Summary: | Biohydrogen production from organic solid waste has shown particular advantages over other methods owing to the combination of waste reduction and bioenergy production. In this study, biohydrogen production from the co-digestion of cattle manure and food waste was optimized in a mesophilic semi-continuous process. To maximize hydrogen production, the effects of the mixing ratio (the proportion of food waste in the substrate), substrate concentration, and hydraulic retention time (HRT) on the co-digestion were systematically analyzed using a Box–Behnken design. The results showed that strong interactive effects existed between the three factors, and they had a direct effect on the responses. Hydrogen was primarily produced via the butyrate pathway, which was accompanied by the competing heterolactic fermentation pathway. Propionate and valerate produced from lipids and proteins, respectively, were obtained along with butyrate. The optimal process parameters included a mixing ratio of 47% to 51%, a substrate concentration of 76 to 86 g L<sup>−1</sup>, and an HRT of 2 d. Under these optimal conditions, the hydrogen production rate and hydrogen yield were higher than 1.00 L L<sup>−1</sup> d<sup>−1</sup> and 30.00 mL g<sup>−1</sup> VS, respectively, and the predicted results were consistent with the experimental data. The results indicate that the co-digestion of cattle manure and food waste is a practical and economically promising approach for biohydrogen production.
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