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In-vitro culture of single gut bacterial strains with added sugars

ABSTRACT: Gut commensal bacterial strains were cultured with or without added sugars (glucose, sucrose and kestose). The base culture medium without sugar added were modified based on the Yeast Casitone Fatty Acids (YCFA) broth. Samples were analyzed for proteomics using LC-MS/MS.

INSTRUMENT(S):

ORGANISM(S): Bacteroides Thetaiotaomicron Bacteroides Uniformis Atcc 8492 Bacteroides Ovatus Atcc 8483 Escherichia Coli Blautia Hydrogenotrophica Dsm 10507 Phocaeicola Vulgatus Atcc 8482

SUBMITTER: Zhibin Ning

LAB HEAD: Daniel Figeys

PROVIDER: PXD037923 | Pride | 2023-06-08

REPOSITORIES: Pride

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			Action	DRS
	Leyuan_20210924_niche_study_101114C.raw	Raw
	Leyuan_20210924_niche_study_101114G.raw	Raw
	Leyuan_20210924_niche_study_101114K.raw	Raw
	Leyuan_20210924_niche_study_101114S.raw	Raw
	Leyuan_20210924_niche_study_10507C.raw	Raw

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Project description:Epithelial cells of the mammalian intestine are covered with a mucus layer that prevents direct contact with intestinal microbes but also constitutes a substrate for mucus-degrading bacteria. To study the effect of mucus degradation on the host response, germ-free mice were colonized with Akkermansia muciniphila. This anaerobic bacterium belonging to the Verrucomicrobia is specialized in the degradation of mucin, the glycoprotein present in mucus, and found in high numbers in the intestinal tract of human and other mammalian species. Efficient colonization of A. muciniphila was observed with highest numbers in the cecum, where most mucin is produced. In contrast, following colonization by Lactobacillus plantarum, a facultative anaerobe belonging to the Firmicutes that ferments carbohydrates, similar cell-numbers were found at all intestinal sites. Whereas A. muciniphila was located closely associated with the intestinal cells, L. plantarum was exclusively found in the lumen. The global transcriptional host response was determined in intestinal biopsies and revealed a consistent, site-specific, and unique modulation of about 750 genes in mice colonized by A. muciniphila and over 1500 genes after colonization by L. plantarum. Pathway reconstructions showed that colonization by A. muciniphila altered mucosal gene expression profiles toward increased expression of genes involved in immune responses and cell fate determination, while colonization by L. plantarum led to up-regulation of lipid metabolism. These indicate that the colonizers induce host responses that are specific per intestinal location. In conclusion, we propose that A. muciniphila modulates pathways involved in establishing homeostasis for basal metabolism and immune tolerance toward commensal microbiota. Keywords: Analysis of target gene regulation by using microarrays Adult germ-free female NMRI-KI mice (45 – 65 days) were used for bacterial mono-association. Two bacterial strains were used in this study, A. muciniphila MucT (ATTC BAA-835) and L. plantarum WCFS1 (NCIMB 8826). A. muciniphila was grown anaerobically in a basal mucin based medium and L. plantarum was grown anaerobically at 37°C in Man-Rogosa-Sharpe broth (MRS; Le Pont de Claix, France). After 7 days of colonization, mice were killed by cervical dislocation and terminal ileum, cecum and ascending colon specimens were sampled.

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In-vitro culture of single gut bacterial strains with added sugars

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