Project description:Comparison of the growth of Bifidobacterium animalis subsp. lactis BB12 in MRS (without carbon source) with either 2% XOS (xylo-oligosaccharides) or 2% glucose using whole-genome transcriptome analysis.
Project description:Bifidobacterium animalis ssp. lactis Bb12 (Bb12) is a probiotic bacteria with several reported benefits, including immunomodulatory capacity. This is due to the bacteria recognition by receptors such as TLR2, which in turn affects the miRNA transcriptome of immune system cells. We aimed to analyze miRNA expression in swine monocytes stimulated with Bb12, and their expression when TLR2 receptor is blocked. A microarray containing 407 porcine unique miRNA probes was employed to identify differences in the miRNA expression pattern in swine monocytes in response to Bb12. A total of 143 were differentially expressed miRNAs (p<0.05); out of which, 87 miRNAs are statistically significant but have lower signals (signal <500).
Project description:Bifidobacterium animalis subsp. lactis BB12 is a widely used probiotic bacterium. However the molecular mechanism by which this bacterium confers positive health effects to the host is largely unknown. The importance of lipoteichoic acids (LTA) in the cross-talk between the bacteria and human enterocyte-like Caco-2 cells was investigated in the present study. A microarray-based analysis of gene expression revealed induction of genes encoding enzymes involved in the synthesis of lipoteichoic acids of BB12. This observation was confirmed in phenotypic experiments, where a specific galactofuranosyl carbohydrate epitope present in the structure of lipoteichoic acids of BB12 was used as a detection marker. An exopolysaccharide EPS/LTA extract isolated from BB12 and commercially available pure LTAs from the human pathogen S. pyogenes were tested in competitive adhesion experiments. The adhesion of B. animalis subsp. lactis BB12 to Caco-2 cells was inhibited in presence of LTA. The strongest inhibition was observed in the concentration range 0.5-10 ug/ml for LTAs from S. pyogenes and 5-10 ug/ml for EPS/LTA extract from BB12. The inhibitory concentrations in both cases likely reflect the physico-chemical properties of LTAs with the highest efficacy below the critical micelle concentration, and therefore we ascribe the main inhibitory effect to LTAs for the BB12 isolated EPS/LTA extract as well. In conclusion we present increased biosynthesis of lipoteichoic acids of BB12 co-incubated with human Caco-2 cells and evidence that LTA serve as important cell wall factors involved in adhesion to these human enterocyte-like cells.
Project description:Bifidobacterium animalis subsp. lactis BB12 is a widely used probiotic bacterium. However the molecular mechanism by which this bacterium confers positive health effects to the host is largely unknown. The importance of lipoteichoic acids (LTA) in the cross-talk between the bacteria and human enterocyte-like Caco-2 cells was investigated in the present study. A microarray-based analysis of gene expression revealed induction of genes encoding enzymes involved in the synthesis of lipoteichoic acids of BB12. This observation was confirmed in phenotypic experiments, where a specific galactofuranosyl carbohydrate epitope present in the structure of lipoteichoic acids of BB12 was used as a detection marker. An exopolysaccharide EPS/LTA extract isolated from BB12 and commercially available pure LTAs from the human pathogen S. pyogenes were tested in competitive adhesion experiments. The adhesion of B. animalis subsp. lactis BB12 to Caco-2 cells was inhibited in presence of LTA. The strongest inhibition was observed in the concentration range 0.5-10 ug/ml for LTAs from S. pyogenes and 5-10 ug/ml for EPS/LTA extract from BB12. The inhibitory concentrations in both cases likely reflect the physico-chemical properties of LTAs with the highest efficacy below the critical micelle concentration, and therefore we ascribe the main inhibitory effect to LTAs for the BB12 isolated EPS/LTA extract as well. In conclusion we present increased biosynthesis of lipoteichoic acids of BB12 co-incubated with human Caco-2 cells and evidence that LTA serve as important cell wall factors involved in adhesion to these human enterocyte-like cells. The experimental design is described under the single sample. Data desribed in the manuscript (as briefly described under Summary/Abstract) were validated by qPCR.
Project description:Modulation of gut microbiota through probiotic supplementation is an interesting strategy to prevent obesity We use microarrays to study the global genome expression of C. elegans fed with the probiotic strain Bifidobacterium animalis sbsp. lactis CECT 8145
Project description:Stress survival tactics in bacteria utilize the up- and down-regulation of stress response genes. In bacteria that lack classical stress response genes for oxidative stress, other cellular systems can be used for cell survival. We used custom microarrays to study the regulation of genes in Bifidobacterium animalis ssp. lactis strains to oxidative stress to elucidate novel stress response mechanisms. Bifidobacterium cells were grown to late log phase then harvested and exposed to a sub-lethal level of hydrogen peroxide. Samples were taken at 5 and 20 mins for RNA extraction and hybridization on Affymetrix microarrays. Controls were prepared for each time point which recieved no hydrogen peroxide treatment.
Project description:Stress survival tactics in bacteria utilize the up- and down-regulation of stress response genes. In bacteria that lack classical stress response genes for oxidative stress, other cellular systems can be used for cell survival. We used custom microarrays to study the regulation of genes in Bifidobacterium animalis ssp. lactis strains to oxidative stress to elucidate novel stress response mechanisms.
Project description:Modulation of gut microbiota through probiotic supplementation is an interesting strategy to prevent obesity We use microarrays to study the global genome expression of C. elegans fed with the probiotic strain Bifidobacterium animalis sbsp. lactis CECT 8145 Wild type strain N2 of C. elegans was cutured in Nematode Growth medium (NGM, control fed) or NGM with a bacterial lawn fed of the strain B. animalis subsp. lactis CECT 8145, until reach young adult stage. Worm population were age-synchronized. RNA was isolated from each populations (control and treated) using RNAasy Kit (Qiagen) and hybridizated on Affymetrix microarrays.