Dependent Anaerobic Methane Oxidation Research Articles

Humic substances as electron acceptor for anaerobic oxidation of methane (AOM) and electron shuttle in Mn (IV)-dependent AOM

Anaerobic methanotrophic archaea (ANME) belonging to the family Methanoperedenaceae are crucial for the global carbon cycle and different biogeochemical processes, owing to their metabolic versatility to couple anaerobic oxidation of methane (AOM) with different electron acceptors. A universal feature of Methanoperedenaceae is the abundant genes encoded in their genomes associated with extracellular electron transfer (EET) pathways. Candidatus. ‘Methanoperedens manganicus’, an archaeon belonging to the family Methanoperedenaceae, was recently enriched in a bioreactor performing AOM coupled with Mn (IV) reduction. Using this EET-capable ANME, we tested the hypothesis in this study that ANME can catalyse the humic-dependent AOM for growth. A two-year incubation showed that AOM activity can be sustained by Ca. ‘M. manganicus’ consortium in a bioreactor fed only with humic acids and methane. An isotopic mass balance batch test confirmed that the observed AOM was coupled to the reduction of humic acids. The increase of relative abundance of Ca. ‘M. manganicus’, and the total archaea population in the microbial community suggested that Ca. ‘M. manganicus’ can grow on methane and humic acids. The observation of humic-dependent AOM led to a subsequent hypothesis that humic acids could be used as the electron shuttle to mediate the EET in dissimilatory Mn (IV) reduction by Ca. ‘M. manganicus’. We tested this hypothesis by adding humic acids to a Ca. ‘M. manganicus’ dominated-culture, which showed that the AOM rate was doubled by the addition of humic acids. X-ray photoelectron spectroscopy (XPS) showed that quinone moieties were consumed when humic acids worked as electron acceptors while remaining stable when functioning as a shuttle for electron transfer. The results of our study suggest that humic acids may serve as electron shuttles to allow ANME to access more electron acceptors through long-range EET.

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.

Soil nitrogen content and key functional microorganisms influence the response of wetland anaerobic oxidation of methane to trivalent iron input

Trivalent iron (Fe3+)-dependent anaerobic oxidation of methane (Fe-AOM), which is mediated by metal-reducing bacteria, is widely recognized as a major sink for the greenhouse gas methane (CH4), and is a key driver of the carbon (C) biogeochemical cycle. However, the effect of Fe3+ addition on AOM in the present investigation is still ambiguous, and the mechanism is vague. In this study, we investigated the mechanism of changes in AOM response to Fe3+ input at different wetlands by using laboratory incubation methods combined with molecular biology techniques. Results indicated that Fe3+ input did not always lead to promoted AOM rates, which may be mediated by complex environmental factors, while lower soil total nitrogen (TN) had a positive effect on the response of AOM subjected to Fe3+ input. Notably, the promoted response of AOM was regulated by higher soil microbial diversity, of which the Shannon index was a key indicator leading to variation in the AOM response. Additionally, several biomarkers, including Planctomycetota and Burkholderiaceae, were key microorganisms responsible for alterations in AOM response. Our results suggest that the capacity of Fe3+ cycling-mediated AOM may gradually decrease in light of increasing anthropogenic N and Fe inputs to global estuarine wetlands, while its reaction processes will become more complex and more strongly coupled with multiple environmental factors. This finding contributes to the enhanced understanding and prediction of the wetland CH4-related C with Fe cycles, as well as provides theoretical support for the underlying mechanisms.

Response of the Anaerobic Methanotrophic Archaeon Candidatus "Methanoperedens nitroreducens" to the Long-Term Ferrihydrite Amendment.

Anaerobic methanotrophic (ANME) archaea can drive anaerobic oxidation of methane (AOM) using solid iron or manganese oxides as the electron acceptors, hypothetically via direct extracellular electron transfer (EET). This study investigated the response of Candidatus “Methanoperedens nitroreducens TS” (type strain), an ANME archaeon previously characterized to perform nitrate-dependent AOM, to an Fe(III)-amended condition over a prolonged period. Simultaneous consumption of methane and production of dissolved Fe(II) were observed for more than 500 days in the presence of Ca. “M. nitroreducens TS,” indicating that this archaeon can carry out Fe(III)-dependent AOM for a long period. Ca. “M. nitroreducens TS” possesses multiple multiheme c-type cytochromes (MHCs), suggesting that it may have the capability to reduce Fe(III) via EET. Intriguingly, most of these MHCs are orthologous to those identified in Candidatus “Methanoperedens ferrireducens,” an Fe(III)-reducing ANME archaeon. In contrast, the population of Ca. “M. nitroreducens TS” declined and was eventually replaced by Ca. “M. ferrireducens,” implying niche differentiation between these two ANME archaea in the environment.

Anaerobic oxidation of methane (AOM) driven by multiple electron acceptors in constructed wetland and the related mechanisms of carbon, nitrogen, sulfur cycles

Anaerobic oxidation of methane (AOM) mediated by microorganisms plays an important role in the global carbon cycle and methane emission control. This study demonstrated that simultaneous multi-electron acceptor-driven AOM existed in the electroactive constructed wetland environment of freshwater, which is crucial to global carbon, sulfur, nitrogen cycles and the manganese, iron, humics reduction. This biochemical process was mediated by two anaerobic methanotrophic archaea (ANME) jointly, ANME-2a and ANME-2d. Better removal efficiency of sulfate (45.65 ± 3.47%), ammonium nitrogen (96.48 ± 3.82%) and COD (94.83 ± 5.89%) was observed in CW-MFC with AOM driven by multi-electron acceptor (CW-MFC_Mn). In addition, the methane emissions from CW-MFC with sulfate and nitrogen dependent AOM were reduced by 57.50% in presence of manganese ore, suggesting that multi-electron acceptors-driven AOM can effectively control methane emissions. The transformation of manganese oxide in wetland manganese ore was studied, and almost no Mn (Ⅱ) residue was detected in the effluent, indicating the potential role of Mn-dependent AOM in manganese pollution removal. This study also revealed the interactions of methanogenesis, electrogenesis, multi-electron acceptor-driven AOM and dissimilatory metal reduction (DMR) involved in CW-MFC, in which Geobacter played a crucial role in the association of various biological reactions. This study not only expands the understanding of AOM in nature, but also proposed a novel sight to simultaneously control wetland methane emissions, manganese, sulfur, and nitrogen pollution.

Insight into SO4(-II)- dependent anaerobic methane oxidation in landfill: Dual-substrates dynamics model, microbial community, function and metabolic pathway

In anaerobic landfill, SO42- could serve as electron receptor for methane oxidation. In theory, concentrations of both methane and SO42- should be related to methane oxidation rate. However, the dynamics process has yet to be discovered, and the understanding of metabolic pathways of the sulfate-dependent anaerobic methane oxidation (S-DAMO) process in landfill remains limited. In this study, S-DAMO dynamics was investigated by observing the CH4 oxidation rates under different CH4/ SO42-counter-gradients. The CH4-SO42- dual-substrate model based on MichaeliseMenten equation was got (maximum substrate degradation rate Vmax [22.9 ± 1.31] µmol/[kg·d], half-saturation constantsKm,CH446.095±0.41ppmv, andKm,so42-(7.26±0.605)g/g). High-throughput sequencing analysis indicated Methanobacterials, Methanosarcinales, and Soil Crenarchaeotic were the main functional microorganisms for S-DAMO in landfill. The metabolic pathway of S-DAMO was speculated as the reverse methanogenesis pathway through Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUST) analysis, while methanogenesis was the methyl nutrition way based on methanol. The enzymes related to the carbon and sulfur cycles and their relative abundances in the microcosms were analyzed to graph the methane metabolic pathway and the sulfur metabolic pathway. The findings provide important parameters for CH4 mitigation in landfills, and give a new insight for understanding S-DAMO metabolic pathway in landfill.

Analysing seasonal variations of methane oxidation processes coupled with denitrification in a stratified lake using stable isotopes and numerical modeling

Since the discovery of nitrite (n-damo) and nitrate dependent anaerobic oxidation of methane in 2006, evidence has arisen about the occurrence of this process in freshwater ecosystems. Anaerobic oxidation of methane (AOM) linked with denitrification has been proposed to be a previously overlooked sink of methane in freshwater environments. However, the occurrence and relevance of AOM coupled with denitrification in methane removal and, therefore, mitigating methane emissions in the environment is so far largely unquantified.We investigated methane oxidation processes in the water column at the seasonally stratified lake Fohnsee located in Southern Germany to determine the seasonal dynamics and to quantitatively describe the contribution of anaerobic and aerobic oxidation of methane removing CH₄ from the water column throughout nearly one year. Vertical concentration profiles and corresponding stable isotope ratios of methane (δ13C) and nitrate (δ15N and δ18O), together with dissolved oxygen concentrations were measured, and a numerical model was developed to evaluate whether anaerobic oxidation of methane coupled with denitrification is a key biogeochemical process at lake Fohnsee. Our data set together with the results of the numerical model revealed a redox zone within the water column where both denitrification and anaerobic oxidation of methane are linked, contributing to approximately 70% of methane removal between June and September. Therefore, AOM linked with denitrification represents the dominant methane sink in the water column of lake Fohnsee during summer stratification.The data patterns also showed that the anaerobic oxidation of methane zone was located near the lake bottom in May, and moved upwards following the seasonal vertical displacement of the oxycline and the availability of nitrate. This redox dynamic within the water column of lake Fohnsee had also an effect on the relevance of aerobic and anaerobic oxidation of methane removing methane from the water column during the year and the formation of a bacterial sulfate reduction zone close to the lake sediments.

The Role of Crystalline Iron Oxides in Methane Mitigation through Anaerobic Oxidation of Methane

Biological Fe(III)-dependent anaerobic oxidation of methane (AOM) is predicted to be a key methane sink in natural and anthropogenic environments. Crystalline Fe(III) is an important environmentall...

The possible occurrence of iron-dependent anaerobic methane oxidation in an Archean Ocean analogue

In the ferruginous and anoxic early Earth oceans, photoferrotrophy drove most of the biological production before the advent of oxygenic photosynthesis, but its association with ferric iron (Fe3+) dependent anaerobic methane (CH4) oxidation (AOM) has been poorly investigated. We studied AOM in Kabuno Bay, a modern analogue to the Archean Ocean (anoxic bottom waters and dissolved Fe concentrations > 600 µmol L−1). Aerobic and anaerobic CH4 oxidation rates up to 0.12 ± 0.03 and 51 ± 1 µmol L−1 d−1, respectively, were put in evidence. In the Fe oxidation–reduction zone, we observed high concentration of Bacteriochlorophyll e (biomarker of the anoxygenic photoautotrophs), which co-occurred with the maximum CH4 oxidation peaks, and a high abundance of Candidatus Methanoperedens, which can couple AOM to Fe3+ reduction. In addition, comparison of measured CH4 oxidation rates with electron acceptor fluxes suggest that AOM could mainly rely on Fe3+ produced by photoferrotrophs. Further experiments specifically targeted to investigate the interactions between photoferrotrophs and AOM would be of considerable interest. Indeed, ferric Fe3+-driven AOM has been poorly envisaged as a possible metabolic process in the Archean ocean, but this can potentially change the conceptualization and modelling of metabolic and geochemical processes controlling climate conditions in the Early Earth.

Nitrate/nitrite dependent anaerobic methane oxidation coupling with anammox in membrane biotrickling filter for nitrogen removal

Combining nitrate/nitrite dependent anaerobic methane oxidation (n-DAMO) and anaerobic ammonium oxidation (Anammox) is a promising sustainable wastewater treatment technology, which simultaneously achieve nitrogen removal and methane emission mitigation. However, the practical application of n-DAMO has been greatly limited by its extremely slow growth-rate and low reaction rate. This work proposes an innovative Membrane BioTrickling Filter (MBTF), which consist of hollow fiber membrane for effective methane supplementation and polyurethane sponge as support media for the attachment and growth of biofilm coupling n-DAMO with Anammox. When steady state with a hydraulic retention time (HRT) of 6.00 h was reached, above 99.9% of nitrogen was removed from synthetic sidestream wastewater at a rate of 3.99 g N L−1 d−1. This system presented robust capacity to withstand unstable partial nitritation effluent, achieving complete nitrogen removal at a varied nitrite to ammonium ratio in the range of 1.10–1.40. It is confirmed that n-DAMO and Anammox microorganisms jointly dominated the microbial community by pyrosequencing technology. The complete nitrogen removal potential at high-rate and efficient biomass retention (12.4 g VSS L−1) of MBTF offers promising alternative for sustainable wastewater treatment by the combination of n-DAMO and Anammox.

Shift in nitrogen transformation in peatland soil by nitrogen inputs

Inputs of nitrogen (N) to peatlands in the form of fertilizers have rapidly increased due to the intensification of agricultural systems, impacting ecological processes, and the carbon storage function of peatland. However, detailed information on the impacts of long-term N inputs on the individual steps of N transformation processes in peatland soils still needs to be fully understood. We investigated N mineralization and nitrification rates as well as nitrite dependent anaerobic methane oxidation (n-damo), anaerobic ammonium oxidation (anammox), denitrification, and dissimilatory nitrate reduction to ammonium (DNRA) in a peatland affected by N inputs for >50 years, using isotope tracing technique and quantitative PCR. Based on the results, N inputs increased N mineralization and nitrification rates by 77 and 43%, respectively. Notably, the contributions of n-damo and anammox to N2 production were enhanced by 242 and 170%, accounting for 30 and 12%, respectively. The contributions of denitrification and DNRA to N2 production decreased by 27 and 52%, accounting for 48 and 10% of N2 production, respectively. Nitrifier abundance increased significantly, with AOA being the dominant prokaryote (from 696 to 1090 copies g−1), but AOB responded more strongly to N inputs (from 5 to 68 copies g−1). The N inputs also promoted the growth of n-damo and anammox bacteria, whose abundances increased by 3.7% (from 565 to 586 copies g−1) and 85.7% (from 305 to 567 copies g−1), respectively, while denitrifier abundance was significantly reduced, with nirK and nirS abundances decreasing by 58% (from 738 to 308 copies g−1) and 50% (from 218 to 109 copies g−1), respectively. Soil pH was the key environmental factor influencing N transformations. We show that n-damo plays important roles in N cycling in peatland subjected to N inputs, providing a scientific basis for improved peatland management.

Niche Differentiation of Sulfate- and Iron-Dependent Anaerobic Methane Oxidation and Methylotrophic Methanogenesis in Deep Sea Methane Seeps.

Methane seeps are widespread seafloor ecosystems shaped by complex physicochemical-biological interactions over geological timescales, and seep microbiomes play a vital role in global biogeochemical cycling of key elements on Earth. However, the mechanisms underlying the coexistence of methane-cycling microbial communities remain largely elusive. Here, high-resolution sediment incubation experiments revealed a cryptic methane cycle in the South China Sea (SCS) methane seep ecosystem, showing the coexistence of sulfate (SO42–)- or iron (Fe)-dependent anaerobic oxidation of methane (AOM) and methylotrophic methanogenesis. This previously unrecognized methane cycling is not discernible from geochemical profiles due to high net methane consumption. High-throughput sequencing and Catalyzed Reporter Deposition-Fluorescence in situ Hybridization (CARD-FISH) results suggested that anaerobic methane-oxidizing archaea (ANME)-2 and -3 coupled to sulfate-reducing bacteria (SRB) carried out SO42–-AOM, and alternative ANME-2 and -3 solely or coupled to iron-reducing bacteria (IRB) might participate in Fe-AOM in sulfate-depleted environments. This finding suggested that ANME could alter AOM metabolic pathways according to geochemical changes. Furthermore, the majority of methylotrophic methanogens belonged to Methanimicrococcus, and hydrogenotrophic and acetoclastic methanogens were likely inhibited by sulfate or iron respiration. Fe-AOM and methylotrophic methanogenesis are overlooked potential sources and sinks of methane in methane seep ecosystems, thus influencing methane budgets and even the global carbon budget in the ocean.

Modeling nitrate/nitrite dependent anaerobic methane oxidation and Anammox process in a membrane granular sludge reactor

The granular bioreactor, characterized by excellent settling velocity, high rate and low cost is an ideal choice for achieving coupled nitrate/nitrite dependent denitrifying anaerobic methane oxidation (n-DAMO) and anaerobic ammonium oxidation (Anammox) process. To fundamentally understand its underlining mechanisms and provide suggestions for process optimization, a granule-based model framework was developed to describe simultaneous anaerobic methane and ammonium oxidation by functional microbes. The proposed model was evaluated based on long-term experimental data from two membrane granular sludge reactors (MGSRs) with different operational conditions. The model possessed of good predictive ability to reproduce removal rates and effluent concentrations of nitrogen species. The predicted biomass abundance in two MGSRs and stratified microbial distribution along granule depth were consistent with experimental observations. The estimated parameter values, with good identifiability and reliability indicated a stimulated growth of DAMO archaea in MGSRs. Both hydraulic retention times (HRTs) and granule sizes have influences on microbial abundance in the MGSR and community distribution inside granules. Within the investigated HRTs from 1.4 h to 20 h and granule sizes from 500 μm to 2900 μm, it was revealed that a proper control of relatively short HRTs and small granule sizes resulted in an increased fraction of DAMO archaea and a reduced DAMO bacteria abundance with AnAOB less impacted, which would lower the required nitrite nitrogen to ammonium nitrogen ratio in the nitritation reactor (prior unit) and thus minimize operational cost in sidestream treatment lines.

Development of granular sludge coupling n-DAMO and Anammox in membrane granular sludge reactor for high rate nitrogen removal

The integration of nitrate/nitrite dependent anaerobic methane oxidation (n-DAMO) and anaerobic ammonium oxidation (Anammox) provides sustainable solution to simultaneously remove nitrate, nitrite and ammonium. This study demonstrated the sludge granulation process coupling n-DAMO and Anammox from mixed inoculum including river sediment, return activated sludge and crushed anaerobic granule sludge in a novel membrane granular sludge reactor (MGSR). Flocculent biomass gradually turned into compact aggregates and retained as granular sludge with an average diameter of 2.2 mm in MGSR after 684 days’ operation. When steady state with a hydraulic retention time of 1.19 days was reached, the MGSR achieved a nitrogen removal rate of 1.77 g N L−1 d−1. Granules with density of 1.043 g mL−1, settling velocity of 72 m h−1 and sludge volume index of 22 mL g−1 leaded to excellent biomass retention (42 g VSS L−1). Pyrosequencing analysis revealed that two dominant microbial groups, n-DAMO archaea and Anammox bacteria, in the microbial community of the granule were enriched to 31.09% and 12.45%. Fluorescence in situ hybridization revealed a homogenous distribution of n-DAMO archaea and Anammox bacteria throughout the granule. The granular sludge coupling n-DAMO and Anammox microorganisms provides significant potential for high rate nitrogen removal from wastewater.

Different responses of nitrite- and nitrate-dependent anaerobic methanotrophs to increasing nitrogen loading in a freshwater reservoir

Nitrite (NO2−)- and nitrate (NO3−)-dependent anaerobic oxidation of methane (AOM) are two new additions in microbial methane cycle, which potentially act as important methane sinks in freshwater aquatic systems. Here, we investigated spatial variations of community composition, abundance and potential activity of NO2−- and NO3−-dependent anaerobic methanotrophs in the sediment of Jiulonghu Reservoir (Zhejiang Province, China), a freshwater reservoir having a gradient of increasing nitrogen loading from upstream to downstream regions. High-throughput sequencing of total bacterial and archaeal 16S rRNA genes showed the cooccurrence of Candidatus Methylomirabilis oxyfera (M. oxyfera)-like and Candidatus Methanoperedens nitroreducens (M. nitroreducens)-like anaerobic methanotrophs in the examined reservoir sediments. The community structures of these methanotrophs differed substantially between the sediments of upstream and downstream regions. Quantitative PCR suggested higher M. oxyfera-like bacterial abundance in the downstream (8.6 × 107 to 2.8 × 108 copies g−1 dry sediment) than upstream sediments (2.4 × 107 to 3.5 × 107 copies g−1 dry sediment), but there was no obvious difference in M. nitroreducens-like archaeal abundance between these sediments (3.7 × 105 to 4.8 × 105 copies g−1 dry sediment). The 13CH4 tracer experiments suggested the occurrence of NO2−- and NO3−-dependent AOM activities, and their rates were 4.7–14.1 and 0.8–2.6 nmol CO2 g−1 (dry sediment) d−1, respectively. Further, the rates of NO2−-dependent AOM in downstream sediment were significantly higher than those in upstream sediment. The NO3− concentration was the key factor affecting the spatial variations of abundance and activity of NO2−-dependent anaerobic methanotrophs. Overall, our results showed different responses of NO2−- and NO3−-dependent anaerobic methanotrophs to increasing nitrogen loading in a freshwater reservoir.

Methane emission suppression in flooded soil from Amazonia

The coupling between ferrous iron and methane production has important global implications, with iron ions acting as electron acceptors for anaerobic oxidation of methane (AOM) and inhibitors of methanogenesis in different environments, including floodplain soils. In this sense, we analyzed the relationship between Fe(II) concentration and methane production in soil layers collected at 0–15 cm and 15–30 cm from flooded-forest and -agroforestry in Amazonian clear water floodplain incubated in anaerobic batch reactors using acetate, formate and glucose as organic sources. High throughput sequencing of archaeal and bacterial 16S rRNA genes was employed to assess the abundance and composition of the active methanogenic and methanotrophic microbial groups potentially involved in Fe(III)-dependent AOM in the soil used as inoculum. Positive correlation was revealed between Fe(II) concentration and methane production, with higher accumulation of Fe(II) in incubated soil layer collected at 0–15 cm in both forest and agroforestry sites for all the three organic sources. The accumulation of Fe(II) in the incubated soil evidenced the oxidation of Fe(III) potentially by Methanobacterium, Desulfobulbus and ‘Candidatus methanoperedens nitroreducens’ living in anaerobic condition at this soil layer. The results point out to the microbial ferric iron reduction as an important potential pathway for anaerobic organic matter decomposition in Amazonian floodplain, evidencing methanogenesis suppression by Fe(III) reduction in flooded-forest and -agroforestry in Amazonian clear water river floodplain.

Anaerobic methane oxidation coupled to manganese reduction by members of the Methanoperedenaceae

Anaerobic oxidation of methane (AOM) is a major biological process that reduces global methane emission to the atmosphere. Anaerobic methanotrophic archaea (ANME) mediate this process through the coupling of methane oxidation to different electron acceptors, or in concert with a syntrophic bacterial partner. Recently, ANME belonging to the archaeal family Methanoperedenaceae (formerly known as ANME-2d) were shown to be capable of AOM coupled to nitrate and iron reduction. Here, a freshwater sediment bioreactor fed with methane and Mn(IV) oxides (birnessite) resulted in a microbial community dominated by two novel members of the Methanoperedenaceae, with biochemical profiling of the system demonstrating Mn(IV)-dependent AOM. Genomic and transcriptomic analyses revealed the expression of key genes involved in methane oxidation and several shared multiheme c-type cytochromes (MHCs) that were differentially expressed, indicating the likely use of different extracellular electron transfer pathways. We propose the names “Candidatus Methanoperedens manganicus” and “Candidatus Methanoperedens manganireducens” for the two newly described Methanoperedenaceae species. This study demonstrates the ability of members of the Methanoperedenaceae to couple AOM to the reduction of Mn(IV) oxides, which suggests their potential role in linking methane and manganese cycling in the environment.

Anaerobic oxidation of methane in paddy soil: Role of electron acceptors and fertilization in mitigating CH4 fluxes

The anaerobic oxidation of methane (AOM) in marine ecosystems is ubiquitous and largely coupled to sulfate reduction. In contrast, the role of AOM in terrestrial environments and the dominant electron acceptors driving terrestrial AOM needs deeper understanding. Submerged rice paddies with intensive CH4 production have a high potential for AOM, which can be important for greenhouse gas mitigation strategies. Here, we used 13CH4 to quantify the AOM rates in paddy soils under organic (Pig manure, Biochar) and mineral (NPK) fertilization. Alternative-to-oxygen electron acceptors for CH4 oxidation, including Fe3+, NO3−, SO42−, and humic acids, were examined and their potential for CH4 mitigation from rice paddies was assessed by 13CH4 oxidation to 13CO2 under anoxic conditions.During 84 days of anaerobic incubation, the cumulative AOM (13CH4-derived CO2) reached 0.15–1.3 μg C g-1 dry soil depending on fertilization. NO3- was the most effective electron acceptor, yielding an AOM rate of 0.80 ng C g-1 dry soil h-1 under Pig manure. The role of Fe3+ in AOM remained unclear, whereas SO42- inhibited AOM but strongly stimulated the production of unlabeled CO2, indicating intensive sulfate-induced decomposition of organic matter. Humic acids were the second most effective electron acceptor for AOM, but increased methanogenesis by 5–6 times in all fertilization treatments. We demonstrated for the first time that organic electron acceptors (humic acids) are among the key AOM drivers and are crucial in paddy soils. The most pronounced AOM in paddy soils occurred under Pig manure, followed by Control and NPK, while AOM was the lowest under Biochar. We estimate that nitrate (nitrite)-dependent AOM in paddy fields globally consumes ~3.9 Tg C–CH4 yr-1, thereby offsetting the global CH4 emissions by ~10–20%. Thus, from a broader agroecological perspective, the organic and mineral fertilizers control an important CH4 sink under anaerobic conditions in submerged ecosystems. Appropriate adjustments of soil fertilization management strategies would therefore help to decrease the net CH4 flux to the atmosphere and hence the global warming.

A methanotrophic archaeon couples anaerobic oxidation of methane to Fe(III) reduction.

Microbially mediated anaerobic oxidation of methane (AOM) is a key process in the regulation of methane emissions to the atmosphere. Iron can serve as an electron acceptor for AOM, and it has been suggested that Fe(III)-dependent AOM potentially comprises a major global methane sink. Although it has been proposed that anaerobic methanotrophic (ANME) archaea can facilitate this process, their active metabolic pathways have not been confirmed. Here we report the enrichment and characterisation of a novel archaeon in a laboratory-scale bioreactor fed with Fe(III) oxide (ferrihydrite) and methane. Long-term performance data, in conjunction with the 13C- and 57Fe-labelling batch experiments, demonstrated that AOM was coupled to Fe(III) reduction to Fe(II) in this bioreactor. Metagenomic analysis showed that this archaeon belongs to a novel genus within family Candidatus Methanoperedenaceae, and possesses genes encoding the "reverse methanogenesis" pathway, as well as multi-heme c-type cytochromes which are hypothesised to facilitate dissimilatory Fe(III) reduction. Metatranscriptomic analysis revealed upregulation of these genes, supporting that this archaeon can independently mediate AOM using Fe(III) as the terminal electron acceptor. We propose the name Candidatus "Methanoperedens ferrireducens" for this microorganism. The potential role of "M. ferrireducens" in linking the carbon and iron cycles in environments rich in methane and iron should be investigated in future research.