A guide for comparing microbial co-occurrence networks.

The article provides a pipeline for comparing microbial co-occurrence networks based on the R microeco package and meconetcomp package. It has high flexibility and expansibility and can help users efficiently compare networks built from different groups of samples or different construction approaches.

Anaerobic methane oxidation coupled to ferrihydrite reduction by Methanosarcina barkeri.

Fe(III) has been recognized as a potential electron sink for the anaerobic oxidation of methane (Fe-AOM) in diverse environments. However, most of previous Fe-AOM processes are limited to ANME archaea and the Fe-AOM mechanism remains unclear. Here we investigate, for the first time, the Fe-AOM performance and mechanisms by a single methanogen Methanosarcina barkeri. The results showed that M. barkeri was capable of oxidizing methane to CO2 and reducing ferrihydrite to siderite simultaneously. The presence of methane enhanced both the abundances of redox-active species (such as cytochromes) and electrochemical activity of M. barkeri. The proteomic analyses revealed that M. barkeri up-regulated the expressions of a number of methanogenic enzymes during Fe-AOM, and significantly enriched metabolic pathways of amino acid synthesis and nitrogen fixation. Metabolic inhibition experiments indicated that membrane-bound redox-active components (cytochromes, methanophenazine and F420H2:quinone oxidoreductase) were probably involved in extracellular electron transfer (EET) from cells to ferrihydrite. Overall, these results provide a deep insight into the single­carbon metabolism and survival strategy for methanogens and suggest that methanogens may play an important role in linking methane and iron cycling in the substrate-limited environments.

Acetate and electricity generation from methane in conductive fiber membrane- microbial fuel cells.

Microbial conversion of methane to electricity, fuels, and liquid chemicals has attracted much attention. However, due to the low solubility of methane, it is not considered a suitable substrate for microbial fuel cells (MFCs). In this study, a conductive fiber membrane (CFM) module was constructed as the bioanode of methane-driven MFCs, directly delivering methane. After biofilm formation on the CFM surface, a steady voltage output of 0.6 to 0.7 V was recorded, and the CFM-MFCs obtained a maximum power density of 64 ± 2 mW/m2. Moreover, methane oxidation produced a high concentration of intermediate acetate (up to 7.1 mM). High-throughput 16S rRNA gene sequencing suggests that the microbial community was significantly changed after electricity generation. Methane-related archaea formed a symbiotic consortium with characterized electroactive bacteria and fermentative bacteria, suggesting a combination of three types of microorganisms for methane conversion into acetate and electricity.

High-rate anaerobic decolorization of methyl orange from synthetic azo dye wastewater in a methane-based hollow fiber membrane bioreactor.

Anaerobic biological techniques are widely used in the reductive decolorization of textile wastewater. However, the decolorization efficiency of textile wastewater by conventional anaerobic biological techniques is generally limited due to the low biomass retention capacity and short hydraulic retention time (HRT). In this study, a methane-based hollow fiber membrane bioreactor (HfMBR) was initially inoculated with an enriched anaerobic methane oxidation (AOM) culture to rapidly form an anaerobic biofilm. Then, synthetic azo dye wastewater containing methyl orange (MO) was fed into the HfMBR. MO decolorization efficiency of ∼ 100 % (HRT = 2 to 1.5 days) and maximum decolorization rate of 883 mg/L/day (HRT = 0.5 day) were obtained by the stepwise increase of the MO loading rate into the methane-based HfMBR. Scanning electron microscopy (SEM) and fluorescence in situ hybridization (FISH) analysis visually revealed that archaea clusters formed synergistic consortia with adjacent bacteria. Quantitative PCR (qPCR), phylogenetic and high-throughput sequencing analysis results further confirmed the biological consortia formation of methane-related archaea and partner bacteria, which played a synergistic role in MO decolorization. The high removal efficiency and stable microbial structure in HfMBR suggest it is a potentially effective technique for high-toxic azo dyes removal from textile wastewater.

Decolorization of Acid Orange 7 by extreme-thermophilic mixed culture.

Although a large amount of textile wastewater is discharged at high temperatures, azo dye reduction under extreme-thermophilic conditions by mixed cultures has gained little attention. In this study, Acid Orange 7 (AO7) was used as the model azo dye to demonstrate the decolorization ability of an extreme-thermophilic mixed culture. The results showed that a decolorization efficiency of over 90% was achieved for AO7. The neutral red (NR, 0.1 mM) could promote AO7 decolorization, in which the group of Cell + NR offered the highest decolorization rate of 1.568 1/h and t1/2 was only 0.44 h, whereas after CuCl2 addition, the decolorization rate (0.141 1/h) was lower and t1/2 (4.92 h) was much longer. Thus, CuCl2 notably inhibited this process. Caldanaerobacter (64.0%) and Pseudomonas (25.4%) were the main enriched bacteria, which were not reported to have the ability for dye decolorization. Therefore, this study extends the application of extreme-thermophilic biotechnology.

Humic substances as electron acceptors for anaerobic oxidation of methane driven by ANME-2d.

Humic substances (humics) are ubiquitous in terrestrial and aquatic environments where they can serve as electron acceptors for anaerobic oxidation of organic compounds. Methane is a powerful greenhouse gas, as well as the least reactive organic molecule. Anaerobic oxidation of methane (AOM) coupled to microbial reduction of various electron acceptors plays a crucial role in mitigating methane emissions. Here, we reported that humics could serve as terminal electron acceptors for AOM using enriched nitrate-reducing AOM microorganisms. AOM coupled to the reduction of humics was demonstrated based on the production of 13C-labelled carbon dioxide, and AOM activity was evaluated with different methane partial pressures and electron acceptor concentrations. After three-cycle reduction, both AOM activity and copy numbers of the archaea 16S rRNA and mcrA genes were the highest when anthraquinone-2,6-disulfonic acid and anthraquinone-2-sulfonic acid were electron acceptors. The high-throughput sequencing results suggested that ANME-2d were the dominant methane oxidation archaea after humics reduction, although the partner bacteria NC10 trended downward, other reported humics reduction bacteria (Geobactor and Anammox) appeared. The potential electron transfer models from ANME-2d to humics were proposed. These results enable a better understanding of available electron acceptors for AOM in natural environments and broaden our insight into the significant role of ANME-2d.

Microbial selenite reduction coupled to anaerobic oxidation of methane.

Denitrifying anaerobic methane oxidation (DAMO) is the process of coupling the anaerobic oxidation of methane (AOM) with denitrification, which plays an important part in controlling the flow of methane in anoxic niches. In this study, we explored the feasibility of microbial selenite reduction using methane by DAMO culture. Isotopic 13CH4 and long-term experiments showed that selenite reduction was coupled to methane oxidation, and selenite was ultimately reduced to Se (0) by the analyses of scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The introduction of nitrate, the original electron acceptor in the DAMO culture, inhibited selenite reduction. Meanwhile, the microbial community of DAMO culture was significantly changed when the electron acceptor was changed from nitrate to selenite after long-term selenite reduction. High-throughput 16S rRNA gene sequencing indicated that Methylococcus (26%) became the predominant microbe performing selenite reduction and methane oxidation and the possible pathways of AOM accompanied with selenite reduction were proposed. This study revealed more potential relation during the biogeochemical cycle of carbon, nitrogen, and selenium.

Nontemplating Porous Carbon Material from Polyphosphamide Resin for Supercapacitors.

The nontemplating preparation of porous carbon materials by using specially designed polymer precursors for supercapacitor is attracting considerable research attention because of the more controllable frame structure and easier processes than templating methods. Herein, a deliberately designed cross-linking polyphosphamide resin with defined N and P structure is synthesized and then carbonized to obtain porous carbon material. The as-obtained porous carbon material has a specific surface area of 2,620 m2 g-1, high porosity of 1.49 cm3 g-1, and well-distributed micro/mesoporous carbon structure. Different from activation by post-added NH4H2PO4, the confined N and P in the polymer frame are confirmed to play an important role in pore structure development by forming in situ highly dispersed NH4H2PO4 during carbonization. When evaluated as the electrode material for supercapacitors, the polyphosphamide-resin-based porous carbon material demonstrates excellent capacitance (440 F g-1 under 0.5 A g-1) and high stability (retention of 93% over 10,000 cycles).

Novel Gas Diffusion Cloth Bioanodes for High-Performance Methane-Powered Microbial Fuel Cells.

Microbial fuel cells (MFCs) are a promising technology that converts chemical energy into electricity. However, up to now only few MFCs have been powered by gas fuels, such as methane, and their limited performance is still challenged by the low solubility and bioavailability of gases. Here, we developed a gas diffusion cloth (GDC) anode to significantly enhance the performance of methane-powered MFCs. The GDC anode was constructed by simply coating waterproof GORE-TEX cloth with conductive carbon cloth in one step. After biofilm enrichment, the GDC anodes obtained a methane-dependent current up to 1130.2 mA m-2, which was 165.2 times higher than conventional carbon cloth (CC) anodes. Moreover, MFCs equipped with GDC anodes generated a maximum power density of 419.5 mW m-2. Illumina high-throughput sequencing revealed that the GDC anode biofilm was dominated mainly by Geobacter, in contrast with the most abundant Methanobacterium in planktonic cells. It is hypothesized that Methanobacterium reversed the methanogenesis process by transferring electrons to the anodes, and Geobacter generated electricity via the intermediates (e.g., acetate) of anaerobic methane oxidation. Overall, this work provides an effective route in preparing facile and cost-effective anodes for high-performance methane MFCs.

Long solid retention time (SRT) has minor role in promoting methane production in a 65°C single-stage anaerobic sludge digester.

In this study, a thermophilic (65°C) single-stage wasted activated sludge (WAS) digester was established and the effects of solid retention time (SRT) on the reactor performance were investigated. The result showed that the optimum SRT was 6days with methane yield of 186.16mL/g VS. It was found that SRT had little effect on the hydrolysis and volatile solids (VS) destruction, and the high temperature employed seemed sufficient to achieve maximum hydrolysis and VS destruction performance. Longer SRT, however, promoted the release of recalcitrant compounds and impaired acidification, leading to the low methane yield. The microbial community analysis revealed that the dominant pathway for methane production was through syntrophic activity of acetate oxidizing bacteria and hydrogenotrophic methanogens while acetoclastic methanogens were absent in the system.

Role of extracellular polymeric substances in efficient chromium(VI) removal by algae-based Fe/C nano-composite.

Subsequent application of the obtained algae by chemical coagulation (e.g. Fe(III) addition) presents a challenge because of various iron compounds in algae. In this study, algae obtained by chemical coagulation were carbonized to yield an algae-based Fe/C nano-composite with a high capacity for hexavalent chromium (Cr(VI)) removal (236.9 mg/g), which is attributed to the high reductive Fe content (e.g., FeS, Fe(0), and FeO) and specific surface area. The optimal conditions-that is, 100 mg/L Fe(III) addition and 800 °C-were determined. Moreover, the role of extracellular polymeric substances (EPS) in carbonization was examined as it affected the product composition and efficiency of Cr(VI) removal, owing to the stabilizing property effect of EPS in algae. Algal EPS induced the homogeneous distribution of Fe compounds on the surface of the algae, and the generated α-FeOOH nanoparticles were wrapped in organic carbon matrix, resulting in a sufficient reaction between Fe compounds and organic carbon during carbonization. X-ray photoelectron spectroscopy showed that reduction and adsorption contributed 83.44% and 16.56% to Cr(VI) removal, respectively. This study provides a new insight into the role of EPS in the efficient Cr(VI) removal by algae-based Fe/C nano-composite and presents a promising application of this Fe/C nano-composite in environmental remediation.

A modeling understanding on the phosphorous removal performances of A2O and reversed A2O processes in a full-scale wastewater treatment plant.

Reversed A2O process (anoxic-anaerobic-aerobic) and conventional A2O process (anaerobic-anoxic-aerobic) are widely used in many wastewater treatment plants (WWTPs) in Asia. However, at present, there are still no consistent results to figure out which process has better total phosphorous (TP) removal performance and the mechanism for this difference was not clear yet. In this study, the treatment performances of both processes were compared in the same full-scale WWTP and the TP removal dynamics was analyzed by a modeling method. The treatment performance of full-scale WWTP showed the TP removal efficiency of the reversed A2O process was more efficient than in the conventional A2O process. The modeling results further reveal that the TP removal depends highly on the concentration and composition of influent COD. It had more efficient TP removal than the conventional A2O process only under conditions of sufficient influent COD and high fermentation products content. This study may lay a foundation for appropriate selection and optimization of treatment processes to suit practical wastewater properties.

Hydrogen production from a thermophilic alkaline waste activated sludge fermenter: Effects of solid retention time (SRT).

This study aims to investigate the effects of solid retention times (SRTs) on hydrogen production via thermophilic alkaline fermentation of waste activated sludge. The reactor was subjected to a SRT from 10 to 6 days during approximately 82 days of operation. The results revealed that SRT had minor influence on hydrolysis and hydrolysis efficiency in different phases were from 48.11% to 50.55%. Nevertheless, the efficiency of acidogenesis process was highly related to SRT and longer SRT could enhance the acidogenesis. On the other hand, acidogenesis efficiency was also related to H2 partial pressure and high H2 partial pressure negatively affected the acidogenesis. Thus, the maximum acidification was achieved in phase 1 (21.29%) resulting in the maximum H2 yield in phase 1 (95.94 mL/g VSS; SRT = 10 days; H2 partial pressure = 0-18%). Phyla Actinobacteria and Proteobacteria, who are highly related to hydrolytic microbial population, were abundant in all phases that resulted in high hydrolysis extent. H2 production was attributed to the relative high abundance of Clostridia. Thus, this study suggested that longer SRT and lower H2 partial pressure was necessary to improve the H2 yield under alkaline pH condition.

Low-level concentrations of aminoglycoside antibiotics induce the aggregation of cyanobacteria.

The interactions between antibiotics and microorganisms have attracted enormous research attentions. In this study, we investigated the effects of two typical aminoglycoside antibiotics on the aggregation of the model cyanobacterium, Synechococcus elongatus, and the dominating strain in algal blooms, Microcystis aeruginosa, via the analysis of zeta potentials, hydrophobicity, and extracellular polymeric substances (EPS) secretion. The results showed that low-level antibiotics promoted the aggregation of S. elongatus and M. aeruginosa by 40 and 18% under 0.10 and 0.02 µg/mL of kanamycin, respectively, which was mainly attributed to the combined effects of increased zeta potentials and the ratio between extracellular proteins and polysaccharides. Tobramycin exerted similar effects. Additionally, we discovered that at low pH (pH 5) and ionic strength (1 mM Na+ and 2 mM Mg2+), the inducing effects of antibiotics would be even larger than those with higher pH and ionic strength. As aggregation is important to cyanobacteria in either the basic physiology of biofilm formation or the algal bloom, our study demonstrated that low-level antibiotics exert ecological impacts via interfered aggregation. We believe this study will shed light on the mechanisms underlying antibiotic-induced biofilm formation and help with the evaluation of the environmental and ecological risks of antibiotics and other emerging pollutants.

Valuable biochemical production in mixed culture fermentation: fundamentals and process coupling.

The mixed culture fermentation is an important environmental biotechnology that converts biodegradable organic wastes to valuable chemicals such as hydrogen, methane, acetate, ethanol, propionate, and so on. For the multistep process of hydrolysis, acidogenesis, acetogenesis/homoacetogensis, and methanogenesis, the typical metabolic reactions are firstly summarized. And then, since the final metabolites are always a mixture, the separation and purification processes are necessary to couple with anaerobic fermentation. Therefore, several typical coupling technologies including biogas upgrading, two-stage fermentation, gas stripping, membrane technology of pervaporation, membrane distillation, electrodialysis, bipolar membrane electrodialysis, and microbial fuel cells are summarized to separate the metabolites and recover energy. At last, the novel technologies such as the controlled metabolite production, medium chain carboxylic acid production, and high temperature ethanol recovery in thermophilic mixed culture fermentation are also reviewed. However, the novel concepts are still needed to meet the demands of better overall performances and lower total costs.

Recent developments of post-modification of biochar for electrochemical energy storage.

Biochar is a common byproduct from thermochemical conversion of biomass to produce bioenergy. However, the biochar features, such as morphology, porosity and surface chemistry, cannot be well controlled in conventional conversion approaches, limiting the wide application of raw biochar. Aiming to meet the specific requirements, post-modification of raw biochar was frequently conducted to improve the quality. In this review, recent developments regarding post-modification methods of biochar are presented and discussed. Progresses on the applications of post modified biochar as electrode materials for supercapacitors are intensively summarized. This review aims to reveal the key factors that affecting the performance of biochar-based supercapacitors, and provide guidance for rationalizing the modification methods to expand the applications of biochar-based functional materials in supercapacitors.

Electricity production and microbial characterization of thermophilic microbial fuel cells.

Thermophilic microbial fuel cell (TMFC) offers many benefits, but the investigations on the diversity of exoelectrogenic bacteria are scarce. In this study, a two-chamber TMFC was constructed using ethanol as an electron donor, and the microbial dynamics were analyzed by high-throughput sequencing and 16S rRNA clone-library sequencing. The open-circuit potential of TMFC was approximately 650mV, while the maximum voltage was around 550mV. The maximum power density was 437mW/m2, and the columbic efficiency in this work was 20.5±6.0%. The Firmicutes bacteria, related to the uncultured bacterium clone A55_D21_H_B_C01 with a similarity of 99%, accounted for 90.9% of all bacteria in the TMFC biofilm. This unknown bacterium has the potential to become a new thermophilic exoelectrogenic bacterium that is yet to be cultured. The development of TMFC-involved biotechnologies will be beneficial for the production of valuable chemicals and generation of energy in the future.

Applying rheological analysis to better understand the mechanism of acid conditioning on activated sludge dewatering.

The pH value is a key parameter and affects sludge dewatering. Comprehensive understanding of the effects and mechanism of pH is important for sludge treatment process and sludge dewatering. The goal of this study was to evaluate the proposed mechanism of acid conditioning on sludge dewatering based on rheological analysis. At lower sludge pH, changes in floc structure, surface properties, and flocculation improved the performance of dewatering. Additionally, lower sludge pH caused the hydrolysis of EPS and intracellular materials, which released greater amounts of bound water. These changes resulted in altered rheological properties, weakening network strength and shrinking the linear viscoelastic regime, making the sludge system sensitive to shear. Thus, both the sludge dewatering rate and moisture reduction efficiency were improved by lowering the pH. These factors demonstrate that rheological analysis can understand the mechanism of acid conditioning on activated sludge dewatering better.

Tracking the activity of the Anammox-DAMO process using excitation-emission matrix (EEM) fluorescence spectroscopy.

Coupling of anaerobic ammonium oxidation (anammox) and denitrifying anaerobic methane oxidation (DAMO) microorganisms in a hollow-fiber membrane biofilm reactor (HfMBR) is a potential strategy for simultaneous anaerobic removal of nitrogen and methane in wastewater streams. In these systems, effluents contain dissolved organic substances from anammox and DAMO microorganisms, but their characteristics and relationships have not been investigated. In the present study, excitation-emission matrix (EEM) fluorescence spectroscopy was used to characterize effluent dissolved organic matter (EfDOM) from an Anammox-DAMO HfMBR. Four component types (Component 1-4) were identified by parallel factor analysis (PARAFAC) of EEM data. Component 1 was produced when anammox and DAMO microorganisms simultaneously starved, whereas Component 4 was only generated through the starving period of DAMO microorganisms, and the longer the starving period, the higher the fluorescence intensity of the components. Components 2 and 3 were generated via active and starving periods of co-cultures. More efficient nitrogen removal was accompanied by a higher fluorescence intensity and microbial activity. Compared to measuring both influent and effluent nitrogen concentrations, monitoring EfDOM can obtain other information about the reactor, such as nitrogen removal activity of the reactor, status of the microbes and the duration of starving period the reactor suffered, which therefore offers a complementary but direct tool for assessing reactor performance in complex co-culture systems.

In-situ sludge pretreatment in a single-stage anaerobic digester.

This study aimed to develop an in-situ sludge pretreatment method by increasing the temperature from thermophilic to extreme thermophilic condition in a single-stage anaerobic digester. The results revealed that a stable performance was obtained within the temperature range of 55-65°C, and the maximum methane yield of 208.51±13.66mL/g VS was obtained at 65°C. Moreover, the maximum extent of hydrolysis (33%) and acidification (27.1%) was also observed at 65°C. However, further increase of temperature to 70°C did not improve the organic conversion efficiency. Microbial community analysis revealed that Coprothermobacter, highly related to acetate oxidisers, appeared to be the abundant bacterial group at higher temperature. A progressive shift in methanogenic members from Methanosarcina to Methanothermobacter was observed upon increasing the temperature. This work demonstrated single-stage sludge digestion system can be successfully established at high temperature (65°C) with stable performance, which can eliminate the need of conventional thermophilic pretreatment step.