In the present study a biological “in vivo” assay, with freshwater isopod Asellus aquaticus, was used to define and evaluate the potential impact of the wastewater treatment plant (WWTP) wastewaters on local wildlife. Samples of both untreated and mechanically and biologically treated WWTP wastewater, were tested in the presence and absence of the formulated sediment for their lethal and sublethal effects. Chronic exposures to wastewater samples caused concentration dependent reduced locomotion, body pigmentation and molting frequency in exposed organisms. The observed effects indicate the overall toxicity and endocrine disruption of the wastewater samples. In contrast stimulations of the feeding rate and growth rate of the test organisms during the chronic exposure to sublethal levels of wastewater samples were observed, indicating an improvement in nutritional quality of the wastewater. The most sensitive exposure endpoint was molting frequency of tested organisms, which indicated the presence of estrogenically active endocrine disrupting compounds (EDCs). Raw wastewater caused up to 42% molting frequency reduction of exposed A. aquaticus when exposed to five times diluted untreated wastewater sample, while undiluted treated wastewater caused a 61% molting frequency reduction. The presence of estrogenically active compounds in the wastewater was confirmed with the yeast estrogen screen assay (YES test), which assigned the highest estrogenic activity to a mechanically and biologically treated wastewater sample, and lower estrogenic activity to all other tested samples. The importance of presence of the formulated sediment was determined, as it lessened the effects of all WWTP wastewater samples in all observed exposures.