The vast majority of processes in the carbon and nitrogen cycles are driven by microorganisms. The nitrite-dependent anaerobic oxidation of methane (N-DAMO) process links carbon and nitrogen cycles, offering a novel approach for the simultaneous reduction of methane emissions and nitrite pollution. However, there is currently no comprehensive summary of the current status of the N-DAMO process in natural aquatic environments. Therefore, our study aims to fill this knowledge gap by conducting a comprehensive review of the global research trends in N-DAMO processes in various aquatic environments (excluding artificial bioreactors). Our review mainly focused on molecular identification, global study sites, and their interactions with other elemental cycling processes. Furthermore, we performed a data integration analysis to unveil the effects of key environmental factors on the abundance of N-DAMO bacteria and the rate of N-DAMO process. By combining the findings from the literature review and data integration analysis, we proposed future research perspectives on N-DAMO processes in global aquatic environments. Our overarching goal is to advance the understanding of the N-DAMO process and its role in synergistically reducing carbon emissions and removing nitrogen. By doing so, we aim to make a significant contribution to the timely achievement of China\'s carbon peak and carbon neutrality targets.

Nitrite-dependent anaerobic methane oxidation (n-DAMO) has been demonstrated to play important roles in the global methane and nitrogen cycle. However, despite diverse n-DAMO bacteria widely detected in environments, little is known about their physiology for microbial niche differentiation. Here, we show the microbial niche differentiation of n-DAMO bacteria through long-term reactor operations combining genome-centered omics and kinetic analysis. With the same inoculum dominated by both species \"Candidatus Methylomirabilis oxyfera\" and \"Candidatus Methylomirabilis sinica\", n-DAMO bacterial population was shifted to \"Ca. M. oxyfera\" in a reactor fed with low-strength nitrite, but shifted to \"Ca. M. sinica\" with high-strength nitrite. Metatranscriptomic analysis showed that \"Ca. M. oxyfera\" harbored more complete function in cell chemotaxis, flagellar assembly, and two-component system for better uptake of nitrite, while \"Ca. M. sinica\" had a more active ion transport and stress response system, and more redundant function in nitrite reduction to mitigate nitrite inhibition. Importantly, the half-saturation constant of nitrite (0.057 mM vs 0.334 mM NO2-) and inhibition thresholds (0.932 mM vs 2.450 mM NO2-) for \"Ca. M. oxyfera\" vs \"Ca. M. sinica\", respectively, were highly consistent with genomic results. Integrating these findings demonstrated biochemical characteristics, especially the kinetics of nitrite affinity and inhibition determine niche differentiation of n-DAMO bacteria.

The denitrifying anaerobic methane oxidation is an ecologically important process for reducing the potential methane emission into the atmosphere. The responsible bacterium for this process was Candidatus Methylomirabilis oxyfera belonging to the bacterial phylum of NC10. In this study, a new pair of primers targeting all the five groups of NC10 bacteria was designed to amplify NC10 bacteria from different environmental niches. The results showed that the group A was the dominant NC10 phylum bacteria from the sludges and food waste digestate while in paddy soil samples, group A and group B had nearly the same proportion. Our results also indicated that NC10 bacteria could exist in a high pH environment (pH9.24) from the food waste treatment facility. The Pearson relationship analysis showed that the pH had a significant positive relationship with the NC10 bacterial diversity (p<0.05). The redundancy analysis further revealed that the pH, volatile solid and nitrite nitrogen were the most important factors in shaping the NC10 bacterial structure (p=0.01) based on the variation inflation factors selection and Monte Carlo test (999 times). Results of this study extended the existing molecular tools for studying the NC10 bacterial community structures and provided new information on the ecological distributions of NC10 bacteria.

Both aerobic methane-oxidizing bacteria (MOB) and nitrite-dependent anaerobic methane oxidation (n-damo) organisms can be important methane sinks in a wetland. However, the influences of the vegetation type on aerobic MOB and n-damo communities in wetland, especially in constructed wetland, remain poorly understood. The present study investigated the influences of the vegetation type on both aerobic MOB and n-damo organisms in a constructed urban landscape wetland. Sediments were collected from eight sites vegetated with different plant species. The abundance (1.19-3.27 × 107 pmoA gene copies per gram dry sediment), richness (Chao1 estimator = 16.3-81.5), diversity (Shannon index = 2.10-3.15), and structure of the sediment aerobic MOB community were found to vary considerably with sampling site. In contrast, n-damo community abundance (8.74 × 105-4.80 × 106 NC10 16S rRNA gene copies per gram dry sediment) changed slightly with the sampling site. The richness (Chao1 estimator = 1-11), diversity (Shannon index = 0-0.78), and structure of the NC10 16S rRNA gene-based n-damo community illustrated slight site-related changes, while the spatial changes of the pmoA gene-based n-damo community richness (Chao1 estimator = 1-8), diversity (Shannon index = 0-0.99), and structure were considerable. The vegetation type could have a profound impact on the wetland aerobic MOB community and had a stronger influence on the pmoA-based n-damo community than on the NC10 16S-based one in urban wetland. Moreover, the aerobic MOB community had greater abundance and higher richness and diversity than the n-damo community. Methylocystis (type II MOB) predominated in urban wetland, while no known type I MOB species was detected. In addition, the ratio of total organic carbon to total nitrogen (C/N) might be a determinant of sediment n-damo community diversity and aerobic MOB richness.

Managing nitrogen and carbon cycles in engineered and natural ecosystems is an environmental challenge. In this manuscript, we report a process which connects these two cycles with immense ecological and engineering significance. Sediments, collected from Jordan River in Salt Lake City, Utah were used as seed to start a laboratory-scale denitrification coupled to anaerobic methane oxidation (n-DAMO) reactor fed with methane (CH4) and nitrite (NO2(-)). Methane (CH4)-dependent denitrification in sediments of a nutrient-impaired river was found to be in the range of 40 nmol kg(-1) d(-1) to 70 nmol kg(-1) d(-1). Post 19 months of operation of the lab scale reactor, the n-DAMO reactor achieved nitrite removal rate of 2.88 mmol L(-1) d(-1). Enrichment of n-DAMO prokaryotes was evident from the increase in 16S rRNA gene copy number of bacteria belonging to the NC10 phylum in the reactor, corroborating with increase in the oxidation rates of CH4 coupled with NO2(-)-N removal from 21 μM to 190 μM of CH4 d(-1). Based on stable isotope experiments by other researchers, nitric oxide dismutase (nod) functional gene was hypothesized to be responsible for splitting nitric oxide to nitrogen and oxygen and this internally generated oxygen is utilized by n-DAMO prokaryotes to oxidize methane gas. Primers targeting the unique nitric oxide dismutase (nod) gene were developed and tested on the enrichment culture for the first time. This revealed that n-DAMO organisms are closely related yet distinct from, the M. oxyfera which had been enriched in earlier studies. The results emphasize tremendous future promise to use these novel organisms for wastewater treatment purposes, especially to take advantage of the dissolved methane present in anaerobic digester effluents.

OBJECTIVE: To better explore the distribution and diversity of Candidatus Methylomirabilis oxyfera (M. oxyfera)-like bacteria of NC10 phylum in soil environments.
RESULTS: The vertical distribution and diversity of NC10 phylum bacteria were investigated in an agricultural field (surface layer, 0-10 cm; subsurface layer, 20-30 cm; deep layers, 50-60 and 90-100 cm) by using Illumina-based 16S rRNA (V3-V4 region) gene sequencing of soil DNA samples and quantitative PCR assays. It was found that the NC10-related reads accounted for 0·8-4·5% of the 16S rRNA pool in each examined core sample, with greater percentage being observed in deep soils than in surface soils and subsurface soils. The recovered NC10-related reads showed 85·1-96·9% identity to the 16S rRNA gene of M. oxyfera. A high diversity of NC10 phylum bacteria was observed in the examined soil cores. A total of 115 operational taxonomic units (OTU) were detected based on 3% sequence divergence in the recovered 16S rRNA genes. Phylogenetic analysis showed that four distinct groups of NC10 phylum bacteria (groups A, B, C and D) were present in the examined soil cores, with group B members being the dominant bacteria. The group A members, which are identified as the dominant bacteria responsible for anaerobic methane oxidation (AMO) coupled to nitrite reduction, can mainly be detected at 50-60 cm. Quantitative PCR further confirmed the presence of NC10 phylum bacteria, ranging from 3·8 × 10(6) to 9·3 × 10(6) copies g(-1) soil.
CONCLUSIONS: The results showed the presence of diverse NC10 phylum bacteria in agricultural soils by using Illumina-based 16S rRNA gene sequencing and qPCR assays.
CONCLUSIONS: The greatest level of diversity of NC10 phylum bacteria was reported to date in this study, which improved our understanding of the distribution of NC10 phylum bacterial communities in soil environments.

The anaerobic oxidation of methane (AOM) is an important sink of methane that plays a significant role in global warming. AOM was first found to be coupled with sulfate reduction and mediated by anaerobic methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB). ANME, often forming consortia with SRB, are phylogenetically related to methanogenic archaea. ANME-1 is even able to produce methane. Subsequently, it has been found that AOM can also be coupled with denitrification. The known microbes responsible for this process are Candidatus Methylomirabilis oxyfera (M. oxyfera) and Candidatus Methanoperedens nitroreducens (M. nitroreducens). Candidatus Methylomirabilis oxyfera belongs to the NC10 bacteria, can catalyze nitrite reduction through an \"intra-aerobic\" pathway, and may catalyze AOM through an aerobic methane oxidation pathway. However, M. nitroreducens, which is affiliated with ANME-2d archaea, may be able to catalyze AOM through the reverse methanogenesis pathway. Moreover, manganese (Mn(4+) ) and iron (Fe(3+) ) can also be used as electron acceptors of AOM. This review summarizes the mechanisms and associated microbes of AOM. It also discusses recent progress in some unclear key issues about AOM, including ANME-1 in hypersaline environments, the effect of oxygen on M. oxyfera, and the relationship of M. nitroreducens with ANME.