Figure 2. Impact of Repeated Controlled Ovarian Hyperstimulation (COH) on Alpha and Beta Diversity of the Gut Microbiota in Female Mice. Female Swiss mice (n=10 per group) were assigned to a treatment group receiving repeated COH for 10 weeks or a placebo-treated control group. (A-D) Alpha diversity was assessed using four metrics: (A) Observed OTUs (Control: 430.5±29.7; Treatment: 418.7±15.6; p=0.7294), (B) Shannon diversity index (Control: 6.02±0.98; Treatment: 6.27±0.67; p=0.5252), (C) Simpson diversity index (Control: 0.96±0.02; Treatment: 0.97±0.01; p=0.3260), and (D) Faith's Phylogenetic Diversity (Control: 17.41±1.73; Treatment: 18.19±1.29; p=0.2702). Data are presented as mean±standard deviation (SD). No significant differences were observed between groups for any of the alpha diversity metrics (unpaired two-tailed t-test). (E) Beta diversity was evaluated using weighted UniFrac distances, and visualized via principal coordinates analysis (PCoA). Each dot represents a fecal sample; ellipses denote 95% confidence intervals for each group. No clear clustering by group was observed (PERMANOVA; p>0.05).