This study investigated and optimized the operational conditions for continuous hydrogen production from sugar beet molasses, co-fermented with liquid swine manure in an anaerobic sequencing batch reactor. Results indicated that pH, HRT and total solids content in the swine manure (TS) had significant impact on all the responses such as biogas production rate (BPR), hydrogen content (HC), hydrogen production rate (HPR), and hydrogen yield (HY), although the highest level of each response was achieved at different combination of the three variables. The maximum BPR, HC, HPR and HY of 32.21 L/d, 30.51%, 2.23 L/d/L and 1.57 mol-H-2/mol-sugar were estimated at the optimal pH, HRT, and TS of 5.55, 15.78 h, and 0.71% for BPR; 5.22, 12.04, and 0.69 for HC; 5.32, 15.62, and 0.78% for HPR; and 5.36, 17.56, and 0.74% for HY, respectively. Good linear relationships of the predicted and tested results for all the parameters were observed. (C) 2013 Elsevier Ltd. All rights reserved.