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One of the most important advancements in harnessing the biological nitrification in the field is enrichment solution of nitrifying microbial consortia. In the current study, we developed an improved multi-step enrichment to amplify a targeted microbial consortium from a sediment sample collected in tropical mangrove, Vietnam. The results showed that it took 122 culturing days with five unique continuous enrichment steps, the microbial consortium consumed total 5665 mgN L−1. Relative substrate removal rate increased rapidly from 0.114 mgN L−1 h−1 at the end of the first-step enrichment up to 3.58 mgN L−1 h−1 at the end of the fifth-step enrichment. High-throughput sequencing revealed that Nitrospirae, Proteobacteria and Bacteroidetes were the dominant taxa at the phylum level while Nitrospira, Marinobacter, Denitromonas and Nitrosomonas were the dominant taxa at the genus level in the enriched consortia. A pilot-scale experiment for shrimp cultivation of L. vannamei in 84 day-period proved the efficiency of Total ammonium nitrogen and nitrite removal in the consortium-activated treatment was much higher than the control. [ABSTRACT FROM AUTHOR]