Aromatic amines (anilines and related derivates) are an important class of environmental pollutants that can be released to the aquatic environment as industrial effluents or as breakdown products of pesticides and dyes. The toxicity of aniline, 2-chloroaniline, 3-chloroaniline, 4-chloroaniline and 3,5-dichloroaniline towards a multitrophic test battery comprised of bacteria Aliivibrio fischeri (formerly Vibrio fischeri), a ciliated protozoan Tetrahymena thermophila and two crustaceans (Daphnia magna and Thamnocephalus platyurus) were investigated. Under the applied test conditions, the toxicity of the anilines notably varied among the test species. The bacteria and protozoa were much less sensitive towards the anilines than the crustaceans: EC50 values 13–403 mg L-1 versus 0.13–15.2 mg L-1. No general tendency between toxicity and the chemical structure of the anilines (the degree of chloro-substitution and the position of the chloro-substituents) was found in the case of all the tested aquatic species. The replacement of the artificial test medium (ATM) by the river water remarkably decreased the toxicity of anilines to crustaceans but not to protozoa. This research is part of the EU 6th Framework Integrated Project OSIRIS, in which ecotoxicogenomic studies of anilines (e.g., for Daphnia magna) will also be performed that may help to clarify the mechanisms of toxicity of different anilines.