Fig. 5: Effects of SPI and GP-SPI on fecal gut bacterial genomes and guilds.

Fecal microbial guilds and Akkermansia muciniphila were altered by SPI or GP-SPI supplementation. A Number of up- or down-regulated fecal microbial genomes detected by shotgun sequencing after SPI supplementation (Day 0 vs. Day –5) or after GP-SPI supplementation (Day 0 vs. Day 2, 4, or 10), based on a threshold of q ≤ 0.25. B Bubble-plot of bacterial strains that were significantly changed by GP-SPI supplementation as determined by screening with Benjamini Hochberg test, q ≤ 0.25. Friedman test followed by Nemenyi post hoc test was then applied to detect significant difference between GP-SPI phase timepoints and day 0 (p < 0.05). C qPCR quantification of total bacteria (top) and relative abundance of A. muciniphila species (bottom) in fecal samples collected at indicated time points. Data are mean ± SD. Different letters denote statistical significance (p < 0.05) determined by Nemenyi post hoc test following Friedman test. D Relative abundance of A. muciniphila 208.20, as detected by shot-gun sequencing, at day 0 and 10, with statistical significance (**p < 0.005) determined by a Wilcoxon t test. E Adjusted PCoA (aPCoA) plot of bacterial guilds based on Bray Curtis dissimilarity. F Subject stratified PERMANOVA test results based on Bray-Curtis dissimilarity for total participant pool, males only, or females only. G Change in relative abundance (%) of bacterial guilds (mean ± SEM) by GP-SPI, as detected by shotgun sequencing. Data are mean ± SD. One-way ANOVA followed by the Benjamini Hochberg post-hoc test was used to screen for a significant effect of SPI or GP-SPI (q ≤ 0.25). Friedman test followed by Nemenyi’s post hoc test was used to detect guilds significantly altered by SPI (day –5 vs. 0) or GP-SPI-supplementation (Day 0 vs. Day 2, 4, or 10).