Microbial community dynamics were examined across different positions within each reactor to explore temporal and spatial variations in microbial composition. Beta-diversity analysis, based on Bray-Curtis distance matrices (Figure ), was used to assess how microbial communities varied with time and by reactor position. The analysis revealed that biofilm communities in R1 and R3 differed significantly from their original inocula over time, while R2 displayed a less pronounced difference. This pattern aligns with earlier observations showing that microbial communities in R1 inoculated with AS and R3 inoculated with CM experienced greater shifts than those in R2. The limited change in R2 may result from the AD inoculum being more pre-adapted to anaerobic digestion conditions, leading to less dramatic community shifts.