Dominant Methanogens Research Articles

A new integrated single-chamber air-cathode microbial fuel cell - Anaerobic membrane bioreactor system for improving methane production and membrane fouling mitigation

A novel integrated single-chamber air-cathode microbial fuel cell - anaerobic membrane bioreactor (ScMFC-AnMBR) system was designed. It involved an anaerobic membrane bioreactor (AnMBR) and a single-chamber air-cathode microbial fuel cell (ScMFC) being constructed in a common reaction chamber to enhance methane production and reduce membrane fouling in the AnMBR. Results indicated that ScMFC-AnMBR delivered a stable micro-bioelectric field environment with a voltage output of 95 ± 4 mV. Compared with conventional AnMBR (C-AnMBR), methane production using this system increased by 35.89%. Soluble microbial product (SMP) and extracellular polymeric substances (EPS) dropped by 65.3% and 43.1%, respectively. Particularly, the transmembrane pressure (TMP) in the operating cycle was in a slow growth status with the maximum value of only 18.5 kPa. The bioelectric field helped aceticlastic methanogens (Methanosaeta) replace hydrogenotrophic methanogens (Methanobacterium) as the dominant methanogens via electron transfer under a closed-circuit scenario. As syntrophic bacteria of methanogens (Syntrophobacter, Smithella and Syner-01) and exoelectrogens of Desulfovibrio were selected by the bioelectric field and gained a stable foothold, bio-foulant (Megasphaera) was significantly reduced. The complex microbial synergism in ScMFC-AnMBR greatly improved the methanogenic performance, thus effectively alleviated membrane fouling and prolonged the operation cycle of the system. Demonstrated here is the feasibility of practical application.

Anaerobic co-digestion of thermo-alkaline pretreated microalgae and sewage sludge: Methane potential and microbial community

To improve methane production from sewage sludge (SS), co-digestion of SS and microalgae (MA) was studied and the application of thermo-alkaline pretreatment to MA was evaluated. The results showed that thermo-alkaline pretreatment at 90°C for 120 min on MA was the optimum pretreatment condition. Furthermore, when the volatile solids (VS) ratio of SS and MA was 1:2, the methane yield reached maximum (368.94 mL/g VS). Fourier transform infrared (FT-IR) and thermogravimetric analysis confirmed the synergetic effects of thermo-alkaline pretreated MA on its co-digestion with SS. The analyses of microbial community indicated that Methanobacterium and Methanosarcina were the dominant methanogens during the co-digestion process. However, the relative abundance of Methanosarcina in thermo-alkaline pretreated groups was higher compared to unpretreated groups. The microbial community structure might be affected by thermo-alkaline pretreatment rather than by the MA dosage in the co-digestion.

Effect of different microbial seeds on batch anaerobic digestion of fish waste

Initial microbial compositions would be the precursor for the efficient anaerobic digestion (AD) of fish waste (FW). A mesophilic batch test was conducted using four seeds collected from different digesters treating various combinations of substrates to investigate their effects on FW degradation. Key microbial groups were identified by 16s rRNA gene-based metagenomics analysis. Among four, the seed from the digester co-digesting livestock manure, food waste, and food wastewater showed the best performance and obtained the highest methane yield (350.5 ± 5.2 mL/gVSadded) and lowest lag phase (0.6 ± 0.1 d). Proteiniphilum, Aminobacterium, dgA-11 gut group, and Syntrophomonas were dominant bacterial genera identified in FW degradation. Methanosaeta was the dominant methanogen in the best performing seed and microbial network analysis revealed its contribution to achieving the highest CH4 yield. Obtained results could be useful in selecting microbial seed sources to avoid system imbalance in full-scale digesters that treat FW.

16S rRNA genes- and metagenome-based confirmation of syntrophic butyrate-oxidizing methanogenesis enriched in high butyrate loading

The understanding and enrichment of consortia formed by syntrophic butyrate-oxidizing bacteria and methanogens in the complex environment are crucial for effectively degrading butyrate and preventing acid inhibition. In this study, the better butyrate-tolerated and highly efficient microbial consortia were domesticated and enriched through adding butyric acid ranging from 0.2 to 4.4 g/(L·d). The volumetric biogas production continuously increased to 1.65 L/(L·d). Microbial community diversity showed that a dramatic shift of bacterial structure occurred at BAL of 1.6 g/(L·d) and the structure presented better stability at high BAL. The syntrophic consortia and the main metabolic pathways were revealed through combination of the 16S rDNA and metagenome sequencing analyses. Syntrophomonas was the major butyrate-oxidizing bacterium and oxidized butyrate mainly through β-oxidaiton. Synergistaceae and Mesotoga acted as the main acetate-oxidizing bacteria. IHT and methanogenesis pathways were strongly enhanced by DMER64 and Methanosarcina as the main H2 carrier and dominant methanogen, respectively.

Impact of Temperature on the Performance and Character of the Methanogenic Community of a Fixed-Bed Anaerobic Reactor at Psychrophilic Temperature

To determine the effects of a gradual temperature decrease on reactor performance and the microbial community, four fixed-bed reactors that were packed with a biofilm carrier were operated for 217 days. The temperature of the reactors was decreased from 30 °C to 3 °C. The reactors initially soured at 3 °C and recovered when they were returned to 4 °C, as indicated by the stabilization of biogas production, methane production, VFA concentration, pH, and the COD removal rate. Our results also revealed that methanomicrobiales were the dominant methanogen, the concentration of the 16S rRNA gene in the carbon-fiber carrier sludge exceeded the same gene concentration in the deposited sludge, and that the carbon-fiber carrier played an important role in methanomicrobiale colonization at low temperatures. We suggest that 4 °C is the low-temperature threshold for optimal reactor performance.

Patterns of syntrophic interactions in methanogenic conversion of propionate.

Methanogenesis is central to anaerobic digestion processes. The conversion of propionate as a key intermediate for methanogenesis requires syntrophic interactions between bacterial and archaeal partners. In this study, a series of methanogenic enrichments with propionate as the sole substrate were developed to identify microbial populations specifically involved in syntrophic propionate conversion. These rigorously controlled propionate enrichments exhibited functional stability with consistent propionate conversion and methane production; yet, the methanogenic microbial communities experienced substantial temporal dynamics, which has important implications on the understanding of mechanisms involved in microbial community assembly in anaerobic digestion. Syntrophobacter was identified as the most abundant and consistent bacterial partner in syntrophic propionate conversion regardless of the origin of the source culture, the concentration of propionate, or the temporal dynamics of the culture. In contrast, the methanogen partners involved in syntrophic propionate conversion lacked consistency, as the dominant methanogens varied as a function of process condition and temporal dynamics. Methanoculleus populations were specifically enriched as the syntrophic partner at inhibitory levels of propionate, likely due to the ability to function under unfavorable environmental conditions. Syntrophic propionate conversion was carried out exclusively via transformation of propionate into acetate and hydrogen in enrichments established in this study. Microbial populations highly tolerant of elevated propionate, represented by Syntrophobacter and Methanoculleus, are of great significance in understanding methanogenic activities during process perturbations when propionate accumulation is frequently encountered. Key points • Syntrophobacter was the most consistent bacterial partner in propionate metabolism. • Diverse hydrogenotrophic methanogen populations could serve as syntrophic partners. • Methanoculleus emerged as a methanogen partner tolerant of elevated propionate.

Methane emissions may be driven by hydrogenotrophic methanogens inhabiting the stem tissues of poplar.

Living trees in forests emit methane (CH4 ) from their stems. However, the magnitudes, patterns, drivers, origins, and biogeochemical pathways of these emissions remain poorly understood. We measured insitu CH4 fluxes in poplar stems and soils using static chambers and investigated the microbial communities of heartwood and sapwood by sequencing bacterial 16S, archaeal 16S, and fungal ITS rRNA genes. Methane emissions from poplar stems occurred throughout the sampling period. The mean CH4 emission rate was 2.7 mg m-2 stem d-1 . Stem CH4 emission rate increased significantly with air temperature, humidity, soil water content, and soil CH4 fluxes, but decreased with increasing sampling height. The CO2 reduction and methylotrophic methanogenesis were the major methanogenic pathways in wood tissues. The dominant methanogen groups detected in stem tissues were Methanobacterium, Methanobrevibacter, Rice Cluster I, Methanosarcina, Methanomassiliicoccus, Methanoculleus, and Methanomethylophilaceae. In addition, three methanotrophic genera were identified in the heartwood and sapwood - Methylocystis, Methylobacterium, and Paracoccus. Overall, stem CH4 emissions can originate directly from the internal tissues or co-occur from soils and stems. The co-existence of methanogens and methanotrophs within heartwood and sapwood highlights a need for future research in the microbial mechanisms underlying stem CH4 exchange with the atmosphere.

Analysis of enzyme activity and microbial community structure changes in the anaerobic digestion process of cattle manure at sub-mesophilic temperatures

Abstract To improve the biogas potential in anaerobic digestion of cattle manure in cold regions, we investigated, in this study, the potential of the anaerobic digestion of cattle manure through sequencing batch anaerobic digestion experiments at sub-mesophilic temperatures (15°C, 20°C, 25°C, and 37°C) for 50 days. Specifically, the changes in the biogas yield, pH, sCOD concentration, enzyme activity, and microbial community structure were examined. The maximum daily yield of biogas was 19.18 mL·gVS−1, which was recorded on day 6 at 37°C, and the final biogas accumulation yield at this temperature was 403.64 mL·gTS−1. The concentration of sCOD gradually increased as enzyme activity increased. The maximum activities of cellulase, hemicellulase, protease, and amylase were recorded in the 37°C experimental group. The decrease in temperature severely limited the activity of different types of enzymes, resulting in a decrease in the activity of microorganisms, which greatly influenced the methanogenic reaction. The dominant bacteria at the phylum level were Bacteroidetes and Proteobacteria, and the dominant methanogen at the genus level was Methanosaeta.

Stable, high-rate anaerobic digestion through vacuum stripping of digestate

This study coupled anaerobic digestion with vacuum stripping to achieve stable digestion at higher organic loading rates. Besides mitigation of ammonia inhibition, vacuum stripping of digestate improves solids solubilization and dewaterability due to vacuum-enhanced low-temperature thermal and mild-alkaline treatment under the vacuum stripping conditions (65 °C, 25–27 kPa, and pH 9). Batch vacuum stripping for 8 h removed 97.4–99.4% of ammonia, increased the dissolved fraction of volatile solids (VS) by 72.5%, and improved dewaterability with 30% decreases in time-to-filter and viscosity. The digesters having 2.9% of digestate replaced daily by vacuum stripped digestate were stable up to organic loading rate of 4.3 g-VS/Lreactor/d with biogas production at 3.15 L/Lreactor/d, while the digesters without stripping attained biogas production of 1.90 L/Lreactor/d at its highest stable organic loading rate of 2.5 g-VS/Lreactor/d. Acetoclastic Methanosaeta were the dominant methanogens, which became more resistant to ammonia stress in the digesters with vacuum stripping.

Long-term performance, membrane fouling behaviors and microbial community in a hollow fiber anaerobic membrane bioreactor (HF-AnMBR) treating synthetic terephthalic acid-containing wastewater

Purified terephthalic acid (PTA) wastewater with properties of poor biodegradation and high toxicity is produced from refining and synthesis of petrochemical products. In this study, a lab-scale hollow fiber membrane bioreactor (HF-AnMBR) fed with synthetic PTA wastewater was operated over 200 days with stepwise decreased hydraulic retention time (HRT) to investigate the long-term performance, membrane fouling mechanism and microbial community evolution. Results showed that a stable chemical oxygen demand (COD) removal rate of 65.8 ± 4.1% was achieved at organic loading rate of 3.1 ± 0.3 g-COD/L-reactor/d and HRT 24 h, under which the methane production rate reached 0.33 ± 0.02 L/L-reactor/d. Further shortening HRT, however, led to the decreased COD removal efficiency and low methane bioconversion. A mild membrane fouling occurred due to the production of colloidal biopolymers and the interaction between increased colloidal substances secreted/cracked by microorganisms and membrane interface. Further 16S rRNA analysis indicated that microbial diversity and richness had changed with the variation of HRT while Methanosaeta, and Methanolinea species were always the dominant methanogens responsible for methane production. The results verify that HF-AnMBR is an alternative technology for PTA wastewater treatment along with energy harvesting, and provide a new avenue toward sustainable petrochemical wastewater management.

Responses of methane production, microbial community and antibiotic resistance genes to the mixing ratio of gentamicin mycelial residues and wheat straw in anaerobic co-digestion process

Anaerobic co-digestion (AcoD) of gentamicin mycelial residues (GMRs), a kind of nitrogen-rich biowaste, and wheat straw (WS) is an attractive technology for the recycling of GMRs. However, the effects of the co-substrate ratio on methane production, system stability and antimicrobial resistance during co-digestion remain unclear. Thus, this study aimed to fill in the blanks through AcoD of GMRs and WS with different mixing ratios (1:0, 2:1, 1:1, 1:2, 0:1, VS basis) via batch tests. Results showed that AcoD facilitated methane production than mono anaerobic digestion and reduced the accumulation of the toxic substances, such as ammonia nitrogen and humic-like substances. The maximum methane production was obtained at the reactors with the mixing ratio of 1:1 and 1:2 (R-1:1 and R-1:2), which matched with the relative abundance of key enzymes related to methanogenesis predicted by PICRUSt. Microbial community analysis indicated that Methanosaeta was the most dominant methanogen in the AcoD reactors. The highest relative abundance of Methanosaeta (45.1%) was obtained at R-1:1 due to the appropriate AcoD conditions, thus, providing greater possibilities for high stability of AcoD system. Additionally, AcoD of the GMRs and WS under the mixing ratio of 1:1 and 1:2 did not prompt the increase of antibiotic resistance genes (ARGs). Not only that, the likelihood of horizontal gene transfer declined in R-1:1 due to the weaker connection and transport between host and recipient bacteria. Findings of this study suggested that the suitable mixing ratio of GMRs and WS contributes to methane production and system stability, and reduces the dissemination risks of ARGs.

Syngas biomethanation: effect of biomass-gas ratio, syngas composition and pH buffer

The concept of syngas biomethanation is attractive, however, it still needs improvement in optimizing the operational conditions. In the present study, syngas fermentations under different carbon monoxide (CO), carbon dioxide (CO2) and hydrogen (H2) compositions were conducted under two different biomass-gas ratio (BGR) systems. The results showed that high BGR enhanced the CO consumption rate, achieving a 60% enhancement with CO as the sole substrate. Stoichiometric H2 addition could successfully convert all the CO and CO2 to pure methane, however, higher H2 partial pressure might decline the CO consumption due to pH inhibition from consumption of bicarbonate. Microbial analysis showed different syngas composition could affect the bacteria community, while, archaea community was only slightly affected with Methanothermobacter as the dominant methanogen. This study provided strategy for efficient syngas biomethanation and deeper insight into effect of H2 addition on CO conversion under different BGR systems.

Effect of magnetite on anaerobic digestion treating saline wastewater: Methane production, biomass aggregation and microbial community dynamics

In this study, the effect of magnetite amendment on anaerobic digestion was investigated at three increasing salinity levels (0.5%, 1% and 2% NaCl). The amendment of magnetite enhanced the methane yield by 36.3%, 33.3% and 16.5% at low salinity (0.5% NaCl) and high salinity (1% and 2% NaCl), respectively. Meanwhile, a larger proportion of granules was obtained in the magnetite amended reactor (48.05% vs 33.16% at the end of operation). Microbial analysis suggested magnetite could induce more methanogenesis partnerships between hydrogenotrophic methanogens and syntrophic bacteria. Methanosaeta and Methanocorpusculum were the alternating dominant methanogens at low salinity and high salinity. While Streptococcus and Mesotoga were two prevalent bacteria that showed totally different transition tendency in two reactors. Additionally, the supplement of magnetite could relieve the suppression of methanogenesis-related gene expression caused by salinity, thus facilitated the higher methane production.

Consecutive batch anaerobic digestion under ammonia stress: Microbial community assembly and process performance

To reveal the effects of the degree and mode of total ammonia nitrogen (TAN) stress on community assembly, composition, and process performance, sequential batch experiments of anaerobic digestion (AD) of food waste were conducted with three different ammonia levels (1800, 3000, and 6000 mg/L) and two exposure modes (stepwise and direct). The selected TAN levels varied in the degree to which they inhibited the AD process. However, deterministic processes may play a larger role in shaping the microbial community than stochastic processes regardless of the level and mode of TAN stress. Under controlled conditions, long-term exposure can lead to the convergence of the community to a specific equilibrium state. The key link affecting community function was acetogenesis. The relative abundance of hydrolytic and acidogenic bacteria under the 1800, 3000, and 6000 mg/L TAN groups was 67.36 ± 3.59%, 68.54 ± 5.73%, and 77. 09 ± 2.69%, respectively. The resistance and redundancy of functional bacteria can maintain the hydrolysis and acidification function of the AD process. Methanogens can also maintain their metabolic performance through the transfer of dominant methanogens from Methanosaeta and Methanospirillum to Methanosarcina and Methanoculleus. Acetogens, especially Pelotomaculum, were inhibited at 6000 mg/L TAN. This led to a decrease in their abundance and the accumulation of longer-chain volatile fatty acids, which was the main reason for the deterioration of the AD process. A new method that promotes acetogenesis under high ammonia stress needs to be developed to achieve efficient AD performance.

Diversity of dimethylsulfide-degrading methanogens and sulfate-reducing bacteria in anoxic sediments along the Medway Estuary, UK.

Methane is a powerful greenhouse gas but the microbial diversity mediating methylotrophic methanogenesis is not well-characterized. One overlooked route to methane is via the degradation of dimethylsulfide (DMS), an abundant organosulfur compound in the environment. Methanogens and sulfate-reducing bacteria (SRB) can degrade DMS in anoxic sediments depending on sulfate availability. However, we know little about the underlying microbial community and how sulfate availability affects DMS degradation in anoxic sediments. We studied DMS-dependent methane production along the salinity gradient of the Medway Estuary (UK) and characterized, for the first time, the DMS-degrading methanogens and SRB using cultivation-independent tools. DMS metabolism resulted in high methane yield (39%-42% of the theoretical methane yield) in anoxic sediments regardless of their sulfate content. Methanomethylovorans, Methanolobus and Methanococcoides were dominant methanogens in freshwater, brackish and marine incubations respectively, suggesting niche-partitioning of the methanogens likely driven by DMS amendment and sulfate concentrations. Adding DMS also led to significant changes in SRB composition and abundance in the sediments. Increases in the abundance of Sulfurimonas and SRB suggest cryptic sulfur cycling coupled to DMS degradation. Our study highlights a potentially important pathway to methane production in sediments with contrasting sulfate content and sheds light on the diversity of DMS degraders.

Metagenomic analysis reveals the methanogenic, ATP, and potassium-transport metabolisms of anaerobic systems with different ammonia concentrations

Excessive ammonia is a well-known inhibitor of anaerobic wastewater treatment systems. In this study, responses of methanogenic activity and pathway, microbial communities, adenosine triphosphatases (ATPase) system, and potassium (K+) transport system to different ammonia concentrations were investigated. The long-term acclimation with 1 g NH4+-N/L of ammonia decreased the methanogen abundance by more than 13% and shifted the dominant methanogen from Methanothrix to Methanosarcina. In the acetate-substrate reactor, genes encoding methanogenesis, ATPase, and K+ transport systems were suppressed. However, in the reactor fed with ethanol and acetate, the long-term acclimation with ammonia enriched ammonia-tolerant syntrophic bacteria (i.e., Geobacter) with a high abundance of ATPase and K+ transport genes. The robust ethanol oxidation and ammonia-detoxicate activities benefited the subsequent methanogenesis (including acetoclastic methanogenesis), increasing 78% of the maximum methane production rate, compared with the control reactor. Direct interspecies electron transfer-capable partners and PilA genes were both detected under 1 g NH4+-N/L of ammonia condition.

Microbiota of a UASB Reactor Treating Palm Oil Mill Effluent Using HiSeq Sequencing

Potential microbial communities in the UASB reactor fed with two different substrates i.e synthetic wastewater and raw palm oil mill effluent (POME) were elucidated by using one of the most popular techniques in molecular methods, viz 16S rDNA cloning. The methodology involved; the extraction of nucleic acids, amplification and cloning of the 16S rRNA genes on sequencing HiSeq platform and finally identification and affiliation of the isolated clone with the aid of phylogenetic software. Results showed that the genus methanosarcina and methanosaeta were dominant methanogens in this study for both substrates types. Overall, microbial population (Bacteria and Archaea) in sample A (POME as substrate) is more diverse compared to sample B (synthetic wastewater as substrate) due to abundance of microorganism population in raw POME which was used as a substrate. However for the methanogenic (Archaea) diversity in both samples, there was not much different between sample A and sample B probably due to similar inoculum was inoculated in the reactor despite of have using different substrate type.

Microbial Communities in Flexible Biomethanation of Hydrogen Are Functionally Resilient Upon Starvation

Ex situ biomethanation allows the conversion of hydrogen produced from surplus electricity to methane. The flexibility of the process was recently demonstrated, yet it is unknown how intermittent hydrogen feeding impacts the functionality of the microbial communities. We investigated the effect of starvation events on the hydrogen consumption and methane production rates (MPRs) of two different methanogenic communities that were fed with hydrogen and carbon dioxide. Both communities showed functional resilience in terms of hydrogen consumption and MPRs upon starvation periods of up to 14 days. The origin of the inoculum, community structure and dominant methanogens were decisive for high gas conversion rates. Thus, pre-screening a well performing inoculum is essential to ensure the efficiency of biomethanation systems operating under flexible gas feeding regimes. Our results suggest that the type of the predominant hydrogenotrophic methanogen (here: Methanobacterium) is important for an efficient process. We also show that flexible biomethanation of hydrogen and carbon dioxide with complex microbiota is possible while avoiding the accumulation of acetate, which is relevant for practical implementation. In our study, the inoculum from an upflow anaerobic sludge blanket reactor treating wastewater from paper industry performed better compared to the inoculum from a plug flow reactor treating cow manure and corn silage. Therefore, the implementation of the power-to-gas concept in wastewater treatment plants of the paper industry, where biocatalytic biomass is readily available, may be a viable option to reduce the carbon footprint of the paper industry.

Effects of different functional strains on key metabolic pathways of methanogenesis in the domestic waste fermentation reactor

The improvement of landfill gas utilization efficiency will produce considerable environmental and economic benefits. This study simulated the fermentation of domestic waste through the addition of functional strains. The related gas-liquid parameters and microbial community diversity were measured, and the Tax4Fun analysis method was used to predict the methanogenic function of the methanogens. The results showed the following: i) Only the addition of Proteiniphilum acetatigenes and Bacteroides thetaiotaomicron produced significant colonization by functional microorganisms and strengthened the decomposition of organic matters in the fermentation system, especially protein, which improved the gas production efficiency. The average gas production by the PB (An experimental group only added Proteiniphilum acetatigenes and Bacteroides thetaiotaomicron) group was 1353.70 mL which was higher than that of the other groups by 19.27%–57.59%. ii) The dominant methanogens were Methanobacterium which mainly produced methane by using hydrogen and carbon dioxide as substrates according to the functional prediction. The results of the study reveal the biological strengthening mechanism of domestic waste landfill gas, which is conducive to improving the energy recycling efficiency of landfills.

Can the addition of biochar improve the performance of biogas digesters operated at 45°C?

Biogas from anaerobic digestion (AD) may contribute towards a green energy scene in the future. Despite many benefits, AD plant operators are still challenged by limitations of the technology. This study aimed at evaluating the effect of biochar addition to AD reactors operated at 45°C to optimize productivity, efficiency and stability of the process. Biochar is characterized by a large surface area and may therefore facilitate the creation of biofilms on the one hand and remove environmental contaminants on the other hand. Moreover, biochar may serve as pH control agent due to its alkaline properties, counteracting reactor acidification. Our results showed that the addition of biochar reduced efficiently the amount of H2S that initially occurred in the headspace during reactor start-up. Nevertheless, no impact on biogas- and methane yields was observed. This might be explained with an already well-working AD system without any severe disturbances, where the balancing nature of biochar did hardly count. Future studies, however, need to evaluate the effect on stressed systems. The microbiome analysis provided a detailed view of the microbial dynamics during the start-up phase, revealing Methanosarcina as dominant methanogen in the end. The addition of biochar did not alter the microbial community composition.