The toxic effect of exposure to arsenic, As(V), at concentrations of 0 to 30 mg L−1, for 49 days, on epipsammic biofilms, was evaluated in a microcosm experiment. The growth and composition of biofilms developed on sediments containing As concentrations of 31 mg kg−1 and 85 mg kg−1 were compared, using photosynthetic parameters and Live/Dead stains as end points. A toxic effect of arsenic could not be demonstrated; however, biofilm growth was higher over the sediment with higher arsenic concentrations, suggesting the development of pollution-induced community induced tolerance (PICT). Nevertheless, PICT was not observed after exposure to high arsenic concentration in the laboratory, as there were no differences in algal growth between the previous 0 and 30 mg L−1 systems exposed to new 30 mg As L−1 dissolution over 29 days. The algal composition was affected by the added arsenic, and brown algae were the most tolerant compared to green algae and cyanophyceae, as their percentage increased from 25 and 33% in the control samples to 57 and 47% in the samples with the highest added As concentration. In turn, the biofilm development influenced arsenic redistribution and speciation. Arsenic concentration in water decreased with time during the incubation experiment, retained by the sediment particles and the biofilm. In the biofilm, extracellular As was significantly higher (up to 11 times) than intracellular arsenic. As(V) was the predominant species in water and in the biofilm, but products of biotic transformation, namely As(III), DMA(V) and MMA(V), were also found in the solution and in the biofilm in some systems, demonstrating reduction and methylation by the organisms. As a conclusion, a toxic effect was not detected for the concentrations evaluated. Biofilms naturally exposed in the river system to high As concentrations acquire pollution-induced tolerance; however, tolerance was not acquired by exposure to 30 mg L−1 for 29 days in the laboratory.