATERIALS AND ASSESSING THE SURVIVABILITY OF MICROENCAPSULATED PROBIOTIC BACTERIA IN DAIRY BASED FOOD PRODUCTS

Purpose of the conducted study was to evaluate the possibility of using underutilized seaweed (Sargassum sp.) to encapsulate prebiotics in dairy food applications. This will enable dairy based industries to utilize local materials instead of imports, whereas direct crude exports of the seaweeds can also be consumed for more valued added purposes. For encapsulation purposes, Lactobacillus acidophilus and Bifidobacterium bifidum strains were selected. When considering the surviving ability of the two strains of bacteria in 0.5% bile solution and gastric solution followed by intestinal juice and inclusive and non-inclusive of bile salt, it was evident that the strains coated with Sodium alginate from Sargassum provide a positive protection and therefore is foreseen as a better natural solution for probiotic encapsulation in dairy based food products. Extraction and utilization of local seaweeds in South Asia, has not been an exploited area so far, and therefore this research will form a base to formulate probiotics in local dairy industry by using Sargassum seaweed.