Microbial and physicochemical monitoring of granular sludge during start-up of thermophilic UASB reactor

Mesophilically-grown granular sludge seeded in thermophilic UASB reactor was monitored to better understand the start-up process of the reactor. The reactor was fed with a synthetic wastewater containing glucose. As COD loading rate increased stepwise, methane production rate increased. Maximum values of COD removal efficiency (95%) and methane production rate (5.3 l/day) were achieved by approximately day-80 and remained constant afterward. However, physicochemical and microbial properties of granules kept changing even after day-80. Specific methanogenic activity (SMA) was initially negligible, and increased continuously until day-153 and remained constant afterward, showing the maximum value of 1.51+/-0.13 g CH4-COD/g VSS/day. Deteriorated settling ability of granules recovered the initial value by day-98 and was maintained afterward, as determined by sludge volume index. Initially reduced granule size increased until day-126, reaching a plateau of 1.1 rum. Combined use of fluorescence in situ hybridization and confocal laser scanning microscopy (CLSM) allowed to localize families of Methanosaetaceae and Methanosarcinaceae in granules with time. Quantitative analyses of CLSM images of granule sections showed abundance patterns of the methanogens and numerical dominance of Methanosaeta spp. throughout the start-up period. The trend of SMA agreed well with abundance patterns of the methanogens.