Biohydrogen production from Thermoanaerobacterium thermosaccharolyticum strain PSU-2 was examined in upflow anaerobic sludge blanket (UASB) reactor and carrier-free upflow anaerobic reactor (UA), both fed with sucrose and operating at 60 degrees C. Heat-pretreated methanogenic granules were used as carrier to immobilize T. thermosaccharolyticum strain PSU-2 in UASB reactor operated at a hydraulic retention time (HRT) ranging from 0.75 to 24h and corresponding sucrose loading rate from 58.5 to 2.4 mmol sucrose l(-1)h(-1). In comparison with hydrogen production rate of 12.1 mmol H(2)l(-1)h(-1) obtained by carrier-free reactor upflow anaerobic (UA) system, a greatly improved hydrogen production rate up to 152 mmol H(2)l(-1)h(-1) was demonstrated by the granular cells in UASB system. The biofilm of T. thermosaccharolyticum strain PSU-2 developed on treated methanogenic granules in UASB reactor substantially enhanced biomass retention (3 times), and production of hydrogen (12 times) compared to carrier-free reactor. It appears to be the most preferred process for highly efficient dark fermentative hydrogen production from sugar containing wastewater under thermophilic conditions. (C) 2008 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.