A thermophilic anaerobic bacterium Clostridium sp. TCW1 was isolated from dairy cow dung and was used to produce hydrogen from cellulosic feedstock. Extracellular cellulolytic enzymes produced from TCW1 strain were identified as endoglucanases (45, 53 and 70 kDa). exoglucanase (70 kDa), xylanases (53 and 60 kDa), and beta-glucosidase (45 kDa). The endoglucanase and xylanase were more abundant. The optimal conditions for H-2 production and enzyme production of the TCW1 strain were the same (60 degrees C, initial pH 7, agitation rate of 200 rpm). Ten cellulosic feedstock, including pure or natural cellulosic materials, were used as feedstock for hydrogen production by Clostridium strain TCW1 under optimal culture conditions. Using filter paper at 5.0 g/L resulted in the most effective hydrogen production performance, achieving a H-2 production rate and yield of 57.7 ml/h/L and 2.03 mol H-2/mol hexose, respectively. Production of cellulolytic enzyme activities was positively correlated with the efficiency of dark-H-2 fermentation. (C) 2011 Elsevier Ltd. All rights reserved.