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Rapid enrichment of anaerobic ammonia oxidation bacteria (AnAOB) is highly associated with the granulation process; however, the interactive mechanism remains unclear, especially for the initial granulation process. A single-stage partial nitritation-anammox (PN/A) bioreactor combined with granular/floc sludge was operated for 400 days. During the experimental period, the nitrogen removal rate increased from 0.60 to 1.21 kg N m-3d-1, and the nitrogen removal capability improved primarily during a transition period (days 200-250), which was accompanied by a particle size increase and AnAOB proliferation (4.9 ± 1.7 days). Moreover, as observed by the biomass physio-morphology, the size distribution, and the microbial community shift, small flocs (< 200 μm) aggregated due to the addition of excess sodium acetate. The emerging floc aggregates represented an early form of granules, providing the initial biological carrier and necessary anaerobic microenvironment for the growth of attached AnAOB, resulting in a high AnAOB growth-rate. These results are the first direct evidence that floc aggregates are essential to AnAOB enrichment, and that they can be affected by operational conditions. This study provides an in-depth understanding of the link between floc aggregations and AnAOB enrichment and broadens the feasibility of optimizing PN/A applications.
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