Methanogenic Pathway Research Articles

Revisiting the biological pathway for methanogenesis in landfill from metagenomic perspective—A case study of county-level sanitary landfill of domestic waste in North China plain

Landfill is the third highest contributor to anthropogenic methane (CH4) emissions, produced primarily by the anaerobic decomposition of organic matter by microbes. However, how various microbial metabolic processes contribute to CH4 production in domestic waste landfill remains elusive. We addressed this problem by investigating the methanogenic communities, methanogenic functional genes, KEGG modules and KEGG pathways in a county-level MSW sanitary landfill in North China Plain, China. Results showed that Methanomicrobiales, Methanobacteriales, Methanosarcinales, Micrococcales, Corynebacteriales and Bacillales were the dominant methanogens. M00357, M00346, M00567 and M00563 were the four major methane metabolic modules. The most abundant genes were ACSS, ackA and fwd with the relative abundance of 19.26–54.54%, 6.14–25.78% and 6.76–16.51%, respectively. The two essential genes of methanogenesis were detected with the relative abundance of 2.66–9.58% (mtr) and 1.63–9.14% (mcr). These findings indicated that acetotrophic and hydrogenotrophic methanogenesis were the major pathways. Methanomicrobiales, Methanosarcinales and Clostridiales were the key microbes to these pathways identified by co-occurrence network. Analysis of relative contribution of species to function further showed that Micrococcales, Corynebacteriales and Bacillales were special contributors to acetotrophic methanogenesis pathway. Redundancy analysis revealed that above functional genes and microbes were mainly controlled by NH4+ and pH. Our results can help to provide develop the fine management strategies for methane utilization and emission reduction in landfill.

Impacts of 2-bromoethanesulfonic sodium on methanogenesis: Methanogen metabolism and community structure

Production of medium-chain carboxylic acids (MCCAs) by chain elongation (CE) presents a competitive alternative to conventional products of methane in anaerobic digestion treating organic waste streams, considering energy recovery, economic, and environmental profits. However, the system stability and performance largely rely on the selective suppression of methanogens while stimulation of CE bacteria. Commercial inhibitors such as 2-bromoethanesulfonic sodium (BES) was shown to be effective, but controversial conclusions exist on its inhibition characteristics and the inhibition mechanism remains unclear. Therefore, this study systematically investigated the responses of methanogenesis in granular sludge to various BES levels, focusing on methane production, methanogenic pathway, dynamic populations, electron transport and energy metabolism. Results showed that compared with the control, 3.0 g/L BES was sufficient to induce a 72.9% reduced level on accumulative methane production by the end of 4 cycles (28 days), which was likely to be attributed to the significantly suppressed metabolic pathways and intracellular regulations. Specifically, BES suppressed the electron transport via unproper electron carriers and reduced electron amount as indicated by the decreased level of enzymes and genes involved such as coenzyme F420, CO dehydrogenase and NADH:ubiquinone reductase (H+-translocating). Moreover, BES regulated the intracellular energy metabolism, leading to the impeded ATP synthesis but enhanced ATP consumption as evidenced by the variations on the activity or abundance of acetate kinase, A1Ao-ATP synthase, nitrogenase and ATP citrate synthase. Additionally, BES enriched hydrogenotrophic methanogenesis over acetoclastic one as supported by variations on the archaeal community structures and regulations of differentially expressed genes involved. Moreover, BES also reduced the contents of both protein and carbohydrate in extracellular polymeric substances (EPS). This study is expected to enhance understanding of BES contribution to methanogenesis inhibition but MCCAs production in CE bioreactors.

Mechanism of action of single and mixed antibiotics during anaerobic digestion of swine wastewater: Microbial functional diversity and gene expression analysis

The mechanism by which antibiotics in swine wastewater affect anaerobic digestion (AD) remains unclear. Herein, we investigated how single and mixed antibiotics affect AD in swine wastewater. Both single and mixed antibiotics stimulated methane production at actual concentrations of 0.5–2 mg/L. Low-dose antibiotics (0.5 mg/L) exerted the most significant stimulatory effect on methane production, which increased by 211.63% (single) and 60.93% (mixed), respectively. However, an increased dose decreased the stimulatory effect on methane production. Overall, single antibiotics were more beneficial for methane production than mixed antibiotics since single antibiotics could promote the conversion of propionic and butyric acid, while mixed antibiotics inhibited the process. Microbial community analysis showed that single and mixed antibiotics could also lead to large changes in functional acidogens, ultimately leading to changes in methanogenic pathways.

Microbial population dynamics and meta-proteomic evaluation for functional insight into an anaerobic membrane bioreactor during startup

Startup of an anaerobic membrane reactor (AnMBR) involves a significant concentration of active and functioning microbial communities participating in the process. Here, we monitored the physical, microbial, and functional performance during the first 90 days of AnMBR operation. The results indicate initiation of anaerobic digestion on days 31–60 (transitional stage) and its stabilization on days 61–90 (stable stage), with an average COD removal efficiency of 93.14 ± 1.41% and biogas production of 0.82 ± 0.15 L/d. Moreover, the microbial population and its associated proteome were studied using 16s rDNA-based PCR-DGGE and SDS-PAGE-LC/MS/MS techniques to acquire a detailed understanding of reactor startup conditions. Later, a conventional library was created through the UniprotKB protein database using the observed community of all organisms identified with 16s rDNA sequencing. The results clearly depicted Gammaproteobacteria as the dominant fermentative during the initial and transitional stages. Firmicutes predominated over Gammaproteobacteria during the stabilization stage. During the transitional stage, methanobacteriales and metheanosarcinales coexisted. In the two-step reaction, a combination of syntrophic acetate oxidation and hydrogenotrophic methanogenesis was observed. During the stabilization stage, diverse metheanosarcinales were detected. The methanogenic pathway seemed to shift to acetoclastic methanogenesis, as indicated by detection of acetyl-coenzyme A synthetase, aminomethyltransferase, and methyl-coenzyme M reductase. Thus, by linking microbial community dynamics with the meta-proteomic functions of AnMBR during startup, a stable reactor performance with up-regulated acetoclastic methanogenesis was observed in 90 days.

Understanding Life at High Temperatures: Relationships of Molecular Channels in Enzymes of Methanogenic Archaea and Their Growth Temperatures

Analyses of protein structures have shown the existence of molecular channels in enzymes from Prokaryotes. Those molecular channels suggest a critical role of spatial voids in proteins, above all, in those enzymes functioning under high temperature. It is expected that these spaces within the protein structure are required to access the active site and to maximize availability and thermal stability of their substrates and cofactors. Interestingly, numerous substrates and cofactors have been reported to be highly temperature-sensitive biomolecules. Methanogens represent a singular phylogenetic group of Archaea that performs anaerobic respiration producing methane during growth. Methanogens inhabit a variety of environments including the full range of temperatures for the known living forms. Herein, we carry out a dimensional analysis of molecular tunnels in key enzymes of the methanogenic pathway from methanogenic Archaea growing optimally over a broad temperature range. We aim to determine whether the dimensions of the molecular tunnels are critical for those enzymes from thermophiles. Results showed that at increasing growth temperature the dimensions of molecular tunnels in the enzymes methyl-coenzyme M reductase and heterodisulfide reductase become increasingly restrictive and present strict limits at the highest growth temperatures, i.e., for hyperthermophilic methanogens. However, growth at lower temperature allows a wide dimensional range for the molecular spaces in these enzymes. This is in agreement with previous suggestions on a potential major role of molecular tunnels to maintain biomolecule stability and activity of some enzymes in microorganisms growing at high temperatures. These results contribute to better understand archaeal growth at high temperatures. Furthermore, an optimization of the dimensions of molecular tunnels would represent an important adaptation required to maintain the activity of key enzymes of the methanogenic pathway for those methanogens growing optimally at high temperatures.

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.

Lab- and pilot-scale anaerobic digestion of municipal bio-waste and potential of digestate for biogas upgrading sustained by microbial analysis

The challenge of handling the enormous amount of bio-waste that is globally produced on an annual basis can become an opportunity by exploiting such residues for bioenergy production in the form of biogas or biomethane. The application of anaerobic digestion to bio-waste is still limited due to their specific characteristics (e.g., recalcitrant matter, high solids content, etc.) that might lead to process imbalances and microbial toxicity issues. The present study aimed to elucidate microbial ecology in the lab- and pilot-scale biogas reactors fed with bio-waste. The results showed that the indigenous microbial community in the bio-waste was almost extinct during the digestion process. The AD microbiome was mainly composed of members belonging to the orders Clostridiales and Bacteroidales, whose relative abundance dynamically changed to adapt to operational conditions. The increase in organic loading rate led to the proliferation of members associated with the phylum Synergistetes and the transition to versatile acetoclastic/hydrogenotrophic methanogenic pathways. Furthermore, digestate is a suitable biological inoculum and a nutrient supplement to support further biogas upgrading process. The results identified functional microbes that could benefit bio-waste digestion, unveiled insights into microbiome reorganization, and proposed a potential application of the digestate.

Regulating effects of Fe/C materials on thermophilic anaerobic digestion of kitchen waste: Digestive performances and methanogenic metabolism pathways

This study aimed to evaluate the effects of typical exogenous materials (Fe and C) addition on the performances of kitchen waste thermophilic (55 °C) anaerobic digestion in semi-continuous system. The expression of key enzymes in methanogenesis were demonstrated by microbiological approach, and the regulating effectsf Fe and C addition on methanogenic metabolism pathway were specially explored. Four anaerobic digestion systems (TAD, TAD + Fe, TAD + C and TAD + Fe + C) were carried out, and the running period lasted for 120 days, which was divided into four stages according to organic loading rates (OLRs) from 1.05 to 4.22 gVS/L/day. From this study, C addition improved soluble polysaccharide (sPolysaccharide), soluble protein (sProtein), soluble total organic carbon (sTOC) concentrations and hydrolase (amylase, protease and lipase) activities, while Fe addition facilitated the subsequent degradation of volatile fatty acids (VFAs). As a result, the average methane yields were promoted from 478.0 (TAD) to 487 (TAD + Fe), 597 (TAD + C) and 647.3 mL/gVS/day (TAD + Fe + C), respectively. For microbial communities, C addition promoted the relative abundances of hydrolytic bacteria (Defluviitoga), while Fe addition realized the directional enrichment of Methanothermobacter, and improved hydrogenotrophic methanogenesis ratios from 30.81 % (TAD) to 52.27 % (TAD + Fe) and 64.40 % (TAD + Fe + C). According to the expression of key enzymes in methanogenic metabolism pathways, Fe addition enhanced the syntrophic acetic acid oxidation and hydrogenotrophic methanogenesis pathway, realizing the conversion of dominated methanogenic metabolism pathway from aceticlastic methanogenesis to hydrogenotrophic methanogenesis. This study is of great significance to exposit the behaviors of conductive materials during thermophilic digestion of kitchen waste.

Metagenomic evidence of suppressed methanogenic pathways along soil profile after wetland conversion to cropland.

Wetland conversion to cropland substantially suppresses methane (CH4) emissions due to the strong suppression of methanogenesis, which consists of various pathways. In this study, we evaluated the cultivation impacts on four predominant CH4 production pathways, including acetate, carbon dioxide (CO2), methylamines, and methanol, in a wetland and cultivated cropland in northeastern China. The results showed significant suppression of CH4 production potential and the abundance of genes for all four methanogenic pathways in cropland. The consistency between CH4 production and methanogenesis genes indicates the robustness of genomic genes in analyzing methanogenesis. The suppression effects varied across seasons and along soil profiles, most evident in spring and 0 to 30 cm layers. The acetate pathway accounted for 55% in wetland vs. 70% in the cropland of all functional genes for CH4 production; while the other three pathways were stronger in response to cultivation, which presented as stronger suppressions in both abundance of functional genes (declines are 52% of CO2 pathway, 68% of methanol pathway, and 62% of methylamines pathway, vs. 19% of acetate pathway) and their percentages in four pathways (from 20 to 15% for CO2, 15 to 9% for methylamines, and 10 to 6% for methanol pathway vs. 55 to 70% for acetate pathway). The structural equation models showed that substrate availability was most correlated with CH4 production potential in the wetland, while the positive correlations of acetate, CO2, and methylamine pathways with CH4 production potential were significant in the cropland. The quantitative responses of four CH4 production pathways to land conversion reported in this study provide benchmark information for validating the CH4 model in simulating CH4 cycling under land use and land cover change.

Enhanced sludge digestion using anaerobic dynamic membrane bioreactor: Effects of hydraulic retention time

Performance of conventional anaerobic digestion is limited due to coupling of solid retention time (SRT) and hydraulic retention time (HRT). In this study, the sludge digestion efficiency was enhanced by decoupling SRT and HRT in a pilot-scale anaerobic dynamic membrane bioreactor, with a particular focus on the effects of HRT. With HRT decreased to 10 d, the total solids concentration reached 43.3 g/L in the membrane tank with the influent sludge concentration 23.4 ± 0.7 g/L, while volatile solids (VS) reduction rate and specific biogas production were 41% and 0.59 L/g VS, respectively, achieving simultaneous thickening and digestion of sludge. Microbial community analysis showed that Cloroflexi and Spirochaetota, which had an increased abundance at low HRT, were beneficial to VS reduction and multiple methanogenesis pathways. Methanosaeta, being a strict acetoclastic methanogen dominant at HRT 30 d, decreased in the abundance from 78.4% to 50.7% with HRT changing from 30 d to 10 d. In contrast, the abundance of hydrogenotrophic and methylotrophic methanogens increased from 17.2% to 39.4% with the decrease of HRT, indicating the influence of HRT on methanogenic pathways. Furthermore, the predicted gene analysis confirmed the transformation from dominant acetoclastic methanogenesis to the coexistence of multiple methanogenesis pathways. Finally, the recovered energy from methane increased from 0.43 kWh/kg VS to 0.56 kWh/kg VS as HRT decreased to 10 d from 30 d, and the net energy consumption at HRT of 10 d was 0.47 kWh/kg VS, only 66% of that at HRT of 30 d.

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.

Effects of different low temperature conditions on anaerobic digestion efficiency of pig manure and composition of archaea community.

To explore the effect of low temperature on the anaerobic digestion of pig manure, the anaerobic digestion experiment was carried out under the conditions of inoculum concentration of 30% and TS of 8%. Five low-temperature gradients of 4, 8, 12, 16 and 20 °C were set to study the activities of gas production, pH, solluted chemical oxygen demand (SCOD), volatile fatty acids (VFAs), coenzymes F420 and archaea community composition in the digestion process. The results were demonstrated: as the temperature decreased, the more unstable the gas production became, the less gas production produced, and the later the gas peak occurred. There were no significant peaks at either 4 °C or 8 °C, and the SCOD was unstable over time. From 12 °C, the SCOD increased over time, and the higher the temperature, the faster the growth trend. The pH was always greater than 7.6. 8, 12, 16, 20 °C had different degrees of VFAs accumulation at the late digestion stage. The higher the temperature, the greater the amount of volatile acid accumulation. When the VFAs of each reactor reached the maximum, the proportion of acetic acid also reached the highest. The digestion system of the five treatment groups was dominated by hydrogen-nutrient methanogenic pathway. The results could provide a further reference for the mechanism of anaerobic digestion of pig manure at low temperatures.

Hormesis-Like Effects of Tetrabromobisphenol A on Anaerobic Digestion: Responses of Metabolic Activity and Microbial Community.

Tetrabromobisphenol A (TBBPA) has extensive applications in various fields; its release into ecosystems and the potential toxic effects on organisms are becoming major concerns. Here, we investigated the effects of TBBPA on anaerobic digestion, whose process is closely related to the carbon cycles under anaerobic conditions. The results revealed that TBBPA exhibited dose-dependent hormesis-like effects on methane production from glucose, i.e., the presence of 0.1 mg/L TBBPA increased the methane production rate by 8.79%, but 1.0-4.0 mg/L TBBPA caused 3.45-28.98% of decrement. We found that TBBPA was bound by the tyrosine-like proteins of the extracellular polymeric substances of anaerobes and induced the increase of reactive oxygen species, whose slight accumulation stimulated the metabolism activities but high accumulation increased the apoptosis of anaerobes. Owing to the differences between individual anaerobes in tolerance, TBBPA at 0.1 mg/L stimulated the acidogenesis and hydrogenotrophic methanogenesis, whereas higher levels (i.e., 1.0-4.0 mg/L) severely restrained all of the processes of acidogenesis, acetogenesis, and methanogenesis. Along with the accumulation of bisphenol A (BPA) produced from TBBPA by Longilinea sp. and Pseudomonas sp., the methanogenic pathway was partly shifted from acetate-dependent to hydrogen-dependent direction, and the activities of carbon monoxide dehydrogenase and acetyl-CoA decarbonylase/synthase were inhibited, while acetate kinase and F420 were hormetically affected. These findings elucidated the mechanism of anaerobic syntrophic consortium responses to TBBPA, supplementing the potential environmental risks of brominated flame retardants.

Impacts of cyanobacterial biomass and nitrate nitrogen on methanogens in eutrophic lakes

Methanogenesis is a key process in carbon cycling in lacustrine ecosystems. Knowledge of the methanogenic pathway is important for creating mechanistic models as well as predicting methane emissions. Due to low concentrations of methyl substrates in freshwater lakes, the proportion of methylotrophic methanogenesis is believed to be negligible in such environments. However, the high abundance of methylotrophic methanogens previously detected in Dianchi Lake suggests that methylotrophic methanogenesis may be underestimated in eutrophic lakes, whereas their influencing factors and mechanisms are not yet clear. In this study, the effects of cyanobacteria biomass (CB) or/and nitrate nitrogen on methanogenesis, especially methylotrophic pathway, in eutrophic lakes were investigated using microcosm simulation experiments combined with chemical analysis and high-throughput sequencing techniques. The results showed that either CB or nitrate nitrogen had significant effects on methane flux, the archaeal diversity and community structure of methanogens. Functional prediction, together with the result of chemical analysis, revealed that CB could promote methylotrophic methanogenesis by providing methyl organic substrates, while nitrate nitrogen increased the relative abundance of obligate methylotrophic methanogens by competitively inhibiting the other two methanogenic pathways. In eutrophic lake where both CB and nitrate present at a high concentration, methylotrophic methanogenesis could play a much more important role than previously believed.

Light triggers green recovery: Boosted biomethane production from ammonia-stressed anaerobic digestion through optimized illuminated bioreactor

Excess ammonia generated during anaerobic digestion (AD) often poses a great challenge on efficient waste-to-energy bioconversion, which requires feasible strategies to improve the performance. Herein, by response surface methodology (RSM), an innovative light-assisted bioreactor was designed for boosting ammonia-stressed AD. The proposed optimal light condition (225 W/m2 with 63 min/day) with incandescent lighting boosted CH4 yield (198 ± 15 mL CH4/g-DOCremoval), which was twofold of the dark reactor under ammonia stress. This light-induced enhancement was further found to be reliable and adaptive at different ammonia levels (2500–8000 mg/L), revealing its effectiveness for treating various ammonia-rich wastes. Additionally, the practical feasibility of sunlight utilization was confirmed by the similar performance obtained under simulated solar lighting (ultraviolet cut). Furthermore, the developed bioreactor exhibited a stable and superior conversion efficiency for 2 months, suggesting its sustainability for long-term operation. The mechanism insights revealed that light acted as stimulus on ammonia-stressed anaerobes, effectively regulated the enzyme activities, the functional microbiome, and the sludge properties to alleviate the ammonia inhibition. The critical coenzymes involved in methanogenic pathways (hydrogenotrophic and acetotrophic) were upregulated by light. Meanwhile, proper light stimulation diversified the ammonia-tolerant fermentative taxa (hydrolytic and acetogenic genera), which jointly collaborated with the enriched Methanosarcina. Under ammonia pressure, those enriched anaerobes aggregated in compact clusters which thermodynamically favored the sludge electroactivity and hydrophobicity, and assisted the syntrophic bioconversion. Therefore, the proposed bioreactor would be a promising technique to solve ammonia inhibition and achieve the sustainable waste-to-energy bioconversion with solar integration.

Improvement of anaerobic digestion of food waste by addition of synthesized allophane

Anaerobic digestion (AD) of food waste (FW) always confronts the challenges of over-acidification in application. This work evaluated the effectiveness of synthesized allophane, a mineral with desirable physicochemical properties (e.g., high pH buffer and organic matter adsorption capacity, and high porosity and specific surface area), in increasing biogas yield during AD of FW as an additive. Results showed that allophane addition (0 to 10 g total solid (TS)) increased the cumulative biogas yield from 409.69 ± 20.77 mL/g TS to 624.06 ± 6.63 mL/g TS, and methane production from 224.12 ± 9.26 mL/g TS to 391.52 ± 0.87 mL/g TS. Improved AD performance was mainly attributed to mitigating over-acidification during the start-up period, and favoring microbial growth, particularly the acetotrophic methanogen of Methanosarcina, indicating an intensified acetoclastic methanogenic pathway. The findings provided a mechanistic insight into the improved AD performance with allophane addition, and offered a potential strategy to stabilize AD of FW in application.

Upgrade the high-load anaerobic digestion and relieve acid stress through the strategy of side-stream micro-aeration: biochemical performances, microbial response and intrinsic mechanisms

In high-load anaerobic digestion such as in kitchen waste, side-stream micro-aeration (SMA) shows excellent operational performance to direct micro-aeration (DMA). It immediately restores the acidification to stability. Methanogenic performance remained stable when organic load ratios (OLR) was further increased to 5.5 g VS/L. Enhanced enzyme activity, microbial aggregation, and proliferation of bacteria and archaea were observed in SMA. The results indicates that SMA enriched Methanosaeta (relative abundance exceeded 93%) and induced the change of the main methanogenic pathway to acetoclastic methanogenesis. Mechanisms was further explored by using metagenomic analysis, and the results show SMA avoids mass formation of ROS (reactive oxygen species) by cycling the aerated slurry, and retains benefits of trace O2 on material and energic metabolism, which poses great application potentials and deserves further investigation.

Insights into the impact of polyethylene microplastics on methane recovery from wastewater via bioelectrochemical anaerobic digestion

Bioelectrochemical anaerobic digestion (BEAD) is a promising next-generation technology for simultaneous wastewater treatment and bioenergy recovery. While knowledge on the inhibitory effect of emerging pollutants, such as microplastics, on the conventional wastewater anaerobic digestion processes is increasing, the impact of microplastics on the BEAD process remains unknown. This study shows that methane production decreased by 30.71% when adding 10 mg/L polyethylene microplastics (PE-MP) to the BEAD systems. The morphology of anaerobic granular sludge, which was the biocatalysts in the BEAD, changed with microbes shedding and granule crack when PE-MP existed. Additionally, the presence of PE-MP shifted the microbial communities, leading to a lower diversity but higher richness and tight clustering. Moreover, fewer fermentative bacteria, acetogens, and hydrogenotrophic methanogens (BEAD enhanced) grew under PE-MP stress, suggesting that PE-MP had an inhibitory effect on the methanogenic pathways. Furthermore, the abundance of genes relevant to extracellular electron transfer (omcB and mtrC) and methanogens (hupL and mcrA) decreased. The electron transfer efficiency reduced with extracellular cytochrome c down and a lower electron transfer system activity. Finally, phylogenetic investigation of communities by reconstruction of unobserved states analysis predicted the decrease of key methanogenic enzymes, including EC 1.1.1.1 (Alcohol dehydrogenase), EC 1.2.99.5 (Formylmethanofuran dehydrogenase), and EC 2.8.4.1 (Coenzyme-B sulfoethylthiotransferase). Altogether, these results provide insight into the inhibition mechanism of microplastics in wastewater methane recovery and further optimisation of the BEAD process.

Methane Production Potential and Methanogenic Pathways in Paddy Soils Under Different Rice-based Cropping Systems

Differences inmethane (CH4) production potential in paddy soils under different rice-based cropping systems and especially in the methanogenic pathways (mainly acetate fermentation and CO2/H2 reduction) remain unclear. Anaerobic incubations of soil with or without fluoromethane (CH3F) inhibitor (2% and 0%) were conducted. With the soils from three typical paddy ecosystems (rice-wheat rotation, RW; rice-fallow, RF; double-rice, DR) in China, the cumulative concentration of CH4 production, CH4 production potential, dissolved organic carbon (DOC) content, and acetic acid content were determined. Meanwhile, the relative contribution of acetate-dependent methanogenesis (fac) was quantified using the stable carbon isotope method. The results showed that the CH4 production potential was 7.18 μg·(g·d)-1 in RF, which was significantly lower than that in RW[10.33 μg·(g·d)-1]and DR[13.42 μg·(g·d)-1] (P<0.05). Correlation analysis showed that CH4 production potential was significantly negatively correlated with soil cation exchange capacity and pH (P<0.01); the addition of CH3F significantly inhibited CH4 production (P<0.05). The content of DOC and acetic acid in DR were 255 mg·kg-1 and 7.34 mg·kg-1, respectively, which were 17%-51% and 22%-23% higher than those in RW and RF, respectively. The δ13CH4 and δ13CO2 values were affected greatly by different rice-based cropping systems, and the highest δ13CH4 value was -43.89‰ in RF, which was more positive than that of RW and DR by 11.06‰ and 8.33‰, respectively (P<0.05). By contrast, the lowest value of δ13CO2 was observed in RF, which was more negative than that of RW (7.63‰) and DR (5.14‰) (P<0.05). The α(CO2/CH4) values of RW and RF were 1.057 and 1.058, respectively, which were significantly lower than 1.062 in DR (P<0.05). The fac values of RF ranged from 84% to 98%, being 34%-38% and 20%-23% higher than those of RW and DR, respectively (P<0.05).

Interpretation of Origin and Methanogenic Pathways of Coalbed Gases from the Asem-Asem Basin, Southeast Kalimantan, Indonesia

Interpretation of Origin and Methanogenic Pathways of Coalbed Gases from the Asem-Asem Basin, Southeast Kalimantan, Indonesia