Microsensor measurements and fluorescence in situ hybridization (FISH) analysis were combined to investigate the microbial populations and activities in a laboratory-scale sequencing batch reactor (SBR) for completely autotrophic nitrogen removal over nitrite (CANON). Fed with synthetic wastewater rich in ammonia, the SBR removed 82.5±5.4% of influent nitrogen and a maximum nitrogen-removal rate of 0.52 kgN·m−3·d−1 was achieved. The FISH analysis revealed that aerobic ammonium-oxidizing bacteria (AerAOB) Nitrosomonas and anaerobic ammonium-oxidizing bacteria (AnAOB) dominated the community. To quantify the microbial activities inside the sludge aggregates, microprofiles were measured using pH, dissolved oxygen (DO), , and microelectrodes. In the outer layer of sludge aggregates (0–700 μm), nitrite-oxidizing bacteria (NOB) showed high activity with 4.1 μmol·cm−3·h−1 of maximum nitrate production rate under the condition of DO concentration higher than 3.3 mg·L−1. Maximum AerAOB activity was detected in the middle layer (depths around 1700 μm) where DO concentration was 1.1 mg·L−1. In the inner layer (2200–3500 μm), where DO concentration was below 0.9 mg·L−1, AnAOB activity was detected. We thus showed that information obtained from microscopic views can be helpful in optimizing the SBR performance.
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