An experiment was conducted to compare the efficiency of biofilm production in natural and artificial substrates and to study their effect on water quality and growth of Etroplus suratensis. Four different substrates were used for biofilm formation: paddy straw (T1), sugarcane bagasse (T2), polyvinylchloride (PVC) pipe (T3) and plastic sheet (T4). The experiment was carried out in mud-bottomed fibre-reinforced plastic tanks (300 L) in triplicates. About 3000 cm 2 surface area (600 g) of each substrate was suspended in water supplemented with fertilizers. Only cow dung and urea were applied in control tanks. The tanks were stocked with 25 fishes with average weight of 9.1 ± 0.22 g. The overall mean value of heterotrophic bacteria in substrate was found higher in straw followed by bagasse, plastic and PVC. The dominant genera of bacteria in the substrate were Bacillus, Pseudomonas and Micrococcus in that order of preponderance. The mean phytoplankton and zooplankton density on the substrates were higher in bagasse followed by straw, plastic and PVC. The biofilm developed on the substrate significantly reduced the ammonia nitrogen and nitrite-nitrogen content of water. The growth of fishes was significantly (P \ 0.05) higher in substrate-based treatments than that in the control with better results in bagasse followed by straw, plastic and PVC. The conclusions of the present study are that biofilm produced on natural substrates, especially on bagasse, enhanced growth of E. suratensis and reduced the necessity of water exchange during the culture, which certainly decreases the cost of Etroplus production.