Dominant Methanogenic Pathway Research Articles

A novel approach to estimate methanogenic pathways in biogas reactors via stable carbon isotope analysis

Two microbial pathways are responsible for most of the methane produced during anaerobic digestion: acetoclastic methanogenesis (AM) and hydrogenotrophic methanogenesis (HM) coupled with syntrophic acetate oxidation (SAO). Identifying the dominant methanogenic pathway active in a system provides the information necessary to manage and optimize productivity, stability, process control, and gas quality in biogas reactors. In this study, a modified method is proposed to estimate methanogenic pathways in different biogas systems via short-term parallel incubations with methyl-labeled acetate (2-13C-acetate). Cavity ring-down spectroscopy was applied to measure the δ13C–CH4 and δ13C–CO2 isotopic signatures of produced biogas. Preliminary experiments demonstrated that longer incubation times led to significant variations in δ13C–CH4 and δ13C–CO2 and consequently interfered with the calculated fraction of CH4 produced from HM (fHM). This variability is likely caused by the dilution of 13CH4 and 13CO2 as 2-13C-acetate is consumed, along with potential changes in organic matter quality and quantity, microbial community composition, and environmental factors such as pH, volatile fatty acid content, and ammonia levels, during longer incubations. We applied this new approach to sludge from six full-scale reactors (three mesophilic and three thermophilic) and validated its potential with consistent estimates of fHM with minimal variation. Mesophilic reactors exhibited AM dominance, while HM was the dominant pathway in thermophilic reactors, aligning with reports in the literature.

Net Methane Production Predicted by Patch Characteristics in a Freshwater Wetland

AbstractMethane (CH4) dynamics in wetlands are spatially variable and difficult to estimate at ecosystem scales. Patches with different plant functional types (PFT) represent discrete units within wetlands that may help characterize patterns in CH4 variability. We investigate dissolved porewater CH4 concentrations, a representation of net CH4 production and potential source of atmospheric flux, in five wetland patches characterized by a dominant PFT or lack of plants. Using soil, porewater, and plant variables we hypothesized to influence CH4, we used three modeling approaches—Classification and regression tree, AIC model selection, and Structural Equation Modeling—to identify direct and indirect influences on porewater CH4 dynamics. Across all three models, dissolved porewater CO2 concentration was the dominant driver of CH4 concentrations, partly through the influence of PFT patches. Plants in each patch type likely had variable influence on CH4 via root exudates (a substrate for methanogens), capacity to transport gas (both O2 from and CH4 to the atmosphere), and plant litter quality which impacted soil respiration and production of CO2 in the porewater. We attribute the importance of CO2 to the dominant methanogenic pathway we identified, which uses CO2 as a terminal electron acceptor. We propose a mechanistic relationship between PFT patches and porewater CH4 dynamics which, when combined with sources of CH4 loss including methanotrophy, oxidation, or plant‐mediated transport, can provide patch‐scale estimates of CH4 flux. Combining these estimates with the distribution of PFTs can improve ecosystem CH4 flux estimates in heterogenous wetlands and improve global CH4 budgets.

Co-enhancing effects of zero valent iron and magnetite on anaerobic methanogenesis of food waste at transition temperature (45 °C) and various organic loading rates

Deoiling of food waste (FW) after hydrothermal pretreatment occurs at high temperatures, and more energy is required for substrate cooling before the anaerobic digestion (AD) process. AD at the transition temperature (for example 45 °C) is good for energy saving and carbon emission reducing when treating deoiling FW. However, the metabolic activity of methanogens must increase at the transition temperatures. This study proposes the use of zero-valent iron (Fe0) and magnetite (Fe3O4) to boost CH4 yield from deoiling FW. The results showed a co-enhancing effect on CH4 yield upgradation when using Fe0 and Fe3O4 simultaneously, and the highest CH4 yield reached 536.23 mLCH4/gVS, which was 67.5 % higher than that of Fe0 alone (320.14 mLCH4/gVS). In addition, a high organic loading was favorable for increasing the CH4 yield from deoiling FW. Microbial diversity analysis suggested that the dominant methanogenic pathway at 45 °C was hydrogenotrophic methanogenesis. Herein, a potential metabolic pathway analysis revealed that the co-enhancing effects of Fe0 and Fe3O4 enhanced syntrophic methanogenesis and possibly boosted electron transfer efficiency.

Hydrogenotrophic pathway dominates methanogenesis along the river-estuary continuum of the Yangtze River

Rivers are considered as an important source of methane (CH4) to the atmosphere, but our understanding for the methanogenic pathway in rivers and its linkage with CH4 emission is very limited. Here, we investigated the diffusive flux of CH4 and its stable carbon isotope signature (δ13C-CH4) along the river-estuary continuum of the Yangtze River. The diffusive CH4 flux was estimated to 27.9 ± 11.4 μmol/m2/d and 36.5 ± 24.4 μmol/m2/d in wet season and dry season, respectively. The δ13C-CH4 values were generally lower than -60‰, with the fractionation factor (αc) higher than 1.055 and the isotope separation factor (εc) ranged from 55 to 100. In situ microbial composition showed that hydrogenotrophic methanogens accounts for over 70% of the total reads. Moreover, the incubation test showed that the headspace CH4 concentration by adding CO2/H2 to the sediment was orders of magnitude higher than that by adding trimethylamine and sodium acetate. These results jointly verified the river-estuary continuum is a minor CH4 source and dominated by hydrogenotrophic pathway. Based on the methanogenic pathway here and previous reported data in the same region, the historical variation of diffusive CH4 flux was calculated and results showed that CH4 emission has reduced 82.5% since the construction of Three Gorges Dam (TGD). Our study verified the dominant methanogenic pathway in river ecosystems and clarified the effect and mechanism of dam construction on riverine CH4 emission.

Insights into the impact of quaternary ammonium disinfectant on sewage sludge anaerobic digestion: Dose-response, performance variation, and potential mechanisms

Wide commercial applications of antimicrobial quaternary ammonium compounds (QACs) inevitably lead to the release into wastewater and enrichment in sewage sludge. This study evaluated the impacts of levels and structures of QACs on sewage sludge properties, microbial community, and methane production during anaerobic digestion. Methane production was stimulated or not affected at low QACs concentrations, but significantly inhibited at high QACs concentrations. Compared with benzyl and alkyltrimethyl QACs, dialkyl QACs showed least toxicity on digestion performance. Meanwhile, microbial community analysis indicated that shifts in bacterial communities mainly depended on QACs doses, but the archaeal communities were affected by both QACs doses and types. The dominant methanogenic pathway shifted from acetotrophic/methylotrophic methanogens to mixotrophic methanogens by low levels of benzyl and alkyltrimethyl QACs but not dialkyl QACs, and further to hydrogenotrophic methanogens at high QACs concentration. Mechanism exploration revealed that the presence of QACs promoted sludge solubilization by the integrated effects of cell lysis, electric neutralization, and hydrophobicity improvement, but inhibited methanogenesis due to the accumulation of volatile fatty acids and susceptibility of methanogens to QACs. These findings provided a reference for potential impacts of different QACs on sludge biological treatment, which had implications for the use and selection of QACs disinfectants.

Enhanced anaerobic digestion of kitchen waste at different solids content by alkali pretreatment and bentonite addition: Methane production enhancement and microbial mechanism

High solid anaerobic digestion (AD) has been considered as a promising and sustainable technology for treating kitchen waste. To enhance AD of kitchen waste, alkali pretreatment and bentonite addition treatment (AP/Be) was performed on kitchen waste, and microbial community was investigated at different total solids (TS) content (10%, 13%, 19%, 22% and 25%). The results indicated that after AP/Be treatment, methane yield was as high as 198 mL CH4/g volatile solid (VS), which increased by 236% as the control. Moreover, microbial community analysis revealed that AP/Be treatment enriched bacterial microbial diversity. At TS of 10%, AP/Be treatment enhanced the hydrogenotrophic methanogens (Methanobacterium) significantly. In addition, the dominant methanogenic pathways changed at different TS content. These results demonstrated AP/Be treatment had a positive effect on methanogenesis during kitchen waste anaerobic digestion process. This study threw new insights towards enhancing kitchen waste anaerobic digestion, as well as the microbial mechanism.

Syntrophic Acetate-Oxidizing Microbial Consortia Enriched from Full-Scale Mesophilic Food Waste Anaerobic Digesters Showing High Biodiversity and Functional Redundancy.

ABSTRACTSyntrophic acetate oxidation (SAO) coupled with hydrogenotrophic methanogenesis (HM) plays a vital role in the anaerobic digestion of protein-rich feedstocks such as food wastes. However, current knowledge of the biodiversity and genetic potential of the involved microbial participants, especially syntrophic acetate-oxidizing bacteria (SAOB), is limited due to the low abundance of these microorganisms and challenges in their isolation. The intent of this study was to enrich and identify potential SAOB. Therefore, we conducted continuous acetate feeding under high ammonia concentrations using two separate inoculum consortia of microorganisms that originated from full-scale mesophilic food waste digesters, which lasted for more than 200 days. Using 16S rRNA gene amplicon and metagenomic analyses, we observed a convergence of the experimental microbial communities during the enrichment regarding taxonomic composition and metabolic functional composition. Stable carbon isotope analyses of biogas indicated that SAO-HM was the dominant methanogenic pathway during the enrichment process. The hydrogenotrophic methanogen Methanoculleus dominated the archaeal community. The enriched SAO community featured high biodiversity and metabolic functional redundancy. By analyzing the metagenome-assembled genomes, the known SAOB Syntrophaceticus schinkii and six uncultured populations were identified to have the genetic potential to perform SAO through the conventional reversed Wood-Ljungdahl pathway, while another six bacteria were found to encode the reversed Wood-Ljungdahl pathway combined with a glycine cleavage system as novel SAOB candidates. These results showed that the food waste anaerobic digesters harbor diverse SAOB and highlighted the importance of the glycine cleavage system for acetate oxidation.IMPORTANCE Syntrophic acetate oxidation to CO2 and H2, together with hydrogenotrophic methanogenesis, contributes to much of the carbon flux in the anaerobic digestion of organic wastes, especially at high ammonia concentrations. A deep understanding of the biodiversity, metabolic genetic potential, and ecology of the SAO community can help to improve biomethane production from wastes for clean energy production. Here, we enriched the SAO-HM functional guild obtained from full-scale food waste anaerobic digesters and recorded dynamic changes in community taxonomic composition and functional profiles. By reconstructing the metabolic pathways, diverse known and novel bacterial members were found to have SAO potential via the reversed Wood-Ljungdahl (WL) pathway alone, or via the reversed WL pathway with a glycine cleavage system (WLP-GCS), and those catalyzing WLP-GCS showed higher microbial abundance. This study revealed the biodiversity and metabolic functional redundancy of SAOB in full-scale anaerobic digester systems and provided inspiration for further genome-centric studies.

Methanogenic archaea in subsurface coal seams are biogeographically distinct: an analysis of metagenomically-derived mcrA sequences.

The production of methane as an end-product of organic matter degradation in the absence of other terminal electron acceptors is common, and has often been studied in environments such as animal guts, soils and wetlands due to its potency as a greenhouse gas. To date, however, the study of the biogeographic distribution of methanogens across coal seam environments has been minimal. Here, we show that coal seams are host to a diverse range of methanogens, which are distinctive to each geological basin. Based on comparisons to close relatives from other methanogenic environments, the dominant methanogenic pathway in these basins is hydrogenotrophic, with acetoclastic being a second major pathway in the Surat Basin. Finally, mcrA and 16S rRNA gene primer biases were predominantly seen to affect the detection of Methanocellales, Methanomicrobiales and Methanosarcinales taxa in this study. Subsurface coal methanogenic community distributions and pathways presented here provide insights into important metabolites and bacterial partners for in situ coal biodegradation.

Deterministic processes drive the microbial assembly during the recovery of an anaerobic digester after a severe ammonia shock

Anaerobic digestion allows to produce sustainable energy but the microbial community involved in this process is highly sensitive to perturbations. In this study, a longitudinal experiment was performed in two sets of triplicate bioreactors to evaluate the influence of ammonia addition on AD microbiome and its recovery. Zeolite was added in three reactors to mitigate the inhibition. Microbial dynamics were monitored with 16S rRNA sequencing at 15 time points. Dominant methanogenic pathways were determined with gas isotopic signature analysis. Zeolite addition did not enable to reduce ammonia inhibition or improve the process under the conditions tested. In all the bioreactors, ammonia inhibition sharply decreased the methane production but the process could restart thanks to the increase of hydrogenotrophic archaea and syntrophic bacteria. Interestingly, similar behaviour was observed in the six reactors. Neutral modelling and null model were used and showed that a deterministic process governed the recovery of AD microbiome after failure.

Hydrogenotrophic Methanogenesis Under Alkaline Conditions.

A cement-based geological disposal facility (GDF) is one potential option for the disposal of intermediate level radioactive wastes. The presence of both organic and metallic materials within a GDF provides the opportunity for both acetoclastic and hydrogenotrophic methanogenesis. However, for these processes to proceed, they need to adapt to the alkaline environment generated by the cementitious materials employed in backfilling and construction. Within the present study, a range of alkaline and neutral pH sediments were investigated to determine the upper pH limit and the preferred route of methane generation. In all cases, the acetoclastic route did not proceed above pH 9.0, and the hydrogenotrophic route dominated methane generation under alkaline conditions. In some alkaline sediments, acetate metabolism was coupled to hydrogenotrophic methanogenesis via syntrophic acetate oxidation, which was confirmed through inhibition studies employing fluoromethane. The absence of acetoclastic methanogenesis at alkaline pH values (>pH 9.0) is attributed to the dominance of the acetate anion over the uncharged, undissociated acid. Under these conditions, acetoclastic methanogens require an active transport system to access their substrate. The data indicate that hydrogenotrophic methanogenesis is the dominant methanogenic pathway under alkaline conditions (>pH 9.0).

Resilience of methane cycle and microbial functional genes to drought and flood in an alkaline wetland: A metagenomic analysis

Alkaline wetlands distributed in arid or semi-arid areas are hotspots of methane (CH4) emissions. Periods of drought and flood, although regular, are stressful events encountered by methanogenic anaerobes in alkaline wetlands. To investigate the response of the CH4 cycle of alkaline wetlands to such stresses, we take Zhalong wetland as an example, then determined the CH4 flux and soil microbiomes in the wetland during wet, dry, and flooded periods. The in-situ CH4 flux in the wet period was 9.55–17.29 mg‧m−2‧h−1, but sharply degraded to 3.37–6.61 mg‧m−2‧h−1 in the dry period. It resumed to 4.51–20.80 mg‧m−2‧h−1 when the wetland was flooded again, which indicated that methanogenesis is quite resilient to drought. Syntrophic acetogenesis, and subsequently aceticlastic methanogenesis, were the dominant methanogenic pathways and resisted drought. Members belonging to Syntrophobacterales were the dominant syntrophic acetogens. They enter a viable but non-culturable (VBNC) state to resist drought. The dominant Methanosarcinales have the ability to repair reactive oxygen species damage during dry periods. The community of CH4 sink was governed by anaerobic methanotrophs, which entered a VBNC state or used repair systems to survive dry periods. This study revealed the responses of the CH4 cycle and microbial functional genes to drought and flood in alkaline wetlands.

Obstacles faced by methanogenic archaea originating from substrate-driven toxicants in anaerobic digestion

Anaerobic digestion (AD) is used to treat waste and produce bioenergy. However, toxicants, which originate from the substrate, can inhibit or damage the digestion process. Methanogenic archaea (MA), which are the executor in the methanogenesis stage, are more sensitive than bacteria to these toxicants. This review discusses the effects of substrate-driven toxicants, namely, antibiotics, H2S and sulfate, heavy metals (HMs), long-chain fatty acids (LCFAs), and ammonia nitrogen, on the activity of MAs, methanogenic pathways, and the inter-genus succession of MAs. The adverse effects of these five toxicants on MA include effects on pH, damages to cell membranes, the prevention of protein synthesis, changes in hydrogen partial pressure, a reduction in the bioavailability of trace elements, and hindrance of mass transfer. These effects cause a reduction in MA activity and the succession of MAs and methanogenic pathways, which affect AD performance. Under the stress of these toxicants, succession occurs among HA (hydrogenotrophic methanogen), AA (acetoclastic methanogen), and MM (methylotrophic methanogen), especially HA gradually replaces AA as the dominant MA. Simultaneously, the dominant methanogenic pathway also changes from the aceticlastic pathway to other methanogenic pathways. A comprehensive understanding of the impact of toxicants on MA permits more specific targeting when developing strategies to mitigate or eliminate the effects of these toxicants.

Highly efficient anaerobic co-digestion of food waste and horticultural waste using a three-stage thermophilic bioreactor: Performance evaluation, microbial community analysis, and energy balance assessment

The thermophilic anaerobic co-digestion of food waste and horticultural waste with a three-stage bioreactor was investigated to produce renewable energy, focusing on performance evaluation, microbial community analysis, and energy balance assessment. The results of this study indicated that the average methane yield (0.42 ± 0.06 L/gVS) in the bench-scale three-stage thermophilic bioreactor was approximately 45% and 31% higher than that of the one and two-stage bioreactors, respectively. The average volatile solid (VS) reduction achieved in the bench-scale three-stage thermophilic bioreactor was 63.0 ± 8.2%, which was 61% and 32% higher than that of the one and two-stage bioreactors, respectively. Pyrosequencing analysis of microbial communities in simulated thermophilic three-stage co-digestion experiments showed that bacterial phyla Firmicutes, Thermotogae and Bacteroidetes dominated in all three types of bioreactors. However, significant differences were observed at the genus level of bacterial communities between the one, two and three-stage bioreactors. Indeed, due to selective enrichment, the dominant methanogenic pathways in the three-stage bioreactor shifted from hydrogenotrophic methanogenesis at a low organic loading rate (OLR) to a balance of hydrogenotrophic and acetoclastic pathways at high OLR. Furthermore, assessments of the energy balance and the economic viability of the bench-scale system demonstrated that the three-stage thermophilic co-digestion could be a promising process for the treatment of food waste and horticultural waste as well as for bioenergy recovery at an industrial scale.

Methyl-compounds driven benthic carbon cycling in the sulfate-reducing sediments of South China Sea.

Methane is a potent greenhouse gas; methane production and consumption within seafloor sediments has generated intense interest. Anaerobic oxidation of methane (AOM) and methanogenesis (MOG) primarily occur at the depth of the sulfate-methane transition zone or underlying sediment respectively. Methanogenesis can also occur in the sulfate-reducing sediments through the utilization of non-competitive methylated compounds; however, the occurrence and importance of this process are not fully understood. Here, we combined a variety of data, including geochemical measurements, rate measurements and molecular analyses to demonstrate the presence of a cryptic methane cycle in sulfate-reducing sediments from the continental shelf of the northern South China Sea. The abundance of methanogenic substrates as well as the high MOG rates from methylated compounds indicated that methylotrophic methanogenesis was the dominant methanogenic pathway; this conclusion was further supported by the presence of the methylotrophic genus Methanococcoides. High potential rates of AOM were observed in the sediments, indicating that methane produced in situ could be oxidized simultaneously by AOM, presumably by ANME-2a/b as indicated by 16S rRNA gene analysis. A significant correlation between the relative abundance of methanogens and methanotrophs was observed over sediment depth, indicating that methylotrophic methanogenesis could potentially fuel AOM in this environment. In addition, higher potential rates of AOM than sulfate reduction rates at in situ methane conditions were observed, making alternative electron acceptors important to support AOM in sulfate-reducing sediment. AOM rates were stimulated by the addition of Fe/Mn oxides, suggesting AOM could be partially coupled to metal oxide reduction. These results suggest that methyl-compounds driven methane production drives a cryptic methane cycling and fuels AOM coupled to the reduction of sulfate and other electron acceptors.

Effects of activated carbon on mesophilic and thermophilic anaerobic digestion of food waste: Process performance and life cycle assessment

Anaerobic digestion (AD) is widely used to recover energy (in the form of methane) and nutrients (in the form of biofertilizer) from organic wastes, and adding conductive materials has been reported to be an effective way to enhance AD efficiency. However, the process performance and environmental sustainability of adding conductive materials into AD under different temperature conditions have not been addressed. In this study, four different groups of lab-scale reactors were operated under both mesophilic and thermophilic conditions and with/without activated carbon (AC) addition. Based on the lab-scale results, a pilot-scale reactor was used to highlight the effectiveness of activated carbon addition, and the derived data was used for a life cycle assessment (LCA) study. The results showed the enhancement of methane production by adding AC, particularly under thermophilic conditions. At the organic loading rate of 4.7 g VS/L/d, the average methane yield of the thermophilic AD reactors with AC was 550 mL CH4/g VS, which was significantly higher than that of the control reactors (i.e. 220 mL CH4/g VS). Adding AC into AD reactors accelerated the shift of dominant methanogenic pathways from aceticlastic methanogenesis to hydrogenotrophic methanogenesis by enriching Methanoculleus (84% of total archaea). The LCA results highlighted the environmental benefits of using activated carbon to enhance AD of food waste at both midpoint and endpoint levels based on ReCiPe2016 methodology.

Methanogenesis and secondary processes in a fractured claystone aquifer influenced by a former landfill site: Implications for the interpretation of dissolved gases

Similar to marine and/or freshwater sediments, dissolved methane and longer chained hydrocarbons are ubiquitous in all natural groundwater environments including highly productive porous aquifers. The isotopic and chemical characteristics of dissolved gases in groundwater related to municipal landfills commonly differ from other non-polluted environments. In the survey presented in this study we investigated the geochemical and isotopic variations of dissolved hydrocarbons in a unique study area with different well defined ecological micro-habitats influenced by a former landfill for industrial and special waste. The dissolved methane concentrations have generally remained stable over the last decade but vary by several orders of magnitude in different zones of the area. The dominant methanogenic pathways can be clearly distinguished in the different ecosystems. Methyl-type fermentation methanogenesis prevails in the shallow groundwater zone (5–15 m depth) in- and outside the landfill. Anaerobic methane oxidation coupled to sulfate reduction occurs in the shallow downstream area within a former leachate plume. At larger depths CO2 reduction is the predominant methane formation pathway except within the physically isolated landfill site. The high ethane concentrations in a small part of the study area are potentially related to the degradation of volatile organic compounds. The background ethane concentrations are higher in the deeper groundwater zones, indicating the coupling to the CO2 reduction methanogenic pathway. The propane concentrations in most wells are below the detection limit, except within the waste site.Secondary methane oxidation, which frequently occurs in the groundwater in all environments can alter the gas composition mimicing the thermogenic gas signature in the standard Bernard-diagram. The isotopic fractionation between co-existing methane and formation water and carbon dioxide, respectively, differs from that suggested in standard interpretational diagrams (e.g. Bernard-Diagram and Whiticar-plots). Thus, the standard evaluation based on methane hydrogen and carbon isotopes is also susceptible to an erroneous interpretation as non-biogenic dissolved gases in other groundwater environments.

Three-stage anaerobic co-digestion of food waste and waste activated sludge: Identifying bacterial and methanogenic archaeal communities and their correlations with performance parameters

A three-stage anaerobic digester setup was configured and evaluated for enhanced methane production during co-digestion of food waste and waste activated sludge and the corresponding bacterial and methanogen communities were characterized. Results showed that the average methane yield (0.496 L/gVS) in the three-stage digester was 13–52% higher than that of one- and two-stage digesters. Compared to controls, an increase of 12–47% in volatile solids reduction was achieved in the three-stage digester (69.3 ± 6.7%). Bacterial phyla Proteobacteria, Firmicutes and Bacteroidetes dominated in one-, two- and three-stage digester while genera Pseudomonas, Tissierella, and Petrimonas were selectively enriched in the three-stage digester due to functional segregation. Taxonomic analysis identified 8 dominant methanogen genera, of which Methanosarcina, Methanosaeta, Methanobacterium and Methanolinea collectively accounted for 80%. With increasing OLR and digester stage number, the dominant methanogenic pathway shifted from hydrogenotrophic pattern to acetoclastic pattern and reached a final synergy of these two.

Hydrogenotrophic methanogenesis dominates at high pH

One potential design for a geological disposal facility (GDF) for intermediate level radioactive waste (ILW) involves the use of a cement base grout which will establish a highly alkaline environment for extended time periods [1]. Methane generation by colonising microbes could impact the long-term performance of the facility by influencing gas pressures and potentially leading to the migration of 14C to the biosphere [1]. Sediments acquired from a wide-range of anthropogenic alkaline sites in the UK were used to develop acetoclastic and hydrogenotrophic methanogen enrichment cultures over a broad range of pH values (7.0–12.0). The generation of methane from hydrogen and acetate was assessed to determine the dominant methanogenic pathways. Archaeal community analysis via Illumina MiSeq was employed to describe the populations involved and the acetoclastic inhibitor methyl fluoride was utilised to confirm the lack of acetate-dependent methane generation under alkaline conditions. High pH (pH>9.0) microcosms employing alkaline sediments were dominated by hydrogen-consuming methanogens of the orders Methanobacteriales and Methanomicrobiales, with no acetate consumption detected under these conditions. In contrast, neutral pH microcosms employing control sediments were dominated by acetoclastic methanogens of the order Methanosarcinales and demonstrated high acetate consumption rates. The rate of acetate consumption and proportion of acetoclastic methanogens decreased in a linear fashion as the pH within cultures was increased, however hydrogen consumption rates remained stable up to pH 11.0. The data shown suggests hydrogenotrophic methanogenesis is the dominant methanogenic pathway at high pH which could have important consequences on gas pressures within a GDF.

The C-biogeochemistry of a Midwestern USA agricultural impoundment in context: Lake Decatur in the intensively managed landscape critical zone observatory

The damming of rivers has created hotspots for organic carbon sequestration and methane production on a global scale as the reservoirs intercept fluvial suspended and dissolved loads. To better understand how the C-biogeochemistry of a reservoir responds to watershed processes and evolves over time, Lake Decatur, located in the Intensively Managed Landscape Critical Zone Observatory (IML-CZO) was studied. Solid phase analyses (% organic C, C/N, δ13C, δ15N) of soils and sediments sampled from stream bank exposures, river suspensions, and the lake bottom were conducted to characterize organic C (OC) sources throughout the sedimentary system. Agriculturally-driven soil erosion rapidly altered lake bathymetry causing an evolution of sedimentary and OC deposition patterns, which in turn shaped where and when methane production occurred. A positive correlation between OC accumulation rate and porewater dissolved inorganic C (DIC) δ13C profiles indicates that methane generation is strongly influenced by OC burial rate. The sources of the lake bed particulate organic C (POC) have also evolved over time. Drowned vegetation and/or shoreline inputs were dominant initially in areas adjacent to the original river channel but were rapidly overwhelmed by the deposition of sediments derived from eroded agricultural soils. Eutrophication of the lake followed with the onset of heavy fertilizer application post-1960. This succession of sources is expected to be commonplace for reservoirs greater than ~ 50–60 years old in agricultural settings because of the relative timing of tillage and fertilizer practices. The 13C/12C ratios of methane from Lake Decatur were more depleted in 13C than what is commonly expected for freshwater sedimentary environments. The 13C-depletion suggests that CO2-reduction is the dominant methanogenic pathway rather than the anticipated acetate dissimilation process. The isotopic observations reveal that commonly held assumptions about methane production and its C-isotopic signature in freshwater systems are over-simplified and not strictly applicable to this system.

Relative importance of methylotrophic methanogenesis in sediments of the Western Mediterranean Sea

Microbial production of methane is an important terminal metabolic process during organic matter degradation in marine sediments. It is generally acknowledged that hydrogenotrophic and acetoclastic methanogenesis constitute the dominant pathways of methane production; the importance of methanogenesis from methylated compounds remains poorly understood. We conducted various biogeochemical and molecular genetic analyses to characterize substrate availability, rates of methanogenesis, and methanogen community composition, and further evaluated the contribution of different substrates and pathways for methane production in deltaic surface and subsurface sediments of the Western Mediterranean Sea. Major substrates representing three methanogenic pathways, including H2, acetate, and methanol, trimethylamine (TMA), and dimethylsulfide (DMS), were detected in the pore waters and sediments, and exhibited variability over depth and between sites. In accompanying incubation experiments, methanogenesis rates from various 14C labeled substrates varied as well, suggesting that environmental factors, such as sulfate concentration and organic matter quality, could significantly influence the relative importance of individual pathway. In particular, methylotrophic and hydrogenotrophic methanogenesis contributed to the presence of micromolar methane concentrations in the sulfate reduction zone, with methanogenesis from methanol accounting for up to 98% of the total methane production in the topmost surface sediment. In the sulfate-depleted zone, hydrogenotrophic methanogenesis was the dominant methanogenic pathway (67–98%), and enhanced methane production from acetate was observed in organic-rich sediment (up to 31%). Methyl coenzyme M reductase gene (mcrA) analysis revealed that the composition of methanogenic communities was generally consistent with the distribution of methanogenic activity from different substrates. This study provides the first quantitative assessment of methylotrophic methanogenesis in marine sediments and has important implications for marine methane cycling. The occurrence of methylotrophic methanogenesis in surface sediments could fuel the anaerobic oxidation of methane (AOM) in the shallow sulfate reduction zone. Release of methane produced from methylotrophic methanogenesis could be a source of methane efflux to the water column, thus influencing the benthic methane budgets.