The objective of this work was to evaluate the effect of the use of an artificial substrate for periphytic biofilm growth on the microbiological composition of the biofilm, water quality, and zootechnical performance of Nile tilapia (Oreochromis niloticus) in rearing tanks. The experiment consisted of two treatments: presence and absence of artificial substrate for the growth of periphyton, with five replicates. Water quality parameters were evaluated throughout the entire experimental period. Every two weeks, samples of the periphyton were collected for the analysis of its microbiological composition and biometric measurements of the fish were performed. The number of heterotrophic bacteria in the biofilm increased during the experimental period, but that of Aeromonas decreased with the use of the artificial substrate. The total ammonia nitrogen in the tanks with periphyton reduced in 30% compared with that of the control. Tilapia reared with the biofilm showed a final weight 2.4 times greater than that of those subjected to the control, as well as an increase of 59.19% in their productivity. The use of an artificial substrate for periphytic biofilm in Nile tilapia rearing tanks favors the maintenance of the quality of the culture water, the protection of the fish against pathogens, and the improvement of the zootechnical performance of the fish.