To improve the nitrogen removal capability of constructed wetlands, the biochar, produced from bamboo, activated with HCl and coated with Fe (FeCl3·6H2O), and then was added as a substrate into the systems. Three horizontal subsurface flow constructed wetlands (HSCWs) was established to treat the low C/N tailwater from the wastewater treatment plant: C-HSCW (quartz sand + soil), B-HSCW (quartz sand + soil + unmodified biochar), and FeB-HSCW (quartz sand + soil + Fe-modified biochar). Under different combinations of hydraulic retention time and nitrogen loading, the FeB-HSCW revealed extremely effective nitrogen removal, compared to the C-HSCW and B-HSCW. The highest removal efficiencies of NO3−-N (95.30%), TN (86.68%), NH4+-N (86.33%), NO2−-N (79.35%) and COD (63.36%) were obtained in FeB-HSCW with the hydraulic retention time of 96 h. and low influent nitrogen loading (C/N of 2.5). Nitrogen mass balance analysis showed that microbial processes played the most important role of nitrogen removal in HSCWs and the Fe-modified biochar significantly enhanced the microbial nitrogen removal. A total of 128.40 g nitrogen was removed by microorganisms in FeB-HSCW (average removal rate of 2.52 g N/(m3·d1)), much higher than that in other two HSCWs. The contributions of microorganisms, substrate storage and plant uptake on the total amount of nitrogen removal in the FeB-HSCW was 92.69%, 2.97% and 4.34%, respectively. Moreover, FeB significantly increased the abundances of genes involved in nitrogen removal. The copy numbers of bacterial 16S rRNA and amx, as well as of genes nirS, nirK, nosZ-I, nosZ-II, and hzsA were 1.3- to 27.8-fold higher in the FeB-HSCW than that in the other two HSCWs. Thus, Fe-modified biochar provides a feasible and effective amendment for constructed wetlands to improve the nitrogen removal, particularly nitrate-N, for low C/N wastewaters by enhancing the microbial nitrogen removal capacity (mainly of the denitrification).
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