An expanded granular sludge bed (EGSB) process with granular activated carbon (GAC) was developed for fermentative hydrogen production from molasses-containing wastewater by mixed microbial cultures. No pH regulation was performed during the whole operation period. Running at the temperature of 35 degrees C, the EGSB reactor presented a high hydrogen production ability as the hydrogen production rate (HPR) maximized at 0.71 L/Lh. At the same time, the hydrogen yield (HY) peaked at 3.47 mol/mol sucrose and the maximum specific hydrogen production rate (SHPR) was found to be 3.16 mmol H-2/g VSS h. Hydrogen volume content was estimated to be 30-53% of the total biogas and the biogas was free of methane throughout the study. Dissolved fermentation products were predominated by acetate and ethanol, with smaller quantities of propionate, butyrate and valerate. It was found that high hydrogen yield was always associated with a high level of ethanol production. When the pH value and alkalinity ranged from 4.2-4.4 ing CaCO3/L to 280-340 mg CaCO3/L, respectively, stable ethanol-type fermentation was formed with the sum of ethanol and acetate concentration ratio of 89.1% to the total liquid products. The average attached biofilm concentration was estimated to be 17.1 g/L, which favored hydrogen production efficiently. With high biomass retention at high organic loading rate (OLR), this EGSB system showed to be a promising high-efficient bioprocess for hydrogen production from high-strength wastewater. (C) 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.