Benthic microalgae have the natural capacity to adhere to a diversity of fixed submerged substrates to form biofilms, which have important roles not only in natural ecosystems, but also in aquaculture systems. An experimental investigation was performed to assess the biofilm-forming capacity of two microalgae (Navicula incerta and Navicula sp.) on three different substrates (plastic net, fabric, and wood) under controlled temperature and light conditions. The substrates were arranged on curtains suspended from a wood stick, into plastic aquariums (45 L in capacity) filled with filtered marine water enriched with F/2 medium. The trial was carried out until the exponential growing phase of the microalgae was reached. After that, the incorporated biomass was gravimetrically calculated, and its biochemical composition was determined by standard methods. The greatest amount of incorporated dry matter was observed for Navicula sp. on fabric and the lowest was observed for wood. The highest number of cells associated with the biofilm was obtained for Navicula sp. on the plastic net (1.24 x 109 cells/m2), while the lowest was recorded for Navicula sp. on the wood (1.43 x 108 cells/m2). Significant differences in organic matter were found among the substrates, with the highest values for N. incerta on the fabric (3.22 g/m2) and the lowest for Navicula sp. on the wood (0.02 g/m2). The best biochemical profiles among the formed biofilms were observed for N. incerta on the plastic net and Navicula sp. on the fabric. The plastic net was considered the best substrate because of the stability of the biofilm and the easiness of harvesting the biomass.Benthic microalgae have the natural capacity to adhere to a diversity of fixed submerged substrates to form biofilms, which have important roles not only in natural ecosystems, but also in aquaculture systems. An experimental investigation was performed to assess the biofilm-forming capacity of two microalgae (Navicula incerta and Navicula sp.) on three different substrates (plastic net, fabric, and wood) under controlled temperature and light conditions. The substrates were arranged on curtains suspended from a wood stick, into plastic aquariums (45 L in capacity) filled with filtered marine water enriched with F/2 medium. The trial was carried out until the exponential growing phase of the microalgae was reached. After that, the incorporated biomass was gravimetrically calculated, and its biochemical composition was determined by standard methods. The greatest amount of incorporated dry matter was observed for Navicula sp. on fabric and the lowest was observed for wood. The highest number of cells associated with the biofilm was obtained for Navicula sp. on the plastic net (1.24 x 109 cells/m2), while the lowest was recorded for Navicula sp. on the wood (1.43 x 108 cells/m2). Significant differences in organic matter were found among the substrates, with the highest values for N. incerta on the fabric (3.22 g/m2) and the lowest for Navicula sp. on the wood (0.02 g/m2). The best biochemical profiles among the formed biofilms were observed for N. incerta on the plastic net and Navicula sp. on the fabric. The plastic net was considered the best substrate because of the stability of the biofilm and the easiness of harvesting the biomass.