An Upflow Anaerobic Sludge Blanket Reactor/Continuous Stirred Tank Reactor was used sequentially to decolorize and mineralize C.I. Direct Black 38 azo dye (3200 mg/L) in a synthetic wastewater containing glucose as co-substrate. At the steady state conditions color was effectively removed under anaerobic condition, while the total aromatic amines and organic fraction could be mainly reduced under aerobic conditions. NO3--N, COD, BOD5, aromatic amine, HPLC and GC analyses showed that Direct Black 38 could be chiefly mineralized by the sequential system. The toxicity levels were determined using three different test organism (ATA-anaerobic toxicity, respiration/inhibition and Daphnia magna tests) through the continuous operation of anaerobic/aerobic sequential system treating Direct Black 38 dye containing synthetic wastewater. Feed and anaerobic effluent had greater toxicity than the aerobic effluent after mineralization of dye. (C) 2004 Elsevier B.V. All rights reserved.