For dark fermentation (DF) to be accepted as a sustainable process for biohydrogen production, the net energy gain should be positive and as high as possible. A theoretical approach is proposed in this study to evaluate the net energy gain possible from hydrogen generated by the DF process as well as from the end products of DF via anaerobic digestion (AD) and microbial fuel cells (MFC). Experimental data on hydrogen evolution and aqueous end products formation from sucrose and from sucrose/dairy manure blends were used to validate the proposed approach for estimating net energy gain via DF, DF + AD, DF + MFC. Good agreement was found between the experimental and predicted net energy gain values, with overall correlation coefficient of 0.998. Based on the results of this study, DF + MFC is recommended as the best combination to maximize net energy gain. Copyright (C) 2011, Hydrogen Energy Publications, LLC. Published by Elsevier Ltd. All rights reserved.