Fermentative hydrogen production from a synthetic wastewater containing 10 g/L of sucrose was studied in two upflow reactors at 26 degrees C for 400 days. One reactor was filled with packing rings (RP) and the other was packing free (RF). The effect of hydraulic retention time (HRT) from 2 h to 24 h was investigated. Results showed that, under steady state, the hydrogen production rate significantly increased from 0.63 L/L/d to 5.35 L/L/d in the RF when HRT decreased from 24 h to 2 h; the corresponding rates were 0.56 L/L/d to 6.17 L/L/d for the RP. In the RF, the hydrogen yield increased from 0.96 mol/mol-sucrose at 24 h of HRT to the maximum of 1.10 mol/mol-sucrose at 8 h of HRT, and then decreased to 0.68 mol/mol-sucrose at 2 h. In the RP, the yield increased from 0.86 mol/mol-sucrose at 24 h of HRT to the maximum of 1.22 mol/mol-sucrose at 14 h of HRT, and then decreased to 0.78 mol/mol-sucrose at 2 h. Overall, the reactor with packing was more effective than the one free of packing. In both reactors, sludge agglutinated into granules. The microbial community of granular sludge in RP was investigated using 16S rDNA based techniques. The distribution of bacterial cells and extracellular polysaccharides in hydrogen-producing granules was investigated by fluorescence-based techniques. Results indicated that most of the N-acetyl-galactosamine/galactose-containing extracellular polysaccharides were distributed on the outer layer of the granules with a filamentous structure.