Metabolomics data analysis

To get more insight into the variability in the amine metabolomics data, PCA was performed in Canoco (V5.0) using the intensities of the identified amine metabolites from plasma sample of pigs fed with either SBM or BSF-based diets (Supplementary file ^S3). Employing the statistical module of MetaboAnalyst 4.0⁸³, the heat map graphic distances were measured using Euclidean distances and the clustering algorithm using a Ward dendrogram. The amine metabolites data was normalized by the pooled sample from group SBM and thereafter log transformation to the data was performed. To retain the most contrasting patterns among the treatment groups, the top 25 metabolites ranked by Student’s T-tests were displayed in the heat-map. Additionally, fold changes of the amine metabolites were calculated using the statistic suits of MetaboAnalyst 4.0. Metabolic pathway analysis module of MetaboAnalyst 4.0⁸³ was employed to determine the amine metabolism that was affected by the BSF diet compared to the SBM diets. All the compound names of the metabolites were matched with the human metabolome database (HMDB). To normalize the data, a log transformation was performed. Thereafter, the human pathway library was selected and a reference metabolome (Supplementary file ^S3) based on our technical platform was uploaded. The analysis includes pathway enrichment analysis and topological analysis. The impact-value threshold calculated from the topology analysis was set at 0.4 and the –log (p)-value calculated from pathway enrichment was set to 5 to identify the most related metabolic pathway²¹.