The aim of this work was to examine the ecotoxicity of reclaimed wastewater by the use of bioassays and the determination of immunological parameters. Secondary and tertiary mucicipal wastewater samples were examined for their physicochemical and microbiological characteristics as well as for their endotoxin concentrations. The ecotoxicological characteristics were assessed by a battery of bioassays, using Vibrio fischeri, Daphnia magna and Tetrahymena thermophilla as test species and phytotoxicity. The mitogenic responses of mouse splenocytes were as well used as bioassay. The cytokines of IL-1, IL-2, IL-10, IFNγ and TNFα, were also determined in the supernatant of splenocyte cultures and served as molecular biomarkers. All bioassays exhibited decrease of the ecotoxicological responses after tertiary treatment. However, mitogenic responses were proved to be more sensitive. IL-1 increased, while IL-2 production was unaffected. The fact that IL-10 production increased in response to secondary treated effluents in conjunction with the increased endotoxin levels, suggest Th2 type immune responses. Although results obtained from the toxicity bioassays after the tertiary treatment showed comparable results to those of controls, cytokine levels indicated the induction of immune response even after tertiary treatment. Consequently, cytokine production could be used as a sensitive biomarker for the evaluation of treatment efficiency of the reclaimed wastewaters intended for reuse.