The estrogenic potency of a wastewater treatment plant (WWTP) was evaluated using chemical and biological analyses, which showed that after the station treatment processes some of the selected endocrine disruptor compounds (EDCs) were still present in the treated effluent (e.g., bisphenol A, alkylphenols, estrone). Thus, the most common endocrine EDCs were identified and quantified and the overall estrogenicity of the treated effluent assessed by integrating the results. Male goldfish (Carassius auratus) were used as biological indicators in a 28-day experiment. Vitellogenin (Vtg), gonadosomatic and hepatosomatic indices, steroids (17beta-estradiol and 11-ketotestosterone) and histopathology were biomarkers used in fish to evaluate WWTP treated effluent estrogenicity, in combination with instrumental analyses. The results showed a significant increase (P < 0.01) in plasma and liver Vtg, which were significantly correlated (r = 0.66; P < 0.01). The gonadosmatic index was significantly (P < 0.01) reduced in exposed fish. The steroid analyses revealed significant elevations in 17beta-estradiol and depressed 11-ketotestosterone concentrations. The histological examinations show changes in exposed fish gonads, such as regressed testes and in some cases (43% to 75%) the development of ovo-testis in fish exposed to 50% and 100% treated effluent.