Screening and Diversity Analysis of Aerobic Denitrifying Phosphate Accumulating Bacteria Cultivated from A2O Activated Sludge

Abstract

The aerobic denitrifying phosphate accumulating bacteria (ADPB) use NO3− as an electron acceptor and remove nitrate by denitrification and concomitant uptake of excessive phosphorus in aerobic conditions. Activated sludge was collected from the A2O aerobic biological pool of the sewage treatment plant at Hezuo Town, Chengdu City. The candidate ADPB strains were obtained by cultivation in the enriched denitrification media, followed by repeated isolation and purification on bromothymol blue (BTB) solid plates. The obtained candidates were further screened for ADPB strains by phosphorus uptake experiment, nitrate reduction test, metachromatic granules staining, and poly-β-hydroxybutyrate (PHB) staining. The 16 sedimentation ribosome deoxyribonucleic acid (16 S rDNA) molecular technique was used to determine their taxonomy. Further, the denitrification and dephosphorization capacities of ADPB strains were ascertained through their growth characteristics in nitrogen-phosphorus-rich liquid media. The results revealed a total of 25 ADPB strains screened from the activated sludge of the A2O aerobic biological pool of the sewage treatment plant at Hezuo Town. These strains belonged to two classes, four orders, and five genera. Among them, the strain SW18NP2 was a potentially new species in the Acinetobacter genus, while the strain SW18NP24 was a potential new species in the Pseudomonas genus at the time of their characterization. The Acinetobacter was the dominant genus. The obtained ADPB strains demonstrated a rich diversity. The ADPB strains had significant variations in denitrification and dephosphorization capacities. Twenty-three strains exhibited a total phosphorus removal rate of above 50%, and 19 strains exhibited a total nitrogen removal rate of above 50%. The strain SW18NP2 showed the best denitrifying phosphorus removal (DPR) capacity, with a dephosphorization rate of 82.32% and a denitrification rate of 73.73%. The ADPB in the A2O aerobic biological pool of the sewage treatment plant at Hezuo Town demonstrated a rich diversity and a strong DPR capacity.