Granular sludge from a full-scale methane reactor treating brewery wastewater was used as a seed for the treatment of glycerol in a laboratory-scale repeated-batch methane reactor, and the change in the microbial community during the acclimation stages was examined. Two types of substrate solutions, a glucose, sodium acetate, and lactic acid mixture, as well as glycerol, were prepared and fed by mixing the two solutions to increase the ratio, in a stepwise manner, of glycerol from 0% to 100%, while keeping a loading of COD at 2.5 kg m(-3) d(-1) throughout the fermentation process. Vigorous methane gas production, approximately 580 dm(3) m(-3) d(-1), was observed during the acclimation stages. Microbial analysis revealed that both bacterial and archaeal communities changed significantly; bacteria (genus Trichococcus and family Syntrophomonadaceae) became dominant rapidly after the start of acclimation, and archaea belonging to the hydrogenotrophic methanogens (genera Methanobacterium and Methanospirillum), increased gradually with the progress of acclimation. (C) 2014 Elsevier Ltd. All rights reserved.