Bacterial Community and Candidatus Accumulibacter Population Dynamics in Laboratory-Scale Enhanced Biological Phosphorus Removal Reactors

Candidatus Accumulibacter and total bacterial community dynamics were studied in two lab-scale enhanced biological phosphorus removal (EBPR) reactors by using a community fingerprint technique, automated ribosomal intergenic spacer analysis (ARISA). We first evaluated the quantitative capability of ARISA compared to quantitative real-time PCR (qPCR). ARISA and qPCR provided comparable relative quantification of the two dominant Ca. Accumulibacter clades (IA and IIA) detected in our reactors. The quantification of total Ca. Accumulibacter 16S rRNA genes relative to that from the total bacterial community was highly correlated, with ARISA systematically underestimating Ca. Accumulibacter abundance, probably due to the different normalization techniques applied. During 6 months of normal (undisturbed) operation, the distribution of the two clades within the total Ca. Accumulibacter population was quite stable in one reactor while comparatively dynamic in the other reactor. However, the variance in the clade distribution did not appear to affect reactor performance. Instead, good EBPR activity was positively associated with the abundance of total Ca. Accumulibacter. Therefore, we concluded that the different clades in the system provided functional redundancy. We disturbed the reactor operation by adding nitrate together with acetate feeding in the anaerobic phase to reach initial reactor concentrations of 10 mg/liter NO3-N for 35 days. The reactor performance deteriorated with a concomitant decrease in the total Ca. Accumulibacter population, suggesting that a population shift was the cause of performance upset after a long exposure to nitrate in the anaerobic phase.