The ecological assessment of a C12-15 AE-3S linear alkylethoxysulfate (AES) anionic surfactant to invertebrates, fish, periphyton, and an aquatic macrophyte was conducted in a 30-d outdoor stream mesocosm study with five replicated concentrations and controls. Alkylethoxysulfate structural integrity and exposure concentrations were maintained during the 30-d treatment period, with average measured concentrations of 0.7, 1.27, 2.2, 4.31, and 10.18 mg/L. No effects were observed on the aquatic macrophyte Myriophyllum aquaticum at the highest concentration tested. A sevenfold increase in periphyton biomass at 10.18 mg/L was observed relative to controls primarily because of increases in density of the filamentous alga Mougeotia sp. Densities of the invertebrates Annelida (Stylaria), Amphipoda, Copepoda, Trichoptera (Hydropsychidae), Cladocera, and Diptera (Chironomidae) significantly decreased in streams treated with AES at 10.18 mg/L. Densities of drifting invertebrates were not observed to be affected at any concentration tested. Reproduction of Pimephales promelas significantly decreased at 1.27 mg/L and growth of juvenile Lepomis macrochirus was significantly affected at 4.31 mg/L. Multivariate cluster analysis and nonmetric multidimensional scaling ordination showed distinct structural effects on the invertebrate communities in the streams treated with AES at 10.18 mg/ L compared to the control and streams treated at < 10.18 mg/L through the 30-d treatment. Convergence of the communities treated at 10.18 mg/L toward control communities in the ordination suggests recovery in these communities after termination of surfactant treatment. The results from this study support an ecosystem value of > 2.0 mg/L, and indicate that the conservative Dutch risk assessment for AESs has at least a fivefold margin of safety.