To explore the availability of native microbes and activated sludge for ammonium removal, the native microbes and activated sludge in Jialu River basin were investigated in terms of ammonium-removing activities and their microbial communities using spectrophotometry and high-throughput sequencing. NH4+-N and total nitrogen (TN) in the targeted river ranged from 2.45 ± 1.76 to 8.56 ± 2.54 mg/L and from 3.42 ± 2.79 to 13.49 ± 5.06 mg/L, respectively. Both the native microbes and activated sludge had strong ammonium-removing activities with the removal efficiencies of more than 94%. High-throughput sequencing results indicated that, after five batches of operation, the class Gammaproteobacteria (28.55%), Alphaproteobacteria (14.55%), Betaproteobacteria (13.89%), Acidobacteria (8.82%) and Bacilli (7.04%) were dominated in native community, and there was a predominance of Gammaproteobacteria (21.57%), Betaproteobacteria (16.33%), Acidobacteria (12.41%), Alphaproteobacteria (10.01%), Sphingobacteriia (6.92%) and Bacilli (6.66%) in activated sludge. These two microbial sources were able to remove ammonium, while activated sludge was more cost-effective.