Treatment of Anaerobic Digester Liquids via Membrane Biofilm Reactors: Simultaneous Aerobic Methanotrophy and Nitrogen Removal

Anaerobic digestion (AD) produces useful biogas and waste streams with high levels of dissolved methane (CH₄) and ammonium (NH₄⁺), among other nutrients. Membrane biofilm reactors (MBfRs), which support dissolved methane oxidation in the same reactor as simultaneous nitrification and denitrification (ME-SND), are a potential bubble-less treatment method. Here, we demonstrate ME-SND taking place in single-stage, AD digestate liquid-fed MBfRs, where oxygen (O₂) and supplemental CH₄ were delivered via pressurized membranes. The effects of two O₂ pressures, leading to different O₂ fluxes, on CH₄ and N removal were examined. MBfRs achieved up to 98% and 67% CH₄ and N removal efficiencies, respectively. The maximum N removal rates ranged from 57 to 94 mg N L^-1 d^-1, with higher overall rates observed in reactors with lower O₂ pressures. The higher-O₂-flux condition showed NO₂^- as a partial nitrification endpoint, with a lower total N removal rate due to low N₂ gas production compared to lower-O₂-pressure reactors, which favored complete nitrification and denitrification. Membrane biofilm 16S rRNA amplicon sequencing showed an abundance of aerobic methanotrophs (especially <i>Methylobacter</i>, <i>Methylomonas,</i> and <i>Methylotenera</i>) and enrichment of nitrifiers (especially <i>Nitrosomonas</i> and <i>Nitrospira</i>) and anammox bacteria (especially Ca. <i>Annamoxoglobus</i> and Ca. <i>Brocadia</i>) in high-O₂ and low-O₂ reactors, respectively. Supplementation of the influent with nitrite supported evidence that anammox bacteria in the low-O₂ condition were nitrite-limited. This work highlights coupling of aerobic methanotrophy and nitrogen removal in AD digestate-fed reactors, demonstrating the potential application of ME-SND in MBfRs for the treatment of AD's residual liquids and wastewater. Sensor-based tuning of membrane O₂ pressure holds promise for the optimization of bubble-less treatment of excess CH₄ and NH₄⁺ in wastewater.