The present work evaluated the ecological risk of glyphosate by its commercial formulation (Roundup Original®) used to control floating aquatic macrophytes. Exposure analysis and ecological effects were performed from microcosm studies. The risk characterization was performed based on the calculation of the risk quotient. The commercial formulation of glyphosate had high toxicity when it was assessed separately. On the other hand, ecotoxicological evaluation of water samples from microcosms did not present toxicity to any tested organisms, however, glyphosate application is recommended exclusively to water bodies that have the surface completely covered by macrophytes.