Bioelectrochemically altering microbial ecology in upflow anaerobic sludge blanket to enhance methanogenesis fed with high-sulfate methanolic wastewater.

A bioelectrochemical upflow anaerobic sludge blanket (BE-UASB) was constructed and compared with the traditional UASB to investigate the role of bioelectrocatalysis in modulating methanogenesis and sulfidogensis involved within anaerobic treatment of high-sulfate methanolic wastewater (COD/SO42- ratio ≤ 2). Methane production rate for BE-UASB was 1.4 times higher than that of the single UASB, while SO42- removal stabilized at 16.7%. Bioelectrocatalysis selectively enriched key functional anaerobes and stimulated the secretion of extracellular polymeric substances, especially humic acids favoring electron transfer, thereby accelerating the electroactive biofilms development of electrodes. Methanomethylovorans was the dominant genus (35%) to directly convert methanol to CH4. Methanobacterium as CO2 electroreduction methane-producing archaea appeared only on electrodes. Acetobacterium exhibited anode-dependence, which provided acetate for sulfate-reducing bacteria (norank Syntrophobacteraceae and Desulfomicrobium) through synergistic coexistence. This study confirmed that BE-UASB regulated the microbial ecology to achieve efficient removal and energy recovery of high-sulfate methanolic wastewater.

Optimizing bioelectromethanosynthesis of CO2 and membrane fouling mitigation in MECs via in-situ biogas recirculation.

The CO2 bioelectromethanosynthesis via two-chamber microbial electrolysis cell (MEC) holds tremendous potential to solve the energy crisis and mitigate the greenhouse gas emissions. However, the membrane fouling is still a big challenge for CO2 bioelectromethanosynthesis owing to the poor proton diffusion across membrane and high inter-resistance. In this study, a new MEC bioreactor with biogas recirculation unit was designed in the cathode chamber to enhance secondary-dissolution of CO2 while mitigating the contaminant adhesion on membrane surface. Biogas recirculation improved CO2 re-dissolution, reduced concentration polarization, and facilitated the proton transmembrane diffusion. This resulted in a remarkable increase in the cathodic methane production rate from 0.4 mL/L·d to 8.5 mL/L·d. A robust syntrophic relationship between anodic organic-degrading bacteria (Firmicutes 5.29%, Bacteroidetes 25.90%, and Proteobacteria 6.08%) and cathodic methane-producing archaea (Methanobacterium 65.58%) enabled simultaneous organic degradation, high CO2 bioelectromethanosynthesis, and renewable energy storage.

Biogas upgrading and membrane anti-fouling mechanisms in electrochemical anaerobic membrane bioreactor (EC-AnMBR): Focusing on spatio-temporal distribution of metabolic functionality of microorganisms.

Electrochemical anaerobic membrane bioreactor (EC-AnMBR) by integrating a composite anodic membrane (CAM), represents an effective method for promoting methanogenic performance and mitigating membrane fouling. However, the development and formation of electroactive biofilm on CAM, and the spatio-temporal distribution of key functional microorganisms, especially the degradation mechanism of organic pollutants in metabolic pathways were not well documented. In this work, two AnMBR systems (EC-AnMBR and traditional AnMBR) were constructed and operated to identify the role of CAM in metabolic pathway on biogas upgrading and mitigation of membrane fouling. The methane yield of EC-AnMBR at HRT of 20 days was 217.1 ± 25.6 mL-CH4/g COD, about 32.1 % higher compared to the traditional AnMBR. The 16S rRNA analysis revealed that the EC-AnMBR significantly promoted the growth of hydrolysis bacteria (Lactobacillus and SJA-15) and methanogenic archaea (Methanosaeta and Methanobacterium). Metagenomic analysis revealed that the EC-AnMBR promotes the upregulation of functional genes involved in carbohydrate metabolism (gap and kor) and methane metabolism (mtr, mcr, and hdr), improving the degradation of soluble microbial products (SMPs)/extracellular polymeric substances (EPS) on the CAM and enhancing the methanogens activity on the cathode. Moreover, CAM biofilm exhibits heterogeneity in the degradation of organic pollutants along its vertical depth. The bacteria with high hydrolyzing ability accumulated in the upper part, driving the feedstock degradation for higher starch, sucrose and galactose metabolism. A three-dimensional mesh-like cake structure with larger pores was formed as a biofilter in the middle and lower part of CAM, where the electroactive Geobacter sulfurreducens had high capabilities to directly store and transfer electrons for the degradation of organic pollutants. This outcome will further contribute to the comprehension of the metabolic mechanisms of CAM module on membrane fouling control and organic solid waste treatment and disposal.

Simultaneous removal of antibiotic resistance genes and improved dewatering ability of waste activated sludge by Fe(II)-activated persulfate oxidation.

Waste activated sludge properties vary widely with different regions due to the difference in living standards and geographical distribution, making a big challenge to developing a universally effective sludge dewatering technique. The Fe(II)-activated persulfate (S2O82-) oxidation process shows excellent ability to disrupt sludge cells and extracellular polymeric substances (EPS), and release bound water from sludge flocs. In this study, the discrepancies in the physicochemical characteristics of sludge samples from seven representative cities in China (e.g., dewaterability, EPS composition, surface charge, microbial community, relative abundance of antibiotic resistance genes (ARGs), etc.) were investigated, and the role of Fe(II)-S2O82- oxidation in enhancing removal of antibiotic resistance genes and dewatering ability were explored. The results showed significant differences between the EPS distribution and chemical composition of sludge samples due to different treatment processes, effluent sources, and regions. The Fe(II)-S2O82- oxidation pretreatment had a good enhancement of sludge dewatering capacity (up to 76 %). Microbial analysis showed that the microbial community in each sludge varied significantly depending on the types of wastewater, the wastewater treatment processes, and the regions, but Fe(II)-S2O82- oxidation was able to attack and rupture the sludge zoogloea indiscriminately. Genetic analysis further showed that a considerable number of ARGs were detected in all of these sludge samples and that Fe(II)-S2O82- oxidation was effective in removing ARGs by higher than 90 %. The highly active radicals (e.g., SO4-·, ·OH) produced in this process caused drastic damage to sludge microbial cells and DNA stability while liberating the EPS/cell-bound water. Co-occurrence network analysis highlighted a positive correlation between population distribution and ARGs abundance, while variations in microbial communities were linked to regional differences in living standards and level of economic development. Despite these variations, the Fe(II)-S2O82- oxidation consistently achieved excellent performance in both ARGs removal and sludge dewatering. The significant modularity of associations between different microbial communities also confirms its ability to reduce horizontal gene transfer (HGT) by scavenging microbes.

Self-assembled electrochemically active biofilms doped with carbon nanotubes: Electron exchange efficiency and cytotoxicity evaluation.

Thick electrochemically active biofilms (EABs) will lead to insufficient extracellular electron transfer (EET) rate because of the limitation of both substrate diffusion and electron exchange. Herein, carbon nanotubes (CNTs)-doped EABs are developed through self-assembly. The highly conductive biofilms (internal resistance of ∼211 Ω) are efficiently enriched at CNTs dosage of 1 g L-1, with the stable power output of 0.568 W m-2 over three months. The embedded CNTs can act as electron tunnel to accelerate the EET rate in thick biofilm. Self-charging/discharging experiments and Nernst-Monod model stimulation demonstrate a higher net charge storage capacity (0.15 C m-2) and more negative half-saturation potential (-0.401 V) for the hybrid biofilms than that of the control (0.09 C m-2, and -0.378 V). Enzyme activity tests and the observation of confocal laser scanning microscopy by live/dead staining show a nearly negligible cytotoxicity of CNTs, and non-targeted metabonomics analysis reveals fourteen differential metabolites that do not play key roles in microbial central metabolic pathways according to KEGG compound database. The abundance of typical exoelectrogens Geobacter sp. is 2-fold of the control, resulting in a better bioelectrocatalytic activity. These finding provide a possible approach to prolong electron exchange and power output by developing a hybrid EABs doped with conductive material.

Impact of sandwich-type composite anodic membrane on membrane fouling and methane recovery from sewage sludge and food waste via electrochemical anaerobic membrane bioreactor.

Membrane fouling presents a big challenge for the real-world implementation of anaerobic membrane bioreactors (AnMBRs) in digesting high-solid biowastes. In this study, an electrochemical anaerobic membrane bioreactor (EC-AnMBR) with a novel sandwich-type composite anodic membrane was designed and constructed for controlling membrane fouling whilst improving the energy recovery. The results showed that EC-AnMBR produced a higher methane yield of 358.5 ± 74.8 mL/d, rising by 12.8% compared to the AnMBR without applied voltage. Integration of composite anodic membrane induced a stable membrane flux and low transmembrane pressure through forming an anodic biofilm while total coliforms removal reached 97.9%. The microbial community analysis further provided compelling evidence that EC-AnMBR enriched the relative abundance of hydrolyzing (Chryseobacterium 2.6%) bacteria and methane-producing (Methanobacterium 32.8%) archaea. These findings offered new insights into anti-biofouling performance and provided significant implications for municipal organic waste treatment and energy recovery in the new EC-AnMBR.

Enhanced co-digestion of sewage sludge and food waste using novel electrochemical anaerobic membrane bioreactor (EC-AnMBR).

Membrane fouling remains a big challenge hindering the wide-application of anaerobic membrane bioreactor (AnMBR) technology. In this study, an electrochemical anaerobic membrane bioreactor (EC-AnMBR) was developed by coupling electrochemical regulation to enhance co-digestion of sewage sludge and food waste and mitigate membrane fouling. The highest methane production (0.12 ± 0.02 L/Lreactor/day) and net energy recovery (31.82 kJ/day) were achieved under the optimum conditions of 0.8 V, hydraulic retention time of 10 days and solids retention time of 50 days. Electrochemical regulation accelerated the mineralization of high-molecular-weight organics and reinforced the membrane antifouling ability by inducing electrostatic repulsive force and electrochemical oxidation. Besides, symbiotic relationships among functional microorganisms (Spirochaetes, Methanolinea, etc.) were enhanced, improving the hydrolysis and methanogenesis processes of complex organics and the long-term stability. This study confirms the technical feasibility of EC-AnMBR in treating high-solid biowastes, and provides the fundamental data to support its application in real-world scenarios.

Disordered mesoporous carbon activated peroxydisulfate pretreatment facilitates disintegration of extracellular polymeric substances and anaerobic bioconversion of waste activated sludge.

The potential of disordered mesoporous carbon (DMC) as catalyst of peroxydisulfate (PDS) to improve sludge solubilization and methane production was investigated. Results showed that DMC activated PDS (DMC/PDS) to produce sulfate radicals (SO4-), facilitating cells rupture and sludge matrix dissociation by degrading the carbonyl and amide groups in organic biopolymers (especially proteins, polysaccharides and humus). At the optimal DMC/PDS dosage of 0.04/1.2 g-mmol/g-VS, SCOD was increased from initial 294.0 to 681.5 mg/L, with the methane production rate of 12.6 mL/g-VS/day. Moreover, DMC could serve as electron mediator to accelerate electron transfer of microorganisms, building a more robust anaerobic metabolic environment. Modelling analysis further demonstrated the crucial role of DMC/PDS pretreatment in biological degradation and methane productivity. This study indicated that DMC/PDS pretreatment can prominently enhance the release of soluble substances and methane production, aiding the utilization of PDS oxidation technology for improving anaerobic bioconversion of sludge.

Magnetite-enhanced bioelectrochemical stimulation for biodegradation and biomethane production of waste activated sludge.

Microbial electrolytic cell (MEC) and magnetite (M) have shown excellent performance in promoting anaerobic digestion (AD) of biowastes. In this study, four types of anaerobic systems (i.e. single AD, M-AD, MEC-AD, and M-MEC-AD) were developed to comprehensively investigate the potential effects of magnetite-enhanced bioelectrochemical stimulation on the biodegradation of waste activated sludge (WAS) and methane (CH4) production. Results showed that M-MEC-AD system produced the highest cumulative CH4 yield, 9.4% higher than that observed in MEC-AD system. Bioelectrochemical stimulation enriched electroactive Geobacter, and classical methanogens (Methanosaeta and Methanobacterium), and the proliferation was further promoted when coupling with magnetite. The relative abundance of Geobacter (6.9%), Methanosaeta (0.3%), and Methanobacterium (12.6%) in M-MEC-AD system was about 10.8, 1.2, and 1.2 times of MEC-AD system, respectively. The integration of magnetite could serve as the conductive materials, and promote inherent indirect electron transfer (IET) and emerging direct electron transfer (DET) between methanogens and fermentative bacteria, building a more energy-efficient route for interspecies electron transfer and methane productivity. This study demonstrated the positive promotion of the coupled bioelectrochemical regulation and magnetite on organic biodegradation, process stability and CH4 productivity, providing some references for the integrated technology in sludge treatment and bioenergy recovery.

Effects of long-term acclimatization on the optimum substrate mixture ratio and substrate to inoculum ratio in anaerobic codigestion of food waste and cow manure.

The effects of long-term acclimatization on the optimum food waste to cow manure ratio (FW/CM) and substrate to inoculum ratio (S/I) in anaerobic codigestion with FW and CM were investigated by batch trials. For the unacclimated sludge, the highest CH4 yields of 646.6 and 653.4 mL/g VS were achieved under the optimum FW/CM (2.5 VS/VS) and S/I (0.07 VS/VS) ratios, respectively. After more than 550 days of acclimatization, the optimum FW/CM and S/I of the acclimated sludge were improved to 3.4 and 0.68 VS/VS with more anaerobic digestion enzymes and lignocellulose, respectively. Based on high-throughput sequencing analysis, the microbial community structures of bacteria, fungi, and archaea were changed, which was the main reason for the change in the optimum FW/CM and S/I. Therefore, the FW/CM and S/I should be periodically optimized during the long-term operation of codigestion to improve the codigestion efficiency for biogas production.

A comparative study of artificial cow and sheep rumen fermentation of corn straw and food waste: Batch and continuous operation.

To optimize the artificial rumen microorganism sources and develop a stable artificial rumen system, batch and continuous operation were investigated with corn straw and food waste as substrates. The batch trials evaluated the volatile fatty acid (VFA) yield, biogas production, and lignocellulose degradation efficiency. The continuous test evaluated the performance of the artificial cow and sheep rumen systems using a dynamic membrane bioreactor (DMBR) with a stepwise organic loading rate at mesophilic temperature. The anaerobic digestion (AD) of the lignocellulose biomass after rumen fermentation pretreatment and of the permeate from the artificial rumen system were also evaluated for CH4 production. The results indicated that the cow rumen microorganisms were more suitable than sheep rumen microorganisms for lignocellulosic biomass pretreatment and maximized the CH4 yield through the AD process without inhibition. After approximately four months of continuous operation, a stable and continuous artificial rumen system for lignocellulosic biomass degradation was achieved with cow rumen fluid as inoculum. Based on analysis of the core lignocellulose-degrading enzyme levels and gel filtration chromatography, the cow rumen microorganisms could secrete more extracellular multienzyme complexes to hydrolyze lignocellulosic biomass than the sheep rumen microorganisms in vitro. During the batch and continuous operations, a high diversity and similar richness of bacteria and fungi demonstrated that the cow rumen microorganisms can be used as a preferred inoculum for the artificial rumen system. The use of an artificial cow rumen system with a DMBR is a promising way to construct a stable and continuous artificial rumen system to biodegrade lignocellulosic biomass for biogas production.

Acclimatization of anaerobic sludge with cow manure and realization of high-rate food waste digestion for biogas production.

Long-term acclimatization of anaerobic sludge was conducted by operating a mesophilic continuously stirred anaerobic reactor (CSTR) with continuous feeding of food wastes (FW) and cow manure (CM). During the long-term acclimatization, continued increase of enzyme activity was revealed, while the microbial structure tended stable as shown by the Shannon index and microbial community. By biomethane potential analysis, the acclimated sludge had a methane yield about 13 times higher than the initial anaerobic sludge. The acclimated sludge was subsequently used for FW digestion with stepwise organic loading rate increase without CM addition. The functional phyla of Bacteroidetes and Proteobacteria, which originated from CM but not very abundant, were significantly enriched not only during sludge acclimatization with CM addition but also in the process of FW digestion without CM addition. A microbe coexistence network was proposed to support an explanation of the metabolic pathways of FW digestion using the acclimated sludge.

Stable and high-rate anaerobic co-digestion of food waste and cow manure: Optimisation of start-up conditions.

Food waste (FW) and cow manure (CM) were co-digested to achieve a stable and high-rate of methane production. The start-up conditions (substrate mixing (FW/CM) ratio, substrate to inoculum ratio, and initial pH) were optimised, and the optimised parameters were experimentally confirmed by batch operation under mesophilic temperatures. To further verify the effects of start-up conditions on the long-term co-digestion process, a semi-continuous dynamic membrane bioreactor was operated for over 300 days with an FW/CM ratio of 2.5. Following the optimised operation scheme, the organic loading rate gradually increased to 11.9 g COD/L/d. Thus, stable anaerobic co-digestion was maintained at FW/CM ratio of 2.5, and a high CH4 production of 2.71 L CH4/L/d and CH4 yield of 441 mL CH4/g VS was achieved. In the long-term operation, the digestate pH was stable at approximately 8.4, which indicated a very favourable anaerobic reaction condition without volatile fatty acid accumulation.

Cosubstrate strategy for enhancing lignocellulose degradation during rumen fermentation in vitro: Characteristics and microorganism composition.

To enhance the degradation of wheat straw (WS) and corn straw (CS) in rumen fermentation, characterization of degradation and ruminal microorganisms of monosubstrate (WS/CS) groups and a cosubstrate strategy with food waste (FW) group was performed. The cellulose, hemicellulose, and lignin degradation efficiency of WS and CS; soluble chemical oxygen demand; volatile fatty acid yields; and activity of ligninolytic, cellulolytic, and hemicellulolytic enzymes for the cosubstrate group were improved compared with those for the corresponding monosubstrate groups. An accurate and a good of fit of the Weibull kinetic model, decreased crystallinity index values, and characteristic absorbance bands in the Fourier transform-infrared spectra further confirmed that cosubstrate addition with FW decreased the resistance of cellulose and hemicellulose to biodegradation. High-throughput sequencing results suggested that the bacterial diversity in CS rumen fermentation and fungal diversity and richness in WS rumen fermentation were promoted with FW as a cosubstrate. The cosubstrate addition with FW significantly affected the composition of the ruminal bacteria and fungi in rumen fermentation. The relative abundances (RAs) of rumen bacteria were increased in the cosubstrate CS/WS and FW fermentation conditions, and the enhancement of CS degradation with FW supplementation was stronger than that of WS rumen fermentation with FW supplementation. The RAs of the ruminal fungal genera Ustilago and Fusarium were promoted in CS and WS fermentation with FW, respectively. Moreover, the fermentation properties and rumen flora in the FW rumen fermentation also provided some evidence to suggest an enhancement of the cosubstrate strategy compared with the monosubstrate strategy.

Persistent action of cow rumen microorganisms in enhancing biodegradation of wheat straw by rumen fermentation.

Rumen fermentation is known to be effective for lignocellulosic-wastes biodegradation to certain extent but it is still unclear if there exists a termination of the microorganisms' action to further degrade the bio-refractory fractions. In order to illuminate the related microbiological characteristics, experiments were conducted in a prolonged duration of rumen fermentation of mechanically ruptured wheat straw, with inoculation of cow rumen microorganisms in vitro. Although the organic wastes could not be biodegraded quickly, continuous conversion of the lignocellulosic contents to volatile fatty acids and biogas proceeded in the duration of more than three months, resulting in 96-97% cellulose and hemicellulose decomposition, and 42% lignin decomposition. X-ray diffraction and Fourier transform infrared spectroscopy further demonstrated the characteristics of lignocellulosic structure decomposition. Under the actions of cow rumen microorganisms, stable pH was maintained in the fermentation liquid, along with a steady NH4+-N, volatile fatty acids accumulation, and a large buffering ability. It was identified by enzyme analysis and Illumina MiSeq sequencing that the rich core lignocellulolytic enzymes secreted by the abundant and diverse rumen bacteria and fungi contributed to the persistent degradation of lignocellulosic wastes. Members of the Clostridiales order and Basidiomycota phylum were found to be the dominant lignocellulolytic bacteria and fungi, respectively. It could thus be inferred that the main lignocellulose degradation processes were a series of catalytic reactions under the actions of lignocellulolytic enzymes secreted from bacteria and fungi. The dominant hydrogenotrophic methanogens (Methanomassiliicoccus, Methanobrevibacter, Methanosphaera, and Methanoculleus) in the rumen could also assist CH4 production if the rumen fermentation was followed with anaerobic digestion.

Cow manure as additive to a DMBR for stable and high-rate digestion of food waste: Performance and microbial community.

Cow manure (CM) was added to a dynamic membrane bioreactor (DMBR) operated under anaerobic condition for enhancing food waste (FW) digestion for over 300 days with stepwise increase of organic loading rates (OLRs) from 1.07 to 11.9 g COD/L/day. At a FW/CM ratio of 3.5:1 (based on volatile solids), the mixed liquor pH was always above 8.0 and no apparent volatile fatty acids (VFAs) accumulation occurred even at the highest OLR of 11.9 g COD/L/day (hydraulic retention time as 10 days and solid retention time as 15.5 days, correspondingly), indicating a very stable operation condition which resulted in an average CH4 yield as high as 250 mL/g COD and CH4 production as high as 2.71 L CH4/L/day. The hardly biodegradable organic components, such as cellulose, hemicellulose, and lignin, were effectively degraded by 78.3%, 58.8%, and 47.5%, respectively. Significantly high anaerobic digestion reaction ratios, especially the hydrolysis ratio which is usually the limiting factor, were calculated based on experimental results. Furthermore, the high lignocellulase contents and coenzyme F420 levels, along with the decrease of cellulose crystallinity from 72.6% to 16.4% in the feedstock, provided strong evidence of an enhanced biological activity by CM addition. By high-throughput sequencing analysis, more abundant and diverse bacterial, archaeal, and fungal genera were identified from the DMBR sludge. With CM addition, the biodegradation of lignocellulose might have produced sufficient H2 and CO2 for the hydrogenotrophic methanogens such as Methanoculleus, Methanomassiliicoccus, and Methanobacterium, which were highly tolerant to ammonium inhibition, and then the elevated ammonium level would have provided high buffering capacity in the DMBR thus ensuring a stable condition for high rate FW digestion and CH4 production.