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• A novel dPAO from aquaculture sediment was isolated. • Strain K14 could utilize ammonium through assimilation rather than nitrification. • The strain can effectively remove phosphorus during ammonium assimilation. • The napA , nirS , norB , nosZ , and ppk genes coexisted in strain K14. Pseudomonas chloritidismutans K14, a novel phosphate-accumulating organism with the capacity to perform ammonium assimilation, aerobic denitrification, and phosphorus removal, was isolated from aquaculture sediments. It produced no hemolysin, and showed susceptibility to most antibiotics. Optimum conditions were achieved with sodium pyruvate as a carbon source, a C/N ratio of 10, pH of 7.5, temperature of 27 °C, P/N ratio of 0.26, and shaking at 140 rpm. Under optimum conditions, the highest removal efficiencies of ammonium, nitrite, and nitrate were 99.82%, 99.11%, and 99.78%, respectively; the corresponding removal rates were 6.27, 4.51, and 4.99 mg/L/h. The strain removed over 98% of phosphorus, and over 87% of chemical oxygen demand. The highest biomass nitrogen during ammonium assimilation was 99.18 mg/L; no gaseous nitrogen was produced. The genes involved in nitrogen and phosphorus removal were amplified by PCR. This study demonstrated the potential application prospects of strain K14 for nitrogen and phosphorus removal. [ABSTRACT FROM AUTHOR]