Methanogenic Substrates Research Articles

Applicability of blue algae as an activator for microbial enhanced coal bed methane technologies

The blue algae can be used as a nitrogen agent for promoting biological coalbed methane, but its applicability and microbial mechanism in different microbial enhanced coalbed methane technologies kept unknown. This study evaluated the methanogenic efficiency of blue algae addition with a mass ratio of 10% under fermentative degradation and microbial electrolytic cell technologies, and studied the changes of coal microstructure, surface functional groups, organic components and microbiome. The results showed that the algae addition affected the micro-concave-convex structure, non-uniform distribution of micro-particles and micro-cracks of coals, and finally increased the methanogenic rate by 1.74–2.66 times. The algae addition mainly affected the coal organic components including hydroxyl structure, hydrocarbon structure, aliphatic oxygen-containing functional groups and aromatic structure, as well increased the humus acids and microbial metabolites in fermentation broth; among them, the increased metabolites showed great differences between different technologies. The algae addition mainly increased the genera belonging to phylum Bacillota (such as Bacillus and Clostridium) and methanogens (Methanosarcina and Methanoculleus). These Bacillota groups could degrade organic matter into acetate and methanol via pathways of glycolysis and benzoate degradation, which provided substrates for such methanogens. This study strengthened the effectiveness of blue algae in enhancing technologies for biological coalbed methane.

Glacier melting promotes methane emission via increased methanogenic activity in the foreland alpine meadow

Annual glacier melting alters hydrothermal conditions of the foreland alpine meadows, and causes significant fluctuations in methane (CH4) flux. Previously we found that Tibetan glacier foreland alpine meadow shifts to CH4 source from sink during the melting season, but the potential mechanisms remain unclear. This study, via combination of in-situ measurement of seasonal CH4 flux and survey of microbial species that may involve in CH4 metabolism, explores the causes of glacier melting on CH4 flux in a glacier foreland alpine meadow on Tibetan Plateau. We determined a pronounced CH4 emission (13.95 μg·m−2·h−1) in August (melting season) but CH4 uptake in June (−3.76 μg·m−2·h−1) and October (−17.77 μg·m−2·h−1), and 1.4-fold higher soil moisture in August than the other two months. This showed a direct correlation of CH4 flux with glacier melting increased soil water. Additionally, glacier melting caused more CH4 fluxes increase in hollows than in hummocks. Amplicon sequencing determined 126-fold higher abundance of mcrA, the methanogenic marker gene, in August than in June and October, and a higher relative abundance of a fungal phylum Mortierellomycota and syntrophic bacteria that convert the fatty acids, the degradation intermediates of organic complexes to CO2 and acetate, the methanogenic substrates like in August. However, no seasonal variation of pmoA, the marker gene of aerobic methanotrophs, was observed. It appears that glacier melting promotes the CH4 producing but not the consuming microorganisms, thus leading to increased CH4 emission. The findings of this work indicate that global warming resulted glacier melting would increase global CH4 emissions, and in turn worsens global warming, so an alarming positive feedback loop.

Bridging energy and microbes: How the anodic potential of microbial electrolysis cell improves anaerobic digestion

Enhanced methane production is typically attributed to cathodic electron transfer in microbial electrolysis cell coupled anaerobic digestion (MEC-AD). This study shifts focus to the overlooked anodic reactions, illuminating their integral role in methane production during MEC-AD. Experimentation at controlled anodic potentials established 0.6 V as optimal; this parameter yielded an 87 % increase in methane and a 53 % reduction in chemical oxygen demand. The anode enriched for electroactive bacteria, notably Pseudomonas, facilitating the oxidation of propionic and butyric acids into acetic acid, thus providing sufficient substrate for methanogens. The modulation of the NADH/NAD+ ratio by the anodic potential proved central to converting carbohydrates and volatile fatty acids into acetic acid. Selective potentials enriched microbes proficient in anaerobic digestion, corroborated by bioinformatics analyses indicating enzymatic upregulation in key pathways. This investigation provides actionable insights into optimizing MEC-AD systems, advancing bioenergy recovery technologies.

Methylotrophic substrates stimulated higher methane production than competitive substrates in mangrove sediments

Although mangrove forests can uptake atmospheric CO2 and store carbon as organic matter called “blue carbon”, it is also an important natural source of greenhouse gas methane. Methanogens are major contributors to methane and play important roles in the global carbon cycle. However, our understanding of the key microbes and metabolic pathways responsible for methanogenesis under specific substrates in mangrove sediments is still very limited. Here, we set an anaerobic incubation to evaluate the responses of methanogens in mangrove sediments from South China to the addition of diverse methanogenic substrates (H2/CO2, acetate, trimethylamine (TMA), and methanethiol (MT)) and further investigated the dynamics of the whole microbial community. Our results showed that diverse substrates stimulated methanogenic activities at different times. The stimulation of methanogenesis was more pronounced at early and late periods by the addition of methylotrophic substrates TMA and MT, respectively. The amplicon sequencing analysis showed that genus Methanococcoides was mainly responsible for TMA-utilized methanogenesis in mangrove sediment, while the multitrophic Methanococcus was most abundant in H2/CO2 and MT treatments. Apart from that, the bacteria enrichments of Syntrophotalea, Clostridium_sensu_stricto_12, Fusibacter in MT treatments might also be associated with the stimulation of methane production. In addition, the metagenomic analysis suggested that Methanosarcinaceae was also one of the key methanogens in MT treatments with different genomic information compared to that in TMA treatments. Finally, the total relative abundances of methanogenesis-related genes were also highest in TMA and MT treatments. These results will help advance our understanding of the contributions of different methanogenesis pathways and methanogens to methane emissions in mangrove sediments.

Insights into mechanisms of red mud promoting biogas production from waste activated sludge anaerobic digestion

Anaerobic digestion of waste activated sludge is usually limited by the low overall methane yield. The addition of exogenous materials is considered to be a potential method to improve anaerobic methane production. In this study, red mud was added into waste activated sludge anaerobic digester and its effects on anaerobic digestion were investigated. Results showed that methane production was improved with the addition of red mud, and almost reached maximum in 10 g/L red mud added reactor (124.4 mL/g VSS), which was about 24.0 % higher than control. Further investigation indicated that the key genes related to methane metabolism, the activity of the corresponding coenzymes and the rate of extracellular electron transfer were also increased in red mud added reactor. Batch experiments on the effects of red mud in each stage of waste activated sludge anaerobic digestion showed that red mud could improve solubilization, hydrolysis, and acidification processes, but inhibit the methanogenesis process. The potential mechanism behind the enhancement of methane production during whole waste activated sludge anaerobic digestion is that the addition of red mud can intensify the hydrolysis and acidification processes, thereby providing more substrates for methanogens and subsequently enhancing the activity of coenzymes related to methane production.

Rice breeding for low methane and high yields

Root exudates serve as a substrate for methanogens. Breeding efforts aimed at maximizing rice photosynthate allocation to grains may reduce methane emissions and increase yields. Kwon et al. report that a loss-of-function rice allele, GS3, mitigates methane emissions and increases yield by allocating more photosynthates to grains rather than roots.

Effect of micro-aeration on syntrophic and methanogenic activity in anaerobic sludge

Micro-aeration was shown to improve anaerobic digestion (AD) processes, although oxygen is known to inhibit obligate anaerobes, such as syntrophic communities of bacteria and methanogens. The effect of micro-aeration on the activity and microbial interaction in syntrophic communities, as well as on the potential establishment of synergetic relationships with facultative anaerobic bacteria (FAB) or aerobic bacteria (AB), was investigated. Anaerobic sludge was incubated with ethanol and increasing oxygen concentrations (0–5% in the headspace). Assays with acetate or H2/CO2 (direct substrates for methanogens) were also performed. When compared with the controls (0% O2), oxygen significantly decreased substrate consumption and initial methane production rate (MPR) from acetate or H2/CO2. At 0.5% O2, MPR from these substrates was inhibited 30–40%, and close to 100% at 5% O2. With ethanol, significant inhibition (>36%) was only observed for oxygen concentrations higher than 2.5%. Oxygen was consumed in the assays, pointing to the stimulation of AB/FAB by ethanol, which helped to protect the syntrophic consortia under micro-aerobic conditions. This highlights the importance of AB/FAB in maintaining functional and resilient syntrophic communities, which is relevant for real AD systems (in which vestigial O2 amounts are frequently present), as well as for AD systems using micro-aeration as a process strategy.Key points•Micro-aeration impacts syntrophic communities of bacteria and methanogens.•Oxygen stimulates AB/FAB, maintaining functional and resilient consortia.•Micro-aeration studies are critical for systems using micro-aeration as a process strategy.

Pyruvate-dependent growth of Methanosarcina acetivorans.

Methanogenesis is a key step during anaerobic biomass degradation. Methanogenic archaea (methanogens) are the only organisms coupling methanogenic substrate conversion to energy conservation. The range of substrates utilized by methanogens is limited, with acetate and H2+CO2 being the ecologically most relevant. The only single methanogenic energy substrate containing more carbon-carbon bonds than acetate is pyruvate. Only the aggregate-forming, freshwater methanogen Methanosarcina barkeri Fusaro was shown to grow on this compound. Here, the pyruvate-utilizing capabilities of the single-celled, marine Methanosarcina acetivorans were addressed. Robust pyruvate-dependent, methanogenic, growth could be established by omitting CO2 from the growth medium. Growth rates which were independent of the pyruvate concentration indicated that M. acetivorans actively translocates pyruvate across the cytoplasmic membrane. When 2-bromoethanesulfonate (BES) inhibited methanogenesis to more than 99%, pyruvate-dependent growth was acetogenic and sustained. However, when methanogenesis was completely inhibited M. acetivorans did not grow on pyruvate. Analysis of metabolites showed that acetogenesis is used by BES-inhibited M. acetivorans as a sink for electrons derived from pyruvate oxidation and that other, thus far unidentified, metabolites are produced.IMPORTANCEThe known range of methanogenic growth substrates is very limited and M. acetivorans is only the second methanogenic species for which growth on pyruvate is demonstrated. Besides some commonalities, analysis of M. acetivorans highlights differences in pyruvate metabolism among Methanosarcina species. The observation that M. acetivorans probably imports pyruvate actively indicates that the capabilities for heterotrophic catabolism in methanogens may be underestimated. The mostly acetogenic growth of M. acetivorans on pyruvate with concomitant inhibition of methanogenesis confirms that energy conservation of methanogenic archaea can be independent of methane formation.

Nitrous oxide inhibition of methanogenesis represents an underappreciated greenhouse gas emission feedback.

Methane (CH4) and nitrous oxide (N2O) are major greenhouse gases that are predominantly generated by microbial activities in anoxic environments. N2O inhibition of methanogenesis has been reported, but comprehensive efforts to obtain kinetic information are lacking. Using the model methanogen Methanosarcina barkeri strain Fusaro and digester sludge-derived methanogenic enrichment cultures, we conducted growth yield and kinetic measurements and showed that micromolar concentrations of N2O suppress the growth of methanogens and CH4 production from major methanogenic substrate classes. Acetoclastic methanogenesis, estimated to account for two-thirds of the annual 1 billion metric tons of biogenic CH4, was most sensitive to N2O, with inhibitory constants (KI) in the range of 18-25μM, followed by hydrogenotrophic (KI, 60-90μM) and methylotrophic (KI, 110-130μM) methanogenesis. Dissolved N2O concentrations exceeding these KI values are not uncommon in managed (i.e. fertilized soils and wastewater treatment plants) and unmanaged ecosystems. Future greenhouse gas emissions remain uncertain, particularly from critical zone environments (e.g. thawing permafrost) with large amounts of stored nitrogenous and carbonaceous materials that are experiencing unprecedented warming. Incorporating relevant feedback effects, such as the significant N2O inhibition on methanogenesis, can refine climate models and improve predictive capabilities.

Methane emissions and the microbial community in flooded paddies affected by the application of Fe-stabilized natural organic matter

Redox chemistry involving the quinone/phenol cycling of natural organic matter (NOM) is known to modulate microbial respiration. Complexation with metals or minerals can also affect NOM solubilization and stability. Inspired by these natural phenomena, a new soil amendment approach was suggested to effectively decrease methane emissions in flooded rice paddies. Structurally stable forms of NOM such as lignin and humic acids (HAs) were shown to decrease methane gas emissions in a vial experiment using different soil types and rice straw as a methanogenic substrate, and this inhibitory behavior was likely enhanced by ferric ion–NOM complexation. A mechanistic study using HAs revealed that complexation facilitated the slow release of the humic components. Interestingly, borohydride-based reduction, which transformed quinone moieties into phenols, caused the HAs to lose their inhibitory capacity, suggesting that the electron-accepting ability of HAs is vital for their inhibitory effect. In rice field tests, the humic–metal complexes were shown to successfully mitigate methane generation, while carbon dioxide emissions were relatively unchanged. Microbial community analysis of the rice fields by season revealed a decrease in specific cellulose-metabolizing and methanogenic genera associated with methane emissions. In contrast, the relative abundance of Thaumarchaeota and Actinomycetota, which are associated with NOM and recalcitrant organics, was higher in the presence of Fe-stabilized HAs. These microbial dynamics suggest that the slow release of humic components is effective in modulating the anoxic soil microbiome, possibly due to their electron-accepting ability. Given the simplicity, cost-effectiveness, and soil-friendly nature of complexation processes, Fe-stabilized NOM represents a promising approach for the mitigation of methane emissions from flooded rice paddies.

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.

Stable Isotope Approach to Assess the Production and Consumption of Methylphosphonate and Its Contribution to Oxic Methane Formation in Surface Waters.

Recent studies in aquatic environments have indicated that microbial methane production is not limited to strictly anoxic conditions and is widespread in the oxic water column. Based on recent investigations proposing linkage between the microbial turnover of methylphosphonate (MPn) and the widespread methane oversaturation in surface waters, we conducted an MPn/13C-MPn tracer approach that combines liquid chromatography-mass spectrometry and gas chromatography-stable isotope ratio mass spectrometry to assess concentrations of the MPn tracer and its contribution to oxic methane formation. In our study, conducted during summer 2020 in the Baltic Sea, we show that MPn is a potent methanogenic substrate in the surface water. However, we found that MPn was produced within the surface and subthermocline water bodies and that its turnover was not limited to the phosphorus-stressed and cyanobacteria-rich surface water. However, our study revealed that most of the MPn was probably degraded via alternative pathways, not releasing methane. Our assessment indicates that the contribution of the MPn degradation pathway only contributed marginally to oxic methane production at the study site in the Baltic Sea and that a variety of methanogenic pathways are probably responsible for the surface-water methane enrichments.

Conversion of natural coastal wetlands to mariculture ponds dramatically decreased methane production by reducing substrate availability

Coastal wetlands are increasingly being converted into aquaculture ponds to meet growing global demand for fish protein. Coastal wetlands conversion has been predicted to result in significant carbon (C) emissions; however, a mechanistic understanding of the effects of coastal wetland conversion on soil’s capacity to produce greenhouse gases remains lacking. Here, integrated biogeochemical investigations on methanogenic substrates, community structures, and activities were conducted in a coastal Spartina alterniflora saltmarsh and three saltmarsh-converted mariculture ponds aged 6, 13, and 20 years. Saltmarsh conversion into mariculture ponds decreased CH4 production potential from 353 to 16.3–78.3 µg CH4 kg−1 d−1. The concentrations of dissolved organic C and non-competitive methanogenic substrate, trimethylamine, were reduced by 84.8% and 79.7%, respectively, whereas methanogens abundance decreased by 43.7%, probably due to the decreased C input following conversion. Among the ponds, CH4 production potential, methanogenic substrates (acetate and trimethylamine), and methanogen abundance decreased with conversion chronosequence. However, the dominant methanogen group did not shift after land conversion, and the potential methylamine-utilizing Methanosarcinaceae accounted for 86.5% and 85.3–89.7% of the total populations in the S. alterniflora saltmarsh and mariculture ponds, respectively, implying that methylotrophic methanogenesis was the predominant pathway of CH4 production in both ecosystems. The results indicated that the conversion of natural coastal saltmarsh to mariculture ponds decreased CH4 production by reducing substrate availability and methanogen abundance, and CH4 production potential in mariculture ponds was more efficiently suppressed with conversion chronosequence.

Genetic and Physiological Probing of Cytoplasmic Bypasses for the Energy-Converting Methyltransferase Mtr in Methanosarcina acetivorans.

Methanogenesis is a unique energy metabolism carried out by members of the domain Archaea. Unlike most other methanogens, which reduce CO2 to methane with hydrogen as the electron donor, Methanosarcina acetivorans is able to grow on methylated compounds, on acetate, and on carbon monoxide (CO). These substrates are metabolized via distinct yet overlapping pathways. For the use of any single methanogenic substrate, the membrane-integral, energy-converting N5-methyl-tetrahydrosarcinapterin (H4SPT):coenzyme M (HS-CoM) methyltransferase (Mtr) is required. It was proposed that M. acetivorans can bypass the methyl transfer catalyzed by Mtr via cytoplasmic activities. To address this issue, conversion of different energy substrates by an mtr deletion mutant was analyzed. No significant methyl transfer from H4SPT to HS-CoM could be detected with CO as the electron donor. In contrast, formation of methane and CO2 in the presence of methanol or trimethylamine was indicative of an Mtr bypass in the oxidative direction. As methane thiol and dimethyl sulfide were transiently produced during methylotrophic methanogenesis in the mtr mutant, involvement in this process of methyl sulfide-dependent methyltransferases (Mts) was analyzed in a strain lacking both the Mts system and Mtr. It could be unequivocally demonstrated that the Mts system is not involved in bypassing Mtr, thereby ruling out previous proposals. Conversion of [13C]methanol indicated that in the absence of Mtr M. acetivorans provides the reducing equivalents for methyl-S-CoM reduction to methane by oxidizing (an) intracellular compound(s) to CO2 rather than disproportioning the source of methyl groups. Thus, no in vivo Mtr bypass appears to exist in M. acetivorans. IMPORTANCE Methanogenic archaea possess only a limited number of chemiosmotic coupling sites in their respiratory chains. Among them, N5-methyl-H4SPT:HS-CoM methyltransferase (Mtr) is the most widely distributed. Previous observations led to the conclusion that Methanosarcina acetivorans is able to bypass this reaction via methyl sulfide-dependent methyltransferases (Mts). However, strains lacking Mtr are not able to produce methane from CO. Also, these strains are unable to oxidize methylated substrates to CO2, in contrast to observations in the close relative Methanosarcina barkeri. The results also highlight the sole function of the Mts system in methyl sulfide metabolism. Thus, no in vivo Mtr bypass appears to exist in M. acetivorans.

A rationale for higher ratios of CH4 to CO2 production in warmer anoxic freshwater sediments and soils

A rationale for higher ratios of <scp>CH₄</scp> to <scp>CO₂</scp> production in warmer anoxic freshwater sediments and soils

Characterisation of Methane Production Pathways in Sediment of Overwashed Mangrove Forests

Methane (CH4) emissions in mangrove ecosystems may complicate the ecosystem’s potential carbon offset for climate change mitigation. Microbial processes and the mass balance of CH4 in mangrove sediment are responsible for the emissions from the ecosystems. This is the follow up of our previous research which found the super saturation of CH4 in the pore water of mangrove sediment compared to atmospheric CH4 and the lack of a correlation between pore water sulphate and CH4 concentrations. This study is going to investigate methane production pathways in the sediment of overwashed mangrove forests. Two approaches were used to study methanogens here: (1) the spread plate count method and the quantitative polymerase chain reaction (qPCR) method, and (2) laboratory experiments with additional methanogenic substrates (methanol, acetate, and hydrogen) to determine which substrates are more conducive to methane production. According to the qPCR method, methanogen abundance ranged from 72 to 6 × 105 CFU g−1 sediment, while SRB abundance ranged from 2 × 102 to 2 × 105 CFU g−1 sediment. According to the plate count method, the abundance of methylotrophic methanogens (the only group of methanogens capable of competing with SRBs) ranged from 8.3 × 102 to 5.1 × 104 CFU g−1, which is higher than the abundance of the other group of methanogens (0 to 7.7 × 102 CFU g−1). The addition of methanol to the sediment slurry, a substrate for methylotropic methanogens, resulted in a massive production of CH4 (up to 9 × 104 ppm) and intriguingly the control treatments with autoclaving did not kill methanogens. These findings suggested that mangrove ecosystems in the marine environment provide favourable conditions for methanogens and further characterisation of the methanogen involved in the process is required. As a result, future research in this ecosystem should include methane production in carbon offset calculations, particularly due to methylotropic methanogenesis.

Destruction of Mycotoxins in Poultry Waste under Anaerobic Conditions within Methanogenesis Catalyzed by Artificial Microbial Consortia.

To reduce the toxicity of modern feeds polluted by mycotoxins, various sorbents are added to them when feeding animals. A part of the mycotoxins is excreted from the body of animals with these sorbents and remains in the manure. As a result, bulk animal wastes containing mixtures of mycotoxins are formed. It is known that it is partially possible to decrease the initial concentration of mycotoxins in the process of anaerobic digestion (AD) of contaminated methanogenic substrates. The aim of this review was to analyze the recent results in destruction of mycotoxins under the action of enzymes present in cells of anaerobic consortia catalyzing methanogenesis of wastes. The possible improvement of the functioning of the anaerobic artificial consortia during detoxification of mycotoxins in the bird droppings is discussed. Particular attention was paid to the possibility of effective functioning of microbial enzymes that catalyze the detoxification of mycotoxins, both at the stage of preparation of poultry manure for methanogenesis and directly in the anaerobic process itself. The sorbents with mycotoxins which appeared in the poultry wastes composed one of the topics of interest in this review. The preliminary alkaline treatment of poultry excreta before processing in AD was considered from the standpoint of effectively reducing the concentrations of mycotoxins in the waste.

Microbiome of High-Rank Coal Reservoirs in the High-Production Areas of the Southern Qinshui Basin.

To study the distribution features of microorganisms in distinct hydrological areas of the southern Qinshui Basin, C-N-S microorganisms were studied using 16S RNA sequencing, metagenome sequencing and geochemical technologies, showing the high sensitivity of microorganisms to the hydrodynamic dynamics of coal. The hydrodynamic intensity of the #3 coal gradually decreased from the runoff areas to the stagnant areas. The stagnant zones have higher reservoir pressure, methane content, δ13CDIC and TDS and lower SO42-, Fe3+ and NO3- concentrations than the runoff areas. C-N-S-cycling microorganisms, including those engaged in methanogenesis, nitrate respiration, fermentation, nitrate reduction, dark oxidation of sulfur compounds, sulfate respiration, iron respiration, chlorate reduction, aromatic compound degradation, denitrification, ammonification and nitrogen fixation, were more abundant in the stagnant areas. The relative abundance of C-N-S functional genes, including genes related to C metabolism (e.g., mcr, mer, mtr, fwd and mtd), N metabolism (e.g., nifDKH, nirK, narGHI, nosZ, amoB, norC and napAB) and sulfur metabolism (e.g., dsrAB and PAPSS), increased in the stagnant zones, indicating that there was active microbiological C-N-S cycling in the stagnant areas. The degradation and fermentation of terrestrial plant organic carbon and coal seam organic matter could provide substrates for methanogens, while nitrogen fixation and nitrification can provide nitrogen for methanogens, which are all favorable factors for stronger methanogenesis in stagnant areas. The coal in the study area is currently in the secondary biogenic gas generation stage because of the rising of the strata, which recharges atmospheric precipitation. The random forest model shows that the abundance of C-N-S microorganisms and genes could be used to distinguish different hydrological zones in coal reservoirs. Since stagnant zones are usually high-gas-bearing zones and high-production areas of CBM exploration, these microbiological indicators can be used as effective parameters to identify high-production-potential zones. In addition, nitrate respiration and sulfate respiration microorganisms consumed NO3- and SO42-, causing a decrease in the content of these two ions in the stagnant areas.

Improving Methane Production from Waste Activated Sludge Assisted by Fe(II)-Activated Peroxydisulfate Pretreatment via Anaerobic Digestion: Role of Interspecific Syntrophism Mediated by Sulfate-Reducing Bacteria

Sulfate radical generated via peroxydisulfate (PDS) activation has been extensively applied to energy recovery from waste activated sludge (WAS) during anaerobic digestion (AD), while the residual sulfate is still a limitation for its further application. In this study, a novel coupling strategy of Fe(II)-activated PDS pretreatment [Fe(II)_PDS] with sulfate-reducing bacteria (SRB) mediated was explored to enhance methane yield from WAS during AD. Results showed that the similar methane yield was obtained in Fe(II)_PDS + SRB (16.6 mL/g VSS) and zero-valent iron (ZVI)_PDS + SRB (16.9 mL/g VSS), which enhanced by 19.9–56.5% than other groups, while the hydrolysis rate constant (0.0065 h–1) in Fe(II)_PDS + SRB was much higher than that observed in ZVI_PDS + SRB (0.0046 h–1). Meanwhile, the concentration of short-chain fatty acids (SCFAs) peaked at 360.9 mg COD/g VSS (5 d) in the Fe(II)_PDS + SRB group, with the acetic acid (HAc) and propionic acid (HPr) accumulative proportion of 73.5%, and the total concentration promoted by 28.5% than that of the ZVI_PDS + SRB group, which provided a more ideal substrate for methanogens. The ultimate utilization efficiency of SCFAs reached 62.4% in the Fe(II)_PDS + SRB group. Moreover, MiSeq sequencing, canonical correspondence analysis, and molecular ecological network analysis revealed the intrinsic interaction of the functional microbial consortia, that is, anaerobic fermentation bacteria, SRB, nitrate-reducing bacterium, and acetotrophic methanogen, with the abundances of 9.52, 2.4, 4.93, and 77.7% in the Fe(II)_PDS + SRB group. Considering the treatment performance and the difficulty of the subsequent disposal, Fe(II) may be the superior activator than ZVI for free radicals (i.e., SO4•– and HO•) generation from PDS and further played the important role for interspecific syntrophism coupling with SRB during AD. Results of this study may provide a promising way for energy recovery from WAS.

Impacts of organic loading rate and hydraulic retention time on organics degradation, interspecies interactions and functional traits in thermophilic anaerobic co-digestion of food waste and sewage sludge

This study provided novel insights into the effects of organic loading rate (OLR) and hydraulic retention time (HRT) on thermophilic anaerobic co-digestion of food waste and sewage sludge. The obtained maximum methane (CH4) yield of 328 ± 4 mL CH4/g CODfed at HRT of 15 days (OLR = 5.8 g VS/L/d) was partly attributable to the enhanced acidogenesis, acetogenesis, and methanogenesis phases. The increased key enzyme activities, particularly acetate kinase (improved by 5.2-fold), providing substantial methanogenic substrates for efficient CH4 production. The functional syntrophs that were related to syntrophic decarboxylation, novel acetate oxidation & reductive acetyl-CoA, and β-oxidation pathways could drive trophic interactions with methanogens. This markedly stimulated hydrogenotrophic Methanoculleus thermophilus metabolism and concomitantly enriched mixotrophic Methanosarcina thermophila. The distinctive cross-feeding interspecies interactions significantly affected the assembly and dynamics of thermophilic consortia. These findings shed light on the physicochemical and microbial mechanisms of HRT- and OLR-dependent enhancement of methanogenesis.