Constructed wetlands (CWs) are widely applied for decentralized wastewater treatment. However, achieving efficient removal of ammonia ( \({\rm{N}}{{\rm{H}}_4}^ + - {\rm{N}}\) ) has proven challenging due to insufficient oxygen. In this study, natural hematite (Fe2O3) was employed as a CW substrate (H-CWs) for the first time to drive anaerobic ammonia oxidation coupled with iron(III) reduction (Feammox). Compared to gravel constructed wetlands (G-CWs), ammonia removal was enhanced by 38.14% to 54.03% and nitrous oxide (N2O) emissions were reduced by 34.60% in H-CWs. The synergistic removal of ammonia and nitrate by H-CWs also resulted in the absence of ammoxidation by-products. Inhibitor and 15N isotope tracer incubations showed that Feammox accounting for approximately 40% of all ammonia removal in the H-CWs. The enrichment of iron phosphate (Fe3Fe4(PO4)6) promoted the accumulation of the Feammox intermediate compound FeOOH. Microbial nanowires were observed on the surface of H-CW substrates as well, suggesting that the observed biological ammoxidation was most likely related to extracellular electron transfer (EET). Microbial and metagenomics analysis revealed that H-CWs elevated the integrity and enhanced the abundance of functional microorganisms and genes associated with nitrogen metabolism. Overall, the efficient ammonia removal in the absence of O2 together with a reduction in N2O emissions as described in this study may provide useful guidance for hematite-mediated anaerobic ammonia removal in CWs.
