Bcoagulans
Description
Colonizing Bacillus coagulans in zebrafish can alter their metabolic pathways. Metabolome results showed that 99 metabolites related to gut microbiota were upregulated to varying degrees after colonization with Bacillus coagulans. Among them, 5-hydroxytryptophan showed the most significant upregulation, indicating that Bacillus coagulans is most likely to regulate a series of reactions in zebrafish through 5-hydroxytryptophan.
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Steps to reproduce
Intestines of zebrafish treated with B.coagulans or PBS were collected for metabolomic analysis, with each group consisting of 3 replicates and each replicate comprising three individuals. The zebrafish intestines (100 mg) were individually ground with liquid nitrogen. The resulting homogenates were then resuspended with prechilled 80% methanol through vigorous vortexing. After incubating the samples on ice for 5 min, they were centrifuged at 4 ℃ and 15,000 × g for 20 min. Some of the supernatant was diluted to a final concentration containing 53% methanol by LC-MS grade water, The samples were transferred to new Eppendorf tubes and centrifuged again at 4 ℃ and 15,000 × g for 20 min. In the end, the supernatants were injected into the liquid chromatography-mass spectrom-etry (LC-MS/MS) system for analysis. The identification of these metabolites was performed using the Human Metabolome Database (HMDB, https://hmdb.ca/metabolites), Kyoto Encyclopedia of Genes and Genomes (KEGG) database (https://www.genome.jp/kegg/pathway.html), and LIPIDMaps database (http://www.lipidmaps.org/). Principal components sanalysis (PCA) was conducted using metaX, a versatile and comprehensive software for processing metabolomics data. Univariate analysis (t-test) was conducted to determine the statistical signifcance (P-value). Metabolites were considered differential if they had a Vip >1, P-value <0.05, and fold change ≥2 or FC ≤0.5. Volcano plots, generated using ggplot2 in the R language, were used to identify metabolites of interest based on the log2 (fold change) and -log10 (p-value) values.
Institutions
- Huazhong Agriculture University