Insights into the role of chitosan in hydrogen production by dark fermentation of waste activated sludge

Chitosan is widely used as a dewatering flocculant, but whether it affects hydrogen production from sludge anaerobic fermentation is unclear. This study aimed to elucidate the role of chitosan in the dark fermentation of waste activated sludge for hydrogen production. The results showed that chitosan had a negative effect on hydrogen production from sludge. Chitosan at 30 g/kg total suspended solids reduced hydrogen accumulation by 56.70 +/- 1.22 % from 3.94 +/- 0.12 to 1.71 +/- 0.10 mL/g volatile suspended solids. Chitosan hindered the solubilization of sludge by flocculation, which reduced the available substrate for anaerobic fermentation. In addition, chitosan interfered with the electron transport system by reducing cytochrome C and caused lipid peroxidation by inducing reactive oxygen species, thereby inhibiting the activity of enzymes involved in anaerobic fermentation. Hydrogen production was reduced because hydrogen-producing processes (i.e., hydrolysis, acidification, and acetification) were inhibited more strongly than hydrogen-consuming processes (i.e., methanogenesis, sulfate reduction, and homoacetogenesis). Furthermore, chitosan enriched the abundance of Spirochaetaceae sp. and Holophagaceae sp., which occupied the survival space of hydrogen-producing microorganisms. This study reveals the potential impact of chitosan on hydrogen production in dark fermentation of sludge and provide direct evidence that chitosan triggers oxidative stress in anaerobic fermentation.