Sulfate-dependent Anaerobic Oxidation Of Methane Research Articles

The diversification and potential function of microbiome in sediment-water interface of methane seeps in South China Sea.

The sediment-water interfaces of cold seeps play important roles in nutrient transportation between seafloor and deep-water column. Microorganisms are the key actors of biogeochemical processes in this interface. However, the knowledge of the microbiome in this interface are limited. Here we studied the microbial diversity and potential metabolic functions by 16S rRNA gene amplicon sequencing at sediment-water interface of two active cold seeps in the northern slope of South China Sea, Lingshui and Site F cold seeps. The microbial diversity and potential functions in the two cold seeps are obviously different. The microbial diversity of Lingshui interface areas, is found to be relatively low. Microbes associated with methane consumption are enriched, possibly due to the large and continuous eruptions of methane fluids. Methane consumption is mainly mediated by aerobic oxidation and denitrifying anaerobic methane oxidation (DAMO). The microbial diversity in Site F is higher than Lingshui. Fluids from seepage of Site F are mitigated by methanotrophic bacteria at the cyclical oxic-hypoxic fluctuating interface where intense redox cycling of carbon, sulfur, and nitrogen compounds occurs. The primary modes of microbial methane consumption are aerobic methane oxidation, along with DAMO, sulfate-dependent anaerobic methane oxidation (SAMO). To sum up, anaerobic oxidation of methane (AOM) may be underestimated in cold seep interface microenvironments. Our findings highlight the significance of AOM and interdependence between microorganisms and their environments in the interface microenvironments, providing insights into the biogeochemical processes that govern these unique ecological systems.

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.

Interlinked marine cycles of methane, manganese, and sulfate in the post-Marinoan Doushantuo cap dolostone

The strongly 13C-depleted calcite in the Doushantuo cap dolostones (∼635 Ma) of South China is considered to form via thermochemical or biological oxidation of methane by sulfate in an environment with sulfate concentrations close to that of modern seawater (∼28 mM). Here we demonstrate that the coeval seawater sulfate levels were indeed low (∼1 to 3 mM) because all bulk δ34S values of pyrites are close to and higher than the coeval seawater, with only a few showing a wide in situ δ34S range from 5 to 62‰. Such low sulfate concentrations were initially consumed by sulfate reducing microorganisms to extremely low levels (<∼0.2 mM), and thus Mn-driven anaerobic oxidation of methane (AOM) by sulfate occurred in pore water to produce H2S, which was in turn oxidized by Mn oxides back to sulfate. Consequently, with increasing Mn reduction and decreasing sulfate concentrations in pore waters, the produced pyrite shows an increase of in situ δ34S values, and is in association with calcite with more negative δ13C values, and an inherited positive Ce anomaly from Mn oxides. Finally, the pyrite with the highest δ34S value may have co-precipitated with the 13C-depelted calcite (with a nadir of −58‰), and thus show similarly high Mn concentrations (∼20000 μg/g). This is the first report of sulfate-dependent AOM driven by Mn- or Fe- oxides from the entire Precambrian record. Such a previously unrecognized biogeochemical cycling during Marinoan global deglaciation may serve as the mechanism for major methane sink during the Proterozoic and Archean with low sulfate concentrations in the oceans.

Application of concentration and 2-dimensional stable isotope measurements of methane to constrain sources and sinks in a seasonally stratified freshwater lake

Methane (CH4) emissions from aquatic systems have recently been comprised to account for up to 50% of global CH4 emissions, with lakes representing one of the largest CH4 sources within this pool. However, there is large uncertainty associated with CH4 emissions from freshwater environments to the atmosphere, because of a lack of understanding in the spatial and temporal dynamics of CH4 sources and sinks, as well as underlying mechanisms and processes. In this study, we investigated the concentrations and stable carbon (δ13C-CH4) and hydrogen (δ2H-CH4) isotope composition of CH4 in a small eutrophic lake (Lake Willersinnweiher) with seasonal stratification and its spatial and temporal variation. We found that while supersaturation of CH4 in the entire water column was present throughout the whole year, the isotopic composition of CH4 in sediment and water column varied depending on lake stratification, physiochemical conditions, and lake depth. During the stratification period, isotopic characteristics of pelagic surface water CH4 differed from littoral and sedimentary CH4, suggesting likely mixing of CH4 from different sources including vertical and lateral input as well as groundwater input and potentially oxic methane production in the mixed surface water layer. Aerobic CH4 oxidation indicated by a strong increase in both δ13C-CH4 and δ2H-CH4 values at the bottom of the oxycline was found to significantly reduce upward migrating CH4 released at the sediment-water interface. In the sediment, stable isotope characteristics of CH4 showed an increasing dominance of the acetoclastic CH4 formation pathway from the pelagic towards the littoral area. Furthermore, the occurrence of sulfate-dependent anaerobic methane oxidation in the sediment was suggested by an increase in δ13C-CH4 and δ2H-CH4 values. During the mixing period, the isotopic CH4 composition of the water column was distinctively less negative than during the stratification period potentially resulting from a greater impact of groundwater CH4 input compared to the stratification period. Our findings implicate that the application of concentrations and dual isotope measurements of CH4 is a promising approach for constraining CH4 sinks and sources in Lake Willersinnweiher and potentially other small lakes to clearly disentangle the complex CH4 dynamics in lakes both spatially and seasonally.

Phylogenetically and catabolically diverse diazotrophs reside in deep-sea cold seep sediments

Microbially mediated nitrogen cycling in carbon-dominated cold seep environments remains poorly understood. So far anaerobic methanotrophic archaea (ANME-2) and their sulfate-reducing bacterial partners (SEEP-SRB1 clade) have been identified as diazotrophs in deep sea cold seep sediments. However, it is unclear whether other microbial groups can perform nitrogen fixation in such ecosystems. To fill this gap, we analyzed 61 metagenomes, 1428 metagenome-assembled genomes, and six metatranscriptomes derived from 11 globally distributed cold seeps. These sediments contain phylogenetically diverse nitrogenase genes corresponding to an expanded diversity of diazotrophic lineages. Diverse catabolic pathways were predicted to provide ATP for nitrogen fixation, suggesting diazotrophy in cold seeps is not necessarily associated with sulfate-dependent anaerobic oxidation of methane. Nitrogen fixation genes among various diazotrophic groups in cold seeps were inferred to be genetically mobile and subject to purifying selection. Our findings extend the capacity for diazotrophy to five candidate phyla (Altarchaeia, Omnitrophota, FCPU426, Caldatribacteriota and UBA6262), and suggest that cold seep diazotrophs might contribute substantially to the global nitrogen balance.

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.

Finding the pieces for the anaerobic methane oxidation jigsaw puzzle in mangrove wetlands

Estuarine mangrove wetlands are generally considered a major source of methane (CH4) emissions. Anaerobic methane oxidation (AMO) is an important pathway for reducing CH4 emissions and is known to be tightly linked to carbon, nitrogen, sulfur and metal cycling processes largely driven by microorganisms. However, these coupled processes and their contributions to mitigating global warming remain unclear in mangrove wetlands. In this study, the depth-specific potential methane oxidation rates of nitrate-, nitrite- and nitrous oxide-dependent anaerobic methane oxidation (nitrate-, nitrite- and N2O-AMO) and sulfate-dependent anaerobic methane oxidation (sulfate-AMO) processes were investigated using stable isotope tracing along with an assessment of the vertical nutrient and CH4 content profiles in mangrove sediments. The results indicated that nitrite-AMO, nitrate-AMO, N2O-AMO and sulfate-AMO were active and exhibited methane oxidation rates varying from 24.1 to 1077.4, 14.1 to 66.1, 14.3 to 40.9 and 4.2–36.7 nmol CO2 g−1 dry soil d−1, respectively. Furthermore, nitrite-AMO and N2O-AMO tended to be more active in the upper sediment layers, while nitrate-AMO and sulfate-AMO displayed higher rates in the deeper layers. The contribution rate of nitrite-AMO to the total of the 4 AMO processes examined in the mangrove sediments reached 28.90–95.80%, followed by nitrate-AMO (2.73–46.46%), sulfate-AMO (1.46–24.65%) and N2O-AMO (1.70–4.47%), suggesting that the nitrite-AMO process is a significant methane sink in mangrove wetland ecosystems. The temporal and spatial variations in the nitrite-AMO bacterial depth and genus-specific distribution, abundance, activity and sensitivity to environmental factors were then investigated. The absolute abundances of the nitrite-AMO bacterial 16S rRNA and pmoA genes were 0.51–6.55 × 107 and 0.79–8.43 × 106 copies g−1 ds, respectively, and these values decreased with sediment depth; most of the samples were characterized as belonging to Group B of NC10. In addition, NO2− is the limiting factor for nitrite-AMO bacteria in mangrove wetlands.

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.

Putative fossils of chemotrophic microbes preserved in seep carbonates from Vestnesa Ridge, off northwest Svalbard, Norway

ABSTRACT The microbial key players at methane seeps are methanotrophic archaea and sulfate-reducing bacteria. They form spherical aggregates and jointly mediate the sulfate-dependent anaerobic oxidation of methane (SD–AOM: CH4 + SO42− → HCO3− + HS− + H2O), thereby inducing the precipitation of authigenic seep carbonates. While seep carbonates constitute valuable archives for molecular fossils of SD–AOM-mediating microbes, no microfossils have been identified as AOM aggregates to date. We report clustered spherical microstructures engulfed in 13C-depleted aragonite cement (δ13C values as low as −33‰) of Pleistocene seep carbonates. The clusters comprise Mg-calcite spheres between ∼5 µm (single spheres) and ∼30 µm (clusters) in diameter. Scanning and transmission electron microscopy revealed a porous nanocrystalline fabric in the core area of the spheres surrounded by one or two concentric layers of Mg-calcite crystals. In situ measured sphere δ13C values as low as −42‰ indicate that methane-derived carbon is the dominant carbon source. The size and concentric layering of the spheres resembles mineralized aggregates of natural anaerobic methanotrophic archaea (ANME) of the ANME-2 group surrounded by one or two layers of sulfate-reducing bacteria. Abundant carbonate-bound 13C-depleted lipid biomarkers of archaea and bacteria indicative of the ANME-2-Desulfosarcina/Desulfococcus consortium agree with SD–AOM-mediating microbes as critical agents of carbonate precipitation. Given the morphological resemblance, in concert with negative in situ δ13C values and abundant SD–AOM-diagnostic biomarkers, the clustered spheres likely represent fossils of SD–AOM-mediating microbes.

Anaerobic oxidation of methane by Mn oxides in sulfate-poor environments

Abstract Strongly 13C-depleted authigenic carbonates (e.g., δ13CVPDB &amp;lt;−30‰; VPDB—Vienna Peedee belemnite) in nature are generally believed to form by sulfate-dependent anaerobic oxidation of methane (AOM). However, we demonstrate using geochemical data and thermodynamic calculation that such calcites are most likely derived from biogenic oxidation of methane in sulfate-poor, nonmarine environments during early diagenesis, as observed in the Triassic sandy conglomerates from the Junggar Basin, northwestern China. This process operated through preferential oxidation of 13C-depleted methane by Mn oxides in closed conditions, producing calcites with higher Mn contents and δ13C values in association with more 13C-enriched residual methane as a result of kinetic isotope fractionation. Thus, the Mn-rich and 13C-depleted carbonates are proposed as tracers of Mn-dependent AOM, which should have served as an important sink of greenhouse methane in low-sulfate early Earth's oceans.

Anthropogenic and Environmental Constraints on the Microbial Methane Cycle in Coastal Sediments.

Large amounts of methane, a potent greenhouse gas, are produced in anoxic sediments by methanogenic archaea. Nonetheless, over 90% of the produced methane is oxidized via sulfate-dependent anaerobic oxidation of methane (S-AOM) in the sulfate-methane transition zone (SMTZ) by consortia of anaerobic methane-oxidizing archaea (ANME) and sulfate-reducing bacteria (SRB). Coastal systems account for the majority of total marine methane emissions and typically have lower sulfate concentrations, hence S-AOM is less significant. However, alternative electron acceptors such as metal oxides or nitrate could be used for AOM instead of sulfate. The availability of electron acceptors is determined by the redox zonation in the sediment, which may vary due to changes in oxygen availability and the type and rate of organic matter inputs. Additionally, eutrophication and climate change can affect the microbiome, biogeochemical zonation, and methane cycling in coastal sediments. This review summarizes the current knowledge on the processes and microorganisms involved in methane cycling in coastal sediments and the factors influencing methane emissions from these systems. In eutrophic coastal areas, organic matter inputs are a key driver of bottom water hypoxia. Global warming can reduce the solubility of oxygen in surface waters, enhancing water column stratification, increasing primary production, and favoring methanogenesis. ANME are notoriously slow growers and may not be able to effectively oxidize methane upon rapid sedimentation and shoaling of the SMTZ. In such settings, ANME-2d (Methanoperedenaceae) and ANME-2a may couple iron- and/or manganese reduction to AOM, while ANME-2d and NC10 bacteria (Methylomirabilota) could couple AOM to nitrate or nitrite reduction. Ultimately, methane may be oxidized by aerobic methanotrophs in the upper millimeters of the sediment or in the water column. The role of these processes in mitigating methane emissions from eutrophic coastal sediments, including the exact pathways and microorganisms involved, are still underexplored, and factors controlling these processes are unclear. Further studies are needed in order to understand the factors driving methane-cycling pathways and to identify the responsible microorganisms. Integration of the knowledge on microbial pathways and geochemical processes is expected to lead to more accurate predictions of methane emissions from coastal zones in the future.

Simultaneous nitrate and sulfate dependent anaerobic oxidation of methane linking carbon, nitrogen and sulfur cycles

ANaerobic MEthanotrophic (ANME) archaea are critical microorganisms mitigating methane emission from anoxic zones. In previous studies, sulfate-dependent anaerobic oxidation of methane (AOM) and nitrate-dependent AOM, performed by different clades of ANME archaea, were detected in marine sediments and freshwater environments, respectively. This study shows that simultaneous sulfate- and nitrate-dependent AOM can be mediated by a clade of ANME archaea, which may occur in estuaries and coastal zones, at the interface of marine and freshwater environments enriched with sulfate and nitrate. Long-term (~1,200 days) performance data of a bioreactor, metagenomic analysis and batch experiments demonstrated that ANME-2d not only conducted AOM coupled to reduction of nitrate to nitrite, but also coupled to the conversion of sulfate to sulfide, in collaboration with sulfate-reducing bacteria (SRB). Sulfide was oxidized back to sulfate by sulfide-oxidizing autotrophic denitrifiers with nitrate or nitrite as electron acceptors, in turn alleviating sulfide accumulation. In addition, dissimilatory nitrate reduction to ammonium performed by ANME-2d was detected, providing substrates to Anammox. Metatranscriptomic analysis revealed significant upregulation of flaB in ANME-2d and pilA in Desulfococcus, which likely resulted in the formation of unique nanonets connecting cells and expanding within the biofilm, and putatively providing structural links between ANME-2d and SRB for electron transfer. Simultaneous nitrate- and sulfate-dependent AOM as observed in this study could be an important link between the carbon, nitrogen and sulfur cycles in natural environments, such as nearshore environments.

Biomarker and Isotopic Composition of Seep Carbonates Record Environmental Conditions in Two Arctic Methane Seeps

Present-day activity of cold seeps in the ocean is evident from direct observations of methane emanating from the seafloor, the presence of chemosynthetic organisms, or the quantification of high gas concentrations in sediment pore waters and the water column. Verifying past cold seep activity and biogeochemical characteristics is more challenging but may be reconstructed from proxy records of authigenic seep carbonates. Here, we investigated the lipid-biomarker inventory, carbonate mineralogy, and stable carbon and oxygen isotope compositions of seep-associated carbonates from two active Arctic methane seeps, located to the northwest (Vestnesa Ridge; ∼1,200 m water depth) and south (Storfjordrenna; ∼380 m water depth) offshore Svalbard. The aragonite-dominated mineralogy of all but one carbonate sample indicate precipitation close to the seafloor in an environment characterized by high rates of sulfate-dependent anaerobic oxidation of methane (AOM). In contrast, Mg-calcite rich nodules sampled in sediments of Storfjordrenna appear to have formed at the sulfate-methane-transition zone deeper within the sediment at lower rates of AOM. AOM activity at the time of carbonate precipitation is indicated by the 13C-depleted isotope signature of the carbonates [−20 to −30‰ Vienna Pee Dee Belemnite (VPDB)], as well as high concentrations of 13C-depleted lipid biomarkers diagnostic for anaerobic methanotrophic archaea (archaeol and sn2-hydroxyarchaeol) and sulfate-reducing bacteria (iso and anteiso-C15:0 fatty acids) in the carbonates. We also found 13C-depleted lipid biomarkers (diploptene and a 4α-methyl sterol) that are diagnostic for bacteria mediating aerobic oxidation of methane (MOx). This suggests that the spatial separation between AOM and MOx zones was relatively narrow at the time of carbonate formation, as is typical for high methane-flux regimes. The seep-associated carbonates also displayed relatively high δ18O values (4.5–5‰ VPDB), indicating the presence of 18O-enriched fluids during precipitation, possibly derived from destabilized methane gas hydrates. Based on the combined isotopic evidence, we suggest that all the seep carbonates resulted from the anaerobic oxidation of methane during intense methane seepage. The seepage likely was associated to gas hydrates destabilization, which led to the methane ebullition from the seafloor into the water column.

Active Anaerobic Archaeal Methanotrophs in Recently Emerged Cold Seeps of Northern South China Sea.

Cold seep ecosystems are developed from methane-rich fluids in organic rich continental slopes, which are the source of various dense microbial and faunal populations. Extensive studies have been conducted on microbial populations in this unique environment; most of them were based on DNA, which could not resolve the activity of extant organisms. In this study, RNA and DNA analyses were performed to evaluate the active archaeal and bacterial communities and their network correlations, particularly those participating in the methane cycle at three sites of newly developed cold seeps in the northern South China Sea (nSCS). The results showed that both archaeal and bacterial communities were significantly different at the RNA and DNA levels, revealing a higher abundance of methane-metabolizing archaea and sulfate-reducing bacteria in RNA sequencing libraries. Site ROV07-01, which exhibited extensive accumulation of deceased Calyptogena clam shells, was highly developed, and showed diverse and active anaerobic archaeal methanotrophs (ANME)-2a/b and sulfate-reducing bacteria from RNA libraries. Site ROV07-02, located near carbonate crusts with few clam shell debris, appeared to be poorly developed, less anaerobic and less active. Site ROV05-02, colonized by living Calyptogena clams, could likely be intermediary between ROV07-01 and ROV07-02, showing abundant ANME-2dI and sulfate-reducing bacteria in RNA libraries. The high-proportions of ANME-2dI, with respect to ANME-2dII in the site ROV07-01 was the first report from nSCS, which could be associated with recently developed cold seeps. Both ANME-2dI and ANME-2a/b showed close networked relationships with sulfate-reducing bacteria; however, they were not associated with the same microbial operational taxonomic units (OTUs). Based on the geochemical gradients and the megafaunal settlements as well as the niche specificities and syntrophic relationships, ANMEs appeared to change in community structure with the evolution of cold seeps, which may be associated with the heterogeneity of their geochemical processes. This study enriched our understanding of more active sulfate-dependent anaerobic oxidation of methane (AOM) in poorly developed and active cold seep sediments by contrasting DNA- and RNA-derived community structure and activity indicators.

Mineralogical and geochemical records of seafloor cold seepage history in the northern Okinawa Trough, East China Sea

Cold seep carbonate represents a faithful record of the ancient methane seepage and provides a nonnegligible contribution to the global carbon reservoir. On the western slope of the Northern Okinawa Trough (NOT), a recent seafloor visualized survey has discovered widespread crust of cold seep carbonate. Here we study mineralogy and geochemistry of these authigenic carbonate to investigate source origin and reconstruct its growth history. Mineralogically, the carbonate crusts are mainly composed of micritic aragonite, with botryoidal aragonite, framboidal pyrite, and microcrystalline authigenic gypsum. Petrographic characteristic unambiguously indicates that this carbonate precipitates in relatively open systems due to a considerable rate of sulfate-dependent anaerobic oxidation of methane (AOM). Regarding geochemistry, strongly 13C-depleted carbon isotope values (as low as −56.1‰, V-PDB) demonstrate that the carbon in the carbonate crusts is mainly derived from biogenic methane coupled with AOM. In contrast, the δ18O enrichment (up to +2.7‰, V-PDB) suggests that the fluid flow from which carbonate precipitated is sourced from dissociation of underlying natural gas hydrates. The U–Th ages of authigenic carbonates fall in the timescale of 22.8–55.7 ka BP, consistent with the period of sea-level lowstand in the late Pleistocene. Overall, several lines of evidence of this study indicate that extensive methane was released by gas hydrate decomposition during sea level fall, consequently resulting in the precipitations of carbonate crust in the NOT. Furthermore, the obviously episodic methane seepages led to the constant accretion from the interior to the exterior within the preformed crust, ultimately inducing the carbonate blocks, slabs and crusts to be exposed on the seafloor. The existence of large-scale carbonate crusts represents a good trapper of the later released carbon especially the isotopically light methane from the deep.

Formation of tubular carbonate conduits at Athina mud volcano, eastern Mediterranean Sea

Abstract Tubular carbonate conduits (TCC) represent the termination of fluid plumbing systems in environments of hydrocarbon seepage and play a relevant role in the discharge of methane from sub-seafloor sediments to the water column. However, the biogeochemical reactions and biological activities involved in their formation are not fully understood. To address this, TCC samples were collected with a remotely operated vehicle from the seabed on the SW flank of the Athina mud volcano in the eastern Mediterranean Sea. Petrographic, mineralogical, stable carbon and oxygen isotope and lipid biomarker analyses were performed to elucidate the formation processes of the tubular carbonates. Clotted and fibrous aragonite form the internal lining of the cavities, while the outer portion of the tubes is formed by micritic Mg-calcite cementing hemipelagic sediment. 13C-depleted Mg-calcite and aragonite (as low as −14.4‰ V-PDB) and lipid biomarkers (archaeol, −89.8‰ V-PDB) indicate that carbonate precipitation was influenced by sulfate-dependent anaerobic oxidation of methane (AOM). AOM locally enhances aragonite precipitation, thereby facilitating early lithification of the conduits within the mud volcano sediments. The size and morphology of the TCC comparable with the buried portion of tubeworm colonies found in the proximity of the sampling site. However, our results suggest that TCC likely formed by the action of burrowing organism rather than being mineralizations of the tubeworm colonies. This study provides new insights into the interpretation and understanding of TCC, highlighting the role of macrofaunal activity in the formation of migration pathways for hydrocarbon-rich fluids on the flank of a mud volcano.

Spatial-Temporal Pattern of Sulfate-Dependent Anaerobic Methane Oxidation in an Intertidal Zone of the East China Sea.

Methane is a primary greenhouse gas which is responsible for global warming. The sulfate-dependent anaerobic methane oxidation (S-AOM) process catalyzed by anaerobic methanotrophic (ANME) archaea and sulfate-reducing bacteria (SRB) is a vital link connecting the global carbon and sulfur cycles, and it is considered to be the overriding methane sink in marine ecosystem. However, there have been few studies regarding the role of S-AOM process and the distribution of ANME archaea in intertidal ecosystem. The intertidal zone is a buffer zone between sea and land and plays an important role in global geochemical cycle. In the present study, the abundance, potential methane oxidation rate, and community structure of ANME archaea in the intertidal zone were studied by quantitative PCR, stable isotope tracing method and high-throughput sequencing. The results showed that the potential S-AOM activity ranged from 0 to 0.77 nmol 13CO2 g-1 (dry sediment) day-1 The copy number of 16S rRNA gene of ANME archaea reached 106 ∼ 107 copies g-1 (dry sediment). The average contribution of S-AOM to total anaerobic methane oxidation was up to 34.5%, while denitrifying anaerobic methane oxidation accounted for the rest, which implied that S-AOM process was an essential methane sink that cannot be overlooked in intertidal ecosystem. The simulated column experiments also indicated that ANME archaea were sensitive to oxygen and preferred anaerobic environmental conditions. This study will help us gain a better understanding of the global carbon-sulfur cycle and greenhouse gas emission reduction and introduce a new perspective into the enrichment of ANME archaea.IMPORTANCE The sulfate-dependent anaerobic methane oxidation (S-AOM) process catalyzed by anaerobic methanotrophic (ANME) archaea and sulfate-reducing bacteria (SRB) is a vital link connecting the global carbon and sulfur cycles. We conducted a research into the spatial-temporal pattern of S-AOM process and the distribution of ANME archaea in coastal sediments collected from the intertidal zone. The results implied that S-AOM process was a methane sink that cannot be overlooked in the intertidal ecosystem. We also found that ANME archaea were sensitive to oxygen and preferred anaerobic environmental conditions. This study will help us gain a better understanding of the global carbon-sulfur cycle and greenhouse gas emission reduction and introduce a new perspective into the enrichment of ANME archaea.

Biogeochemical evidence of anaerobic methane oxidation on active submarine mud volcanoes on the continental slope of the Canadian Beaufort Sea

Abstract. In this study, we report lipid biomarker patterns and phylogenetic identities of key microbial communities mediating anaerobic oxidation of methane (AOM) in active mud volcanoes (MVs) on the continental slope of the Canadian Beaufort Sea. The carbon isotopic compositions (δ13C) of sn-2- and sn-3-hydroxyarchaeol showed the highly 13C-depleted values (−114 ‰ to −82 ‰) associated with a steep depletion in sulfate concentrations within 0.7 m of sediment depths. This suggested the presence of methanotrophic archaea involved in sulfate-dependent AOM, albeit in a small amount. The ratio of sn-2-hydroxyarchaeol to archaeol (&gt; 1) and operational taxonomic units (OTUs) indicated that the anaerobic methanotrophic archaea (ANME) clades ANME-2c and ANME-3 were involved in AOM. Higher δ13C values of archaeol and biphytanes (BPs; -55.2±10.0 ‰ and -39.3±13.0 ‰, respectively) suggested that archaeal communities were also assimilating AOM-derived inorganic carbon. Furthermore, the distinct distribution patterns of methanotrophs in the three MVs appears to be associated with varying intensities of ascending gas fluids. Consequently, our results suggest that the niche diversification of active mud volcanoes has shaped distinct archaeal communities that play important roles in AOM in the Beaufort Sea.

Anaerobic methane oxidation inducing carbonate precipitation at abiogenic methane seeps in the Tuscan archipelago (Italy).

Seepage of methane (CH4) on land and in the sea may significantly affect Earth’s biogeochemical cycles. However processes of CH4 generation and consumption, both abiotic and microbial, are not always clear. We provide new geochemical and isotope data to evaluate if a recently discovered CH4 seepage from the shallow seafloor close to the Island of Elba (Tuscany) and two small islands nearby are derived from abiogenic or biogenic sources and whether carbonate encrusted vents are the result of microbial or abiotic processes. Emission of gas bubbles (predominantly CH4) from unlithified sands was observed at seven spots in an area of 100 m2 at Pomonte (Island of Elba), with a total rate of 234 ml m-2 d-1. The measured carbon isotope values of CH4 of around -18‰ (VPDB) in combination with the measured δ2H value of -120‰ (VSMOW) and the inverse correlation of δ13C-value with carbon number of hydrocarbon gases are characteristic for sites of CH4 formation through abiogenic processes, specifically abiogenic formation of CH4 via reduction of CO2 by H2. The H2 for methanogenesis likely derives from ophiolitic host rock within the Ligurian accretionary prism. The lack of hydrothermal activity allows CH4 gas to become decoupled from the stagnant aqueous phase. Hence no hyperalkaline fluid is currently released at the vent sites. Within the seep area a decrease in porewater sulphate concentrations by ca. 5 mmol/l relative to seawater and a concomitant increase in sulphide and dissolved inorganic carbon (DIC) indicate substantial activity of sulphate-dependent anaerobic oxidation of methane (AOM). In absence of any other dissimilatory pathway, the δ13C-values between -17 and -5‰ in dissolved inorganic carbon and aragonite cements suggest that the inorganic carbon is largely derived from CH4. The formation of seep carbonates is thus microbially induced via anaerobic oxidation of abiotic CH4.

Denitrifying Anaerobic Methane Oxidation: A Previously Overlooked Methane Sink in Intertidal Zone.

The intertidal zone is an open ecosystem rich in organic matter and plays an important role in global biogeochemical cycles. It was previously considered that methane was mainly removed by sulfate-dependent anaerobic methane oxidation (sulfate-AOM) process in marine ecosystems while other anaerobic methane oxidation processes were ignored. Recent researches have demonstrated that denitrifying anaerobic methane oxidation (DAMO), consisting of nitrite-dependent anaerobic methane oxidation (nitrite-AOM) and nitrate-dependent anaerobic methane oxidation (nitrate-AOM), can also oxidize methane. In this work, the community structure, quantity and potential methane oxidizing rate of DAMO archaea and bacteria in the intertidal zone were studied by high-throughput sequencing, qPCR and stable isotope tracing method. The results showed that nitrate-AOM and nitrite-AOM were both active in the intertidal zone and showed approximate methane oxidation rates. The copy number of 16S rRNA gene of DAMO archaea and DAMO bacteria were 104 ∼ 105 copies g-1 (dry sediment), whereas NC10 bacteria were slightly higher. The contribution rate of DAMO process to total anaerobic methane removal in the intertidal zone reached 65.6% ∼ 100%, which indicates that DAMO process is an important methane sink in intertidal ecosystem. Laboratory incubations also indicated that DAMO archaea were more sensitive to oxygen and preferred a more anoxic environment. These results help us draw a more complete picture of methane and nitrogen cycles in natural habitats.