Effect of Pretreatment of Anaerobic Acid Fermentation Sludge from Mixed Swine Wastewater and Food Waste Leachate on Biohydrogen Generation

Objectives:This study aims to investigate the effects of heat shock and 2-bromoethanesulfonate (2-BES) injection on hydrogen production and methanogens inhibition in anaerobic acid fermenter sludge and methane fermenter sludge using mixed swine wastewater and food waste leachate as substrates. Additionally, the analysis of biogas, fermentation products such as volatile fatty acids (VFAs) and alcohols, and microbial community changes were conducted to derive efficient pretreatment conditions for anaerobic hydrogen generation.Methods:Sludge samples were collected from two-phase anaerobic digesters treating swine wastewater and food waste leachate. Heat shock (90°C, 15 min) and 2-BES injection (1, 5, 10, 50, and 100 mmol/L) pretreatments were applied to the each sludge, along with control reactor without pretreatment before anaerobic digestion. Batch experiments were conducted at 30°C to assess biogas, organic compounds (COD, VS, VFAs, and alcohols), and microbial community composition.Results and Discussion:In methanogenic sludge, hydrogen gas was not produced under all conditions, but heat shock method inhibited methane production more effectively than 2-BES injection method. As the concentration of 2-BES increased, methane production decreased. Conversely, hydrogen was produced from the all acid fermentation sludges, mainly attributed to the butyric acid production by dominant Clostridium spp., but the pattern of biogas production varied over time. In the experimental group injected with 2-BES, even at a low concentration of 1 mmol/L, methane gas did not produce due to the inhibition of methanogens. However, in the control group, produced hydrogen was entirely converted to methane by hydrogenotrophic methanogens such as Methanobrevibacter and Methanosphaera. The heat shock group showed delayed hydrogen production due to prolonged lag phase of hydrogen-producing bacteria, with hydrogenotrophic methanogen metabolism starting after 12 days. Heat shock and 2-BES injection reduced bacterial diversity in the acid fermentation process compared to the initial stage.Conclusion:For efficient anaerobic hydrogen production from swine wastewater and food waste leachate, 2-BES injection into the acid fermentation sludge was found to be a preferable method over heat shock, considering hydrogen production efficiency and sustained methanogen inhibition. Further research is needed to find more economical and efficient pretreatment methods and operational strategies considering subsequent methane production processes for practical application in the field.