Abstract

This study is the first to reveal the important roles of c-di-GMP on the formation of autotrophic granular sludge (AUGS) and autotrophic biofilm (AUB) in integrated fixed-film activated sludge (IFAS) system. The results revealed that the partial nitrification/anammox (PN/A) performance was successfully developed in the IFAS system and high nitrogen removal efficiency was obtained. Notably, it was clearly indicated that the intracellular c-di-GMP content was highly linearly correlated with the concentration of protein (PN) in tightly bound extracellular polymeric substances (TB-EPS) during AUGS formation. It was also found that the c-di-GMP content was highly linearly correlated with the PN concentration during AUB formation, and also had a good linear correlation with the polysaccharide (PS) content in TB-EPS. This proves that intracellular c-di-GMP played a vital role in regulating the TB-EPS secretion, thereby further affecting the aggregation ability of AUGS and AUB. In addition, when the c-di-GMP regulates the content of PN in TB-EPS, the secondary structure of the PN also changes significantly correspondingly, affecting the hydrophobicity of AUGS and AUB. In long-term operation, Candidatus_Kuenenia, Nitrosomonas and Flavobacterium gradually became dominant species, which were considered to favor the formation of AUGS and AUB by promoting the production of c-di-GMP and TB-EPS.

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