Aim: Biofilm-based aquaculture systems, proven cost-effective, reduce the need for expensive supplementary feed. The biofilm acts as a natural planktonic food source for reared organisms. Previous research highlighted the influence of fish stocking density and biomass on plankton diversity in different aquaculture settings, underscoring the significance of these factors in biofilm-based systems. Hence, the present study attempted to understand how varying stocking densities of pearlspot (Etroplus suratensis) influence plankton diversity indices in the biofilm-based aquaculture system. Methodology: The present experiment was designed in 5 Practical Salinity Unit (PSU) brackishwater with soil-bottomed FRP circular (500 L) tanks for 60 days. The sugarcane bagasse was used as a substrate forbiofilm production after fertilization with cow dung, urea, and lime. Fries of pearlspot (4.67 ± 0.04 mm 1.71 ± 0.03 g-1) were stocked at four different stocking densities in triplicates viz T1 (50 nos); T2 (100 nos), T3 (150 nos) and T4 (200 nos) fish fry m-3 in the biofilm-based rearing system. Plankton samples were collected from water and biofilm deposited on the substrate and analyzed for diversity indices. The Shannon Diversity Index, Simpson Diversity Index, Simpson Dominance Index, and Margalef Richness Index were used to compute the plankton diversity indices in different treatments using standard equations. Results: In the current investigation, lower stocking densities with lower fish biomass were associated with significantly higher (P≤0.05) Shannon Diversity Index, Simpson Diversity Index, and Margalef Richness Index in water. In contrast, the Simpson Dominance Index in water showed a significantly lower (P≤0.05) value for treatments with lower fish biomass than for treatments with higher biomass. Interpretation: The values obtained for various diversity indices indicated that a biofilm-rearing system with lower E. suratensis biomass produced more planktonic abundance, evenness, and species richness. Key words: Biofilm, Pearlspot, Plankton diversity, Stocking density