1.3.1. Alpha diversity.
From the Illumina paired-end raw reads of samples, the reads having a V3 to V4 primer sequence and high-quality bases were filtered. All the samples were individually analyzed for microbiome diversity and species abundance using QIIME analysis. Rarefaction analysis allows comparisons between communities based on the richness of microbial diversity. Alpha rarefaction was performed at a level of 22,000 reads to include all samples, which is used as a measure of the depth of sequencing and the total data. The rarefaction plot in Figure (A) Rarefaction plot for 52 samples of V3 to V4 region at a depth of 24,000. The image was plotted against the number of sequences per sample on the X-axis versus diversity index on Y-axis. The samples are colored by their respective names. As the sequencing depth increased, the number of observed species (OTUs) also increased. Eventually, the curves began to plateau, indicating that as the number of extracted sequences increased, the number of OTUs detected decreased; Box plot representing alpha diversity measured by (B) Shannon, (C) Chao1, (D) Simpson for the Placebo and Active. The line inside the box represents the median, while the whiskers represent the lowest and highest values within the 1.5 interquartile range (IQR); (E and F) The dots represent outliers principal coordinate analysis (PCoA) plots of the active (n = 12), versus placebo group (n = 14) specific taxa with significant (Alpha diversity indexes are composite indexes reflecting abundance and consistency. We observed a minimum of 375 and a maximum of 1481 species in all the samples. The richness of annotated species in V3 to V4, Shannon, Simpson, and Chao1, indexes are represented in Figures 