A two-stage process combining dark/photo fermentation was used to increase the overall hydrogen yield from sucrose and also to reduce the chemical oxygen demand (COD) in the effluent. Dark-H-2 fermentation was conducted using Clostridium pasteurianum CH4, giving a maximum H-2 production yield of 3.80 mol H-2/mol sucrose. The soluble metabolites resulting from dark fermentation, consisting of butyric and acetic acid, were further used for H-2 production in the subsequent photo fermentation. Using soluble products from dark fermentation as substrate, Rhodopseudomonas palustris WP3-5 could produce H-2 phototrophic ally, elevating the total hydrogen yield from 3.80 (dark fermentation) to 10.02 mol H-2/mol sucrose (dark/photo fermentation). Meanwhile, a 72.0% COD removal was also achieved. When the photobioreactor was illuminated with side-light optical fibers and was supplemented with 2.0% (w/v) of clay carriers, the overall H-2 yield of the two-stage process was further enhanced to 14.2 mol H-2/Mol sucrose with a nearly 90% COD removal. Continuous photo fermentation was also carried out at 96 h HRT using effluent from dark fermentation as the feed. The continuous culture maintained stable for nearly 10 days with an average H-2 yield of 10.21 mol H-2/mol sucrose. This demonstrates the feasibility of using the two-stage process combining dark and photo fermentation for simultaneous hydrogen production and COD removal. (c) 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.