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The aim of this study was to determine the optimal conditions for dark fermentation using agro-industrial liquid wastewaters mixed with sweet sorghum stalks (i.e., 55% sorghum, 40% cheese whey, and 5% liquid cow manure). Batch experiments were performed to investigate the effect of controlled pH (5.0, 5.5, 6.0, 6.5) on the production of bio-hydrogen and volatile fatty acids. According to the obtained results, the maximum hydrogen yield of 0.52 mol H2/mol eq. glucose was measured at pH 5.5 accompanied by the highest volatile fatty acids production, whereas similar hydrogen productivity was also observed at pH 6.0 and 6.5. The use of heat-treated anaerobic sludge as inoculum had a positive impact on bio-hydrogen production, exhibiting an increased yield of 1.09 mol H2/mol eq. glucose. On the other hand, the pretreated (ensiled) sorghum, instead of a fresh one, led to a lower hydrogen production, while the organic load decrease did not affect the process performance. In all experiments, the main fermentation end-products were volatile fatty acids (i.e., acetic, propionic, butyric), ethanol and lactic acid.

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