Anaerobic Methane Oxidation Process Research Articles

Behaviours of methane metabolism and community dynamics of methane anaerobic oxidation microbes on carbonate rocks with long-term cultivation in cold seep environment

The anaerobic oxidation of methane (AOM) and sulfate reduction processes in cold seep environments can control methane emission sources and thus mitigate the pressure of increasing global greenhouse gas concentrations. The surfaces of carbonate rocks in cold seep host an abundance of microorganisms that participate in AOM reactions. Investigating the metabolism and conversion of methane by these microbes is instrumental in advancing the exploration and utilization of deep-sea methane energy. Previous studies primarily focused on in-situ investigations of microbial communities on carbonate rocks in cold seep environments, while the dynamic balance of carbonate mineralization caused by microbial community changes and the characteristics of methane consumption in carbonate samples remains unclear. In this study, we used methane as the sole carbon source, enriched and cultured carbonate samples under high pressure, and monitored the community dynamics regularly. The results demonstrated that methane consumption and metabolic pathways played a crucial role in influencing community succession and carbonate mineralization. AOM processes, coupled with sulfate reduction and nitrate reduction, which are dominated by ANME-2c, facilitate the precipitation of calcium carbonate. Conversely, the acetate production process, dominated by ANME-1, may hinder the efficiency of calcium carbonate mineralization. Additionally, the impact of bacterial groups in the enrichment process, through the production of extracellular enzymes, organic acid pathways, and the expression of carbonic anhydrase pathways on carbonate mineralization, should not be disregarded. These findings highlight the significance of diverse pathway methane metabolism in the dynamics of methane consumption, deep-sea microbial communities, carbonate kinetics, and marine biological carbon sequestration processes.

Equal importance of humic acids and nitrate in driving anaerobic oxidation of methane in paddy soils

Methane (CH4) is both generated and consumed in paddy soils, where anaerobic oxidation of methane (AOM) serves as a crucial process for mitigating CH4 emissions. Although the participation of humic acids (HA) and nitrate in AOM has been recognized, their relative roles and significance in paddy soils remain insufficiently investigated. In this study, we explored the potential activity of AOM driven by HA and nitrate, as well as the composition of archaeal communities in paddy soils across different rice growth periods and fertilization treatments. AOM activity ranged from 0.81 to 1.33 and 1.26 to 2.38 nmol of 13CO2 g−1 (dry soil) day−1 with HA and nitrate, respectively. No significant differences (p < 0.05) were observed between the AOM activity driven by HA and nitrate across the three fertilization treatments. According to AOM activity, the annual consumption of CH4 was estimated at approximately 0.49 ± 0.06 and 0.83 ± 0.19 Tg for AOM processes driven by HA and nitrate in Chinese paddy soils. Nitrate-driven AOM activity exhibited a positive (p < 0.05) correlation with the abundance of the ANME-2d mcrA gene but a negative (p < 0.05) correlation with the content of dissolved organic carbon. Intriguingly, HA-driven AOM activity was only correlated positively with the nitrate-driven AOM activity. Soil water content, soil organic carbon, nitrate and nitrite contents were significantly correlated with the relative abundance of methanogenic and methanotrophic archaea. These results identified the potential importance of HA and nitrate in driving AOM processes within paddy soils, providing a comprehensive understanding of the complex microbial processes regulating greenhouse gas emissions from paddy soils.

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.

Horizontal and vertical distribution of nitrate-driven anaerobic methane oxidation process in coastal wetlands under different plant species cover across southeast China

Coastal wetlands are an important methane emission source. Nitrate-driven anaerobic oxidation of methane (AOM), catalyzing via a novel clade of anaerobic methanotrophic archaea (ANME-2d), is a potentially important process for methane emission mitigation in coastal wetlands. However, the influence of regional and local environmental conditions, as well as plant species cover on this AOM process, remains unclear. Here, we investigated the horizontal and vertical distribution of nitrate-driven AOM activity and ANME-2d archaeal community in four coastal wetlands under both native and invasive plant species across southeast China. It was estimated that this AOM process could potentially reduce 5.7–9.5% methane emissions from the studied wetlands. Generalized linear mixed-effects model showed that sampling locations and sediment layers had greater impact on nitrate-driven AOM activity than plant species cover, and all these factors significantly affected the abundance of ANME-2d archaea. Nitrate-driven AOM activity was higher in low-latitude coastal wetlands than in high-latitude areas, and the activity positively correlated with mean annual temperature. Furthermore, both nitrate-driven AOM activity and ANME-2d archaeal abundance showed significant vertical variations along sediment profiles, with higher values in the slightly alkaline surface sediment. The plant species cover was found to impact the ANME-2d archaeal abundance and diversity by regulating the availability of substrates for the archaea. Overall, our study provides valuable insights into effects of regional and local environmental parameters and plant species cover on the horizontal and vertical distribution of nitrate-driven AOM activity and ANME-2d archaeal community in coastal wetlands.

Effects of magnetite on nitrate-dependent anaerobic oxidation of methane in an anaerobic membrane biofilm reactor: Metatranscriptomic analysis and mechanism prediction

Magnetite has been widely reported to accelerate methane production in anaerobic digestion. However, the effects of magnetite on the process of methane consumption, specifically nitrate-dependent anaerobic oxidation of methane (AOM), and its underlying mechanisms remain poorly understood. In this study, the addition of magnetite significantly improved the removal of nitrate in an anaerobic membrane biofilm reactor (AnMBfR) (25.2 ± 2.5 mg NO3−-N/L/d) compared to that in an AnMBfR without magnetite (8.7 ± 4.2 mg NO3−-N/L/d). Metatranscriptomic analysis showed that genes encoding type IV pilus assembly proteins were highly expressed in nitrate/nitrite-reducing bacteria in the biofilms of the two AnMBfRs. Specifically, the transcript abundance of these genes was observed to be higher in the presence of magnetite (gene expression levels, log2FPKM = 13.08) compared to the absence of magnetite (12.04). Biofilms with magnetite exhibited a linear increase in electron transfer resistance with increasing temperature, and the decrease in pH significantly increased their conductivity, indicating a metallic-like conductivity. Contrarily, an increase in temperature or decrease in pH did not influence the resistance or conductivity of biofilms without magnetite. Some of these cytochrome c (CYC)-like proteins were electrically active in the electrochemical Fourier transform infrared spectra and were closely associated with the outer-membrane multi-heme c-type cytochromes (OMCs). The intensities of the OMC-like bands in the surface-enhanced resonance Raman spectra with biofilms in the presence of magnetite were higher than those without magnetite. This study suggests that magnetite promotes the nitrate-dependent AOM process because of the formation of methanotrophic consortia based on direct interspecies electron transfer (DIET).

Spatio-temporal variations of activity of nitrate-driven anaerobic oxidation of methane and community structure of Candidatus Methanoperedens-like archaea in sediment of Wuxijiang river

Nitrate-driven anaerobic oxidation of methane (AOM), catalyzing by Candidatus Methanoperedens-like archaea, is a new addition in the global CH4 cycle. This AOM process acts as a novel pathway for CH4 emission reduction in freshwater aquatic ecosystems; however, its quantitative importance and regulatory factors in riverine ecosystems are nearly unknown. Here, we examined the spatio-temporal changes of the communities of Methanoperedens-like archaea and nitrate-driven AOM activity in sediment of Wuxijiang River, a mountainous river in China. These archaeal community composition varied significantly among reaches (upper, middle, and lower reaches) and between seasons (winter and summer), but their mcrA gene diversity showed no significant spatial or temporal variations. The copy numbers of Methanoperedens-like archaeal mcrA genes were 1.32 × 105–2.47 × 107 copies g−1 (dry weight), and the activity of nitrate-driven AOM was 0.25–1.73 nmol CH4 g−1 (dry weight) d−1, which could potentially reduce 10.3% of CH4 emissions from rivers. Significant spatio-temporal variations of mcrA gene abundance and nitrate-driven AOM activity were found. Both the gene abundance and activity increased significantly from upper to lower reaches in both seasons, and were significantly higher in sediment collected in summer than in winter. In addition, the variations of Methanoperedens-like archaeal communities and nitrate-driven AOM activity were largely impacted by the sediment temperature, NH4+ and organic carbon contents. Taken together, both time and space scales need to be considered for better evaluating the quantitative importance of nitrate-driven AOM in reducing CH4 emissions from riverine ecosystems.

Implication From Mineralogical and Geochemical Characteristics of Authigenic Micronodules in the Haima Cold Seeps for Understanding the Manganese Geochemistry in Active Seepage

AbstractThe multi‐valent manganese is ubiquitous in various marine environments and is involved in many biogeochemical processes with electron transfers between different oxidation states. The involvement of Mn (IV) in the anaerobic oxidation of methane (AOM) in cold seeps has long been proposed by experimental simulation and was suggested to serve as an important sink of methane in the paleo‐oceanic environment. Nonetheless, metal‐dependent AOM processes still lack explicit geological evidence. Meanwhile, research on micronodules gained momentum in recent years, but the authigenic manganese micronodules in active cold seeps were yet not well studied. In this study, Mn‐rich sediments were discovered at the active Haima cold seeps area in the South China Sea, from which Mn‐micronodules were then extracted. These micronodules are irregular or subspheroidal with rough and porous surfaces and are mainly composed of 7 Å and 10 Å phyllomanganates. They are enriched in Mn, and deficient in Fe, Co, Ni, Cu, and REYs. Characteristic microbial‐like mineralization structures are also observed. Based on these mineralogical/geochemical features and the abnormally high growth rate (calculated by the cobalt chronometer algorithm), these authigenic manganese micronodules were probably formed rapidly via biomineralization and chemical precipitation and were supplied with sufficient Mn by the metal‐rich seepage fluids. The occurrence of these micronodules demonstrates manganese bacterial activities, and indirectly reflects metal‐dependent AOM.

The role of anaerobic methane oxidation on the carbonate authigenesis in sediments of the subtropical Beibu Gulf, South China Sea: A reactive–transport modelling approach

The formation and burial of authigenic carbonate in marine sediment significantly affect the sedimentary carbon cycle and its isotopic mass balance in geological history. Anaerobic oxidation of methane (AOM) is the primary driver of authigenic carbonate precipitation within the sulfate-methane transition zone (SMTZ). Quantitative estimations of the role of AOM on the authigenic carbonate precipitation and its carbon isotope under non-steady-state processes (e.g., changes in methane fluxes at the bottom sediment, sedimentation rates or organic fluxes in the surface sediment), however, are still limited. In this study, we use geochemical data from porewater (e.g., the concentration of sulfate, calcium, magnesium, strontium, dissolved inorganic carbon, total alkalinity) and solid sediment (e.g., organic matter content, and carbonate content) in different depositional environments of the subtropical Beibu Gulf, South China Sea, combined with a diagenetic reactive-transport modelling approach, to estimate the mineralogy of authigenic carbonate, the relationship between AOM and authigenic carbonate precipitation, and the impact of AOM rate on carbon isotope of sediment carbonate (δ13CCar). The results show that high-Mg carbonates (high-Mg calcite and dolomite) are the main type of authigenic carbonate (∼80%) formed in the methane-bearing sediments, leading to higher porewater Sr2+/Ca2+ (>0.02) and Mg2+/Ca2+ (>20) within the SMTZ. Our modelling analysis highlights that the non-steady-state induced by increased methane flux from the underlying sediments can significantly accelerate the authigenic carbonates formation within the SMTZ. Using parametric sensitivity analysis, we observed that even a 1% increase in the authigenic carbonate fraction of sediment carbonates results in significant changes in δ13CCar within the SMTZ (from −1‰ to −2‰), mainly due to lighter carbon isotopes produced by more intensive AOM processes. Noteworthily, the terrestrial-to-marine transition was identified by the sediment and porewater geochemical profiles at site SO-8. Although lower authigenic carbonate precipitation occurs in terrestrial sedimentary environments, the proportion of authigenic carbonate in terrestrial environments (11%) is much higher than that in marine environments (1%), resulting in carbon isotopes of carbonate in terrestrial sediments becoming more negative (−5‰).

Soil nitrogen substances and denitrifying communities regulate the anaerobic oxidation of methane in wetlands of Yellow River Delta, China.

Anaerobic oxidation of methane (AOM) in wetland soils is widely recognized as a key sink for the greenhouse gas methane (CH4). The occurrence of this reaction is influenced by several factors, but the exact process and related mechanism of this reaction remain unclear, due to the complex interactions between multiple influencing factors in nature. Therefore, we investigated how environmental and microbial factors affect AOM in wetlands using laboratory incubation methods combined with molecular biology techniques. The results showed that wetland AOM was associated with a variety of environmental factors and microbial factors. The environmental factors include such as vegetation, depth, hydrogen ion concentration (pH), oxidation-reduction potential (ORP), electrical conductivity (EC), total nitrogen (TN), nitrate (NO3-), sulfate (SO42-), and nitrous oxide (N2O) flux, among them, soil N substances (TN, NO3-, N2O) have essential regulatory roles in the AOM process, while NO3- and N2O may be the key electron acceptors driving the AOM process under the coexistence of multiple electron acceptors. Moreover, denitrification communities (narG, nirS, nirK, nosZI, nosZII) and anaerobic methanotrophic (ANME-2d) were identified as important functional microorganisms affecting the AOM process, which is largely regulated by the former. In the environmental context of growing global anthropogenic N inputs to wetlands, these findings imply that N cycle-mediated AOM processes are a more important CH4 sink for controlling global climate change. This studying contributes to the knowledge and prediction of wetland CH4 biogeochemical cycling and provides a microbial ecology viewpoint on the AOM response to global environmental change.

Inorganic sulfur cycles in sediments of the Pearl River Estuary: Processes, mechanisms, and isotopic indicators

The burial of inorganic sulfur is one of the important components in the global sulfur cycle. In this study, the cycling of inorganic sulfide species and the underlying control mechanisms were examined in sediments found in distinct sub-regions of the Pearl River Estuary (PRE). Specifically, the content and isotopic composition of SO42−, acid-volatile sulfides (AVS), and pyrite were determined, along with sedimentary sulfate reduction rates (SRR). The results show that the formation of iron sulfides (such as AVS and pyrite) was mainly controlled by the content of available reactive iron in the sediments of the PRE, and therefore formed mainly in the shallow sediments, with little variation in the deeper parts of the sediments. The δ34S values of AVS and SO42− increased synchronously with depth in the middle and lower part of the sediment, indicating a closed diagenetic environment. In contrast, the δ34S value of pyrite only increased slowly with depth in the case of a small portion of AVS converted to pyrite, which was controlled by the burial time. There are obvious differences in the formation and burial of pyrite in sediments of different sub-regions of the PRE. The surface layer of stations QA and HQ in the upper estuary and brackish coast was an open environment due to sedimentary reworking, and the contents of AVS and pyrite in the sediment was extremely low due to re-oxidation, while the oxidation of 32S-rich H2S also resulted in lower δ34S values of sulfate in the sediment pore water than in the overlying seawater. Station GS in the estuarine mouth had a much higher percentage of AVS converted to pyrite in the sediments than that at stations QA and HQ due to the relatively long burial time. In addition, near the sulfate methane transition zone (SMT), only the estuarine mouth (station GS) produced iron sulfides (AVS) synchronously with the anaerobic oxidation of methane (AOM), and then converted to pyrite gradually; in contrast, the effect of AOM on the formation of iron sulfides in the sediments of the upper estuary and the brackish coast was almost negligible. The results show that the contribution of AOM processes to the final burial of inorganic sulfur is generally small in PRE.

Isotopic Evidence for Anaerobic Oxidation of Methane in the Freshwater Sediments of Reservoirs: The Impact of Selected Environmental Factors

This paper presents the results of research conducted in 2018–2019 on the anaerobic oxidation of methane (AOM) in reservoir sediments. Located in SE Poland, Maziarnia, Nielisz and Rzeszów Reservoirs were selected for the purposes of the research. Rates of AOM were determined via 50-day incubation of sediment from the 0–5, 5–10 and 10–15 cm layers, to which a 13CH4 isotope tracer was added. The sediments had been collected from a single station at each reservoir in places that had earlier reported high levels of emission of CH4 to the atmosphere. Results demonstrate ongoing AOM processes in the kinds of freshwater ecosystem represented by reservoirs, further implying the existence of an important sink for CH4. More specifically, however, AOM rates were found to differ among both the reservoirs, and the layers of sediment, studied. Preliminary analysis of selected environmental factors capable of affecting AOM failed to suggest the availability of electron acceptors (NO3−, SO42−, Fe3+) as key controlling factors. Important factors also proved to be sediment pH, the quality of organic matter (especially the content of organic electron acceptors), the salinity of pore water, and—primarily—the presence of the microorganisms actually responsible for AOM. The results here are important, given the low level of knowledge of AOM process in reservoirs. They therefore help supply key information on the functioning of these ecosystems and the role in global climate change they play.

The denitrifying anaerobic methane oxidation process and microorganisms in the environments: A review

Methane (CH4) is an important greenhouse gas with a global warming potential 28 – 34 times that of CO2 over the 100-year horizon. Denitrifying anaerobic methane oxidation (DAMO) is a recently discovered process that potentially represents an important CH4 sink globally. This process involves two possible pathways: the nitrite-dependent DAMO mediated by NC10 bacteria and the nitrate-dependent DAMO by ANME-2d archaea. Both are widely detected in freshwater and coastal habitats using molecular tools. However, the distributions of these two processes and the functional microorganisms and their interactions with other N cycling pathways are far from clear. In this review, we conducted a scientometric analysis on a co-citation network consisting of 835 references derived from 354 citing articles closely related to the distribution of DAMO in the environment. Through this analysis, we found that current studies focus more on freshwater systems than coastal systems, and ANME-2d archaea are generally under-studied compared to NC10 bacteria. The emerging research topics in this area include AMO processes coupled to alternative electron acceptors and their role as CH4 sinks. We further reviewed papers focusing on DAMO distribution in freshwater and coastal environments guided by the result of the scientometric analysis. Finally, we identified several areas that require further research and proposed future research including comparisons of DAMO with other N cycling pathways and environmental conditions in the context of the river-estuary-sea continuum.

Anoxia-Related Biogeochemistry of North Indian Ocean

Complex interactions between microbial communities and geochemical processes drive the major element cycles and control the function of marine sediments as a dynamic reservoir of organic matter. Sulfate reduction is globally the dominant pathway of anaerobic mineralisation and is the main source of sulfide. The effective re-oxidation of this sulfide at the direct or indirect expense of oxygen is a prerequisite for aerobic life on our planet. Although largely hidden beneath the oxic sediment surface, the sulfur cycle is therefore critical for Earth’s redox state. This Geochemical Perspectives begins with a brief primer on the sulfur cycle of marine sediments and a description of my own scientific journey through nearly fifty years of studies of sulfur geochemistry and microbiology. Among the main objectives of these studies were to quantify the main processes of the sulfur cycle and to identify the microbial communities behind them. Radiotracers in combination with chemical analyses have thereby been used extensively for laboratory experiments, supported by diverse molecular microbiological methods. The following sections discuss the main processes of sulfate reduction, sulfide oxidation and disproportionation of the inorganic sulfur intermediates, especially of elemental sulfur and thiosulfate. The experimental approaches used enable the analysis of how environmental factors such as substrate concentration or temperature affect process rates and how concurrent processes of sulfate reduction and sulfide oxidation drive a cryptic sulfur cycle. The chemical energy of sulfide is used by chemolithotrophic bacteria, including fascinating communities of big sulfur bacteria and cable bacteria, and supports their dark CO2 fixation, which produces new microbial biomass. During the burial and aging of marine sediments, the predominant mineralisation processes change through a cascade of redox reactions, and the rate of organic matter degradation drops continuously over many orders of magnitude. The main pathways of anaerobic mineralisation and the age control of the organic matter turnover are discussed. In the deep methanic zone, only a few percent of the entire degradation process remains, which provides a small boost of substrate for sulfate reduction through the process of anaerobic methane oxidation. The stable isotopes of sulfur provide an additional tool to understand these diagenetic processes, whereby the combination of microbial isotope fractionation and open system diagenesis generate a differential diffusion flux of the isotopes. In relation to the organic carbon cycle of the seabed and the contribution of methane, the paper discusses the global sulfur budget and the role of sulfate reduction for organic matter mineralisation in different depth regions of the ocean - from coast to deep sea. The published estimates of these parameters are evaluated and compared. Finally, the paper looks at future perspectives with respect to gaps in our current understanding and the need for further studies.

Iron-based passivator mitigates the coupling process of anaerobic methane oxidation and arsenate reduction in paddy soils

Arsenic (As) is a toxic metalloid that is ubiquitous in paddy soils, where passivation is the most widely used method for remediating As contamination. Recently, anaerobic methane oxidation coupled with arsenate (As(V)) reduction (AOM-AsR) has been shown to act as a critical driver for As release in paddy fields. However, the effect and mechanism of the passivators on the AOM-AsR process remain unclear. In this study, we incubated arsenate-contaminated paddy soils under anaerobic conditions. Using isotopically labelled methane and different passivators, we found that an iron-based passivator containing calcium sulfate and iron oxide (9:1, m/m) named IBP showed a much better performance than the other passivators. Adding IBP decreased the arsenite (As(III)) concentration in the soil solution by 78% and increased the AOM rate by 55%. Furthermore, we employed high-throughput sequencing and real-time quantitative polymerase chain reaction (qPCR) to investigate the ability of IBP to control As release mediated by AOM-AsR in paddy fields, as well as its underlying mechanism. Our results showed that IBP addition significantly increased anaerobic methanotrophic (ANME) archaea (ANME-2a-c, ANME-2d, and ANME-3) by 91%, and increased the methane-oxidizing bacterium Methylobacter by 262%. Similarly, IBP addition significantly increased the Fe(III) concentration in soil solution by 39% and increased the absolute abundance of Fe(III)-reducing bacteria (Geobacteraceae) by 21 times in soil. Adding IBP may significantly promote AOM coupled with Fe(III) reduction, significantly reducing electron transfer from AOM to As(V) reduction. Hence, IBP may be used as an efficient passivator to remediate As-contaminated soil using an active AOM-AsR process. These results provide a novel insight into controlling soil As release by regulating an active and critical As mobilization pathway in the environment.

Effect of gradual increase of atmospheric CO2 concentration on nitrate-dependent anaerobic methane oxidation in paddy soils

Nitrate-dependent anaerobic oxidation of methane (AOM) is a new pathway to reduce methane emissions from paddy ecosystems. The elevated atmospheric CO2 concentration can affect methane emissions from paddy ecosystems, but its impact on the process of nitrate-dependent AOM is poorly known. Based on the automatic CO2 control platform with open top chambers and the 13CH4 stable isotope experiments, the responses of the activity of nitrate-dependent AOM, abundance and community composition of Candidatus Methanoperedens nitroreducens (M. nitroreducens)-like archaea to the gradual increase of CO2 concentration were investigated in paddy fields. We set up two CO2 concentration treatments, including an ambient CO2 and a gradual increase of CO2(increase of 40 μL·L-1 per year above ambient CO2 concentration until 160 μL·L-1). The results showed the nitrate-dependent AOM rate of 0.7-11.3 nmol CO2·g-1·d-1 in the studied paddy fields, and quantitative PCR showed the abundance of M. nitroreducens-like archaeal mcrA genes of 2.2×106-8.5×106 copies·g-1. Compared to the ambient CO2 treatment, the slow elevated CO2 treatment enhanced the nitrate-dependent AOM rate and stimulated the abundance of M. nitroreducens-like archaea, particularly in 5-10 cm soil layer. The gradual increased CO2 concentration treatment did not change the community composition of M. nitroreducens-like archaea, but significantly decreased their diversity. The soil organic carbon content was an important factor influencing the nitrate-dependent AOM process. Overall, our results showed that the gradual increase of CO2 concentration could promote the nitrate-dependent AOM, suggesting its positive role in mitigating methane emissions from paddy ecosystems under future climate change.

First evidence for anaerobic oxidation of methane process in landfill cover soils: Activity and responsible microorganisms

Landfill cover soils (LCS) play important roles in mitigating methane emissions from landfills. Anaerobic oxidation of methane (AOM) has been demonstrated as a potential methane removal process in aquatic ecosystems. However, whether AOM could occur in LCS is largely unknown. Here, microcosm incubations with 13CH4 were applied to track the potential activities of AOM and quantitative PCR was used to identify the responsible microorganisms. AOM was found to be active in the bottom and middle layers of LCS. In the bottom layer, sulfate-AOM was the most active process, mainly dominated by ANME archaea (without ANME-2d). Meanwhile, in the middle layer, nitrate and nitrite were the major electron acceptors involved in AOM with high abundances of ANME-2d archaea and NC10 bacteria. Our results implied a spatial segregation of methane oxidizing microbes in LCS and might be helpful for future control of methane emissions by the enhancement of AOM.

Long-term incubations provide insight into the mechanisms of anaerobic oxidation of methane in methanogenic lake sediments

Abstract. Anaerobic oxidation of methane (AOM) is among the main processes limiting the release of the greenhouse gas methane from natural environments. Geochemical profiles and experiments with fresh sediments from Lake Kinneret (Israel) indicate that iron-coupled AOM (Fe-AOM) sequesters 10 %–15 % of the methane produced in the methanogenic zone (&gt;20 cm sediment depth). The oxidation of methane in this environment was shown to be mediated by a combination of mcr-gene-bearing archaea and pmoA-gene-bearing aerobic bacterial methanotrophs. Here, we used sediment slurry incubations under controlled conditions to elucidate the electron acceptors and microorganisms that are involved in the AOM process over the long term (∼ 18 months). We monitored the process with the addition of 13C-labeled methane and two stages of incubations: (i) enrichment of the microbial population involved in AOM and (ii) slurry dilution and manipulations, including the addition of several electron acceptors (metal oxides, nitrate, nitrite and humic substances) and inhibitors (2-bromoethanesulfonate, acetylene and sodium molybdate) of methanogenesis, methanotrophy and sulfate reduction and sulfur disproportionation. Carbon isotope measurements in the dissolved inorganic carbon pool suggest the persistence of AOM, consuming 3 %–8 % of the methane produced at a rate of 2.0 ± 0.4 nmol per gram of dry sediment per day. Lipid carbon isotopes and metagenomic analyses point towards methanogens as the sole microbes performing the AOM process by reverse methanogenesis. Humic substances and iron oxides, although not sulfate, manganese, nitrate or nitrite, are the likely electron acceptors used for this AOM. Our observations support the contrast between methane oxidation mechanisms in naturally anoxic lake sediments, with potentially co-existing aerobes and anaerobes, and long-term incubations, wherein anaerobes prevail.

Metabolic potential of anaerobic methane oxidizing archaea for a broad spectrum of electron acceptors.

Methane (CH4) is a potent greenhouse gas significantly contributing to the climate warming we are currently facing. Microorganisms play an important role in the global CH4 cycle that is controlled by the balance between anaerobic production via methanogenesis and CH4 removal via methanotrophic oxidation. Research in recent decades advanced our understanding of CH4 oxidation, which until 1976 was believed to be a strictly aerobic process. Anaerobic oxidation of methane (AOM) coupled to sulfate reduction is now known to be an important sink of CH4 in marine ecosystems. Furthermore, in 2006 it was discovered that anaerobic CH4 oxidation can also be coupled to nitrate reduction (N-DAMO), demonstrating that AOM may be much more versatile than previously thought and linked to other electron acceptors. In consequence, an increasing number of studies in recent years showed or suggested that alternative electron acceptors can be used in the AOM process including FeIII, MnIV, AsV, CrVI, SeVI, SbV, VV, and BrV. In addition, humic substances as well as biochar and perchlorate (ClO4-) were suggested to mediate AOM. Anaerobic methanotrophic archaea, the so-called ANME archaea, are key players in the AOM process, yet we are still lacking deeper understanding of their metabolism, electron acceptor preferences and their interaction with other microbial community members. It is still not clear whether ANME archaea can oxidize CH4 and reduce metallic electron acceptors independently or via electron transfer to syntrophic partners, interspecies electron transfer, nanowires or conductive pili. Therefore, the aim of this review is to summarize and discuss the current state of knowledge about ANME archaea, focusing on their physiology, metabolic flexibility and potential to use various electron acceptors.

Anaerobic oxidation of methane coupled with sulphate reduction: high concentration of methanotrophic archaea might be responsible for low stable isotope fractionation factors in methane

ABSTRACT The changes in δ 13CH4 and δ 12C1H3 2H during sulphate-dependent anaerobic oxidation of methane (AOM) were described using dynamic modelling. The batch sulphate-dependent AOM at the nearly linear dynamics of methane oxidation with different enriched cultures originating from three marine sediments was simulated. The traditional Rayleigh equation for carbon and hydrogen stable isotopes in methane was derived from the basic dynamic isotope equation. The general and reduced models, taking into account the reaction stoichiometry and based on balances of chemical elements and their isotopes, describes a redistribution of stable isotope values in the sulphate-dependent AOM process. It was shown that AOM is the first and rate-limiting step in the whole AOM + SR (sulphate reduction) process. The different fractionation factors of carbon and hydrogen isotopes in methane were obtained for three marine sediments. It was concluded that during incubation the highest concentration of methanotrophic archaea might be responsible for the lowest fractionation factors of stable isotopes of carbon and hydrogen in methane. The interpretation of this phenomenon was suggested. Different concentrations of methanotrophic archaea can lead to variations of isotope fractionation factors.

Sulfur Biogeochemical Cycle of Marine Sediments

Complex interactions between microbial communities and geochemical processes drive the major element cycles and control the function of marine sediments as a dynamic reservoir of organic matter. Sulfate reduction is globally the dominant pathway of anaerobic mineralisation and is the main source of sulfide. The effective re-oxidation of this sulfide at the direct or indirect expense of oxygen is a prerequisite for aerobic life on our planet. Although largely hidden beneath the oxic sediment surface, the sulfur cycle is therefore critical for Earth’s redox state. This Geochemical Perspectives begins with a brief primer on the sulfur cycle of marine sediments and a description of my own scientific journey through nearly fifty years of studies of sulfur geochemistry and microbiology. Among the main objectives of these studies were to quantify the main processes of the sulfur cycle and to identify the microbial communities behind them. Radiotracers in combination with chemical analyses have thereby been used extensively for laboratory experiments, supported by diverse molecular microbiological methods. The following sections discuss the main processes of sulfate reduction, sulfide oxidation and disproportionation of the inorganic sulfur intermediates, especially of elemental sulfur and thiosulfate. The experimental approaches used enable the analysis of how environmental factors such as substrate concentration or temperature affect process rates and how concurrent processes of sulfate reduction and sulfide oxidation drive a cryptic sulfur cycle. The chemical energy of sulfide is used by chemolithotrophic bacteria, including fascinating communities of big sulfur bacteria and cable bacteria, and supports their dark CO2 fixation, which produces new microbial biomass. During the burial and aging of marine sediments, the predominant mineralisation processes change through a cascade of redox reactions, and the rate of organic matter degradation drops continuously over many orders of magnitude. The main pathways of anaerobic mineralisation and the age control of the organic matter turnover are discussed. In the deep methanic zone, only a few percent of the entire degradation process remains, which provides a small boost of substrate for sulfate reduction through the process of anaerobic methane oxidation. The stable isotopes of sulfur provide an additional tool to understand these diagenetic processes, whereby the combination of microbial isotope fractionation and open system diagenesis generate a differential diffusion flux of the isotopes. In relation to the organic carbon cycle of the seabed and the contribution of methane, the paper discusses the global sulfur budget and the role of sulfate reduction for organic matter mineralisation in different depth regions of the ocean – from coast to deep sea. The published estimates of these parameters are evaluated and compared. Finally, the paper looks at future perspectives with respect to gaps in our current understanding and the need for further studies.