N-damo Process Research Articles

Probing Denitrifying Anaerobic Methane Oxidation via Antimicrobial Intervention: Implications for Innovative Wastewater Management.

Methane emissions present a significant environmental challenge in both natural and engineered aquatic environments. Denitrifying anaerobic methane oxidation (N-DAMO) has the potential for application in wastewater treatment plants. However, our understanding of the N-DAMO process is primarily based on studies conducted on environmental samples or enrichment cultures using metagenomic approaches. To gain deeper insights into N-DAMO, we used antimicrobial compounds to study the function and physiology of 'Candidatus Methanoperedens nitroreducens' and 'Candidatus Methylomirabilis oxyfera' in N-DAMO enrichment cultures. We explored the effects of inhibitors and antibiotics and investigated the potential application of N-DAMO in wastewater contaminated with ammonium and heavy metals. Our results showed that 'Ca. M. nitroreducens' was susceptible to puromycin and 2-bromoethanesulfonate, while the novel methanogen inhibitor 3-nitrooxypropanol had no effect on N-DAMO. Furthermore, 'Ca. M. oxyfera' was shown to be susceptible to the particulate methane monooxygenase inhibitor 1,7-octadiyne and a bacteria-suppressing antibiotic cocktail. The N-DAMO activity was not affected by ammonium concentrations below 10 mM. Finally, the N-DAMO community appeared to be remarkably resistant to lead (Pb) but susceptible to nickel (Ni) and cadmium (Cd). This study provides insights into microbial functions in N-DAMO communities, facilitating further investigation of their application in methanogenic, nitrogen-polluted water systems.

Coupling Partial Nitritation, Anammox and n-DAMO in a membrane aerated biofilm reactor for simultaneous dissolved methane and nitrogen removal

Anaerobic technologies with downstream autotrophic nitrogen removal have been proposed to enhance bioenergy recovery and transform a wastewater treatment plant from an energy consumer to an energy exporter. However, approximately 20–50 % of the produced methane is dissolved in the anaerobically treated effluent and is easily stripped into the atmosphere in the downstream aerobic process, contributing to the release of greenhouse gas emissions. This study aims to develop a solution to beneficially utilize dissolved methane to support high-level nitrogen removal from anaerobically treated mainstream wastewater. A novel technology, integrating Partial Nitritation, Anammox and Methane-dependent nitrite/nitrate reduction (i.e. PNAM) was demonstrated in a membrane-aerated biofilm reactor (MABR). With the feeding of ∼50 mg NH4+-N/L and ∼20 mg/L dissolved methane at a hydraulic retention time of 15 h, around 90 % of nitrogen and ∼100 % of dissolved methane can be removed together in the MABR. Microbial community characterization revealed that ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation microorganisms (n-DAMO bacteria and archaea) and aerobic methanotrophs co-existed in the established biofilm. Batch tests confirmed the active microbial pathways and showed that AOB, anammox bacteria and n-DAMO microbes were jointly responsible for the nitrogen removal, and dissolved methane was mainly removed by the n-DAMO process, with aerobic methane oxidation making a minor contribution. In addition, the established system was robust against dynamic changes in influent composition. The study provides a promising technology for the simultaneous removal of dissolved methane and nitrogen from domestic wastewater, which can support the transformation of wastewater treatment from an energy- and carbon-intensive process, to one that is energy- and carbon-neutral.

Role of n-DAMO in Mitigating Methane Emissions from Intertidal Wetlands Is Regulated by Saltmarsh Vegetations.

Coastal wetlands are hotspots for methane (CH4) production, reducing their potential for global warming mitigation. Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) plays a crucial role in bridging carbon and nitrogen cycles, contributing significantly to CH4 consumption. However, the role of n-DAMO in reducing CH4 emissions in coastal wetlands is poorly understood. Here, the ecological functions of the n-DAMO process in different saltmarsh vegetation habitats as well as bare mudflats were quantified, and the underlying microbial mechanisms were explored. Results showed that n-DAMO rates were significantly higher in vegetated habitats (Scirpus mariqueter and Spartina alterniflora) than those in bare mudflats (P < 0.05), leading to an enhanced contribution to CH4 consumption. Compared with other habitats, the contribution of n-DAMO to the total anaerobic CH4 oxidation was significantly lower in the Phragmites australis wetland (15.0%), where the anaerobic CH4 oxidation was primarily driven by ferric iron (Fe3+). Genetic and statistical analyses suggested that the different roles of n-DAMO in various saltmarsh wetlands may be related to divergent n-DAMO microbial communities as well as environmental parameters such as sediment pH and total organic carbon. This study provides an important scientific basis for a more accurate estimation of the role of coastal wetlands in mitigating climate change.

Active pathways of anaerobic methane oxidization in deep-sea cold seeps of the South China Sea.

Anaerobic oxidation of methane (AOM) is critical for controlling methane emissions from deep-sea cold seeps. Microbial groups associated with different AOM processes have been briefly investigated, but the AOM activities associated with the utilization of different electron acceptors in the deep-sea methane seeps remain largely unknown. Here, surface sediments (0-4 cm) from two cold seeps (Site F/Haima) and the Xisha trough in the South China Sea (SCS) were incubated to measure AOM potentials driven by nitrite, nitrate, and sulfate as different electron acceptors with isotopically labeled methane gas (13CH4), and the population shifts of microbial communities along the incubation were investigated by high-throughput sequencing and quantitative polymerase chain reaction based on both the 16S rRNA gene and functional genes. Nitrite and nitrate were preferentially used prior to sulfate during the incubation, and nitrate-/nitrite-dependent anaerobic methane oxidation (Nr-/N-DAMO) contributed more to the methane flux in both the cold seeps and the trough. The highest rates of DAMO and sulfate-dependent anaerobic methane oxidation (SAMO) were detected at Site F, consistent with the transcript abundance of mcrA and dsrB genes during the incubation, and might be explained by the in situ active functional groups. The former process might be explained by Campylobacteria, which could couple sulfide oxidation and nitrate consumption to further participate in the N-DAMO process, while the latter process might be due to the higher proportions of Desulfobacterota. This study elucidates the diversity and potential activities of major microbial groups in the DAMO and SAMO processes in the deep-sea cold seeps using isotopic tracing incubations and provides direct evidence for the occurrence of Nr-/N-DAMO as a previously overlooked microbial methane sink in the deep-sea hydrate-bearing sediments in the SCS. IMPORTANCE Cold seeps occur in continental margins worldwide and are deep-sea oases. Anaerobic oxidation of methane is an important microbial process in the cold seeps and plays an important role in regulating methane content. This study elucidates the diversity and potential activities of major microbial groups in dependent anaerobic methane oxidation and sulfate-dependent anaerobic methane oxidation processes and provides direct evidence for the occurrence of nitrate-/nitrite-dependent anaerobic methane oxidation (Nr-/N-DAMO) as a previously overlooked microbial methane sink in the hydrate-bearing sediments of the South China Sea. This study provides direct evidence for occurrence of Nr-/N-DAMO as an important methane sink in the deep-sea cold seeps.

How efficiently does a metabolically enhanced system with denitrifying anaerobic methane oxidizing microorganisms remove antibiotics?

In this work, the novel N-damo (Nitrite dependent anaerobic methane oxidation) process was investigated at high biomass activities for its potential to remove simultaneously nitrite and methane, as well as selected antibiotics commonly found in sewage in trace amounts. For this purpose, two MBRs were operated at three high nitrite loading rates (NLRs), namely 76 ± 9.9, 161.5 ± 11.4 and 215.2 ± 24.2 mg N–NO⁻2 L−1 d−1, at long-term operation. The MBRs performance achieved a significantly high nitrite removal activity for an N-damo process (specific denitrifying activity of up to 540 mg N–NO⁻2 g−1 VSS d−1), even comparable to heterotrophic denitrification values. In this study, we have implemented a novel operational strategy that sets our work apart from previous studies with similar bioreactors. Specifically, we have introduced Cerium as a trace element in the feeding medium, which serves as a key differentiating factor. It allowed maintaining a stable reactor operation at high NLRs. Microbial community composition evidenced that both MBRs were dominated with N-damo bacteria (67–87% relative abundance in period III and I, respectively). However, a decrease in functional N-damo bacteria (Candidatus Methylomirabilis) abundance was observed during the increase in biomass activity and concentration, concomitantly with an increase of the other minor families (Hypomicrobiaceae and Xanthobacteraceae). Most of the selected antibiotics showed high biotransformation such as sulfamethoxazole, trimethoprim, cefalexin and azithromycin, whereas others such as roxithromycin and clarithromycin were only partially degraded (20–35%). On the contrary, ciprofloxacin showed almost no removal. Despite the metabolic enhancement, no apparent increase on the antibiotic removal was observed throughout the operation, suggesting that microbiological composition was of greater influence than its primary metabolic activity on the removal of antibiotics.

Heterotrophic denitrification: An overlooked factor that contributes to nitrogen removal in n-DAMO mixed culture

Nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) has been recognized as a sustainable process for simultaneous removal of nitrogen and methane. The metabolisms of denitrifying anaerobic methanotrophs, including Candidatus Methanoperedens and Candidatus Methylomirabilis, have been well studied. However, potential roles of heterotrophs co-existing with these anaerobic methanotrophs are generally overlooked. In this study, we pulse-fed methane and nitrate into an anaerobic laboratory sequencing batch bioreactor and enriched a mixed culture with stable nitrate removal rate (NRR) of ∼28 mg NO3−-N L−1 d−1. Microbial community analysis indicates abundant heterotrophs, e.g., Arenimonas (5.3%–18.9%) and Fimbriimonadales ATM1 (6.4%), were enriched together with denitrifying anaerobic methanotrophs Ca. Methanoperedens (10.8%–13.2%) and Ca. Methylomirabilis (27.4%–34.3%). The results of metagenomics and batch tests suggested that the denitrifying anaerobic methanotrophs were capable of generating methane-derived intermediates (i.e., formate and acetate), which were employed by non-methanotrophic heterotrophs for denitrification and biomass growth. These findings offer new insights into the roles of heterotrophs in n-DAMO mixed culture, which may help to optimize n-DAMO process for nitrogen removal from wastewater.

Response of potential activity, abundance and community composition of nitrite-dependent anaerobic methanotrophs to long-term fertilization in paddy soils.

The process of nitrite-dependent anaerobic methane oxidation (n-damo) catalysed by Candidatus Methylomirabilis oxyfera (M. oxyfera)-like bacteria is a novel pathway in regulating methane (CH4 ) emissions from paddy fields. Nitrogen fertilization is essential to improve rice yields and soil fertility; however, its effect on the n-damo process is largely unknown. Here, the potential n-damo activity, abundance and community composition of M. oxyfera-like bacteria were investigated in paddy fields under three long-term (32 years) fertilization treatments, i.e. unfertilized control (CK), chemical fertilization (NPK) and straw incorporation with chemical fertilization (SNPK). Relative to the CK, both NPK and SNPK treatments significantly (p < 0.05) increased the potential n-damo activity (88%-110%) and the abundance (52%-105%) of M. oxyfera-like bacteria. The variation of soil organic carbon (OrgC) content and inorganic nitrogen content caused by the input of chemical fertilizers and straw returning were identified as the key factors affecting the potential n-damo activity and the abundance of M. oxyfera-like bacteria. However, the community composition and diversity of M. oxyfera-like bacteria did not change significantly by the input of fertilizers. Overall, our results provide the first evidence that long-term fertilization greatly stimulates the n-damo process, indicating its active role in controlling CH4 emissions from paddy fields.

Versatility of nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO): First demonstration with real wastewater

Nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) processes have been proven effective for nitrogen removal from synthetic wastewater. However, the demonstration using real wastewater has not been achieved yet. To this end, this study investigated the versatile applications of n-DAMO process in real wastewater treatment for the first time. Two methane-based membrane biofilm reactors (MBfRs) were employed to combine anammox and n-DAMO microorganisms, targeting nitrogen removal in mainstream (i.e., domestic sewage) and sidestream (i.e., anaerobic digestion liquor), respectively. Considering various technologies in sewage treatment, three different technical routes, including nitritation + methane-based MBfR, partial nitritation + methane-based MBfR and partial nitritation + anammox + methane-based MBfR, were investigated comprehensively, all producing effluent quality with total nitrogen (TN) at 5 mg N/L or less. Regarding the sidestream treatment, the methane-based MBfR also removed up to 96% TN from the partially nitrified anaerobic digestion liquor at a practically useful rate of 0.5 kg N/m3/d. Microbial communities revealed by 16S rRNA gene amplicon sequencing indicated the dominance of n-DAMO archaea in both reactors, along with the existence of anammox bacteria and n-DAMO bacteria. As the first demonstration of n-DAMO process in real wastewater, this study comprehensively confirmed the applicability of using methane as carbon source to remove nitrogen from both mainstream and sidestream wastewater, supporting their adoption by industries in practice.

Enhancement of nitrite-dependent anaerobic methane oxidation via Geobacter sulfurreducens

Nitrite-dependent anaerobic methane-oxidation (n-damo) is a potential novel technology for nitrogen removal in anaerobic wastewater treatment. In this study, Geobacter sulfurreducens (G) was applied to stimulate n-damo activity. Conductive materials such as nano-magnetite (M) or aggregating agents such as hydroxylapatite (H) were co-added with G. sulfurreducens to further investigate the enhancement effect. Results showed that the nitrite reduction activity of the n-damo culture was promoted by G. sulfurreducens, with 1.71–2.38 times higher in treatment G, G + M, and G + H than that in the control, but was inhibited by the single addition of hydroxylapatite. N-damo bacterial abundances based on the qPCR of the n-damo-specific pmoA gene increased in treatments with G. sulfurreducens, compared with that of the control. High-throughput sequencing analysis revealed the enrichment of uncultured phylum WPS-2 in treatments with G. sulfurreducens. Fluorescence in situ hybridization verified the co-occurrence pattern of n-damo bacteria (NC10), G. sulfurreducens, and type-I aerobic methanotrophs (Methylomonas spp.). The above results corroborated the microbial interspecies electron transfer (MIET) potentiality of the n-damo enrichment. Our study provides a novel pathway for enhancing MIET to stimulate n-damo process.

Enhancement of nitrite reduction and enrichment of Methylomonas via conductive materials in a nitrite-dependent anaerobic methane oxidation system

Nitrite-dependent anaerobic methane-oxidizing (n-damo) process has a promising prospect in anaerobic wastewater treatment, utilizing methane as the sole electron source to remove nitrite. However, the metabolic activity of n-damo bacteria is too low for practical application. This study aimed to stimulate n-damo process by introducing conductive nano-magnetite and/or electron shuttle anthraquinone-2,6-disulfonate (AQDS), and also set a comparative treatment of adding insulated ferrihydrite. The results showed that the nitrite reduction rate was enhanced the most significantly in treatment with nano-magnetite, approximately 1.6 times higher than that of the control without any supplement. While ferrihydrite application showed an adverse effect on n-damo process. The well-known aerobic methane oxidizer Methylomonas spp. was found to be enriched under n-damo condition with the supplementation of nano-magnetite and/or AQDS, but abundance of n-damo bacteria did not exhibit significant increase. It was hypothesized that Methylomonas spp. could be survived under anaerobic n-damo condition using oxygen produced by n-damo bacteria for the self-growth, and the nitrite reduction could be promoted through the enhancement of microbial interspecies electron transfer triggered by the introduction of conductive materials. It opens a new direction for the stimulation of n-damo activity, which needs more evidences to verify the hypothetic mechanism.

Anaerobic methane oxidation coupled to denitrification is an important potential methane sink in deep-sea cold seeps

Microbes play a crucial role in mediating the methane flux in deep-sea cold seep ecosystems, where only methane-related microbes have been well studied, while the whole microbial community and their ecological functions were still largely unknown. Here, we utilized metagenomic data to investigate how the structure and metabolism of microbial community shift in the reduced sediment habitats along the spatial scales. Microbial communities in cold seeps and troughs formed two distinct clades likely driven by environmental factors, such as total sulfur, total phosphate and NO3−, rather than geographical proximity. The predominance of Methanosarcinales reflected a high potential for methane production. In addition to the already well-reported ANME-1/SRB consortia, prevalence of bacterial Methylomirabilis and archaeal Methanoperedens as important performers in the n-damo process with respective of nitrite and nitrate as respective electron acceptor was observed in deep-sea hydrate-bearing regions as well. Aerobic methane oxidization was conducted mainly by type I methanotrophs at Site F (Formosa Ridge), but also via the n-damo process by Methanoperedens and Methylomirabilis in the Haima seep and Xisha Trough, respectively. Based on the high abundance of those denitrifying-dependent methane oxidizers and their related functional genes, we concluded that the previously overlooked n-damo process might be a major methane sink in cold seeps or in gas hydrate-bearing sediments if nitrate is available in the anoxic zones. The signature of isotopic labeling would be essential to confirm the contribution of different anaerobic methane oxidizing pathways in deep-sea cold seep ecosystems.

Cometabolic removal of organic micropollutants by enriched nitrite-dependent anaerobic methane oxidizing cultures

The innovative and recently discovered n-damo process, based on anaerobic methane oxidation with nitrite, was developed in a membrane-based bioreactor and evaluated in terms of organic micropollutants (OMPs) removal. The main singularity of this study consisted in the evaluation of organic micropollutants (OMPs) removal in the biological reactor. A strategy consisting on progressively increasing the nitrogen loading rate in order to increase the specific denitrification activity was followed to check if the selected OMPs were co-metabolically biotransformed. Significant nitrite removal rate (24.1 mg N L−1 d−1) was achieved after only 30 days of operation. A maximum specific removal of 186.3 mg N gVSS−1 d−1 was obtained at the end of the operation, which is one of the highest previously reported. A successfully n-damo bacteria enrichment was achieved, being Candidatus Methylomirabilis the predominant bacteria during the whole operation attaining a maximum relative abundance of about 40 %. The natural hormones (E1 and E2) were completely removed in the bioreactor. The specific removal rates of erythromycin (ERY), fluoxetine (FLX), roxithromycin (ROX) and sulfamethoxazole (SMX) were successfully correlated with the specific nitrite removal rates, suggesting a co-metabolic biotransformation.

Depth-specific distribution and significance of nitrite-dependent anaerobic methane oxidation process in tidal flow constructed wetlands used for treating river water.

Tidal flow constructed wetlands (TF CWs) have been considered an effective approach to treat contaminated river water, as well as a significant role in global matter cycles, especially for carbon and nitrogen. Notably, it has been thought that methane oxidation was completely catalyzed by the aerobic process, ignoring the anaerobic methane oxidation, such as the nitrite-dependent anaerobic methane oxidation (n-damo) process. In our current work, therefore, we used molecular and stable isotopes to investigate the biodiversity, quantity and potential rate of n-damo bacteria in the TF CWs located in the Xisha Wetland Park in the Yangtze River estuary, China. The results revealed that n-damo process was active in the collected soil cores, with a decreasing trend along water depths and rates ranging from 8.48 to 23.45nmol CO2 g-1 dry soil d-1. The n-damo bacterial contributions to CH4 oxidation and N2 production in TF CWs reached 9.49-26.26% and 20.73-47.11%, respectively, suggesting that n-damo bacteria was an important nitrogen and methane sink in the TF CWs, but had been previously overlooked. The copy numbers of total bacterial 16S rRNA and pmoA genes were 1.84-11.21×109 and 0.59-2.72×106 copies g-1 ds, respectively, as the higher abundance was found in the soil at lower water levels during tidal submergence. Diverse n-damo bacterial 16S rRNA gene sequences belonged to group B, C and D were measured, and it was found that group B and C were the most frequently measured n-damo clusters in the TF CWs. In addition, nitrite was the key factor regulating the n-damo bacterial distribution in the TF CWs. This study would broaden our horizons and help us better understand the nitrogen and methane cycles in tidal ecosystems.

Molecular Existence and Diversity of Nitrite-Dependent Anaerobic Methane Oxidizing (n-Damo) Bacteria in the Lakes of Badain of the Gobi Desert

Nitrite-dependent anaerobic methane oxidation (n-damo) process, mediated by Candidatus Methylomirabilis oxyfera of the candidate phylum NC10, was discovered recently which plays an important role in coupling the global nitrogen and carbon cycles. However, the distribution and diversity of this new anaerobic methane-oxidizing microorganism have not been investigated in desert lakes yet. The present study successfully retrieved n-damo bacterial 16S rRNA and pmoA gene sequences using PCR technique from lakes in Badain Jaran Desert of China. Phylogenetic analyses showed that n-damo bacteria widely occurred in brine and freshwater lakes on the desert with high diversity, including both sediment and water samples. The results of quantitative PCR indicated that the abundance of the 16S rRNA gene in lake sediments varied from 1.12 ± 0.68 × 105 to 1.64 ± 0.70 × 105 copies g−1 (dry weight), while that in water samples per milliliter was generally one order of magnitude lower than sediments. Correlation analyses suggested that n-damo bacterial abundance and diversity strongly depended on salinity. In lake sediments, the distribution, abundance, and diversity of n-damo bacteria were significantly associated with depth due to the concentration gradient of the NOx- and ammonium. This study provided new insights into both the n-damo community patterns and its interaction with ambient environmental factors in the desert lake ecosystem.

Optimization of a nitrite-dependent anaerobic methane oxidation (n-damo) process by enhancing methane availability

The nitrite-dependent anaerobic methane oxidation (n-damo) process is believed to have good application prospects, but its development is limited by the low growth rate and low specific activity of n-damo bacteria. In this study, immobilized biologically activated carbon (IC) and high static pressure (HP) were investigated to improve the performance of the n-damo process by enhancing methane availability. Both IC and HP improved the activity of n-damo bacteria, and the highest n-damo specific activity was obtained in the HP-IC system, which reached 47.65 ± 0.21 µmol N L−1 g−1 h−1. The larger specific surface area of biomass in the HP system was conducive to the transfer of methane, whereas IC increased the methane absorption capacity by approximately 10-fold and facilitated the methane supply to n-damo bacteria. Furthermore, q-PCR analysis revealed that both HP and IC improved the growth rate of n-damo bacteria, while HP could also enrich the diversity of n-damo bacteria.

Anaerobic methane oxidation potential and bacteria in freshwater lakes: Seasonal changes and the influence of trophic status

Nitrite-dependent anaerobic methane oxidation (n-damo), mainly carried out by n-damo bacteria, is an important pathway for mitigating methane emission from freshwater lakes. Although n-damo bacteria have been detected in a variety of freshwater lakes, their potential and distribution, and associated environmental factors, remain unclear. Therefore, the current study investigated the potential and distribution of anaerobic methanotrophs in sediments from Erhai Lake and Dianchi Lake, two adjacent freshwater lakes in the Yunnan Plateau with different trophic status. Both lakes showed active anaerobic methane oxidation potential and harbored a high density of n-damo bacteria. Based on the n-damo pmoA gene, sediment n-damo bacterial communities mainly consisted of Candidatus Methylomirabils oxyfera and Candidatus Methylomirabils sinica, as well as novel n-damo organisms. Sediment anaerobic methane oxidation potential and the n-damo bacterial community showed notable differences among seasons and between lakes. The environmental variables associated with lake trophic status (e.g. total nitrogen, ammonia nitrogen, nitrate nitrogen, and total organic carbon) might have significant impacts on the anaerobic methane oxidation potential, as well as the abundance and community structure of n-damo bacteria. Therefore, trophic status could determine the n-damo process in freshwater lake sediment.

Molecular and stable isotopic evidence for the occurrence of nitrite-dependent anaerobic methane-oxidizing bacteria in the mangrove sediment of Zhangjiang Estuary, China.

Nitrite-dependent anaerobic methane oxidation (n-damo), which is mediated by "Candidatus Methylomirabilis oxyfera-like" bacteria, is unique in linking the carbon and nitrogen cycles. However, the niche and activity of n-damo bacteria in the mangrove ecosystem have not been confirmed. Here, we report the occurrence of the n-damo process in the mangrove wetland of the Zhangjiang Estuary, China. The widespread occurrence of n-damo bacteria in mangrove wetland was confirmed using real-time quantitative polymerase chain reaction (qPCR) assay, which showed that the abundance of Methylomirabilis oxyfera-like bacterial 16S rRNA and pmoA genes ranged from 2.43 × 106 to 2.09 × 107 and 2.07 × 106 to 3.38 × 107copies per gram of dry soil in the examined sediment cores. The highest amount of targeting genes was all detected in the upper layer (0-20cm). Phylogenetic analyses of n-damo bacterial 16S rRNA and pmoA genes illustrated the depth-specific distribution and high diversity of n-damo bacteria in the mangrove wetland. Stable isotope experiments further confirmed the occurrence of n-damo in the examined mangrove sediments, and the potential n-damo rates ranged from 25.93 to 704.08nmol CO2 per gram of dry soil per day at different depths of the sediment cores, with the n-damo being more active in the upper layer of the mangrove sediments. These results illustrate the existence of active M. oxyfera-like bacteria and indicate that the n-damo process is a previously overlooked microbial methane sink in the mangrove wetlands.

Mitigation of Greenhouse Gas Emission during Nitrite-dependent Methane Anaerobic Oxidation Process by Increasing Static Pressure

Nitrite-dependent anaerobic methane oxidation (n-damo) process is a novel wastewater treatment technology, but the greenhouse gas (GHG) emission during the process is lack of report. N-damo bacteria was enriched and the GHG emission during n-damo process was measured in this study, and the feasibility of GHG emission mitigation by increasing static pressure was also investigated. Both reactors in this study obtained high efficiencies of nitrite removal which were 5.92 mg N L-1 d-1 g-1and 7.49 mg N L-1 d-1 g-1, respectively, the nitrite removal rate in reactor under static pressure was improved 26.52% than reactor under ambient pressure. With similar N2O emission, lower total global warming potential (GWP) in reactor under static pressure was obtained because improved n-damo process led to the higher CH4 sink. According to microbial analysis, improved n-damo process was resulted from higher abundance of n-damo bacteria in reactor under static pressure. Furthermore, static pressure could enhance the screening of non-target bacteria such as denitrifying bacteria and make n-damo bacteria predominate and further enhance n-damo process.

Reduction of greenhouse gases emissions during anoxic wastewater treatment by strengthening nitrite-dependent anaerobic methane oxidation process

Nitrite-dependent anaerobic methane oxidation (n-damo) is a recently discovered process performed by NC10 phylum, which plays an important role in greenhouse gases (GHG) reduction. In this study, co-existence of n-damo bacteria and methanogens was successfully achieved by using upflow anaerobic sludge blanket (UASB) reactor. Reactor with inorganic carbon source (CO2/H2) showed the highest abundance of n-damo bacteria and the highest n-damo potential activity, resulted in its highest nitrogen removal rate. Significant reduction in GHG was obtained after introduction of n-damo process, especially for N2O. Furthermore, GHG emissions decreased with the increase of n-damo bacteria abundance. Community structure analysis found carbon source could influence the diversity of n-damo bacteria indirectly. And phylogenetic analysis showed that all the obtained sequences were assigned to group B, mainly due to in situ production and consumption of CH4.

Spatial change of reservoir nitrite-dependent methane-oxidizing microorganisms

Nitrite-dependent anaerobic methane oxidation (n-damo), catalyzed by microorganisms affiliated with bacterial phylum NC10, can have an important contribution to the reduction of the methane emission from anoxic freshwater sediment to the atmosphere. However, information on the variation of sediment n-damo organisms in reservoirs is still lacking. The present study monitored the spatial change of sediment n-damo organisms in the oligotrophic freshwater Xinfengjiang Reservoir (South China). Sediment samples were obtained from six different sampling locations and two sediment depths (0–5 cm, 5–10 cm). Sediment n-damo bacterial abundance was found to vary with sampling location and layer depth, which was likely influenced by pH and nitrogen level. The presence of the n-damo pmoA gene was found in all these samples. A remarkable shift occurred in the diversity and composition of sediment n-damo pmoA gene sequences. A variety of distinctively different n-damo pmoA clusters existed in reservoir sediments. The pmoA sequences affiliated with Candidatus Methylomirabilis oxyfera formed the largest group, while a significant proportion of the obtained n-damo pmoA gene sequences showed no close relationship to those from any known NC10 species. In addition, the present n-damo process was found in reservoir sediment, which could be enhanced by nitrite nitrogen amendment.