Anaerobic Methanotrophs Research Articles

Discharge of deeply rooted fluids from submarine mud volcanism in the Taiwan accretionary prism

Qualitative and quantitative assessments of fluid cycling are essential to address the role and transport of deeply sourced fluids in subduction systems. In this study, sediment cores distributed across a submarine mud volcano (SMV) offshore southwestern Taiwan were investigated to determine the characteristics of fluids generated through the convergence between the Eurasian and Phillippine Sea Plates. The low dissolved chloride concentration combined with the enrichment of 18O, and depletion of 2H of pore fluids suggest the discharge of deep freshwater formed by smectite dehydration at an equilibrium temperature of 100 to 150 °C. The upward fluid velocities, decreasing from 2.0 to 5.0 cm yr−1 at the center to a negligible value at margin sites, varied with the rate and efficiency of anaerobic methanotrophy, demonstrating the impact of fluid migration on biogeochemical processes and carbon cycling. By extrapolating the velocity pattern, the flux of fluids exported from 13 SMVs into seawater amounted up to 1.3–2.5 × 107 kg yr−1, a quantity accounting for 1.1–28.6% of the smectite-bound water originally stored in the incoming sediments. Our results imply that SMVs could act as a conduit to channel the fluids produced from great depth/temperature into seafloor environments in a subduction system of the western Pacific Ocean.

Microbial communities from Arctic marine sediments respond slowly to methane addition during ex situ enrichments.

Anaerobic methanotrophic archaea (ANME) consume methane in marine sediments, limiting its release to the water column, but their responses to changes in methane and sulfate supplies remain poorly constrained. To address how methane exposure may affect microbial communities and methane- and sulfur-cycling gene abundances in Arctic marine sediments, we collected sediments from offshore Svalbard that represent geochemical horizons where anaerobic methanotrophy is expected to be active, previously active, and long-inactive based on reaction-transport biogeochemical modelling of porewater sulfate profiles. Sediment slurries were incubated at in situ temperature and pressure with different added methane concentrations. Sediments from an active area of seepage began to reduce sulfate in a methane-dependent manner within months, preceding increased relative abundances of anaerobic methanotrophs ANME-1 within communities. In previously active and long-inactive sediments, sulfur-cycling Deltaproteobacteria became more dominant after 30 days, though these communities showed no evidence of methanotrophy after nearly 8 months of enrichment. Overall, enrichment conditions, but not methane, broadly altered microbial community structure across different enrichment times and sediment types. These results suggest that active ANME populations may require years to develop, and consequently microbial community composition may affect methanotrophic responses to potential large-scale seafloor methane releases in ways that provide insight for future modelling studies.

Physiology and Distribution of Archaeal Methanotrophs That Couple Anaerobic Oxidation of Methane with Sulfate Reduction.

In marine anaerobic environments, methane is oxidized where sulfate-rich seawater meets biogenic or thermogenic methane. In those niches, a few phylogenetically distinct microbial types, i.e., anaerobic methanotrophs (ANME), are able to grow through anaerobic oxidation of methane (AOM). Due to the relevance of methane in the global carbon cycle, ANME have drawn the attention of a broad scientific community for 4 decades. This review presents and discusses the microbiology and physiology of ANME up to the recent discoveries, revealing novel physiological types of anaerobic methane oxidizers which challenge the view of obligate syntrophy for AOM. An overview of the drivers shaping the distribution of ANME in different marine habitats, from cold seep sediments to hydrothermal vents, is given. Multivariate analyses of the abundance of ANME in various habitats identify a distribution of distinct ANME types driven by the mode of methane transport. Intriguingly, ANME have not yet been cultivated in pure culture, despite intense attempts. Further advances in understanding this microbial process are hampered by insufficient amounts of enriched cultures. This review discusses the advantages, limitations, and potential improvements for ANME laboratory-based cultivation systems.

Carbon isotopic heterogeneity of coenzyme F430 and membrane lipids in methane‐oxidizing archaea

Archaeal ANaerobic MEthanotrophs (ANME) facilitate the anaerobic oxidation of methane (AOM), a process that is believed to proceed via the reversal of the methanogenesis pathway. Carbon isotopic composition studies indicate that ANME are metabolically diverse and able to assimilate metabolites including methane, methanol, acetate, and dissolved inorganic carbon (DIC). Our data support the interpretation that ANME in marine sediments at methane seeps assimilate both methane and DIC, and the carbon isotopic compositions of the tetrapyrrole coenzyme F430 and the membrane lipids archaeol and hydroxy-archaeol reflect their relative proportions of carbon from these substrates. Methane is assimilated via the methyl group of CH3 -tetrahydromethanopterin (H4 MPT) and DIC from carboxylation reactions that incorporate free intracellular DIC. F430 was enriched in 13 C (mean δ13 C=-27‰ for Hydrate Ridge and -80‰ for the Santa Monica Basin) compared to the archaeal lipids (mean δ13 C=-97‰ for Hydrate Ridge and -122‰ for the Santa Monica Basin). We propose that depending on the side of the tricarboxylic acid (TCA) cycle used to synthesize F430, its carbon was derived from 76% DIC and 24% methane via the reductive side or 57% DIC and 43% methane via the oxidative side. ANME lipids are predicted to contain 42% DIC and 58% methane, reflecting the amount of each assimilated into acetyl-CoA. With isotope models that include variable fractionation during biosynthesis for different carbon substrates, we show the estimated amounts of DIC and methane can result in carbon isotopic compositions of-73‰ to-77‰ for F430 and-105‰ for archaeal lipids, values close to those for Santa Monica Basin. The F430 δ13 C value for Hydrate Ridge was 13 C-enriched compared with the modeled value, suggesting there is divergence from the predicted two carbon source models.

Anaerobic methane oxidation coupled to sulfate reduction in a biotrickling filter: Reactor performance and microbial community analysis

The aim of this work was to evaluate the performance of a biotrickling filter (BTF) packed with polyurethane foam and pall rings for the enrichment of microorganisms mediating anaerobic oxidation of methane (AOM) coupled to sulfate reduction (SR) by activity tests and microbial community analysis. A BTF was inoculated with microorganisms from a known AOM active deep sea sediment collected at a depth of 528 m below the sea level (Alpha Mound, Gulf of Cadiz). The microbial community analysis was performed by catalyzed reporter deposition - fluorescence in situ hybridization (CARD-FISH) and 16S rRNA sequence analysis. The AOM occurrence and rates in the BTF were assessed by performing batch activity assays using 13C-labelled methane (13CH4). After an estimated start-up time of ∼20 days, AOM rates of ∼0.3 mmol l−1 day−1 were observed in the BTF, values almost 20 times higher than previously reported in a polyurethane foam packed BTF. The microbial community consisted mainly of anaerobic methanotrophs (ANME-2, 22% of the total number of cells) and sulfate reducing bacteria (SRB, 47% of the total number of cells). This study showed that the BTF is a suitable reactor configuration for the enrichment of microbial communities involved in AOM coupled to SR at ambient pressure and temperature with a relatively short start-up time.

Microbial chromate reduction coupled with anaerobic oxidation of methane in a membrane biofilm reactor

It has been reported that microbial reduction of sulfate, nitrite/nitrate and iron/manganese could be coupled with anaerobic oxidation of methane (AOM), which plays a significant role in controlling methane emission from anoxic niches. However, little is known about microbial chromate (Cr(VI)) reduction coupling with AOM. In this study, a microbial consortium was enriched via switching nitrate dosing to chromate feeding as the sole electron acceptor under anaerobic condition in a membrane biofilm reactor (MBfR), in which methane was continuously provided as the electron donor through bubble-less hollow fiber membranes. According to long-term reactor operation and chromium speciation analysis, soluble chromate could be reduced into Cr(III) compounds by using methane as electron donor. Fluorescence in situ hybridization and high-throughput 16S rRNA gene amplicon profiling further indicated that after feeding chromate Candidatus ‘Methanoperedens’ (a known nitrate-dependent anaerobic methane oxidation archaeon) became sole anaerobic methanotroph in the biofilm, potentially responsible for the chromate bio-reduction driven by methane. Two potential pathways of the microbial AOM-coupled chromate reduction were proposed: (i) Candidatus ‘Methanoperedens’ independently utilizes chromate as electron acceptor to form Cr(III) compounds, or (ii) Candidatus ‘Methanoperedens’ oxidizes methane to generate intermediates or electrons, which will be utilized to reduce chromate to Cr(III) compounds by unknown chromate reducers synergistically. Our findings suggest a possible link between the biogeochemical chromium and methane cycles.

Key Physiology of a Nitrite-Dependent Methane-Oxidizing Enrichment Culture.

Nitrite-dependent methane-oxidizing bacteria couple the reduction of nitrite to the oxidation of methane via a unique oxygen-producing pathway. This process is carried out by members of the genus Methylomirabilis that belong to the NC10 phylum. Contrary to other known anaerobic methane oxidizers, they do not employ the reverse methanogenesis pathway for methane activation but instead a canonical particulate methane monooxygenase similar to those used by aerobic methanotrophs. Methylomirabilis-like bacteria are detected in many natural and manmade ecosystems, but their physiology is not well understood. Here, using continuous cultivation techniques, batch activity assays, and state-of-the-art membrane-inlet mass spectrometry, we determined growth rate, doubling time, and methane and nitrite affinities of the nitrite-dependent methane-oxidizing bacterium "Candidatus Methylomirabilis lanthanidiphila." Our results provide insight into understanding the interactions of these microorganisms with methanotrophs and other nitrite-reducing microorganisms, such as anaerobic ammonium-oxidizing bacteria. Furthermore, our data can be used in modeling studies as well as wastewater treatment plant design.IMPORTANCE Methane is an important greenhouse gas with a radiative forcing 28 times that of carbon dioxide over a 100-year time scale. The emission of methane to the atmosphere is controlled by aerobic and anaerobic methanotrophs, which are microorganisms that are able to oxidize methane to conserve energy. While aerobic methanotrophs have been studied for over a century, knowledge on the physiological characteristics of anaerobic methanotrophs is scarce. Here, we describe kinetic properties of "Candidatus Methylomirabilis lanthanidiphila," a nitrite-dependent methane-oxidizing microorganism, which is ecologically important and can be applied in wastewater treatment.

Exchange catalysis during anaerobic methanotrophy revealed by 12CH2D2 and 13CH3D in methane

The anaerobic oxidation of methane (AOM) is a crucial component of the methane cycle, but quantifying its role in situ under dynamic environmental conditions remains challenging. We use sediment samples collected during IODP Expedition 347 to the Baltic Sea to show that relative abundances of 12CH2D2 and 13CH3D in methane remaining after microbial oxidation are in internal, thermodynamic isotopic equilibrium, and we attribute this phenomenon to the reversibility of the initial step of AOM. These data suggest that 12CH2D2 and 13CH3D together can identify the influence of anaerobic methanotrophy in environments where conventional bulk isotope ratios are ambiguous, and these findings may lead to new insights regarding the global significance of enzymatic back reaction in the methane cycle.

Enrichment of Anaerobic Methanotrophs in Biotrickling Filters Using Different Sulfur Compounds as Electron Acceptor

Abstract Anaerobic oxidation of methane (AOM) coupled to sulfate reduction is a biological process regulating the methane cycle with potential application in desulfurization of industrial wastewate...

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.

Contrasting Pathways for Anaerobic Methane Oxidation in Gulf of Mexico Cold Seep Sediments.

Gulf of Mexico sediments harbor numerous hydrocarbon seeps associated with high sedimentation rates and thermal maturation of organic matter. These ecosystems host abundant and diverse microbial communities that directly or indirectly metabolize components of the emitted fluid. To investigate microbial function and activities in these ecosystems, metabolic potential (metagenomic) and gene expression (metatranscriptomic) analyses of two cold seep areas of the Gulf of Mexico were carried out. Seeps emitting biogenic methane harbored microbial communities dominated by archaeal anaerobic methane oxidizers of phylogenetic group 1 (ANME-1), whereas seeps producing fluids containing a complex mixture of thermogenic hydrocarbons were dominated by ANME-2 lineages. Metatranscriptome measurements in both communities indicated high levels of expression of genes for methane metabolism despite their distinct microbial communities and hydrocarbon composition. In contrast, the transcription level of sulfur cycle genes was quite different. In the thermogenic seep community, high levels of transcripts indicative of syntrophic anaerobic oxidation of methane (AOM) coupled to sulfate reduction were detected. This syntrophic partnership between the dominant ANME-2 and sulfate reducers potentially involves direct electron transfer through multiheme cytochromes. In the biogenic methane seep, genes from an ANME-1 lineage that are potentially involved in polysulfide reduction were highly expressed, suggesting a novel bacterium-independent anaerobic methane oxidation pathway coupled to polysulfide reduction. The observed divergence in AOM activities provides a new model for bacterium-independent AOM and emphasizes the variation that exists in AOM pathways between different ANME lineages. IMPORTANCE Cold seep sediments are complex and widespread marine ecosystems emitting large amounts of methane, a potent greenhouse gas, and other hydrocarbons. Within these sediments, microbial communities play crucial roles in production and degradation of hydrocarbons, modulating oil and gas emissions to seawater. Despite this ecological importance, our understanding of microbial functions and methane oxidation pathways in cold seep ecosystems is poor. Based on gene expression profiling of environmental seep sediment samples, the present work showed that (i) the composition of the emitted fluids shapes the microbial community in general and the anaerobic methanotroph community specifically and (ii) AOM by ANME-2 in this seep may be coupled to sulfate reduction by Deltaproteobacteria by electron transfer through multiheme cytochromes, whereas AOM by ANME-1 lineages in this seep may involve a different, bacterium-independent pathway, coupling methane oxidation to elemental sulfur/polysulfide reduction.

Anaerobic Methane Oxidation in High-Arctic Alaskan Peatlands as a Significant Control on Net CH4 Fluxes

Terrestrial consumption of the potent greenhouse gas methane (CH4) is a critical aspect of the future climate, as CH4 concentrations in the atmosphere are projected to play an increasingly important role in global climate forcing. Anaerobic oxidation of methane (AOM) has only recently been considered a relevant control on methane fluxes from terrestrial systems. We performed in vitro anoxic incubations of intact peat from Utqiaġvik (Barrow), Alaska using stable isotope tracers. Our results showed an average potential AOM rate of 15.0 nmol cm3 h−1, surpassing the average rate of gross CH4 production (6.0 nmol cm3 h−1). AOM and CH4 production rates were positively correlated. While CH4 production was insensitive to additions of Fe(III), there was a depth:Fe(III) interaction in the kinetic reaction rate constant for AOM, suggestive of stimulation by Fe(III), particularly in shallow soils (<10 cm). We estimate AOM would consume 25–34% of CH4 produced under ambient conditions. Soil genetic surveys showed phylogenetic links between soil microbes and known anaerobic methanotrophs in ANME groups 2 and 3. These results suggest a prevalent role of AOM to net CH4 fluxes from Arctic peatland ecosystems, and a probable link with Fe(III)-reduction.

Microbiological Study of Yamal Lakes: A Key to Understanding the Evolution of Gas Emission Craters

Although gas emission craters (GECs) are actively investigated, the question of which landforms result from GECs remains open. The evolution of GECs includes the filling of deep hollows with atmospheric precipitation and deposits from their retreating walls, so that the final stage of gas emission crater (GEC) lake development does not differ from that of any other lakes. Microbial activity and diversity may be indicators that make it possible to distinguish GEC lakes from other exogenous lakes. This work aimed at a comparison of the activity and diversity of microbial communities in young GEC lakes and mature background lakes of Central Yamal by using a radiotracer analysis and high-throughput sequencing of the 16S rRNA genes. The radiotracer analysis revealed slow-flowing microbial processes as expected for the cold climate of the study area. GEC lakes differed from background ones by slow rates of anaerobic processes (methanogenesis, sulfate reduction) as well as by a low abundance and diversity of methanogens. Other methane cycle micro-organisms (aerobic and anaerobic methanotrophs) were similar in all studied lakes and represented by Methylobacter and ANME 2d; the rates of methane oxidation were also similar. Actinobacteria, Bacteroidetes, Betaproteobacteria, and Acidobacteria were predominant in both lake types. Thus, GEC lakes may be identified by their scarce methanogenic population.

Active pathways of anaerobic methane oxidation across contrasting riverbeds

Anaerobic oxidation of methane (AOM) reduces methane emissions from marine ecosystems but we know little about AOM in rivers, whose role in the global carbon cycle is increasingly recognized. We measured AOM potentials driven by different electron acceptors, including nitrite, nitrate, sulfate, and ferric iron, and identified microorganisms involved across contrasting riverbeds. AOM activity was confined to the more reduced, sandy riverbeds, whereas no activity was measured in the less reduced, gravel riverbeds where there were few anaerobic methanotrophs. Nitrite-dependent and nitrate-dependent AOM occurred in all sandy riverbeds, with the maximum rates of 61.0 and 20.0 nmol CO2 g−1 (dry sediment) d−1, respectively, while sulfate-dependent and ferric iron-dependent AOM occurred only where methane concentration was highest and the diversity of AOM pathways greatest. Diverse Candidatus Methylomirabilis oxyfera (M. oxyfera)-like bacteria and Candidatus Methanoperedens nitroreducens (M. nitroreducens)-like archaea were detected in the sandy riverbeds (16S rRNA gene abundance of 9.3 × 105 to 1.5 × 107 and 2.1 × 104 to 2.5 × 105 copies g−1 dry sediment, respectively) but no other known anaerobic methanotrophs. Further, we found M. oxyfera-like bacteria and M. nitroreducens-like archaea to be actively involved in nitrite- and nitrate/ferric iron-dependent AOM, respectively. Hence, we demonstrate multiple pathways of AOM in relation to methane, though the activities of M. oxyfera-like bacteria and M. nitroreducens-like archaea are dominant.

Extreme 13C-depletions and organic sulfur content argue for S-fueled anaerobic methane oxidation in 2.72 Ga old stromatolites

The extreme 13C-depletions recorded in Neoarchean organic matter (OM) have commonly been interpreted as markers of methanotrophy. This methane oxidation metabolism could have been performed using oxidants such as dioxygen, sulfate, nitrite/nitrate, and/or Fe3+- and Mn4+-minerals. Acetogenesis using acetyl-CoA metabolism may produce similar 13C-depletions. We investigated δ13C and S/C values of OM in 2.72 Ga old lacustrine stromatolites of the Tumbiana Formation (Australia) using Secondary Ion Mass Spectrometry (SIMS), coupled with X-ray spectroscopy. Type-A OM is embedded in quartz and/or chlorite. We show that mixtures of chlorite bias negatively the δ13Corg of associated OM measured by SIMS, likely through recombination of hydrogen from chlorite with carbon from OM during analysis. Type-A OM, associated with quartz or interleaved with chlorites (but remote enough to avoid H-recombination), displays δ13Corg between −56.1 and −50.6‰ VPDB (mean: −53.5‰, SD ± 1.8‰). The intimate association of Type-A OM with Fe-rich chlorite and its main occurrence in pyritic layers coupled with 13C-depletions is most parsimoniously interpreted as anaerobic methanotrophy using Fe3+-minerals and/or oxidized S. Type-B OM comprises <2-µm organic globules embedded in calcite. A mass balance correction using the δ13C values measured on Type-B globules mixed with calcite and those measured on pure calcite yielded corrected δ13C∗org values for the globules ranging between −65.2 and −52.5‰ (mean −58.8‰, SD ± 3.6‰). In a context where Fe and S reduction could outcompete acetogenesis for a fraction of any available H2, these extremely low δ13Corg values are difficult to explain with chemoautotrophic acetogenesis. Atomic S/C ratios in Type-B globules reach up to 0.042, i.e. up to tenfold the values (0.003–0.004) in Type A-OM. These organic S concentrations, combined with the extremely low δ13C∗org values, support anaerobic oxidation of methane coupled to sulfur oxidation. Finally, Type-C OM, interpreted as migrated pyrobitumen nodules, displays δ13Corg of −48 to −39.1‰. The most extreme low-δ13Corg values of the Precambrian are thus best interpreted as the result of lacustrine methanotrophy fueled by oxidized S and/or Fe species, which likely derived from oxygenic and/or anoxygenic photosynthesis. Photosynthesis, followed by methanogenesis and methanotrophy may have been important in lakes at a time of supercontinent growth and eruption of large subaerial igneous provinces. In this context, anaerobic methanotrophy could have played a part in regulating atmospheric methane.

Regulation of coastal methane sinks by a structured gradient of microbial methane oxidizers

Coastal wetlands are widely recognized as atmospheric methane sources. However, recent field studies suggest that some coastal wetlands could also act as methane sinks, but the mechanism is not yet clear. Here, we investigated methane oxidation with different electron acceptors (i.e., oxygen, nitrate/nitrite, sulfate, Fe(III) and Mn(IV)) in four coastal wetlands in China using a combination of molecular biology methods and isotopic tracing technologies. The geochemical profiles and in situ Gibbs free energies suggest that there was significant nitrite-dependent anaerobic oxidation of methane (nitrite-AOM) in the sub-surface sediments; this was subsequently experimentally verified by both the microbial abundance and activity. Remarkably, the methanotrophic communities seemed to exist in the sediments as layered structures, and the surface aerobic methane-oxidizing bacteria were able to take up atmospheric methane at a rate of 0.10–0.18 nmol CH4 day−1 cm−2, while most, if not all, sedimentary methane was being completely consumed by anaerobic methanotrophs (23–58% by methane oxidizers in phylum NC10). These results suggest that coastal methane sinks might be governed by diverse microbial communities where NC10 methane oxidizers contributed significantly. This finding helps to better understand and predict the coastal methane cycle and reduce uncertainties in the estimations of the global methane flux.

Triple sulfur isotope relationships during sulfate-driven anaerobic oxidation of methane

Sulfate-driven anaerobic oxidation of methane (SD-AOM) plays a critical role in regulating the global methane budget. Determination of the diagnostic triple isotope exponent θ33 (≡ln33⁡α/ln34⁡α) for SD-AOM can help to identify and quantify microbial sulfate reduction via SD-AOM in the environment. The history of Earth's surface redox conditions can also be examined through the measurement of triple sulfur isotope compositions in sedimentary rocks. Due to difficulties in both culturing anaerobic methanotrophs and sampling pore-water sulfate in SD-AOM-dominated environments, however, the θ33 values for the processes of SD-AOM have not been constrained. We propose that a set of modern cold-seep associated barite samples with low Δδ18O/Δδ34S values bear a record of residual pore-water sulfate during SD-AOM, and therefore the triple sulfur isotope composition of these barites can be used to deduce θ33 values. We applied a 1-D diagenetic reaction–transport model to fit Δ33S and δ′34S results from modern cold seep barites collected from five sites in the Gulf of Mexico. Based on revealed negative correlations (R2=0.77) between Δ33S and δ′34S values we calculated an upper-limit θ33 value of 0.5100 to 0.5112 (±0.0005) given a 1000ln34⁡α value of −30‰ to −10‰. This θ33 value is distinctively lower than that of organoclastic sulfate reduction (OSR) in marine environments where the diagnostic isotope fractionation (1000ln34⁡α) is typically more negative than that of SD-AOM. In addition, cold seep barite data display a negative Δ33S–δ′34S correlation whereas pore-water sulfates of all OSR-dominated settings show a positive one. Therefore, the diagnostic triple-sulfur isotope exponent and associated negative Δ33S–δ′34S correlation may allow for the identification of SD-AOM in sedimentary records.

Insight into anaerobic methanotrophy from 13C/12C- amino acids and 14C/12C-ANME cells in seafloor microbial ecology

Oceanic methane from global deep-sea sediment is largely consumed through microbially mediated sulfate-coupled oxidation, resulting in 13C-depleted cell biomass of anaerobic methanotrophic archaea (ANME). The general ecological importance of subseafloor ANME has been well recognized in the last two decades. However, the crucial biochemical pathways for the overall anaerobic oxidation of methane (AOM) still remain enigmatic. Here, methanotrophic pathways were analyzed to trace 13C-depleted amino acid biosynthesis in two clades of ANME (ANME-1 and ANME-2) from the Black Sea. Compound-specific analysis of ANME-dominated microbial mats showed a significant 13C-depletion trend in association with increasing carbon numbers in protein-derived amino acid families (e.g., the pyruvate family in the order of alanine, valine, isoleucine and leucine was down to −114‰). This result indicates a stepwise elongation of 13C-depleted carbon during amino acid biosynthesis. The overall results suggest that intracellular protein amino acids and the most 13C-depleted signature of leucine, which has a specific branched-chain structure, are potentially propagated as isoprenoid precursor molecules into archaeal biosynthesis, resulting in the extremely 13C- and 14C-depleted nature of ANME cells in the deep microbial oasis.

Steep redox gradient and biogeochemical cycling driven by deeply sourced fluids and gases in a terrestrial mud volcano.

Mud volcanoes provide an accessible channel through which deep subsurface environments can be observed. The manner in which deeply sourced materials shape biogeochemical processes and microbial communities in such geological features remains largely unknown. This study characterized redox transitions, biogeochemical fluxes and microbial communities for samples collected from a methane-rich mud volcano in southwestern Taiwan. Our results indicated that oxygen penetration was confined within the upper 4 mm of fluids/muds and counteracted by the oxidation of pyrite, dissolved sulfide, methane and organic matter at various degrees. Beneath the oxic zone, anaerobic sulfur oxidation, sulfate reduction, anaerobic methanotrophy and methanogenesis were compartmentalized into different depths in the pool periphery, forming a metabolic network that efficiently cycles methane and sulfur. Community members affiliated with various Proteobacteria capable of aerobic oxidation of sulfur, methane and methyl compounds were more abundant in the anoxic zone with diminished sulfate and high methane. These findings suggest either the requirement of alternative electron acceptors or a persistent population that once flourished in the oxic zone. Overall, this study demonstrates the distribution pattern for a suite of oxidative and reductive metabolic reactions along a steep redox gradient imposed by deep fluids in a mud volcano ecosystem.

Microbial communities involved in aerobic and anaerobic methane cycling in a meromictic ferruginous subarctic lake

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 82:1-18 (2018) - DOI: https://doi.org/10.3354/ame01878 Microbial communities involved in aerobic and anaerobic methane cycling in a meromictic ferruginous subarctic lake Anna Kallistova1,*, Vitaly Kadnikov2,*, Igor Rusanov1, Natalia Kokryatskaya3, Alexey Beletsky2, Andrey Mardanov2, Alexander Savvichev1, Nikolai Ravin2, Nikolai Pimenov1,** 1Winogradsky Institute of Microbiology, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia 2Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow 119071, Russia 3N. Laverov Federal Center for Integrated Arctic Research, Arkhangelsk 163000, Russia *These authors contributed equally to this work**Corresponding author: npimenov@mail.ru ABSTRACT: Since boreal lakes are now considered an important source of atmospheric methane, new information concerning the activity and diversity of methane cycle microorganisms is critical for understanding the causes of methane emission from these lakes. This study investigated the diversity of microbial communities in the water column of subarctic Lake Svetloe, belonging to a rare type of freshwater sulfate-poor meromictic basins with high concentrations of Fe2+ and methane in the permanently anoxic hypolimnion. A combination of physicochemical and radiotracer analysis, high-throughput sequencing of the 16S rRNA genes and incubation experiments was used to link microbial community profile and methane cycle processes. It was shown that methane was produced by aceticlastic Methanothrix and hydrogenotrophic Methanoregula, which were also detected in the oxygenated epilimnion, together with a small increase in methane concentration. Radiotracer analysis revealed methane oxidation (MO) in oxic and anoxic zones with 2 maxima at the chemocline. The first MO peak was attributed to aerobic Methylobacter trophically interacting with cyanobacteria, which was confirmed by obtaining light-dependent MO. The highest MO activity matched the lower chemocline layer where aerobic methanotrophs were less abundant; this suggested that other microorganisms contributed to MO together with Methylobacter. Known anaerobic methanotrophs were not detected, and incubations with Fe3+ did not reveal methane consumption under anoxic conditions. Thus, further investigations are required to determine the microorganisms and electron acceptors driving anaerobic MO. Although some questions remain open, our study may provide insight into the methane cycle microbial communities in boreal lakes. KEY WORDS: Meromictic freshwater lakes · Ferruginous lakes · Lake Svetloe · Methane cycle · Methane oxidation · Methanogenesis · 16S rRNA profiling Full text in pdf format NextCite this article as: Kallistova A, Kadnikov V, Rusanov I, Kokryatskaya N and others (2018) Microbial communities involved in aerobic and anaerobic methane cycling in a meromictic ferruginous subarctic lake. Aquat Microb Ecol 82:1-18. https://doi.org/10.3354/ame01878 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 82, No. 1. Online publication date: August 31, 2018 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2018 Inter-Research.