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[Display omitted] • AA-TFCW accelerated NH 4 +-N transformation with 6.68 mg/(L·h) in start-up phases. • Heterotroph and autotroph synergistically removed 74.2% TN at low temperature. • HN-AD bacteria achieved direct NH 4 +-N removal by nitrate assimilation pathway. • Ferribacterium and Nitrosomonas carried high-weight key genes for N removal. This study explored the quantitative mechanisms of heterotrophic nitrification-aerobic denitrification (HN-AD) in a pilot-scale two-stage tidal flow constructed wetland (TFCW). The TFCW packed shale ceramsite (SC) and activated alumina (AA) at each stage, respectively, and aimed to improve decentralized wastewater treatment efficiency. In start-up phases, AA-TFCW accelerated NH 4 +-N decline, reaching transformation rates of 6.68 mg NH 4 +-N/(L·h). In stable phases, SC-AA-TFCW resisted low-temperatures (<13 °C), achieving stable NH 4 +-N and TN removal with effluents ranging 6.36–8.13 mg/L and 9.43–14.7 mg/L, respectively. The dominant genus, Ferribacterium, was the core of HN-AD bacteria, simultaneously removing NH 4 +-N and NO 3 –-N by nitrate assimilation and complete denitrification (NO 3 –-N → N 2), respectively. The quantitative associations highlighted importance of nitrification, nitrate assimilation, and denitrification in nitrogen removal. HN-AD bacteria (e.g., Lactococcus , Thauera , and Aeromonas) carried high-weight genes in quantitative associations, including nap AB, nas A and glt BD, implying that HN-AD bacteria have multiple roles in SC-AA-TFCW operation. [ABSTRACT FROM AUTHOR]

• Simultaneous N and P removal was achieved in SC-AA-TFCW at low temperature. • SC-TFCW contributed 85.6% of particulate phosphorous removal. • ADNB were highest abundance denitrifiers compared with FADNB and AUDNB. • ADNB showed positive interactions with key genes related to N and P metabolism. • The synergy of porous substrates and ADNB was proposed in SC-AA-TFCW. A pilot-scale two-stage tidal flow constructed wetland (TFCW) with working volume of 0.46 m3/d packing with shale ceramsite (SC) and activated alumina (AA) was constructed (named as SC-AA-TFCW) for nutrients removal at low temperature (<15 °C). SC-AA-TFCW achieved stable removals of 78.1% nitrogen and 98.3% phosphorous. SC-TFCW contributed to 55.2% of organics and 85.6% of particulate phosphorous removal. Among 17 denitrifiers, the absolute abundance of aerobic denitrification bacteria (ADNB) was highest, followed by facultative anaerobic denitrification bacteria (FADNB) and autotrophic denitrification bacteria (AUDNB). Nitrogen assimilating into organic nitrogen, dissimilatory and assimilatory nitrate reduction and complete denitrification may be main nitrogen metabolic pathways. Some ADNB (e. g. Zoogloea , Pseudomonas and Acidovorax) showed positive interactions with various key functional genes related to nutrients removal. Dissolved oxygen and reducing elements were main environmental factors in changing ADNB compositions. This study highlights the importance of ADNB and their synergy to porous substrates in SC-AA-TFCW. [ABSTRACT FROM AUTHOR]

• High C/N ratios and effluent recirculation promoted SND process for N removal. • HNAD was the main nitrogen transformation pathway in TFCWs. • HNADB were the dominant functional genera related to nitrogen removal in TFCWs. • Some core HNADB showed positive interactions with other functional genera. • Tidal strategies contributed more to shifts in functional genera than C/N ratios. This study investigated the influence of C/N ratios and tidal strategies on nitrogen removal and bacterial communities in two pilot-scale tidal flow constructed wetlands (TFCWs) with simultaneous nitrification–denitrification process. Heterotrophic nitrification aerobic denitrification (HNAD) was the main nitrogen transformation pathway in both TFCWs. High C/N ratios and effluent circulation at low temperature promoted HNAD in TFCWs with high nitrogen removal efficiencies (72.6%–95.5% for NH 4 +-N and 70.9%~91.8% for TN). Effluent circulation had more influence on bacterial community structure and diversity than C/N ratios. Among 16 detected genera related to nitrogen removal, HNAD bacteria (HNADB) were abundant. Especially, some dominant HNADB (e.g. Aeromonas , Hydrogenophage and Gemmobacter) were core genera, showing positive interactions with other genera related to nitrogen removal. Tidal strategies had more contribution to the shifts in these genera than C/N ratios. This study highlights the importance of HNADB in pilot-scale TFCWs and their responses to C/N ratios and tidal strategies. [ABSTRACT FROM AUTHOR]