We examined how long-term operation of anaerobic-oxic and anaerobic-anoxic sequencing batch reactors (SBRs) affects the enhanced biological phosphorus removal (EBPR) performance and sludge characteristics. The microbial characteristics of phosphorus accumulating organism (PAO) and denitrifying PAO (DPAO) sludge were also analyzed through a quantitative analysis of microbial community structure. Compared with the initial stage of operation characterized by unstable EBPR, both PAO and DPAO SBR produced a stable EBPR performance after about 100-day operation. From day 200 days (DPAO SBR) and 250 days (PAO SBR) onward, sludge granulation was observed, and the average granule size of DPAO SBR was approximately 5 times larger than that of PAO SBR. The DPAO granular sludge contained mainly rod-type microbes, whereas the PAO granular sludge contained coccus-type microbes. Fluorescence in situ hybridization analysis revealed that a high ratio of Accumulibacter clade I was found only in DPAO SBR, revealing the important role of this organism in the denitrifying EBPR system. A pyrosequencing analysis showed that Accumulibacter phosphatis was present in PAO sludge at a high proportion of 6%, whereas it rarely observed in DPAO sludge. Dechloromonas was observed in both PAO sludge (3.3%) and DPAO sludge (3.2%), confirming that this organism can use both O and NO as electron acceptors. Further, Thauera spp. was identified to have a new possibility as denitrifier capable of phosphorous uptake under anoxic condition.
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