Key function of Kouleothrix in stable formation of filamentous aerobic granular sludge at low superficial gas velocity with polymeric substrates.

Forming and maintaining stable aerobic granular sludge (AGS) at a low superficial gas velocity (SGV) is challenging, particularly with polymeric substrates. This study cultivated filamentous aerobic granular sludge (FAGS) with filamentous Kouleothrix (Type 1851) at low SGV (0.15 cm/s) utilizing mixed acetate-soluble starch. Within approximately 260 days, notable increases in the relative abundance of Kouleothrix (from 4 % to 10 %) and Ca. Competibacter (from 1 % to 26 %) were observed through 16S rRNA gene analysis. Metagenomic analysis revealed increased expression of functional genes involved in volatile fatty acid (VFA) production (e.g., ackA and pta) and polyhydroxyalkanoate synthesis (e.g., phbB and phbC). Kouleothrix acted as a skeleton for bacterial attachment and was the key fermenting bacteria promoting granulation and maintaining granule stability. This study provides insight into the formation of FAGS with low-energy and non-VFA substrates.