Acetoclastic Methanogens Research Articles

Proteomic profiling of robust acetoclastic methanogen in chrysene-altered anaerobic digestion: Global dissection of enzymes

Methanogen is a pivotal player in pollution treatment and energy recovery, and emerging pollutants (EPs) frequently occur in methanogen-applied biotechnology such as anaerobic digestion (AD). However, the direct effect and underlying mechanism of EPs on crucial methanogen involved in its application still remain unclear. The positive effect of chrysene (CH) on semi-continuous AD of sludge and the robust methanogen was dissected in this study. The methane yield in the digester with CH (100 mg/kg dry sludge) was 62.1 mL/g VS substrate, much higher than that in the control (46.1 mL/g VS substrate). Both methane production from acetoclastic methanogenesis (AM) and the AM proportion in the methanogenic pathway were improved in CH-shaped AD. Acetoclastic consortia, especially Methanosarcina and functional profiles of AM were enriched by CH in favor of the corresponding methanogenesis. Further, based on pure cultivation exposed to CH, the methanogenic performance, biomass, survivability and activity of typical Methanosarcina (M. barkeri) were boosted. Notably, iTRAQ proteomics revealed that the manufacturing (transcription and translation), expression and biocatalytic activity of acetoclastic metalloenzymes, particularly tetrahydromethanopterin S-methyltransferase and methyl-coenzyme M reductase with cobalt/nickel-cofactor (F430 and cobalamin), and acetyl-CoA decarbonylase/synthase with cobalt/nickel-active site, of M. barkeri were upregulated significantly with fold changes in the range of 1.21–3.20 due to the CH presence. This study shed light on EPs-affecting industrially crucial methanogen at the molecular biology level during AD and had implications in the technical relevance of methanogens.

Cross-Feedings, Competition, and Positive and Negative Synergies in a Four-Species Synthetic Community for Anaerobic Degradation of Cellulose to Methane.

Complex interactions exist among microorganisms in a community to carry out ecological processes and adapt to changing environments. Here, we constructed a quad-culture consisting of a cellulolytic bacterium (Ruminiclostridium cellulolyticum), a hydrogenotrophic methanogen (Methanospirillum hungatei), an acetoclastic methanogen (Methanosaeta concilii), and a sulfate-reducing bacterium (Desulfovibrio vulgaris). The four microorganisms in the quad-culture cooperated via cross-feeding to produce methane using cellulose as the only carbon source and electron donor. The community metabolism of the quad-culture was compared with those of the R. cellulolyticum-containing tri-cultures, bi-cultures, and mono-culture. Methane production was higher in the quad-culture than the sum of the increases in the tri-cultures, which was attributed to a positive synergy of four species. In contrast, cellulose degradation by the quad-culture was lower than the additive effects of the tri-cultures which represented a negative synergy. The community metabolism of the quad-culture was compared between a control condition and a treatment condition with sulfate addition using metaproteomics and metabolic profiling. Sulfate addition enhanced sulfate reduction and decreased methane and CO2 productions. The cross-feeding fluxes in the quad-culture in the two conditions were modeled using a community stoichiometric model. Sulfate addition strengthened metabolic handoffs from R. cellulolyticum to M. concilii and D. vulgaris and intensified substrate competition between M. hungatei and D. vulgaris. Overall, this study uncovered emergent properties of higher-order microbial interactions using a four-species synthetic community. IMPORTANCE A synthetic community was designed using four microbial species that together performed distinct key metabolic processes in the anaerobic degradation of cellulose to methane and CO2. The microorganisms exhibited expected interactions, such as cross-feeding of acetate from a cellulolytic bacterium to an acetoclastic methanogen and competition of H2 between a sulfate reducing bacterium and a hydrogenotrophic methanogen. This validated our rational design of the interactions between microorganisms based on their metabolic roles. More interestingly, we also found positive and negative synergies as emergent properties of high-order microbial interactions among three or more microorganisms in cocultures. These microbial interactions can be quantitatively measured by adding and removing specific members. A community stoichiometric model was constructed to represent the fluxes in the community metabolic network. This study paved the way toward a more predictive understanding of the impact of environmental perturbations on microbial interactions sustaining geochemically significant processes in natural systems.

Physiological Responses of Methanosarcina barkeri under Ammonia Stress at the Molecular Level: The Unignorable Lipid Reprogramming.

Acetotrophic methanogens' dysfunction in anaerobic digestion under ammonia pressure has been widely concerned. Lipids, the main cytomembrane structural biomolecules, normally play indispensable roles in guaranteeing cell functionality. However, no studies explored the effects of high ammonia on acetotrophic methanogens' lipids. Here, a high-throughput lipidomic interrogation deciphered lipid reprogramming in representative acetoclastic methanogen (Methanosarcina barkeri) upon high ammonia exposure. The results showed that high ammonia conspicuously reduced polyunsaturated lipids and longer-chain lipids, while accumulating lipids with shorter chains and/or more saturation. Also, the correlation network analysis visualized some sphingolipids as the most active participant in lipid-lipid communications, implying that the ammonia-induced enrichment in these sphingolipids triggered other lipid changes. In addition, we discovered the decreased integrity, elevated permeability, depolarization, and diminished fluidity of lipid-supported membranes under ammonia restraint, verifying the noxious ramifications of lipid abnormalities. Additional analysis revealed that high ammonia destabilized the structure of extracellular polymeric substances (EPSs) capable of protecting lipids, e.g., declining α-helix/(β-sheet + random coil) and 3-turn helix ratios. Furthermore, the abiotic impairment of critical EPS bonds, including C-OH, C═O-NH-, and S-S, and the biotic downregulation of functional proteins involved in transcription, translation, and EPS building blocks' supply were unraveled under ammonia stress and implied as the crucial mechanisms for EPS reshaping.

Study on performance and mechanisms of anaerobic oxidation of methane-microbial fuel cells (AOM-MFCs) with acetate-acclimatizing or formate-acclimatizing electroactive culture

Anaerobic oxidation of methane-microbial fuel cells with acetate-acclimatizing or formate-acclimatizing electroactive culture (A-AOM-MFC and F-AOM-MFC) were designed and operated at room temperature in this study to evaluate and explore the electrochemical performance and mechanisms of methane conversion and electricity generation. The results indicated that A-AOM-MFC output a higher voltage (0.526 ± 0.001 V) and F-AOM-MFC started up in a shorter time (51 d), resulting from different mechanisms of methane-electrogen caused by discrepant microbial alliances. Specifically, in A-AOM-MFC, acetoclastic methanogens (e.g., Methanosaeta) converted methane into intermediates (e.g., acetate) through reversing methanogenesis and carried out the direct interspecific electron transfer (DIET) with Geobacter-predominated electricigens which can oxidize the intermediates to carbon dioxide and transfer electrons to the electrodes. Differently, the intermediate-dependent extracellular electron transfer (EET) existed in F-AOM-MFC between hydro-methanogens (e.g., Methanobacterium) and electricigens (e.g., Geothrix), which was more difficult than DIET. Additionally, hydro-methanogens metabolized methane to produce formate-dominant intermediates more quickly.

Insights into methanogenesis of mesophilic-psychrophilic varied anaerobic digestion of municipal sludge with antibiotic stress

Methane production through anaerobic digestion (AD) of municipal sludge is economic and eco-friendly, which is commonly affected by temperature and pollutants residues. However, little is known about methanogenesis in psychrophilic AD (PAD) with temperature variations that simulating seasonal variations and with antibiotic stress. Here, two groups of AD systems with oxytetracycline (OTC) were operated with temperature maintained at 35 °C and 15 °C or variation to explore the influence to methanogenesis. The acetic acid was noticeably accumulated in OTC groups initially (P < 0.001). Methane production was noticeably inhibited initially in PAD with OTC, but had been stimulated later at 35 °C. The dominant acetoclastic methanogens Methanosaeta gradually decreased to 15.48% and was replaced by methylotrophic Methanomethylovorans (73.43%) in PAD with OTC. Correspondingly, the abundances of functional genes related to methylotrophic methanogenesis were also higher in these groups. Besides, genes involving in methane oxidation had over 50 times higher abundances in PAD with OTC groups in the second phase. Further investigation is essential to understand the main dynamics of methanogenesis in PAD and to clear the related molecular mechanism for future methane production regulation in sludge systems.

Proteiniphilum and Methanothrix harundinacea became dominant acetate utilizers in a methanogenic reactor operated under strong ammonia stress.

Microorganisms in anaerobic digestion (AD) are easily affected by ammonia, especially acetoclastic methanogens. Thus, in ammonia-suppressed AD systems, acetate degradation is reported to be carried out mainly by the cooperation of syntrophic acetate oxidizers and hydrogenotrophic methanogens. Previous studies have revealed ammonia inhibition on microbial flora by AD performance, but the effect mechanism of ammonia on microbial metabolism remains poorly understood. In this study, we constructed a mesophilic chemostat fed with acetate as the sole carbon source, gradually increased the total ammonia nitrogen (TAN) concentration from 1 g L-1 to 6 g L-1, and employed the 16S rRNA gene, metagenomics, and metatranscriptomics analysis to characterize the microbial community structure and metabolic behavior. The results showed that even at the TAN of 6 g L-1 (pH 7.5), the methanogenesis kept normal, the biogas production was approximately 92% of that at TAN of 1 g L-1 and the acetate degradation ratio reached 99%, suggesting the strong TAN tolerance of the microbial community enriched. 16S rRNA gene analysis suggested that the microbial community structure changed along with the TAN concentration. Methanothrix predominated in methanogens all the time, in which the dominant species was gradually replaced from M. soehngenii to M. harundinacea with the increased TAN. Dominant bacterial species also changed and Proteiniphilum showed a significant positive correlation with increased TAN. Meta-omics analysis showed that the absolute dominant microorganisms at TAN of 6 g L-1 were M. harundinacea and Proteiniphilum, both of which highly expressed genes for anti-oxidative stress. M. harundinacea and the second dominant methanogen Methanosarcina highly expressed both acetate cleavage and CO2 reduction pathways, suggesting the possibility that these two pathways contributed to methanogenesis together. Proteiniphilum and some other species in Firmicutes and Synergistetes were likely acetate oxidizers in the community as they highly expressed genes for syntrophic acetate oxidization, H2 generation, and electron transfer. These results suggested that Proteiniphilum as well as M. harundinacea have strong ammonia tolerance and played critical roles in acetate degradation under ammonia-suppressed conditions. The achievements of the study would contribute to the regulation and management of the AD process.

Response of methanogenic system to long-term polycyclic aromatic hydrocarbon exposure: Adsorption and biodegradation, performance variation, and microbial function assessment

Phenanthrene (PHE) as a typical polycyclic aromatic hydrocarbon (PAH) is prevalent and harmful to organisms in petroleum-polluted sites. The effects of PHE concentration levels on performance, microbial community and functions in methanogenic system were comprehensively investigated by an operation of UASB reactor (198 days) and a series of batch tests. The results found that PHE was prone to accumulate in reactor by sludge adsorption (Final concentration = 12.53 mg/g TS Sludge), which posed significant influences on methanogenic system. The removal of chemical oxygen demand (COD), NH4+-N and volatile fatty acids (VFAs) in reactor were reduced with PHE accumulation. Meanwhile, microbes with higher ATPase secrete more EPS activity to self-protect against PHE toxicity. Sequencing analysis showed that PHE interfered significantly diversity and structure of microbial community. For bacteria, PHE was toxic to Bacteroidetes and Latescibacteria, while syntrophs (f_Syntrophaceae, Syntrophorhabdus, etc.) involved in VFAs oxidation and aromatic organics degradation were tolerant of PHE stress. For archaea, acetoclastic methanogens (Methanosaeta) abundance was continuously diminished by 45.1% under long-term PHE exposure. Further functions analysis suggested that microbial community accelerated amino acid metabolism, energy metabolism and xenobiotics biodegradation & metabolism to satisfy physiological demanding under PHE stress. Combining batch tests of methanogenic metabolism proved that acetoclastic methanogenesis was negatively affected by PHE due to inhibition of functional enzymes (acetate kinase, phosphate acetyltransferase, etc.) expression. These findings may provide the basis for enhancing bioremediation of PAH pollution in anaerobic environment.

Chemical characterization and anaerobic treatment of bitumen fume condensate using a membrane bioreactor

Bitumen fume condensate (BFC) is a hazardous wastewater generated at asphalt reclamation and production sites. BFC contains a wide variety of potentially toxic organic pollutants that negatively affect anaerobic processes. In this study, we chemically characterized BFC produced at an industrial site and evaluated its degradation under anaerobic conditions. Analyses identified about 900 compounds including acetate, polycyclic aromatic hydrocarbons, phenolic compounds, and metal ions. We estimated the half maximal inhibitory concentrations (IC50) of methanogenesis of 120, 224, and 990 mgCOD·L−1 for three types of anaerobic biomass, which indicated the enrichment and adaptation potentials of methanogenic biomass to the wastewater constituents. We operated an AnMBR (7.0 L, 35 °C) for 188 days with a mixture of BFC, phenol, acetate, and nutrients. The reactor showed a maximum average COD removal efficiency of 87.7 ± 7.0 %, that corresponded to an organic conversion rate of 286 ± 71 mgCOD−1·L−1d−1. The microbial characterization of the reactor’s biomass showed the acetoclastic methanogen Methanosaeta as the most abundant microorganism (43 %), whereas the aromatic and phenol degrader Syntrophorhabdus was continuously present with abundances up to 11.5 %. The obtained results offer the possibility for the application of AnMBRs for the treatment of BFC or other petrochemical wastewater.

Methanogenic consortia from thermophilic molasses-fed structured-bed reactors: microbial characterization and responses to varying food-to-microorganism ratios

The heterogeneous character of fixed-film reactors may create highly specialized zones with a stratified distribution of microbial groups and varying capabilities to withstand high organic loads in anaerobic digestion (AD) systems. The microbial distribution and methane-producing potential of biomass from different regions (feeding zone and structured bed) of two second-stage thermophilic (55 ºC) fixed-film reactors were assessed. Three levels of food-to-microorganism (F/M) ratio (0.4, 1.0 and 3.0 g-COD g−1VS) using fermented (two-stage AD) and fresh (single-stage AD) sugarcane molasses were tested in batch reactors, simulating low to high organic loads. Specific methane production rates increased as the F/M increased when using fermented molasses, maintaining efficient methanogenesis at substrate availability levels threefold higher than single-stage schemes (3.0 vs. 1.0 g-COD g−1VS). Success in methane production derived from the homogenous establishment (similar in both feeding zone and bed) of syntrophic associations between acetogens (Pelotomaculum, Syntrophothermus, Syntrophomonas and Thermodesulfovibrio), acetate oxidizers (Thermoacetogenium, Mesotoga and Pseudothermotoga) and hydrogenotrophic methogens (Methanothermobacter and Methanoculleus) replacing acetoclastic methanogens (Methanosaeta). Phase separation under thermophilic conditions was demonstrated to boost methane production from sugar-rich substrates, because the process depends on microbial groups (hydrogenotrophs) that grow faster and are less susceptible to low pH values compared to acetotrophs.

Anaerobic microbial cocktail of lignocellulolytic fungi and bacteria with methanogens for boosting methane production from unpretreated rice straw

Rice straw (RS) has been recognized as a sustainable renewable energy resource for converting into sugars and volatile fatty acids (acetate, propionate, and butyrate) and subsequently to produce biogas. Enhanced production of these intermediates from RS by the different combinations of two consortia was investigated. Anaerobic microbial cocktails of fungi, bacteria, and methanogens were evaluated for performance and stability in the anaerobic digestion of untreated RS. The best-defined anaerobic microbial cocktail for high RS degradation and methane production, consisting of anaerobic bacteria (mainly Proteiniphilum acetatigenes, Pyramidobacter piscolens, and Mesotoga prima) and anaerobic lignocellulolytic/fermentative fungi (uncultured Neocallimastigales, Orpinomyces, Anaeromyces, and Feramyces sp.) at a copy number ratio of 103-105 copies/mL, including hydrogenotrophic and acetoclastic methanogens (Methanosarcina mazei, Methanoculleus marisnigri, Methanofollis liminatans, Methanoculleus bourgensis, and Methanosaeta harundinacea) concentration of 106 copies/mL, was successfully constructed. The system performance was 80% VS (volatile solids) RS degradation, 34mL/day methane production rate, 318mL/g VSadded methane yield, and a pH range of 6.90-7.70 within a short time of 14days. A defined microbial cocktail has been proven as a potential alternative process for lignocellulose hydrolysis and methane production.

Understanding the effect of carbamazepine on the recovery of methane from lactic acid wastewater by anaerobic digestion

Anaerobic digestion is a promising technology for recovering methane from organic wastewater and contributing to carbon neutrality in wastewater plants or industrial parks. However, the interference of pharmaceuticals and personal care products with anaerobic digestion is commonly overlooked. Carbamazepine, a typical antiepileptic drug, is frequently detected in sewage sludge, but its influence on anaerobic digestion has not been systematically reported. In this work, the effect of carbamazepine on the anaerobic digestion of lactic acid synthesis wastewater was revealed comprehensively. The results showed that carbamazepine could seriously disturb anaerobic digestion performance, the methane yield decreased from 275.12 to 182.04 mL/g lactic acid as the carbamazepine dosage rose from 0 to 0.14 mM. To better understand how the drug affected anaerobic digestion, model tests were conducted on four stages of hydrolysis, acidification, acetogenesis and methanogenesis. The model tests indicated that such drug could promote the solubilization of inoculum, thereby releasing polysaccharides and proteins. But the hydrolysis stage, commonly recognized as the rate-limiting step was restricted. Meanwhile, the enhanced acidification contributed to the accumulation of volatile fatty acids. Both acetogenesis and methanogenesis were severely disturbed eventually causing a significant decrease in methane yield. The level of interference of carbamazepine on anaerobic digestion was concluded as follows: acetogenesis > methanogenesis > hydrolysis > acidification > solubilization. The high-throughput sequencing revealed the abundance of Chloroflexi increased from 21.5% to 26.7% as a main responsibility for the volatile acid accumulation. The acetoclastic methanogen Methanosaeta decreased from 59.86% to 44.78%, while the hydrogenotrophic methanogens Methanobacterium and Methanomassiliicoccus increased from 28%, 0.5%–37.9% and 4.1%, respectively. Although the recovery of methane was limited for clean production, it was more recommendable to recover the volatile fatty acids from organic wastewater or sludge with high dosage of carbamazepine.

Binary-component anaerobic co-digestion: Synergies and microbial profiles

One of the key issues to create synergistic effect in anaerobic co-digestion (AcoD), is how to make the appropriate choice about co-substrate and determine the optimal mixing ratio. Adjusting organic component, namely carbohydrate (C), protein (P) and lipid (L), is expected to be a dependable strategy for obtaining synergistic effect in anaerobic co-digestion. In this study, the AcoD performances of organic components at different load levels were investigated in terms of methane yield, metabolic transformation, and microbial response. The synergistic interval at different loading levels showed considerable stability, and the significant synergy was achieved at C3P7 to C8P2 except for C6P4, C4L6 to C6L4, and P3L7 to P5L5 (based on mass ratio), respectively. Under high feeding load (4%VS), groups within synergistic interval could significantly reduce the toxic effects of intermediate metabolic inhibitors and improve methane production. However, AcoD of protein and lipid would induce antagonistic effects, and high proportion of protein was more serious. In the face of adversity, bacterial community diversity was weakened and the relative abundance of dominated phylum Firmicutes was strengthened. Furthermore, acetoclastic methanogenesis pathway was transformed to hydrogenotrophic pathway since hydrogenotrophic methanogens replaced acetoclastic methanogens as the dominant methanogenic archaea with the increase of syntrophic acetate oxidation bacteria. Suitable organic components proportion can effectively reduce inhibitor accumulation and maintain a well methanogens structure to ensure high methane production.

Effect of bioaugmentation on psychrotrophic anaerobic digestion: Bioreactor performance, microbial community, and cellular metabolic response

Most anaerobic digestion reactors are operated under mesophilic and thermophilic conditions, requiring high levels of heat energy to maintain high temperatures. If an optimized method existed for psychrotrophic anaerobic digestion (PAD), it would have outstanding economic and environmental benefits as an alternative to mesophilic or thermophilic reactors. Therefore, this study aimed to investigate the conditions and corresponding mechanism to optimize PAD for producing high yields of methane. We found that intermittent bioaugmentation with mesophilic propionate-degrading consortia showed enhanced methane production back to uninhibited levels in a psychrotrophic anaerobic reactor under volatile fatty acid stress, with long-term effectiveness for 14 hydraulic retention times. The increase in acetoclastic methanogens (Methanothrix harundinacea and Methanosarcina flavescens) stimulated methanogenesis in the psychrotrophic system. Metagenomic analysis showed that bacteria carried more diverse cold-regulating gene sets and had higher DNA replication rates than methanogens, explaining the higher cold tolerance of bacterial community than that of methanogen community. By coupling reactor performance with microbial metabolic analysis, this study is the first to provide comprehensive findings that intermittent bioaugmentation with methanogenic consortia is a promising strategy to boost the PAD process.

Improved methane production from anaerobic wastewater treatment by conductive zero-valent iron@carbon@polyaniline

In this study, a core-shell type ZVI@C@PANI nanoparticle was prepared and used as a novel conductive medium for electron transfer between bacteria and archaea of the anaerobic system. The synthesized nanocomposite had favorable biocompatibility and a conductivity of 4.89 S/cm, which made the nanocomposite a good transfer medium to achieve efficient methane production. And the enhanced methane production by adding ZVI@C@PANI in the anaerobic sludge had been investigated. The ZVI@C@PANI enhanced methane yield peaked at doses of 2 g/L. The result showed that methane production increased by 71.36% from the anaerobic system added with the ZVI@C@PANI compared to the control system. Moreover, information on the microorganism community and enzymes in the anaerobic sludge was detected. The results showed that the Geobacter, Syntrophus, and Dechlorobacter bacteria and the Methanosaeta and Methanobacterium archaea were significantly enriched for facilitating direct interspecies electron transfer in the anaerobic system by added ZVI@C@PANI. Moreover, the ZVI@C@PANI can strengthen the production of acetic acid, which facilitated the acetoclastic methanogens to generate methane from the anaerobic system.

Elucidating interactive effects of sulfidated nanoscale zero-valent iron and ammonia on anaerobic digestion of food waste

In our previous study, anaerobic digestion of food waste could be effectively enhanced by adding sulfidated nanoscale zero-valent iron (S-nZVI) under high-strength ammonia concentrations. In this study, in order to further elucidate the specific interactive effects of S-nZVI and ammonia on anaerobic digestion of nitrogen-rich food waste, the methanogenic performance of anaerobic digestion systems respectively added with nanoscale zero-valent iron (nZVI) and S-nZVI were compared and monitored under different ammonia stress conditions. Both nZVI and S-nZVI could effectively stimulate the methanogenesis process among ammonia concentrations ranging from 0 to 3500mg/L. However, the enhancing effects of S-nZVI and nZVI on anaerobic digestion of food waste were different, in which anaerobic digestion systems added with S-nZVI and nZVI performed best under 2500mg/L of ammonia and 1500mg/L of ammonia, respectively. Furthermore, the analysis of microbial communities suggested that ammonia stress enriched acetoclastic methanogens, while adding nZVI and S-nZVI into anaerobic digestions stimulated the process of hydrogenotrophic methanogenesis. Moreover, S-nZVI performed better in promoting the evolution of DIET-related microorganisms than nZVI, resulting in enhanced methane production under high ammonia-stressed conditions. This work provided fundamental knowledge about the interactive effects of S-nZVI and ammonia on the anaerobic digestion of food waste.

Aquaculture drastically increases methane production by favoring acetoclastic rather than hydrogenotrophic methanogenesis in shrimp pond sediments

Emissions of methane (CH4), a major greenhouse gas, should be cut by at least 30% by 2030 according to the last Conference of the Parties, CoP26. Aquaculture pond is a major CH4 emitter, yet the microbial mechanisms ruling methanogenesis by degradation of organic matter in sediments remain unclear. In particular, the respective roles of hydrogenotrophic and acetoclastic methanogenesis, and the impact of aquaculture farming practices are unknown. We studied methanogenesis in the surface sediments from a freshwater and an oligohaline pond before, during, and after shrimp farming. Hydrogenotrophic and acetoclastic contributions were distinguished by acetoclastic inhibition with methylfluoride (CH3F), and by 13C-analysis of CO2 and CH4. We also monitored the methanogenic community structure, dissolved organic carbon (DOC) levels, carbon to nitrogen (C/N) ratios, and humification indices derived from Fourier transform infrared spectroscopy. The results reveal that aquaculture farming practices increased methanogenesis rates, and these increases were explained by higher levels of DOC and lower C/N ratios during farming. Of the total methane produced, 51%–78% was by hydrogenotrophic methanogenesis. However, the total methane contribution from acetoclastic methanogenesis increased from approximately 22% before farming to approximately 45% during and after farming, with a decreasing isotope fractionation factor αc and an increasing relative abundance of Methanosaeta acetoclastic methanogen. All humification indices decreased during and after farming compared to before farming due to the input of polysaccharide-rich aquafeed. The close relationship between the humification indices and methanogenesis pathways indicates that the changes in sediment substrate quality drove the variation in the methanogenesis pathways. Increases in salinity decreased the methanogenesis rates but did not change the methanogenesis pathways. Overall, our findings reveal that aquaculture farming practices increase methanogenesis rates and favor acetoclastic over hydrogenotrophic methanogenesis, and that adjusting shrimp diets, increasing salinity, and removing residual aquafeed could reduce methanogenesis.

The role of iron nanoparticles on anaerobic digestion: mechanisms, limitations, and perspectives.

Anaerobic digestion (AD) is the most widely used technology for organic matter treatment. However, multiple types of research have reported on improving the process because different operation inhibition factors and limitations affect the performance of AD process. Owing to the increasing use of iron-nanoparticles (Fe-NP) on AD, this review addresses the knowledge gaps and summarizes the finding from academic articles based on (i) the AD upgrading operations: limitations and upgrade techniques, (ii) Fe-NPs mechanisms on AD, (iii) Fe-NP effect on microbial communities associated to AD systems, and (iv) perspectives. The selected topics give the Fe-NP positive effects on the AD methane-production process in terms of gas production, effluent quality, and process optimization. The main results of this work indicate that (i) Fe-NP addition can be adapted among different feedstocks and complement other pretreatments, (ii) Fe-NP physicochemical characteristics enhance biogas production via direct interspecies electron transfer (DIET) mechanisms, and Fe-ion release due to their structure and their conductivity capability, and (iii) syntrophic bacteria and acetoclastic methanogens have been reported as the communities that better uptake Fe-NPs on their metabolisms. Finally, our research perspectives and gaps will be discussed to contribute to our knowledge of using Fe-NPs on AD systems.

The effect of anode potential on electrogenesis, methanogenesis and sulfidogenesis in a simulated sewer condition

Methane emissions from the sewer system are considered to be a non-negligible source of aggravating the greenhouse effect. Meanwhile, the sewer system has long been plagued by sulfide-induced corrosion problems. This study explored the possibility of using a bioelectrochemical system to intensify the competition between electroactive bacteria, methanogens and sulfate-reducing bacteria, thereby reducing the production of methane and sulfide. Dual-chamber bioelectrochemical reactors were constructed and operated in fed-batch mode with the coexistence of Electroactive bacteria, Methanogenic archaea and Sulfate-reducing bacteria. Acetate was supplied as the sole carbon source. The results indicated that electrogenesis induced by the anode potentials of -0.42 V and -0.2 V (vs. Ag/AgCl) had advantages over methanogenesis and sulfidogenesis in consuming acetate. The stimulated electrogenesis by anode potentials resulted in a decrease in pH. Methane production was suppressed in the reactors with anode potentials of -0.42 and -0.2 V compared to open circuit controls. In contrast to methane, the capacity for sulfide production was facilitated in the reactors with the anode potentials of -0.42 V and -0.2 V compared to open circuit controls. 16s rRNA gene analysis showed that Geobacter was the most abundant genus on the anode biofilm in the anode potential-controlled reactor, while acetoclastic methanogens dominated in open circuit controls. Methanosaeta and Methanosarcina were the most abundant methanogens in open circuit controls. Collectively, our study demonstrates that the use of electrodes with anode potential control can help to control methane emissions, but could not yet prevent sulfide production, which requires further research.

Deeper insight into the effect of salinity on the relationship of enzymatic activity, microbial community and key metabolic pathway during the anaerobic digestion of high strength organic wastewater

The threshold salt concentration to inhibit the anaerobic digestion (AD) has been intensively investigated, but its insight mechanism is not fully revealed. Therefore, this study systematically investigated the effect of salinity on acidogenesis and methanogenesis and its mechanism. Results showed that low salinity level (i.e. 0.6%) had stimulatory effect on volatile fatty acids (VFA) and methane production, while significant inhibition was observed with further increased salinity. Moreover, high salinity limited the butyric acid degradation at acidogenesis process. The decreases of enzymes (AK and PTA) activity and functional genes (ackA, pta and ACOX) expression that related to β-oxidation explained the butyric acid accumulation at high salinity levels. Microbial community analysis revealed high salinity levels significantly inhibited the proliferation of Syntrophomonas sp., which are known to be associated with butyric acid degradation. Similarly, the relative abundance of acetoclastic methanogen (Methanothrix sp.) and methylotrophic methanogen (Methanolinea sp.) significantly decreased at salinity condition.

Ball-milled Fe0/FeS2 enhanced interaction of Shewanella oneidensis MR-1 with anaerobic microbial community: Impact on 2,4-dichloronitrobenzene reduction and methane yield

The technology of integrating sulfidated zero-valent iron and functional anaerobic bacteria has been extensively explored to improve the removal efficiency of pollutants and enhance the performance of anaerobic microbial system. In this study, the simultaneous DCNB(2,4-dichloronitrobenzene) reduction and methane production by ball-milled 1:1 Fe0/FeS2 (briefly, 1:1 Fe0/FeS2), Shewanella oneidensis MR-1 and microbial consortium (MC) were firstly investigated. The result showed that the DCNB reduction was increased by 2.6 and 1.7 times in 1:1 Fe0/FeS2 + MR-1 group compared to 1:1 Fe0/FeS2 and MR-1, respectively, while it further increased by almost 2 times in 1:1 Fe0/FeS2 + MC + MR-1 group as compared to that of 1:1 Fe0/FeS2 + MR-1 group. Moreover, the methane production was drastically facilitated via the reciprocal interaction among FeS as the reaction product, MR-1 and MC. Dechlorination was not found, mainly due to the inability of 1:1 Fe0/FeS2 and the inexistence of dechlorinating bacteria in original inoculum. Meanwhile, the microbial community was also changed, with genera like Eubacterium capable of reducing nitroaromatic compound and Methanosarcina as acetoclastic methanogen enriched by 1:1 Fe0/FeS2 and MR-1. Therefore, fast DCNB reduction and methane production using 1:1 Fe0/FeS2 in anaerobic microbial system, gives useful reference for both establishment of pollutant removal and biogas production. In conclusion, Fe0/FeS2, as a kind of sulfidated ZVI, is a good candidate for enhancing both pollutant degradation and CH4 production in anaerobic wastewater treatment.