The denitrification process was studied in two granular biomass denitrifying reactors (USB1 and USB2). In USB1 large quantities of biomass were accumulated (9.5 gVSS L-1) allowing for the treatment of high nitrogen loads (3.5 g NO3--N L-1 d(-1)). As the biomass granulation process is not immediate the effects of different upflow velocities (0.12-5.5 m h(-1)) and calcium contents (5-200 mg Ca2+ L-1) were studied in order to speed up the process. Obtained results indicate that the optimum values for these parameters, which allow for the stable operation of USB1, are of 0.19 m h(-1) and 60 mg Ca2+ L-1. Then these optimum conditions were applied to USB2 where the effects of concentrations from 335 to 1000 mg NO3--N L-1 were tested. In these conditions nitrate concentrations of 1000 mg NO3--N L-1 are required for denitrifying granular biomass formation. Summarizing denitrifying granules can be formed at low upflow velocities and in hard or extremely hard water composition conditions if sufficient high nitrogen loads are treated. (C) 2015 Elsevier Ltd. All rights reserved.