Biohydrogen production from xylose by fresh and digested activated sludge at 37, 55 and 70 °C

Two heat-treated inocula, fresh and digested activated sludge from the same municipal wastewater treatment plant, were compared for their H₂ production via dark fermentation at mesophilic (37 °C), thermophilic (55 °C) and hyperthermophilic (70 °C) conditions using xylose as the substrate. At both 37 and 55 °C, the fresh activated sludge yielded more H₂ than the digested sludge, whereas at 70 °C, neither of the inocula produced H₂ effectively. A maximum yield of 1.85 mol H₂ per mol of xylose consumed was obtained at 55 °C. H₂ production was linked to acetate and butyrate production, and there was a linear correlation (R² = 0.96) between the butyrate and H₂ yield for the fresh activated sludge inoculum at 55 °C. Approximately 2.4 mol H₂ per mol of butyrate produced were obtained against a theoretical maximum of 2.0, suggesting that H₂ was produced via the acetate pathway prior to switching to the butyrate pathway due to the increased H₂ partial pressure. Clostridia sp. were the prevalent species at both 37 and 55 °C, irrespectively of the inoculum type. Although the two inocula originated from the same plant, different thermophilic microorganisms were detected at 55 °C. Thermoanaerobacter sp., detected only in the fresh activated sludge cultures, may have contributed to the high H₂ yield obtained with such an inoculum.