Novel insight into the inhibitory effects and mechanisms of Fe(II)-mediated multi-metabolism in anaerobic ammonium oxidation (anammox)

Fe(II) participates in complex Fe-N cycles and effects on the microbial metabolism in the anaerobic ammonium oxidation (anammox) dominated system. In this study, the inhibitory effects and mechanisms of Fe(II)-mediated multi-metabolism in anammox were revealed, and the potential role of Fe(II) in the nitrogen cycle was evaluated. The results showed that the long-term accumulation of high Fe(II) concentrations (70-80 mg/L) led to a hysteretic inhibition of anammox. High Fe(II) concentrations induced the generation of high levels of intracellular ·O2-, whereas the antioxidant capacity was insufficient to eliminate the excess ·O2-, thus causing ferroptosis to anammox cells. In addition, Fe(II) was oxidized via nitrate-dependent anaerobic ferrous-oxidation (NAFO) process, and mineralized to coquimbite and phosphosiderite. They formed crusts on the surface of the sludge, leading to mass transfer obstruction. The results of the microbial analysis showed that the addition of appropriate Fe(II) increased the abundance of Candidatus Kuenenia, and served as a potential electron donor to enrich Denitratisoma, promoting anammox and NAFO coupled with nitrogen removal, while high Fe(II) concentrations reduced the enrichment level. In this study, the understanding of Fe(II)-mediated multi-metabolism in the nitrogen cycle was deepened, providing the basis for the development of Fe(II)-based anammox technologies.