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[Display omitted] • A single stage PN/A process treating low strength wastewater was started rapidly. • The biofilm growth process on porous polyurethane hydrogel carrier was studied. • Biomass entrapment by interconnected micropores dominated the initial colonization. • AOB was surface-attached growth, anammox colonies proliferated as embedded growth. Media-supported biofilm is a powerful strategy for growth and enrichment of slow-growing microorganisms. In this study, a single-stage nitritation-anammox process treating low-strength wastewater was successfully started to investigate the biofilm development on porous polyurethane hydrogel carrier. Suspended biomass migration into the carrier and being entrapment by its internal interconnected micropores dominated the fast initial colonization stage. Both surface-attached growth and embedded growth of microbes occurred during the following accumulation stage. Fluorescence in situ hybridization analysis of mature biofilm indicated that ammonium-oxidizing bacteria located at the outer layers featured a surface-attached growth, while anammox microcolonies housed in the inner layers proliferated as an embedded-like growth. In this way, the growth rate of anammox bacteria (predominated by Candidatus Kuenenia) could be 0.079 d-1. The anammox potential of the biofilm reactor reached 1.65 ± 0.3 kg/m3/d within two months. This study provides novel insights into nitritation-anammox biofilm formation on the porous polyurethane hydrogel carrier. [ABSTRACT FROM AUTHOR]