Mixed-culture dark fermentation is an environmentally friendly bio-hydrogen production process. In this work we study the potential for directing microbial anaerobic mixed communities towards improved hydrogen production. Strategies applied for promoting the selection of hydrogen-producing bacteria in anaerobic granular sludge consisted of heat treatment and chemical treatment with 2-bromo-ethane sulfonate (BES) and with BES+Chloroform. Three expanded granular sludge bed reactors, RHeat, RBES and RBES+Chlo, where inoculated with the treated sludges and fed with synthetic sugar-based wastewater ...