Regulatory mechanisms of biochar alleviating ammonia inhibition to methanogenesis during long-term operation of anaerobic membrane bioreactor treating swine wastewater

Anaerobic Membrane Bioreactor (AnMBR) is regarded as an advanced technology for the anaerobic digestion (AD) of swine wastewater. However, the high ammonia level in swine wastewater is a crucial issue to inhibit methane production. This study aimed to establish a biochar-assisted AnMBR (BC) to mitigate ammonia inhibition during AD of swine wastewater. As the gradual ammonia loading rate (ALR) increased from 3.0 g/L to 6.0 g/L, BC maintained methane yields ranging from 74.9 % to 48.1 %, which were 9.1 % to 33.3 % higher than those of a regular AnMBR (CT). Moreover, the COD removal efficiency steadily surpassed 80 %. Although the increase in ALR had negative effects on both BC and CT, the presence of biochar enhanced the methanogenic activity of bulk sludge, specifically accelerating processes involved in syntrophic acetate oxidation and CO 2 - reducing methanogenesis under high ALR conditions. Integrated characterizations using PARAFAC-based EPS analysis, INT-ETS, and WO 3 probe -based extracellular electron transfer (EET) activities indicated that biochar stimulated the secretion of humic-like and aromatic protein -like compounds in the TB-EPS, which likely established a redox-active matrix with biochar between VFA oxidation bacteria and CO 2 -reducing methanogens to facilitate EET. This potentially made the syntrophic VFA oxidation with favorably thermodynamic kinetics under high ammonia stress. Furthermore, the analysis of microbial community coupling prediction of functional genes associated with methanogenic pathways revealed that as ALR increased, the presence of biochar re -shaped the microbial communities by enriching electro-active Syntrophomonas and Methanosarcina / Methanospirillum , which mainly triggering EET-based syntrophic methanogenesis by accelerating the intracellular enzymatic transformation from Formylmethanofuran to 5,10-Methylenetetrahydromethanopterin process. We expect the major findings of this study provide new insights to understand the mechanisms of biochar alleviating ammonia inhibition during long-term operation of AnMBR.