Toxicity tests with aquatic organisms constitute an effective tool in the evaluation, prediction and detection of the potential effect of pollutants from environmental samples in living organisms. Vinasse, a highly colored effluent, is a sub-product rich in nutrients, mainly organic matter, with high pollutant potential when disposed in the environment. Assays for vinasse decolorization were performed using the fungus Pleurotus sajor-caju CCB020 in vinasse biodegradation study, were occurred reductions of 82.8% in COD, 75.3% in BOD, 99.2% in the coloration and 99.7% in turbidity. The vinasse toxicity reduction was determined by the exposition to the following organisms: Pseudokirchneriella subcapitata, Daphnia magna, Daphnia similis and Hydra attenuata. This work concluded that the systematic combination of P. sajor-caju and vinasse can be applied in the bioprocess of color reduction and degradation of complex vinasse compounds, with reduction in the toxicity and improving its physical-chemical properties.