Enhancing total nitrogen removal in constructed wetlands: A Comparative study of iron ore and biochar amendments

Efficient nitrogen removal in constructed wetlands (CWs) remains challenging when treating agricultural runoff with a low carbon-to-nitrogen ratio (C/N). However, using biochar, iron ore, and FeCl₃-modified biochar (Fe-BC) as amendments could potentially improve total nitrogen (TN) removal efficiency in CWs, but the underlying mechanisms associated with adding these substrates are unclear. In this study, five CWs: quartz sand constructed wetland (Control), biochar constructed wetland, Fe-BC constructed wetland, iron ore constructed wetland, and iron ore + biochar constructed wetland, were built to compare their treatment performance. The rhizosphere microbial community compositions and their co-occurrence networks were analyzed to reveal the underlying mechanisms driving their treatment performance. The results showed that iron ore was the most efficient amendment, although all treatments increased TN removal efficiency in the CWs. Ammonia-oxidizing, heterotrophic denitrifying, nitrate-dependent anaerobic ferrous oxidizing (NAFO), and Feammox bacteria abundance was higher in the iron ore system and led to the simultaneous removal of NH₄⁺-N, NO₃^--N, and NO₂^--N. Visual representations of the co-occurrence networks further revealed that there was an increase in cooperative mutualism (the high proportion of positive links) and more complex interactions among genera related to the nitrogen and iron cycle (especially ammonia-oxidizing bacteria, heterotrophic denitrifying bacteria, NAFO bacteria, and Feammox bacteria) in the iron ore system, which ultimately contributed to the highest TN removal efficiency. This study provides critical insights into how different iron ore or biochar substrates could be used to treat agricultural runoff in CWs.