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To improve the low pollutant removal efficiency of traditional biological methods for treating livestock and poultry breeding wastewater under a relatively low temperature, a rotating biological contactor (RBC) inoculated with heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria was designed. A quick start-up process and a well removal performance had been achieved in the novel RBC. To elucidate the anti-load shock ability of the novel RBC, the effects of C/N ratio on nitrogen removal and microbial assemblage were focused under a low temperature (12 ± 2 °C). Results showed that the highest NH 4 +-N and TN removal efficiency were 99.57 ± 0.31% and 68.41 ± 0.52%. Microbial diversity analysis based on high throughput sequencing technique showed that Arcobacter and Flavobacterium with an increasing relative abundance were the key to ensure high nitrogen removal efficiently at a low C/N ratio and temperature. Moreover, nitrogen transferring pathways of the novel RBC was revealed and dissimilatory nitrate reduction and denitrification were the main pathways. The excellent pollutant removal performance demonstrates that the novel RBC is a promising process to effectively treat wastewater with low C/N ratio and low temperature. [Display omitted] • Rotating biological contactor (RBC) started up by HN-AD bacteria has been proposed. • A short start-up time and a high nitrogen removal efficiency were achieved. • Impact of C/N ratio on pollutant removal and microbial assemblage were evaluated. • Arcobacter and Flavobacterium played a key role at low C/N ratio and temperature. • DNRA and denitrification were the main nitrogen transferring pathways in the H-RBC. [ABSTRACT FROM AUTHOR]