Impact of a Lifestyle Intervention on Gut Microbiome Composition: A Quasi-Controlled Before-and-After Analysis

Figures

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  Figure 1. Alpha diversity grouped by treatment (untreated MetS, before MHP, and after MHP). (a,b) Shannon’s diversity index; (c,d) Fisher’s diversity index. The horizontal line inside each box indicates the median. Whiskers extend to the minimum and maximum values within 1.5 times the interquartile range (IQR) from the 1st and 3rd quartiles. The boxes represent the IQR, spanning from the 1st to the 3rd quartile. Solid dots (●) outside the whiskers indicate values exceeding 1.5 times but less than 3 times the IQR. Plots were created from raw, untrimmed data. Samples of MetS untreated participants are shown as squares, before MHP as circles, and after MHP as triangles.

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  Figure 2. Principal coordinate analysis (PCoA)—beta diversity. Bray–Curtis, ordination plot showing the differences in gut microbiota composition from the MetS untreated group and intervention participants before and after the MHP. The microbiota profiles of intervention participants before and after the program are represented as follows: colors indicate individual participants, with matching colors for paired samples (e.g., participant MHP2 before and after are both blue), the MetS untreated group is also assigned distinct colors, separate from the intervention participants. Samples of MetS untreated participants are shown as squares, before MHP as circles, and after MHP as triangles.

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  Figure 3. Relative abundance. Stacked bar charts showing the relative abundance of amplicon sequence variants (ASVs) at the phylum (a,b) and family (c,d) taxonomic levels. (a) Group-level data by treatment (untreated MetS, before MHP, and after MHP). (b) Paired data for each intervention participant and MetS untreated participants. (c) Family-level data grouped by treatment (before vs. after MHP) for intervention participants and MetS untreated participants; (d) Paired data per intervention participant and MetS untreated participants.

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  Figure 4. Key differentially abundant taxa. (a) Differentially abundant genera in the untreated MetS group and (b) in the after-MHP group; (c) Differentially abundant species in the untreated MetS group and (d) in the after-MHP group. Only genera and species that were significantly different in relative abundance (FDR ≤ 0.05) are shown in the plot, with log2 fold change determined using DESeq2. Dot size reflects the magnitude of change (|log2FC|), with larger dots indicating greater shifts in abundance.

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  Figure 5. Correlation between gut microbiota and clinical parameters. Heatmap of Spearman correlations between bacterial genera (Y-axis) and clinical variables (systolic blood pressure (SBP), diastolic blood pressure (DBP), high-density lipoprotein (HDL), urinary albumin-to-creatinine ratio (uACR), aspartate aminotransferase (AST), body weight, triglycerides (TGs), glycated hemoglobin (HbA1c), and alanine aminotransferase (ALT)) (X-axis), annotated with log2 fold change values. Color key: Blue indicates negative correlations, white indicates no correlation, and red indicates positive correlations. The color scale ranges from –0.6 to 0.6, representing the strength and direction of Spearman correlation coefficients (Rho). Asterisks on the heatmap indicate significance levels based on Spearman correlation p-values: * p < 0.05, ** p < 0.01.