Effect of bioaugmentation using Clostridium butyricum on the start-up and the performance of continuous biohydrogen production

This study aimed to mitigate the instability in the start-up and continuous performance of dark fermentative biohydrogen production using heat-treated sludge by the addition of an exogenous H₂-producing strain. Continuous fermentation augmented with Clostridium butyricum showed the highest average biohydrogen production rate (HPR) as 50.35 ± 2.56 and 58.57 ± 5.03 L/L-d with H₂-producing butyric and acetic acid pathways, whereas the fermenters without bioaugmentation showed the termination of biohydrogen production in 3 days of continuous operation with non H₂-producing lactic acid pathway and H₂-consuming propionic acid pathway. The bioaugmentation blocked the growth of the competitors for hexose such as Streptococcus, Lactobacillus and Megasphaera, and provided H₂-producer dominated microbiome with not only Clostridium butyricum, but also Clostridium puniceum and Clostridium neuense originated from heat-treated sludge. Bioaugmentation of a H₂-producing strain would be a reliable dissemination strategy for dark fermentative biohydrogen production by minimizing the influence of seed sludge population.