Operational mode affects the role of organic matter in granular anammox process.

Abstract In the presence of organic matter, the granular anammox system under sequencing batch mode showed more robust anammox performance than that under completely mixed mode, which was attributed to the better biomass retention with high settling ability and stability of granular sludge. Based on the specific anammox activity test, stratified and mixed distribution of heterotrophic bacteria was found under completely mixed and sequencing batch mode, respectively. The stratified microbial distribution resulted in low enzyme activity of anammox bacteria and sludge disintegration by hindering substrate transfer with a large accumulation of EPS on the granular surface. Whereas the heterotrophic bacteria mixed in granules (mixed microbial distribution) act as a “skeleton”, which increased the particle size, density, and stability of granular sludge. Compared with biokinetic-based selection, diffusion-based selection with high substrate penetration depth more likely resulted in the mixed granular structure and strong resistance to organic inhibition under sequencing batch mode.