Hydrogenotrophic Methanogenesis Research Articles

Comprehensive insights into the effects of acidogenic off-gas utilization on successive biogas production, microbial community structure and metabolite distribution during two-stage anaerobic digestion

Although two-stage anaerobic digestion (TSAD) technology has been investigated, the mechanisms regarding the impact of acidogenic off-gas (AOG) on successive methane production have not been well addressed. In this study, a novel TSAD system was designed. Food waste, as the main substrate, was co-digested with chicken manure and corn straw. The acidogenic gas beyond atmospheric pressure was introduced into the bottom of the methanogenesis reactor through a stainless steel diffuser. Results showed the addition of AOG increased the methane yield from 435.2 to 597.1 mL/g VSin in successive methanogenesis stage, improved by 37.2%, and increased the energy yield from 9.0 to 11.3 kJ/g VSsubstrate. However, the theoretical contribution of hydrogenotrophic methanogenesis using H2 contained in AOG was only 15.2% of the increased methane yield. After the addition of AOG, the decreased levels of ammonia nitrogen and butyrate indicate that the stability of the AD system was improved. The electron transfer system and co-enzyme F420 activity were enhanced; however, the decrease in acetate kinase activity indicates aceticlastic methanogenesis may have been weakened. The microbial diversity and species richness were improved by the added AOG. Methanosarcina was more competitive than Methanothermobacter, enhancing the syntrophic effect. The relative abundance of protein degradation bacteria norank_f__Anaerolineaceae and lipid degradation bacteria Syntrophomonas was increased. Metabolite analysis confirmed that the addition of AOG promoted amino acid metabolism, the biosynthesis of other secondary metabolism and lipid metabolism. The improved degradation of recalcitrant organic components (lipids and proteins) in food waste was responsible for the increased methane yield. This study provides an in-depth understanding of the impact of AOG utilization on successive methane production and has practical implications for the treatment of food waste.

Alkaline hydrogenotrophic methanogenesis in Methanococcus vannielii at low carbon dioxide concentrations

Capture of carbon dioxide from the atmosphere is challenging and thermodynamically expensive because of its dilute concentration (421 ppm, December 2023). Utilizing the concentrating effect of dissolving atmospheric CO2 in alkaline solution to form dissolved inorganic carbon (DIC), we explored the potential of an alkalotolerant hydrogenotrophic methanogen, Methanococcus vannielii, to capture and convert CO2 at low partial pressures to methane. Kinetic constants for CO2 reduction to CH4 were determined from experiments in serum bottles at low pCO2 at 30˚C. At pH 7, 8, and 9, the apparent KM is 0.4, 1.2, and 1.2 mM DIC and the apparent vmax is 5.6, 7.8, and 2.9 mmol CH4 L−1 OD−1 hr−1, respectively. Using atmosphere-equilibrated DIC concentrations at pH 7, 8, and 9, methane formation rates were 1.1, 2.3, and 2.1 mmol CH4 L−1 OD−1 hr−1, respectively. Our data show that alkaline hydrogenotrophic methanogenesis is an alternative to photosynthetic CO2 fixation for biological capture and conversion of CO2 at atmospheric concentrations at reasonable rates.

The Selenoproteome as a Dynamic Response Mechanism to Oxidative Stress in Hydrogenotrophic Methanogenic Communities.

Methanogenesis is a critical process in the carbon cycle that is applied industrially in anaerobic digestion and biogas production. While naturally occurring in diverse environments, methanogenesis requires anaerobic and reduced conditions, although varying degrees of oxygen tolerance have been described. Microaeration is suggested as the next step to increase methane production and improve hydrolysis in digestion processes; therefore, a deeper understanding of the methanogenic response to oxygen stress is needed. To explore the drivers of oxygen tolerance in methanogenesis, two parallel enrichments were performed under the addition of H2/CO2 in an environment without reducing agents and in a redox-buffered environment by adding redox mediator 9,10-anthraquinone-2,7-disulfonate disodium. The cellular response to oxidative conditions is mapped using proteomic analysis. The resulting community showed remarkable tolerance to high-redox environments and was unperturbed in its methane production. Next to the expression of pathways to mitigate reactive oxygen species, the higher redox potential environment showed an increased presence of selenocysteine and selenium-associated pathways. By including sulfur-to-selenium mass shifts in a proteomic database search, we provide the first evidence of the dynamic and large-scale incorporation of selenocysteine as a response to oxidative stress in hydrogenotrophic methanogenesis and the presence of a dynamic selenoproteome.

A Carbon Source in a Carbon Sink: Carbon Dioxide and Methane Dynamics in Open‐Water Peatland Pools

AbstractPeatlands store organic carbon available for decomposition and transfer to neighboring water bodies, which can ultimately generate carbon dioxide (CO2) and methane (CH4) emissions. The objective of this study was to clarify the biogeochemical functioning of open‐water peatland pools and their influence on carbon budgets at the ecosystem and global scale. Continuously operated automated equipment and monthly manual measurements were used to describe the CO2 and CH4 dynamics in boreal ombrotrophic peatland pools and porewater (Québec, Canada) over the growing seasons 2019 and 2020. The peat porewater stable carbon isotope ratios (δ13C) for both CO2 (median δ13C‐CO2: −3.8‰) and CH4 (median δ13C‐CH4: −64.30‰) suggested that hydrogenotrophic methanogenesis was the predominant degradation pathway in peat. Open‐water pools were supersaturated in CO2 and CH4 and received most of these dissolved carbon greenhouse gases (C‐GHG) from peat porewater input. Throughout the growing season, higher CO2 concentrations and fluxes in pools were measured when the water table was low—suggesting a steady release of CO2 from deep peat porewater. Higher CH4 ebullition and diffusion occurred in August when bottom water and peat temperatures were the highest. While this study demonstrates that peatland pools are chimneys of CO2 and CH4 stored in peat, it also shows that the C‐GHG concentrations and flux rates in peat pools are comparable to other aquatic systems of the same size. Although peatlands are often considered uniform entities, our study highlights their biogeochemical heterogeneity, which, if considered, substantially influences their net carbon balance with the atmosphere.

Effect of hydrothermal-acid pretreatment on methane yield and microbial community in anaerobic digestion of rice straw

Hydrothermal pretreatment has been proposed to enhance straw methane yield during anaerobic digestion recently. However, the combined effect of hydrothermal and organic acid pretreatment (HTOAP) needs further investigation. This study identified optimal pretreatment at 120 °C with 3 % acetic acid for 24 h by orthogonal design method. The HTOAP increased the reducing sugar content by destroying the lignocellulosic structure. A 79 % increment of methane production after HTOAP was observed compared to the untreated group. Microbial analysis showed that HTOAP enriched the relative abundance of lignocellulose-degraders, such as W5053, Thermanaerovibrio, Caldicoprobacter, as well as the syntrophic acetate oxidizing bacteria Syntrophaceticus. Moreover, Methanobacterium conducted hydrogenotrophic methanogenesis dominantly. Furthermore, the potential function analysis showed that HTOAP stimulated the expression of key enzymes in the hydrogenotrophic pathway, including carbon-monoxide dehydrogenase (EC 1.2.7.4) and coenzyme F420 hydrogenase (EC 1.12.98.1). This investigation illustrated the potential of HTOAP of rice straw to facilitate methane production.

In-situ methane enrichment in anaerobic digestion of food waste slurry by nano zero-valent iron: Long-term performance and microbial community succession

In this work, nano zero-valent iron (nZVI) was added to a lab-scale continuous stirring tank reactor (CSTR) for food waste slurry treatment, and the effect of dosing rate and dosage of nZVI were attempted to be changed. The results showed that anaerobic digestion (AD) efficiency and biomethanation stability were optimum under the daily dosing and dosage of 0.48 g/gTCOD. The average daily methane (CH4) yield reached 495.38 mL/gTCOD, which was 43.65% higher than that at control stage, and the maximum CH4 content reached 95%. However, under single dosing rate conditions, high nZVI concentrations caused microbial cell rupture and loosely bound extracellular polymeric substances (LB-EPS) precipitation degradation. The daily dosing rate promoted the hydrogenotrophic methanogenesis pathway, and the activity of coenzyme F420 increased by 400.29%. The microbial analysis indicated that daily addition of nZVI could promote the growth of acid-producing bacteria (Firmicutes and Bacteroidetes) and methanogens (Methanothrix).

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.

Metabolic and physiological adaptations of microalgal growth-promoting bacterium Azospirillum brasilense growing under biogas atmosphere: a microarray-based transcriptome analysis.

Using microalgal growth-promoting bacteria (MGPB) to improve the cultured microalga metabolism during biotechnological processes is one of the most promising strategies to enhance their benefits. Nonetheless, the culture condition effect used during the biotechnological process on MGPB growth and metabolism is key to ensure the expected positive bacterium growth and metabolism of microalgae. In this sense, the present research study investigated the effect of the synthetic biogas atmosphere (75% CH4-25% CO2) on metabolic and physiological adaptations of the MGPB Azospirillum brasilense by a microarray-based transcriptome approach. A total of 394 A. brasilense differentially expressed genes (DEGs) were found: 201 DEGs (34 upregulated and 167 downregulated) at 24h and 193 DEGs (140 upregulated and 53 downregulated) under the same conditions at 72h. The results showed a series of A. brasilense genes regulating processes that could be essential for its adaptation to the early stressful condition generated by biogas. Evidence of energy production is shown by nitrate/nitrite reduction and activation of the hypothetical first steps of hydrogenotrophic methanogenesis; signal molecule modulation is observed: indole-3-acetic acid (IAA), riboflavin, and vitamin B6, activation of Type VI secretion system responding to IAA exposure, as well as polyhydroxybutyrate (PHB) biosynthesis and accumulation. Moreover, an overexpression of ipdC, ribB, and phaC genes, encoding the key enzymes for the production of the signal molecule IAA, vitamin riboflavin, and PHB production of 2, 1.5 and 11 folds, respectively, was observed at the first 24h of incubation under biogas atmosphere Overall, the ability of A. brasilense to metabolically adapt to a biogas atmosphere is demonstrated, which allows its implementation for generating biogas with high calorific values and the use of renewable energies through microalga biotechnologies.

Time-shifted expression of acetoclastic and methylotrophic methanogenesis by a single Methanosarcina genomospecies predominates the methanogen dynamics in Philippine rice field soil

BackgroundThe final step in the anaerobic decomposition of biopolymers is methanogenesis. Rice field soils are a major anthropogenic source of methane, with straw commonly used as a fertilizer in rice farming. Here, we aimed to decipher the structural and functional responses of the methanogenic community to rice straw addition during an extended anoxic incubation (120 days) of Philippine paddy soil. The research combined process measurements, quantitative real-time PCR and RT-PCR of particular biomarkers (16S rRNA, mcrA), and meta-omics (environmental genomics and transcriptomics).ResultsThe analysis methods collectively revealed two major bacterial and methanogenic activity phases: early (days 7 to 21) and late (days 28 to 60) community responses, separated by a significant transient decline in microbial gene and transcript abundances and CH4 production rate. The two methanogenic activity phases corresponded to the greatest rRNA and mRNA abundances of the Methanosarcinaceae but differed in the methanogenic pathways expressed. While three genetically distinct Methanosarcina populations contributed to acetoclastic methanogenesis during the early activity phase, the late activity phase was defined by methylotrophic methanogenesis performed by a single Methanosarcina genomospecies. Closely related to Methanosarcina sp. MSH10X1, mapping of environmental transcripts onto metagenome-assembled genomes (MAGs) and population-specific reference genomes revealed this genomospecies as the key player in acetoclastic and methylotrophic methanogenesis. The anaerobic food web was driven by a complex bacterial community, with Geobacteraceae and Peptococcaceae being putative candidates for a functional interplay with Methanosarcina. Members of the Methanocellaceae were the key players in hydrogenotrophic methanogenesis, while the acetoclastic activity of Methanotrichaceae members was detectable only during the very late community response.ConclusionsThe predominant but time-shifted expression of acetoclastic and methylotrophic methanogenesis by a single Methanosarcina genomospecies represents a novel finding that expands our hitherto knowledge of the methanogenic pathways being highly expressed in paddy soils.7k5euw-hvBknz_jds4sXXfVideo

Synergistic effect of hydrothermal sludge and food waste in the anaerobic co-digestion process: microbial shift and dewaterability.

While thermal hydrolysis technology is commonly employed for sewage sludge treatment in extensive wastewater treatment facilities, persistent challenges remain, including issues such as ammonia-induced digestive inhibition and reduced productivity stemming from nutrient deficiency within the hydrothermal sludge. In this study, the effects of hydrothermal sludge-to-food waste mixing ratios and fermentation temperatures on anaerobic co-digestion were systematically investigated through a semi-continuous experiment lasting approximately 100days. The results indicated that anaerobic co-digestion of hydrothermal sludge and food waste proceeded synergistically at any mixing ratio, and the synergistic effect is mainly attributed to the improvement of carbohydrate removal and digestive system stability. However, thermophilic digestion did not improve the anaerobic performance and methane yield. On the contrary, mesophilic digestion performed better in terms of organic matter removal, especially in the utilization of soluble carbohydrates, soluble proteins, and VFAs. Microbial community analysis revealed that the transition from mesophilic to thermophilic anaerobic co-digestion prompts changes in the methane-producing pathways. Specifically, the transition entails a gradual shift from pathways involving acetoclastic and hydrogenotrophic methanogenesis to a singular hydrogenotrophic methanogenesis pathway. This shift is driven by thermodynamic tendencies, as reflected in Gibbs free energy, as well as environmental factors like ammonia nitrogen and volatile fatty acids. Lastly, it is worth noting that the introduction of food waste did lead to a reduction in cake solids following dewatering. Nevertheless, it was observed that thermophilic digestion had a positive impact on dewatering performance.

Switching from wet to dry anaerobic digestion of food waste with different dilution times under no mechanical mixing condition

Previous research on anaerobic digestion of food waste has primarily focused on either wet or dry anaerobic digestion (AD), typically accompanied by continuous mechanical mixing. However, the necessary dilution rates and the extent of mixing required have yet to be addressed. In this study, we investigated switching from wet to dry AD of food waste without mechanical mixing, employing different dilution rates. Lab-scale anaerobic reactors were operated with dilution rates of 10, 5, and 2 times during Phases I (0–56 days), II (57–121 days), and III (122–209 days), respectively. The methane production rates were not significantly different (p > 0.05) across the dilution rates decreased from 10 to 2 times. Remarkably, the methane production in the anaerobic reactors exhibited fluctuations due to variations in feeding, with the methane production rate ranging from 2.0 to 2.7 g CH4-COD/(L d), without mechanical mixing, as the solids content transitioned from wet to near-dry digestion conditions (15 %, food waste). The distribution of sludge volatile solids concentrations remained uniform in the reactor, even at high solids concentrations of up to 15 %. A dynamic microbial community response to changes in dilution rates, with a shift from aceticlastic to hydrogenotrophic methanogenesis pathways. Syntrophic acetate oxidization bacteria (the genus Syner-01 (4.2–8.9 %) and f_Synergistaceae (3.6–4.2 %)) were highly enriched as switching from wet AD to dry AD. The study's findings provide crucial operational insights for anaerobic food waste treatment, potentially resulting in decreased water usage and operational costs, particularly in scenarios with low dilution rates and without mechanical mixing.

Anaerobic methane oxidation is quantitatively important in deeper peat layers of boreal peatlands: Evidence from anaerobic incubations, in situ stable isotopes depth profiles, and microbial communities

Boreal peatlands store most of their carbon in layers deeper than 0.5 m under anaerobic conditions, where carbon dioxide and methane are produced as terminal products of organic matter degradation. Since the global warming potential of methane is much greater than that of carbon dioxide, the balance between the production rates of these gases is important for future climate predictions. Herein, we aimed to understand whether anaerobic methane oxidation (AMO) could explain the high CO2/CH4 anaerobic production ratios that are widely observed for the deeper peat layers of boreal peatlands. Furthermore, we quantified the metabolic pathways of methanogenesis to examine whether hydrogenotrophic methanogenesis is a dominant methane production pathway for the presumably recalcitrant deeper peat. To assess the CH4 cycling in deeper peat, we combined laboratory anaerobic incubations with a pathway-specific inhibitor, in situ depth patterns of stable isotopes in CH4, and 16S rRNA gene amplicon sequencing for three representative boreal peatlands in Western Siberia. We found up to a 69 % reduction in CH4 production due to AMO, which largely explained the high CO2/CH4 anaerobic production ratios and the in situ depth-related patterns of δ13C and δD in methane. The absence of acetate accumulation after inhibiting acetotrophic methanogenesis and the presence of sulfate- and nitrate-reducing anaerobic acetate oxidizers in the deeper peat indicated that these microorganisms use SO42− and NO3− as electron acceptors. Acetotrophic methanogenesis dominated net CH4 production in the deeper peat, accounting for 81 ± 13 %. Overall, anaerobic oxidation is quantitatively important for the methane cycle in the deeper layers of boreal peatlands, affecting both methane and its main precursor concentrations.

Enhancement of hydrogenotrophic methanogenesis for methane production by nano zero-valent iron in soils

Nanoscale zero-valent iron (nZVI) is attracting increasing attention as the most commonly used environmental remediation material. However, given the high surface area and strong reducing capabilities of nZVI, there is a lack of understanding regarding its effects on the complex anaerobic methane production process in flooded soils. To elucidate the mechanism of CH4 production in soil exposed to nZVI, paddy soil was collected and subjected to anaerobic culture under continuous flooding conditions, with various dosages of nZVI applied. The results showed that the introduction of nZVI into anaerobic flooded rice paddy systems promoted microbial utilization of acetate and carbon dioxide as carbon sources for methane production, ultimately leading to increased methane production. Following the introduction of nZVI into the soil, there was a rapid increase in hydrogen levels in the headspace, surpassing that of the control group. The hydrogen levels in both the experimental and control groups were depleted by the 29th day of culture. These findings suggest that nZVI exposure facilitates the enrichment of hydrogenotrophic methanogens, providing them with a favorable environment for growth. Additionally, it affected soil physicochemical properties by increasing pH and electrical conductivity. The metagenomic analysis further indicates that under exposure to nZVI, hydrogenotrophic methanogens, particularly Methanobacteriaceae and Methanocellaceae, were enriched. The relative abundance of genes such as mcrA and mcrB associated with methane production was increased. This study provides important theoretical insights into the response of key microbes, functional genes, and methane production pathways to nZVI during anaerobic methane production in rice paddy soils, offering fundamental insights into the long-term fate and risks associated with the introduction of nZVI into soils.

Insight into using hydrochar to alleviate ammonia nitrogen inhibition during anaerobic digestion of waste activated sludge: Performance, metagenomic and metabolomic signatures

In this study, hydrochar (HCR) was used to alleviate high ammonia inhibition to the anaerobic digestion (AD) of waste activated sludge (WAS) and to elucidate the inner microorganism mechanism. After HCR addition, the cumulative methane yield increased by 73.6 % and 35.6 % under ammonia inhibition levels of 3000 and 6000 mg/L, respectively. Metagenomic analysis showed that HCR enriched the diversity of hydrogenotrophic methanotrophs, and the relative abundances of functional microorganisms with electron transfer capabilities (Geobacteraceae bacterium etc.) were 1.5–7.8 times higher than those without HCR addition. Metabolomics analysis implied that metabolites related to fatty acid degradation, such as glutaric acid and hexadecanal, were downregulated (2.9–15.7 %) under ammonia inhibition conditions and that HCR regulates metabolites in the methane metabolic pathway. Moreover, HCR changed the methanogenic pathway from hydrogenotrophic methanogenesis to multiple pathways under ammonia inhibition conditions, especially methanolic and methylotrophic methanogenesis, which facilitated the methane yield. This study provides valuable information for understanding the inner microbial mechanism of HCR addition on alleviating high ammonia inhibition to AD of WAS, and gives basic knowledge for the application of AD of WAS under ammonia inhibition conditions.

Energy yields for acetylenotrophy on Enceladus and Titan

Saturn's moons Titan and Enceladus appear to satisfy three key criteria for life: presence of liquid water, nutrient availability, and metabolic energy sources. Consequently, these ocean worlds are the focus of astrobiology research investigations that aim to assess the potential for life beyond Earth. One plausible source of metabolic energy on these moons is acetylene (C2H2), a simple organic compound that is the second most abundant photochemical product in Titan's atmosphere and has been identified as a likely constituent of Enceladus' ocean. Acetylenotrophy, or the microbial fermentation of acetylene, is utilized by microbes on Earth in a number of environments. Here, we have calculated the energy yield of acetylenotrophy to be 69–78 kJ/mol C within the oceans of Titan and Enceladus, greater than the widely considered, hydrogenotrophic methanogenesis, at 25–65 kJ/mol C. Therefore, we propose acetylenotrophy as a potential metabolism on these moons that should be considered in future astrobiology studies on worlds with abundant acetylene.

Enhancing energy recovery and microbial stability in blackwater anaerobic digestion with graphite felt

Maximizing energy recovery from blackwater (toilet wastewater) helps the sustainable development of human society. This study introduces a novel approach by integrating graphite felt in anaerobic baffled reactor for enhanced blackwater treatment under room temperature. During the 382-day operation, incorporating graphite felt effectively mitigated hydrolysis and methanogenesis constraints, resulting in a 1.26 to 3.53-fold increase in methane yield compared to the control. The maximum energy recovery was attained at a hydraulic retention time of 1.59 d and an organic loading rate of 2.68 kg COD/m3reactor/d, with a methane production rate of 371 ± 55 NL CH4/m3reactor/d and a methanization rate of 39.44 ± 5.58 % in graphite felt amended reactor. Our findings revealed that graphite felt augmented the efficiency of microbial and enzymatic interactions with the substrate, and enriched hydrolysis fermentation-associated bacteria for efficient solid matter degradation. Further analysis unveiled that graphite felt strengthened the activity of hydrogenotrophic methanogens and fortified the syntrophic relationship between bacteria and archaea. This led to the promotion of a combined syntrophic acetate oxidation and hydrogenotrophic methanogenesis pathway, which guaranteed a stable and efficient methane production process. Overall, this approach offers a cost-effective and efficient strategy to support sustainable development in less-developed, resource-constrained regions.

Effect of iron sources on methane production and phosphorous transformation in an anaerobic digestion system of waste activated sludge

The iron materials are commonly employed to enhance resource recovery from waste activated sludge through anaerobic digestion (AD). The influence of different iron sources, such as Fe2O3, Fe3O4, and FeCl3 on methane production and phosphorus transformation in AD systems with thermal hydrolyzed sludge as the substrate was assessed in this study. The results indicated that iron oxides effectively promote methane yield and methane production rate in AD systems, resulting in a maximum increase in methane production by 1.6 times. Soluble FeCl3 facilitated the removal of 92.3% of phosphorus from the supernatant through the formation of recoverable precipitates in the sludge. The introduction of iron led to an increase in the abundance of bacteria responsible for hydrolysis and hydrogenotrophic methanogenesis. However, the enrichment of microbial communities varied depending on the specific irons used. This study provides support for AD systems that recover phosphorus and produce methane efficiently from waste sludge.

Acyl homoserine lactone-based regulation strategy for improved methane production in anaerobic digestion of agricultural wastes

The acyl homoserine lactone (AHL)-based regulation strategy presents a considerable potential in improving anaerobic digestion (AD) efficiency of complex substrates. However, the reinforcement mechanisms are still unclear. In this study, the effects of different AHL types (C4-HSL, C7-HSL, C12-HSL) and different adding times (0 d, 5 d, 15 d) on AD performance of agricultural wastes (corn straw and cattle manure) were investigated. The results indicated that all types of AHLs exerted positive effects on AD, and the adding time rather than the AHL type was the key factor affecting AD processes. The C12-5d group obtained the highest accumulated methane production (310.7 ± 2.56 mL/g VS), which increased by 47.71% than that of the Control group. The exogenous AHLs facilitated the hydrolysis and acidification of complex substrates and the utilization of released low-molecular-weight organic acids in AD systems. Furthermore, the role of methanogenic archaea was upregulated and the dominance of bacteria was relatively weakened in AD systems, which favored the establishment of their balanced and symbiotic metabolic relationship. The enriched Methanobacterium strengthened the hydrogenotrophic methanogenesis pathway. This study provides new insights into the regulation strategy of AD for agricultural waste treatment based on AHL addition.

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.

Effects of Fe–N co-modified biochar on methanogenesis performance, microbial community, and metabolic pathway during anaerobic co-digestion of alternanthera philoxeroides and cow manure

The performance of anaerobic digestion (AD) is susceptible to disturbances in feedstock degradation, intermediates accumulation, and methanogenic archaea activity. To improve the methanogenesis performance of the AD system, Fe–N co-modified biochar was prepared under different pyrolysis temperatures (300,500, and 700 °C). Meanwhile, pristine and Fe-modified biochar were also derived from alternanthera philoxeroides (AP). The aim was to compare the effects of Fe–N co-modification, Fe modification, and pristine biochar on the methanogenic performance and explicit the responding mechanism of the microbial community in anaerobic co-digestion (coAD) of AP and cow manure (CM). The highest cumulative methane production was obtained with the addition of Fe-N-BC500 (260.38 mL/gVS), which was 42.37 % higher than the control, while the acetic acid, propionic acid, and butyric acid concentration of Fe-N-BC were increased by 147.58 %, 44.25 %, and 194.06 % compared with the control, respectively. The co-modified biochar enhanced the abundance of Chloroflexi and Methanosarcina in the AD system. Metabolic pathway analysis revealed that the increased methane production was related to the formation and metabolism of volatile fatty acids and that Fe-N-BC500 enhanced the biosynthesis of coenzyme A and the cell activity of microorganisms, accelerating the degradation of propionic acid and enhancing the hydrogenotrophic methanogenesis pathway. Overall, Fe–N co-modified biochar was proved to be an effective promoter for accelerated methane production during AD.