Nitrate-N in wastewaters is hard to be recovered because it is difficult to volatilize with an opposite charge to ammonium. Here, we have proved the feasibility of dissimilatory nitrate reduction to ammonia (DNRA) by the easy-acclimated mixed electroactive bacteria, achieving the highest DNRA efficiency of 44%. It was then coupled with microbial electrolysis to concentrate ammonium by a factor of 4 in the catholyte for recovery. The abundance of electroactive bacteria in the biofilm before nitrate addition, especially Geobacter spp., was found to determine the DNRA efficiency. As the main competitors of DNRA bacteria, the growth of denitrifiers was more sensitive to C/N ratios. The DNRA microbial community contrarily showed a stable and recoverable ammoniation performance over C/N ratios ranging from 0.5 to 8.0. A strong competition of the electrode and nitrate on electron donors was observed at the early stage (15 d) of electroactive biofilm formation, which can be weakened when the biofilm was mature on 40 d. Quantitative PCR showed a significant increase in nirS and nrfA transcripts in the ammoniation process. nirS was inhibited significantly after nitrate depletion while nrfA was still upregulated. These findings provided a novel way to recover nitrate-N using organic wastes as both electron donor and power, which has broader implications on the sustainable wastewater treatment and the ecology of nitrogen cycling.

Phosphorus undergoes a one-way flow from minerals to soil to water, which creates a phosphorus crisis as well as aquatic eutrophication. Dissimilatory metal reduction bacterial (DMRB)induced vivianite recovery from wastewater is a promising route to solve these problems synthetically. In this study, phosphorus competition between biomass growth and bioinduced vivianite mineralization was investigated at the batch scale. Biomass growth leads to phosphorus utilization over vivianite mineralization. Geobacter was selected as the main functional microorganism and presented higher vivianite recovery rates (20−48%) than sewage biomass (7−33%). An optimal Fe/P stoichiometric ratio of 1:1 was observed for both sewage biomass and Geobacter-inoculated batches. The highest vivianite yield of 4.3 mM was obtained in Geobacter-inoculated batches at a Fe:P of 1:1, with values 59% higher than those at a Fe:P of 1:0.67 (equal to the Fe/P molar ratio in vivianite). Sufficient PO 4 3− stimulated cell growth and yielded a higher Fe 3+ reduction rate and vivianite yield. Nevertheless, excessive PO 4 3− facilitated the precipitation of KFe 3 (PO 4 ) 2 (OH)• 8H 2 O and Fe 7 (PO 4 ) 6 , which inhibited vivianite synthesis. In the optimal Geobacter batch, the μ−S curve indicated a mixed order reaction (0 < x < 1) for both vivianite formation and biomass growth. The vivianite growth series proceeded as follows: tiny blue particles, plain pieces, dark blue nodules, and large spherical crystals.

As electrons generated through substrate oxidation compete with electrodes, dissimilatory nitrate reduction to ammonium (DNRA), denitrification in bioelectrochemical systems in the presence of nitrate, and nitrate reduction through an electroactive biofilm (EAB) are unpredictable. We find that pathways of nitrate reduction are related to EAB thickness and that 76 ± 2 μm is the critical thickness of a biofilm at which both the inner and outer layers simultaneously include DNRA, leading to a maximum level of DNRA efficiency of 42%. Fractions of electrons flowing during nitrate reduction are relatively stable, but their distributions between DNRA and denitrification vary with biofilm thickness. Electrons prefer denitrification in an EAB that is 66 ± 2 μm, while DNRA reversely surpasses denitrification when the thickness increases in the range of 76 ± 2 to 210 ± 2 μm. Biofilm thickening enhances the DNRA of all biofilms close to solution, where nirK remains constant and nrfA is significantly upregulated. However, nrfA is downregulated in layers close to the electrode when the biofilm is thicker than 76 ± 2 μm. These findings reveal the spatially heterogeneous reduction of nitrate in thick EABs, highlighting the importance of biofilm thickness to the regulation of end products of nitrate reduction.

Geobacter spp. are well-known exoelectrogenic microorganisms that often predominate acetate-fed biofilms in microbial fuel cells (MFCs) and other bioelectrochemical systems (BESs). By using an amplicon sequence variance analysis (at one nucleotide resolution), we observed a succession between two closely related species (98% similarity in 16S RNA), Geobacter sulfurreducens and Geobacter anodireducens, in the long-term studies (20 months) of MFC biofilms. Geobacter spp. predominated in the near-electrode portion of the biofilm, while the outer layer contained an abundance of aerobes, which may have helped to consume oxygen but reduced the relative abundance of Geobacter. Removal of the outer aerobes by norspermidine washing of biofilms revealed a transition from G. sulfurreducens to G. anodireducens. This succession was also found to occur rapidly in cocultures in BES tests even in the absence of oxygen, suggesting that oxygen was not a critical factor. G. sulfurreducens likely dominated in early biofilms by its relatively larger cell size and production of extracellular polymeric substances (individual advantages), while G. anodireducens later predominated due to greater cell numbers (quantitative advantage). Our findings revealed the interspecies competition in the long-term evolution of Geobacter genus, providing microscopic insights into Geobacter's niche and competitiveness in complex electroactive microbial consortia.

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