Methane-supported nitrate removal from groundwater in a membrane biofilm reactor

The discovery of denitrifying anaerobic methane oxidation (DAMO) has not only improved our understanding of global methane and nitrogen cycles, but also provided new technology options for removal of nitrate from nitrate-contaminated water. Previous studies have demonstrated DAMO organisms could remove nitrate and nitrite from wastewater under strictly anaerobically conditions. In the study, we investigate the feasibility of nitrate removal from groundwater, which contains dissolved oxygen in addition to nitrate. A membrane biofilm reactor (MBfR), inoculated with DAMO co-culture, was capable of treating synthetic groundwater containing highly contaminated nitrate (50  mg N/L) and oxygen (7-9 mg O₂/L), with a maximum volumetric nitrate removal rate of 45 mg N/L-d. Accumulations of acetate and propionate were observed in some transient periods, indicating the possible involvement of acetate and propionate as intermediates in methane oxidation. The 16 S rRNA gene amplicon sequencing revealed that Candidatus Methylomirabilis, a known bacterial DAMO organism able to couple nitrite reduction with anaerobic oxidation of methane (AOM), was the dominant population. No archaeal DAMO organisms that are capable of coupling nitrate to AOM were observed, however, considerable amount of denitrifiers were developed in this system. Based on known metabolisms of these microorganisms and a series of batch studies, it was assumed that methane was oxidized into volatile fatty acids (VFAs) under oxygen-limiting conditions, then the generated VFAs served as carbon sources for these heterotrophic denitrifiers to remove nitrate. This study offers a potential technology for nitrate removal from groundwater by DAMO process in MBfR.