BACKGROUND: Among the methods to produce hydrogen biologically, dark fermentation stands out mainly due to its low operating requirements. Organic wastes, such as brewing industry waste slurries and glycerol, can provide a cost-effective feedstock with the additional potential of generating value-added byproducts, while addressing a wastewater treatment issue.RESULTS: The hydrogen production potential in dark fermentation of a high-strength brewery waste slurry was optimized with a selected seed sludge, initial COD concentration of 50-60 g L −1 , pH 6.4 and fermentation time of 30 h. The main end product was butyric acid, accounting for over 50% of the carboxylic acids. The efficiency of the process on the basis of volume of H 2 obtained per gram of COD converted into organic acids was 393 ± 5 and 430 ± 6 mL without and with glycerol, respectively, and the molar ratio of H 2 per mole of substrate was 71% of the theoretical molar yield when the fermentation is dominated by butyrate as the end product.CONCLUSIONS: A proposed brewery sludge treatment system comprising dark fermentation followed by anaerobic digestion is promising and can be more advantageous than anaerobic digestion alone with an increase of 18.5% in energy potential. Alternatively, with recovery of valuable butyrate, a reduction in 4.5 kg of CO 2 emissions per cubic meter of sludge treated can be achieved, with a 27% net loss in energy potential.