Aceticlastic Methanogenesis Research Articles

Regulating the porous properties of polyaniline enhances the electron transfer process and methane production during anaerobic wastewater treatment

Conductive materials are known to enhance methane production in anaerobic treatment process by facilitating direct interspecies electron transfer (DIET). However, the role of the porous structure of conductive materials in electron transfer remains underexplored. Polyaniline (PANI) was used as conductive materials in this study due to its easily controlled porosity, investigating how surface area and pore size affect methane production. It was demonstrated that the porosity of PANI is an important factor affecting methane production by anaerobic sludge. Methane production rate of 41.2 mL/h was achieved with the high-porosity PANI, which was ~73.2% higher than the control group. The porous PANI enhanced the electric field in the anaerobic sludge, facilitating the enrichment of electrogenic bacteria and archaea. In the anaerobic system supplemented with porous PANI, the maximum accumulation of acetic acid reached 3.72 mM. The abundance of electroactive bacteria Clostridium involved in DIET increased by 2.13-fold, while the abundance of electroactive archaea Methanosaeta and Methanobacterium, which also participate in DIET, rose by 1.55-fold. The porous structure of PANI promotes DIET and enhances aceticlastic methanogenesis.

Carbon monoxide inhibition on acidogenic glucose fermentation and aceticlastic methanogenesis

Syngas and CO-rich off-gases are key chemical platforms to produce biofuels and bioproducts. From the perspective of optimizing and up-scaling CO co-digestion with organic waste streams, this study aims at assessing and quantifying the inhibitory effects of CO on acidogenic glucose fermentation and aceticlastic methanogenesis. Mesophilic cultures were fed in two sets of batch assays, respectively, with glucose and acetate while being exposed to dissolved CO in equilibrium with partial pressures in the range of 0.25–1.00 atm. Cumulative methane production and microbial monitoring revealed that aceticlastic methanogenic archaea were significantly inhibited (2–20 % of the methane production of CO non-exposed cultures). The acidogenic glucose degrading community was also inhibited by CO, although, thanks to its functional redundancy, shifted its metabolism towards propionate production. Future work should assess the sensitivity of hereby estimated CO inhibition parameters, e.g., on the simulation output of a continuous syngas co-digestion process with organic substrates.

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.

Fractionation of stable carbon isotopes during formate consumption in anoxic rice paddy soils and lake sediments

Abstract. Formate is energetically equivalent to hydrogen and thus is an important intermediate during the breakdown of organic matter in anoxic rice paddy soils and lake sediments. Formate is a common substrate for methanogenesis, homoacetogenesis and sulfate reduction. However, how much these processes contribute to formate degradation and fractionate carbon stable isotopes is largely unknown. Therefore, we measured the conversion of formate to acetate, CH4 and CO2 and the δ13C of these compounds in samples of paddy soils from Vercelli, Italy, and the International Rice Research Institute (IRRI) in the Philippines and of sediments from the NE and SW basins of Lake Fuchskuhle, Germany. The samples were suspended in a phosphate buffer (pH 7.0) in both the absence and presence of sulfate (gypsum) and of methyl fluoride (CH3F), an inhibitor of aceticlastic methanogenesis. In the paddy soils, formate was mainly converted to acetate under both methanogenic and sulfidogenic conditions. Methane was only a minor product and was mainly formed from the acetate. In the lake sediments, the product spectrum was similar but only under methanogenic conditions. In the presence of sulfate, however, acetate and CH4 were only minor products. The isotopic enrichment factors (εform) of formate consumption, determined by Mariotti plots, were in the low range of −8 ‰ to −2.5 ‰ when sulfate was absent, and formate was mainly converted to acetate and CH4. However, no enrichment factor was detectable when formate was degraded with sulfate to mainly CO2. The δ13C of acetate was by about 25 ‰–50 ‰ more negative than that of formate, indicating acetate production by chemolithotrophic homoacetogenesis. Hence, formate seems to be an excellent substrate for homoacetogenesis in anoxic soils and sediments, so that this process is competing well with methanogenesis and sulfate reduction.

Comparing the mechanisms of syntrophic volatile fatty acids oxidation and methanogenesis recovery from ammonia stress in regular and biochar-assisted anaerobic digestion: Different roads lead to the same goal

Biochar has been recognized as a promising additive to mitigate ammonia inhibition during syntrophic methanogenesis, while the key function of biochar in this process is still in debates. This study clarified the distinct mechanisms of syntrophic volatile fatty acids -oxidizing and methanogenesis recovery from ammonia inhibition in regular and biochar-assisted anaerobic digestion. Under 5 g/L ammonia stress, adding biochar shortened the methanogenic lag time by 10.9% and dramatically accelerated the maximum methane production rate from 60.3 to 94.7 mLCH4/gVSsludge/d. A photometric analysis with a nano-WO3 probe revealed that biochar enhanced the extracellular electron transfer (EET) capacity of suspended microbes (Pearson's r = −0.98), confirming that biochar facilitated methanogenesis by boosting EET between syntrophic butyrate oxidizer and methanogens. Same linear relationship between EET capacity and methanogenic rate was not observed in the control group. Microbial community integrating functional genes prediction analysis uncovered that biochar re-shaped syntrophic partners by enriching Constridium_sensu_stricto/Syntrophomonas and Methanosarcina. The functional genes encoding Co-enzyme F420 hydrogenase and formylmethanofuran dehydrogenase were upregulated by 1.4–2.3 times, consequently enhanced the CO2-reduction methanogenesis pathway. Meanwhile, the abundances of gene encoding methylene-tetrahydrofolate transformation, a series of intermediate processes involved in acetate oxidation, in the biochar-assisted group were 28.2–63.7% higher than these in control group. Comparatively, Methanosaeta played a pivotal role driving aceticlastic methanogenesis in the control group because the abundance of gene encoding acetyl-CoA decarbonylase/synthase complex increased by 1.9 times, suggesting an aceticlastic combining H2-based syntrophic methanogenesis pathway was established in control group to resist ammonia stress. A 2nd period experiment elucidated that although depending on distinct mechanisms, the volatile fatty acid oxidizers and methanogens in both groups developed sustained and stable strategies to resist ammonia stress. These findings provided new insights to understand the distinct methanogenic recovery strategy to resist toxic stress under varied environmental conditions.

Methane and nitrous oxide emissions and related microbial communities from mangrove stems on Qi'ao Island, Pearl River Estuary in China

Mangrove forests, crucial carbon-rich ecosystems, are increasingly vulnerable to soil carbon loss and greenhouse gas (GHG) emissions due to human disturbance. However, the contribution of mangrove trees to GHG emissions remains poorly understood. This study monitored CO2, CH4, and N2O fluxes from the stems of two mangrove species, native Kandelia obovata (KO) and exotic Sonneratia apetala (SA), at three heights (0.7 m, 1.2 m, and 1.7 m) during the dry winter period on Qi'ao Island, Pearl River Estuary, China. Heartwood samples were analyzed to identify potential functional groups related to gas fluxes. Our study found that tree stems acted as both sinks and sources for N2O (ranging from −9.49 to 28.35 μg m−2 h−1 for KO and from −6.73 to 28.95 μg m−2 h−1 for SA) and CH4. SA exhibited significantly higher stem CH4 flux (from −26.67 to 97.33 μg m−2 h−1) compared to KO (from −44.13 to 88.0 μg m−2 h−1) (P < 0.05). When upscaled to the community level, both species were net emitters of CH4, contributing approximately 4.68 % (KO) and 0.51 % (SA) to total CH4 emissions. The decrease in stem CH4 flux with increasing height, indicates a soil source. Microbial analysis in the heartwood using the KEGG database indicated aceticlastic methanogenesis as the dominant CH4 pathway. The presence of methanogens, methanotrophs, denitrifiers, and nitrifiers suggests microbial involvement in CH4 and N2O production and consumption. Remarkably, the dominance of Cyanobacteria in the heartwood microbiome (with the relative abundance of 97.5 ± 0.6 % for KO and 99.1 ± 0.2 % for SA) implies roles in carbon and nitrogen fixation for mangroves coping with nitrogen limitation in coastal wetlands, and possibly in CH4 production. Although the present study has limitations in sampling duration and area, it highlights the significant role of tree stems in GHG emissions which is crucial for a holistic evaluation of the global carbon sequestration capability of mangrove ecosystems. Future research should broaden spatial and temporal scales to enhance the accuracy of upscaling tree stem gas fluxes to the mangrove ecosystem level.

Phenotypic and genomic characterization of the first alkaliphilic aceticlastic methanogens and proposal of a novel genus Methanocrinis gen.nov. within the family Methanotrichaceae.

Highly purified cultures of alkaliphilic aceticlastic methanogens were collected for the first time using methanogenic enrichments with acetate from a soda lake and a terrestrial mud volcano. The cells of two strains were non-motile rods forming filaments. The mud volcano strain M04Ac was alkalitolerant, with the pH range for growth from 7.5 to 10.0 (optimum at 9.0), while the soda lake strain Mx was an obligate alkaliphile growing in the pH range 7.7-10.2 (optimum 9.3-9.5) in the presence of optimally 0.2-0.3 M total Na+. Genomes of both strains encoded all enzymes required for aceticlastic methanogenesis and different mechanisms of (halo)alkaline adaptations, including ectoine biosynthesis, which is the first evidence for the formation of this osmoprotectant in archaea. According to 16S rRNA gene phylogeny, the strains possessed 98.3-98.9% sequence identity and belonged to the obligately aceticlastic genus Methanothrix with M. harundinaceae as the most closely related species. However, a more advanced phylogenomic reconstruction based on 122 conserved single-copy archaeal protein-coding marker genes clearly indicated a polyphyletic origin of the species included in the genus Methanothrix. We propose to reclassify Methanothrix harrundinacea (type strain 8AcT) into a new genus, Methanocrinis gen. nov., with the type species Methanocrinis harrundinaceus comb. nov. We also propose under SeqCode the complete genome sequences of strain MxTs (GCA_029167045.1) and strain M04AcTs (GCA_029167205.1) as nomenclatural types of Methanocrinis natronophilus sp. nov. and Methanocrinis alkalitolerans sp. nov., respectively, which represent other species of the novel genus. This work demonstrates that the low energy aceticlastic methanogenesis may function at extreme conditions present in (halo)alkaline habitats.

Acetate and Acetyl-CoA Metabolism of ANME-2 Anaerobic Archaeal Methanotrophs.

Acetyl-CoA synthetase (ACS) and acetate ligase (ACD) are widespread among microorganisms, including archaea, and play an important role in their carbon metabolism, although only a few of these enzymes have been characterized. Anaerobic methanotrophs (ANMEs) have been reported to convert methane anaerobically into CO2, polyhydroxyalkanoate, and acetate. Furthermore, it has been suggested that they might be able to use acetate for anabolism or aceticlastic methanogenesis. To better understand the potential acetate metabolism of ANMEs, we characterized an ACS from ANME-2a as well as an ACS and an ACD from ANME-2d. The conversion of acetate into acetyl-CoA (Vmax of 8.4 μmol mg-1 min-1 and Km of 0.7 mM acetate) by the monomeric 73.8-kDa ACS enzyme from ANME-2a was more favorable than the formation of acetate from acetyl-CoA (Vmax of 0.4 μmol mg-1 min-1 and Km of 0.2 mM acetyl-CoA). The monomeric 73.4-kDa ACS enzyme from ANME-2d had similar Vmax values for both directions (Vmax,acetate of 0.9 μmol mg-1 min-1 versus Vmax,acetyl-CoA of 0.3 μmol mg-1 min-1). The heterotetrameric ACD enzyme from ANME-2d was active solely in the acetate-producing direction. Batch incubations of an enrichment culture dominated by ANME-2d fed with 13C2-labeled acetate produced 3 μmol of [13C]methane in 7 days, suggesting that this anaerobic methanotroph might have the potential to reverse its metabolism and perform aceticlastic methanogenesis using ACS to activate acetate albeit at low rates (2 nmol g [dry weight]-1 min-1). Together, these results show that ANMEs may have the potential to use acetate for assimilation as well as to use part of the surplus acetate for methane production. IMPORTANCE Acetyl-CoA plays a key role in carbon metabolism and is found at the junction of many anabolic and catabolic reactions. This work describes the biochemical properties of ACS and ACD enzymes from ANME-2 archaea. This adds to our knowledge of archaeal ACS and ACD enzymes, only a few of which have been characterized to date. Furthermore, we validated the in situ activity of ACS in ANME-2d, showing the conversion of acetate into methane by an enrichment culture dominated by ANME-2d.

Microbial response behavior to powdered activated carbon in high-solids anaerobic digestion of kitchen waste: Metabolism and functional prediction analysis

Anaerobic digestion (AD) can not only treat organic waste, but also recycle energy. However, high-solids AD of kitchen waste usually failed due to excessive acidification. In this study, the effect of activated carbon (AC) on kitchen waste AD performance was investigated under high-solids conditions (total solids contents = 15%). The results showed that efficiencies of acidogenesis and methanogenesis were promoted in presence of moderate concentration (50 g/L > AC >5 g/L), but high concentration (AC >70 g/L) weakened AD performance. Moreover, AC addition enhanced the methane production rate from 66.0 mL/g VS to 231.50 mL/g VS, i.e., up to 250.7%. High-throughput sequencing results demonstrated that the abundance of electroactive DMER64 increased from less than 1%–29.7% (20 g/L AC). As AC gradually increased，aceticlastic methanogenesis changed to hydrogenotrophic pathway. Predicted functional analysis indicated that AC can enhance abundances of energy and inorganic ion metabolism, resulting in high methane production.

Microbial Diversity, Community Turnover, and Putative Functions in Submarine Canyon Sediments under the Action of Sedimentary Geology.

Sampling challenges in deep-sea ecosystems lead to a lack of knowledge about the distribution of microbes in different submarine canyons. To study microbial diversity and community turnover under different ecological processes, we performed 16S/18S rRNA gene amplicon sequencing for sediment samples from a submarine canyon in the South China Sea. Bacteria, archaea, and eukaryotes made up 57.94% (62 phyla), 41.04% (12 phyla), and 1.02% (4 phyla) of the sequences, respectively. Thaumarchaeota, Planctomycetota, Proteobacteria, Nanoarchaeota, and Patescibacteria are the five most abundant phyla. Heterogeneous community composition was mainly observed in vertical profiles rather than horizontal geographic locations, and microbial diversity in the surface layer was much lower than that in deep layers. According to the null model tests, homogeneous selection dominated community assembly within each sediment layer, whereas heterogeneous selection and dispersal limitation dominated community assembly between distant layers. Different sedimentation processes of sediments, i.e., rapid deposition caused by turbidity currents or slow sedimentation, seem to be primarily responsible for these vertical variations. Finally, functional annotation through shotgun-metagenomic sequencing found that glycosyl transferases and glycoside hydrolases are the most abundant carbohydrate-active enzyme categories. The most likely expressed sulfur cycling pathways include assimilatory sulfate reduction, the link between inorganic and organic sulfur transformation, and organic sulfur transformation, while the potentially activated methane cycling pathways include aceticlastic methanogenesis and aerobic and anaerobic oxidation of methane. Overall, our study revealed high levels of microbial diversity and putative functions in canyon sediments and the important influence of sedimentary geology on microbial community turnover between vertical sediment layers. IMPORTANCE Deep-sea microbes have received growing attention due to their contribution to biogeochemical cycles and climate change. However, related research lags due to the difficulty of collecting samples. Based on our previous study, which revealed the formation of sediments under the dual action of turbidity currents and seafloor obstacles in a submarine canyon in the South China Sea, this interdisciplinary research provides new insights into how sedimentary geology influences microbial community assembly in sediments. We proposed some uncommon or new findings, including the following: (i) microbial diversity was much lower on the surface than in deeper layers (ii) archaea and bacteria dominated the surface and deep layers, respectively; (iii) sedimentary geology played key roles in vertical community turnover; and (iv) the microbes have great potential to catalyze sulfur, carbon, and methane cycling. This study may lead to extensive discussion of the assembly and function of deep-sea microbial communities in the context of geology.

Long-term performance study applying a tandem AnSBR-ASBR to treat wastewater containing N, N-dimethylformamide: Sludge physicochemical properties, microecology, and functional genes

The effective treatment of industrial pollutant N, N-dimethylformamide (DMF) containing wastewater is a matter of worldwide concern. This study investigated the performance and microecology of anaerobic granular sludge (AGS) and aerobic sludge (AS) in treating DMF wastewater. It was also explored the degradation pathways of DMF in an anaerobic sequencing batch reactor series aerobic sequencing batch reactor (AnSBR-ASBR). Results showed that 5 mg/L DMF facilitated the degradation of chemical oxygen demand (COD) by AGS with AS, while 100 mg/L DMF reduced the efficiency of AS. The physicochemical properties of the sludge showed that the polysaccharide and protein levels in extracellular polymeric substances and key enzyme activity (acetate kinase, coenzyme F420) deteriorated when DMF increased from 25 to 100 mg/L. As the DMF dose increased (0–100 mg/L), the Euryarchaeota abundance in the AGS decreased from 28.41% to 11.06%, and the predominant methanogenic pathway shifted from a combination of aceticlastic methanogenesis (AM) and hydrogenotrophic methanogenesis (HM) to HM. The metabolic pathway to effectively remove DMF was as follows: hydrolysis of DMF to dimethylamine, methylamine, and finally mineralisation to CO2. The abundance of dmd-tmd, a functional gene encoding dimethylamine/trimethylamine dehydrogenase, increased as the DMF increased in the AS. This study systematically examined the operating efficiency and stability of the AnSBR-ASBR, and summarized the AGS and AS stress response, microbial community evolution, key metabolic pathways to DMF, also provided useful information for the treatment of industrial wastewater containing DMF.

Thermodynamic restrictions determine ammonia tolerance of methanogenic pathways in Methanosarcina barkeri

Ammonia is a ubiquitous potential inhibitor of anaerobic digestion processes, mainly exhibiting inhibition towards methanogenic activity. However, knowledge as to how ammonia affects the methanogens is still limited. In this study, we cultured a multitrophic methanogen, Methanosarcina barkeri DSM 800, with acetate, H2/CO2, and methanol to evaluate the influence of ammonia on different methanogenic pathways. Aceticlastic methanogenesis was more sensitive to increased ammonia concentrations than hydrogenotrophic and methylotrophic methanogenesis. Theoretical maximum NH3 tolerances of M. barkeri fed with acetate, H2/CO2, and methanol were calculated to be 39.1 ± 9.0, 104.3 ± 7.4, and 85.7 ± 1.0 mg/L, respectively. The order of the ΔG range of M. barkeri under three methanogenic pathways reflected the order of ammonia tolerance of M. barkeri. Our results provide insights into the role of the thermodynamic potential of methanogenesis on the tolerance of ammonia stress; and shed light on the mechanism of ammonia inhibition on anaerobic digestion.

Effect of Inoculum Microbial Diversity in Ex Situ Biomethanation of Hydrogen.

The effects of the inoculum origin, temperature or operational changes on ex situ biomethanation by complex microbial communities have been investigated; however, it remains unclear how the diversity of the inoculum influences the process and its stability. We explored the effect of microbial diversity of four inocula (coded as PF, WW, S37 and Nrich) on methane production, process stability and the formation of volatile fatty acids as by-products. The highest methane amounts produced were 3.38 ± 0.37 mmol, 3.20 ± 0.07 mmol, 3.07 ± 0.27 mmol and 3.14 ± 0.06 mmol for PF, WW, S37 and Nrich, respectively. The highest acetate concentration was found in less diverse cultures (1679 mg L-1 and 1397 mg L-1 for S37 and Nrich, respectively), whereas the acetate concentrations remained below 30 mg L-1 in the more diverse cultures. The maximum concentration of propionate was observed in less diverse cultures (240 mg L-1 and 37 mg L-1 for S37 and Nrich cultures, respectively). The highly diverse cultures outperformed the medium and low diversity cultures in the long-term operation. Methanogenic communities were mainly composed of hydrogenotrophic methanogens in all cultures. Aceticlastic methanogenesis was only active in the highly diverse sludge community throughout the experiment. The more diverse the inocula, the more methane was produced and the less volatile fatty acids accumulated, which could be attributed to the high number of microbial functions working together to keep a stable and balanced process. It is concluded that the inoculum origin and its diversity are very important factors to consider when the biomethanation process is performed with complex microbial communities.

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.

Improved methane production from the two-phase anaerobic digestion and dewaterability of anaerobically digested sludge by β-cyclodextrin pretreatment

Cyclodextrin-based technologies have drawn worldwide attention for wastewater and sludge treatment due to its extraordinary solubilization of hydrophobic organics. β-cyclodextrin (β-CD) has been applied for enhancing sludge solubilization for production of volatile fatty acids. However, the impact of β-CD on methane production in the anaerobic digestion seemingly was unnoticed in the past, which was likely due to its inhibition effect on the methanogenesis in single-phase digestion system. Herein, a new pretreatment strategy using β-CD was firstly explored to enhance methane yield in two-phase anaerobic digestion of waste activated sludge (WAS) and simultaneously improve anaerobically digested sludge (ADS) dewaterability. Results illustrated that the total methane yield in the two phase increased from 101.43 to 277.67 mL/g VS with β-CD level raising from 0 to 0.3 g/g TS. A better ADS dewaterability was also obtained (24.14% capillary suction time increase). Mechanism investigations suggested that WAS solubilization and acidification were significantly enhanced in the first phase (i.e., acidogenic phase), with 8.97-fold soluble chemical oxygen demand (SCOD) and 17.37-fold short-chain fatty acids (SCFAs) increases. This implied that more available substrates after β-CD pretreatment could be used for methanogenesis. Further calculation indicated the utilization of β-CD was not the main reason for the enhanced methane. Model-based explorations revealed hydrolysis rate and methane potential were improved by 153.17% and 175.36% after β-CD pretreatment. The activities of enzymes associated with hydrolysis/acidification/methanogenesis in methanogenic phase were improved even if those were suppressed in acidogenic phase. Moreover, β-CD increased the abundance of key hydrolysis-acidification microbes and strengthened both the hydrogenotrophic and aceticlastic methanogenesis pathways. Finally, a “multifaceted benefits” operation concept based on β-CD treatment was proposed, providing an implication for wastewater treatment and sludge disposal.

Enhanced methane production by using phytoremediated Halogeton glomeratus as substrate via anaerobic digestion

The phytoremediated plant contains rich carbon resources, but the anaerobic digestion (AD) process can be inhibited due to high Heavy metals (HMs). In this study, the phytoremediated Halogeton glomeratus (H. glomeratus) was utilized in AD. Results showed that suitable HMs improved the AD efficiency, and the HMs were positively correlated to methane production and microbiota. Compared to CK (without HMs contaminated), H. glomeratus containing HMs (TG) enhanced methane production (increased 85.5%). Also, the highest VS removal rate of 66.5% were achieved. Furthermore, the increased of hydrolytic and acidogenic Acinetbacter and Hydrogenophaga improved substrates degradation. Besides, the methanogenic Methanosarcina increased from 6.5 to 89.5%, and the methanogenic pathway shifted from hydrogenotrophic methanogenesis to aceticlastic methanogenesis. Thus, substrates conversion to methane was improved. Finally, the HMs in digestion slurry were successfully removed and decreased its secondary pollution. This study provided a potential for the resourceful utilization of the phytoremediated plants with suitable HMs concentration.

Studies on the inhibition of methanogenesis and dechlorination by (4-hydroxyphenyl) chloromethanesulfonate.

The purpose of this study was to demonstrate the inhibitory effect of chemicals on methane emissions in paddy soil. We found that (4-hydroxyphenyl) chloromethanesulfonate (C-1) has a methanogenic inhibition activity, and we studied its inhibition mechanism using laboratory tests. The study found that C-1 treatment of flooded soil did not significantly affect the bacterial community but rather the archaeal community; particularly, Methanosarcina spp. C-1 strongly inhibited the aceticlastic methanogenesis route. It was suggested that the inhibitory target of C-1 was different from the well-known methanogenic inhibitor 2-bromoethanesulfonate, which targets methyl-coenzyme M reductase of methanogen. In addition, C-1 had a secondary effect of inhibiting the dechlorination of chlorophenols. Although field trials are required as the next development step, C-1 can be used to reduce methane emissions from paddy fields, one of the largest sources in the agricultural sector.

Microorganisms as bio-filters to mitigate greenhouse gas emissions from high-altitude permafrost revealed by nanopore-based metagenomics.

The distinct climatic and geographical conditions make high-altitude permafrost on the Tibetan Plateau suffer more severe degradation than polar permafrost. However, the microbial responses associated with greenhouse gas production in thawing permafrost remain obscured. Here we applied nanopore-based long-read metagenomics and high-throughput RNA-seq to explore microbial functional activities within the freeze-thaw cycle in the active layers of permafrost at the Qilian Mountain. A bioinformatic framework was established to facilitate phylogenetic and functional annotation of the unassembled nanopore metagenome. By deploying this strategy, 42% more genera could be detected and 58% more genes were annotated to nitrogen and methane cycle. With the aid of such enlarged resolution, we observed vigorous aerobic methane oxidation by Methylomonas, which could serve as a bio-filter to mitigate CH4 emissions from permafrost. Such filtering effect could be further consolidated by both on-site gas phase measurement and incubation experiment that CO2 was the major form of carbon released from permafrost. Despite the increased transcriptional activities of aceticlastic methanogenesis pathways in the thawed permafrost active layer, CH4 generated during the thawing process could be effectively consumed by the microbiome. Additionally, the nitrogen metabolism in permafrost tends to be a closed cycle and active N2O consumption by the topsoil community was detected in the near-surface gas phase. Our findings reveal that although the increased thawed state facilitated the heterotrophic nitrogen and methane metabolism, effective microbial methane oxidation in the active layer could serve as a bio-filter to relieve the overall warming potentials of greenhouse gas emitted from thawed permafrost.

Inhibition mechanisms of ammonia and sulfate in high-solids anaerobic digesters for food waste treatment: Microbial community and element distributions responses

The horizontal flow anaerobic digester indicated that high ammonia (2923 mg/L) and SO42− (3653 mg/L) would influence the performance of methane production with food waste as substrates. Therefore, bottle anaerobic digestion reactors were carried out to investigate the effect of ammonia/sulfate concentrations on the methane production. Experimental results manifested that the anaerobic digesters with an ammonia concentration of 3500 mg/L or sulfate of 1600 mg/L showed the best performance of methane production, with an average methane yield of 0.32 and 0.33 L (g VS)−1 d−1, respectively. Specifically, a higher ammonia (6500 mg/L) or sulfate (1600-3500 mg/L) level hindered the bioconversion of C from liquid to gas phase (2.68% or 1.73% CH4-Gas, respectively), while insignificantly for the hydrolyzation of C and N from solid to liquid phase. Similar to sulfate, high ammonia nitrogen seriously inhibited the methanation process, leading to a significant carbon accumulation in the anaerobic reactor, especially for propionic acid. The predominant archaea Methanosarcina at genus level indicated that aceticlastic methanogenesis was the major methanogenic pathway. Meanwhile, high ammonia level suppressed the activity of Methanosarcina, while modest sulfate improved H2-consuming methanogens activity. A large fraction of unclassified bacteria within the Firmicutes (43.78%-63.17%) and Bacteroidetes (24.20%-33.30%) phylum played an important role in substrates hydrolysis.

High nitrite–nitrogen stress intensity drives nitrite anaerobic oxidation to nitrate and inhibits methanogenesis

Nitrite is an important intermediate in nitrogen metabolism. We explored the effect of nitrite–nitrogen stress intensity (NNSI) on nitrite metabolism and methanogenesis in anaerobic digestion. The results showed that the NNSI regulated microbial diversity, composition, and functions, and microbial community assembly was primarily shaped by stochastic processes. Moreover, the NNSI was negatively correlated with α-diversity and positively correlated with non-metric multi-dimensional scaling distance. Denitrification gradually increased with increasing NNSI; however, methanogenesis was gradually inhibited, which was primarily due to the inhibition of the aceticlastic methanogenesis pathway (i.e., Methanosaeta) and methylotrophic methanogenesis pathway (i.e., Candidatus_Methanofastidiosum). High NNSI (1882 ± 98.99 mg/L NO2−-N) promoted nitrite anaerobic oxidation to nitrate and was favorable for dissimilatory nitrate reduction to ammonia (DNRA). We present evidence for the microbial transformation of nitrite under anaerobic conditions, with potential geochemical and evolutionary importance. As nitrogen oxides were already present on early Earth, our finding presents the possibility of a nitrogen cycle before the evolution of oxygenic photosynthesis.