This study aimed to investigate the effectiveness of combining zero-valent iron (Fe0) with an electric-magnetic field (EMF) with an intensity of 48-mT generated by an alternating current (AC) of 6 V and 50 Hz applied deep within a reactor using actual wastewater obtained from a secondary stage of a wastewater treatment plant (WWTP). The lab-scale experiment was conducted using the UMSR reactor, maintaining a controlled temperature of 25 +/- 2 degrees C and a hydraulic retention time (HRT) of 5-hour, with a dissolved oxygen (DO) level below 1.5 mg/L. Fine Fe0 particles with a size of <= 0.3 mu m were added into the reactor at a concentration of 3 g/L. The actual wastewater had an approximate nitrate nitrogen (NO3--N) concentration of 30 +/- 2 mg/L, and the substrate mass ratio of carbon to nitrogen (C:N) was at 1:1. The results examined the performance of the EMF-Fe0 system in terms of nitrate removal efficiency and the enrichment of denitrifying bacteria, achieving a nitrate removal rate of 67 % with an effluent NO3--N concentration of 10 mg/L and a chemical oxygen demand (COD) concentration of 11.8 mg/L at removal rate of 60 %. Illumina sequencing revealed various enriched denitrifying bacteria after coupling EMF with Fe0, including autotrophic Nitrospira (NOB) at 4 %, denitrification bacteria like Hydrogenophaga at 1.3 %, Hyphomicrobium at 1 %, and Sphingobium at 5 %, along with thermophilic bacteria Thermus at 1.1 % and Meiothermus at 3.5 %. PICRUSt predicted bacterial functions and analyzed interactions, while KEGG and GO analyses linked nitrogen removal pathways to specific species, genes, and enzymes in the EMF-Fe0 system.
