Chemical and biological effects of zero-valent iron (ZVI) concentration on in-situ production of H2 from ZVI and bioconversion of CO2 into CH4 under anaerobic conditions.

The conversion of carbon dioxide (CO2) to methane (CH4) is a strategy for sequestering CO2. Zero-valent iron (ZVI) has been proposed as an alternative electron donor for the CO2 reduction to CH4. In this study, the effects of ZVI concentrations on the abiotic production of H2 (without the action of microorganisms) in the first part and on the biological conversion of CO2 to CH4 using ZVI as a direct electron donor in the second part were examined. In the abiotic H2 production, the increase in the ZVI concentration from 16 to 32, 64, and 96 g/L was found to have positive effects on both the amounts of H2 generated and the rates of H2 production because the extent of ZVI oxidation positively correlates with increasing surface area. Nevertheless, the increase in ZVI concentration from 96 to 224 g/L did not benefit the H2 production because the ZVI dissolution was suppressed by the increasing aqueous pH above 10. In the bioconversion of CO2 to CH4 using ZVI as an electron donor, the main methanogenesis pathway occurred via hydrogenotrophic methanogenesis at pH 8.7-9.5 driven by the genus Methanobacterium of the class Methanobacteria. At ZVI concentrations of 64 g/L and above, the production of volatile fatty acid (VFA) became clear. Acetate was the main VFA, indicating the induction of homoacetogenesis at ZVI concentrations of 64 g/L and above. In addition, the presence of propionate as the second major VFA suggests the production of propionate from CO2 and acetate under conditions with high H2 partial pressure. The results indicated that the pathway for ZVI/CO2 conversion to CH4 was competitive between hydrogenotrophic methanogenesis and homoacetogenesis.

In-situ methane enrichment in anaerobic digestion of food waste slurry by nano zero-valent iron: Long-term performance and microbial community succession.

In this work, nano zero-valent iron (nZVI) was added to a lab-scale continuous stirring tank reactor (CSTR) for food waste slurry treatment, and the effect of dosing rate and dosage of nZVI were attempted to be changed. The results showed that anaerobic digestion (AD) efficiency and biomethanation stability were optimum under the daily dosing and dosage of 0.48 g/gTCOD. The average daily methane (CH4) yield reached 495.38 mL/gTCOD, which was 43.65% higher than that at control stage, and the maximum CH4 content reached 95%. However, under single dosing rate conditions, high nZVI concentrations caused microbial cell rupture and loosely bound extracellular polymeric substances (LB-EPS) precipitation degradation. The daily dosing rate promoted the hydrogenotrophic methanogenesis pathway, and the activity of coenzyme F420 increased by 400.29%. The microbial analysis indicated that daily addition of nZVI could promote the growth of acid-producing bacteria (Firmicutes and Bacteroidetes) and methanogens (Methanothrix).

Combined disposal of methyl orange and corn straw via stepwise adsorption-biomethanation-composting.

Agriculture wastes have been proved to be the potential adsorbents to remove azo dye from textile wastewater, but the post-treatment of azo dye loaded agriculture waste is generally ignored. A three-step strategy including sequential adsorption-biomethanation-composting was developed to realize the co-processing of azo dye and corn straw (CS). Results showed that CS represented a potential adsorbent to remove methyl orange (MO) from textile wastewater, with the maximum MO adsorption capacity of 10.00 ± 0.46 mg/g, deriving from the Langmuir model. During the biomethanation, CS could serve as electron donor for MO decolorization and substrate for biogas production simultaneously. Though the cumulative methane yield of CS loaded with MO was 11.7 ± 2.28% lower than that of blank CS, almost complete de-colorization of MO could be achieved within 72 h. Composting could achieve the further degradation of aromatic amines (intermediates during the degradation of MO) and decomposition of digestate. After 5 days' composting, 4-aminobenzenesulfonic acid (4-ABA) was not detectable. The germination index (GI) also indicated that the toxicity of aromatic amine was eliminated. The overall utilization strategy gives novel light on the management of agriculture waste and textile wastewater.

Inoculum-to-substrate ratio and solid content effects over in natura spent coffee grounds anaerobic digestion.

Coffee is the second most traded commodity worldwide, and its production is associated with the generation of a large number of residues, which are underused and disposed of in landfills. Notably, the coffee industry annually generates approximately 6 million tons of industrial spent coffee ground (ISCG) when extracting coffee flavorings to produce soluble coffee. That resource loss scenario has been highlighted in sustainable waste management contexts as an opportunity to improve the coffee circular economy. Despite ISCG bioconversion to methane potentially meets the waste-to-energy purposes of reducing residues disposal in landfills, decreasing greenhouse gas (GHG) emissions, and increasing renewable energy sources, data about anaerobic digestion (AD) of ISCG remains quite restricted. That limitation becomes more apparent owing to the lack of data focusing on AD key parameters for ISCG as substrate. This study assessed the influence of inoculum-to-substrate ratio (ISR) and the solid content influences on mesophilic (37 °C) ISCG-AD throughout the Response Surface Methodology (RSM) and Central Composite Design (CCD). Results revealed that both factors, ISR and solid content, should be kept above a certain threshold of 0.5 and 6.0 gTVS L-1 to ensure experimental reliability, as well as reproductively and above 1.0 and 8.0 gTVS L-1 to avoid underestimation on the MY potential achieved. Concerning ISCG-AD kinetics, the quadratic model optimum condition was at 1.36 and 14.83 gTVS L-1 for ISR and solid content, respectively. This optimum range for ISR and solid content could guide further development of process configurations for mono- and co-digestion of ISCG, avoiding underestimation of the MY potential and extended incubation periods.

Anaeropeptidivorans aminofermentans gen. nov., sp. nov., a mesophilic proteolytic salt-tolerant bacterium isolated from a laboratory-scale biogas fermenter, and emended description of Clostridium colinum.

An anaerobic bacterial strain, designated strain M3/9T, was isolated from a laboratory-scale biogas fermenter fed with maize silage supplemented with 5 % wheat straw. Cells were straight, non-motile rods, which stained Gram-negative. Optimal growth occurred between 30 and 40°C, at pH 7.5-8.5, and up to 3.9 % (w/v) NaCl was tolerated. When grown on peptone from casein and soymeal, strain M3/9T produced mainly acetic acid, ethanol, and isobutyric acid. The major cellular fatty acids of the novel strain were C16 : 0 and C16 : 0 DMA. The genome of strain M3/9T is 3757  330 bp in size with a G+C content of 38.45 mol%. Phylogenetic analysis allocated strain M3/9T within the family Lachnospiraceae with Clostridium colinum DSM 6011T and Anaerotignum lactatifermentans DSM 14214T being the most closely related species sharing 57.86 and 56.99% average amino acid identity and 16S rRNA gene sequence similarities of 91.58 and 91.26 %, respectively. Based on physiological, chemotaxonomic and genetic data, we propose the description of a novel species and genus Anaeropeptidivorans aminofermentans gen. nov., sp. nov., represented by the type strain M3/9T (=DSM 100058T=LMG 29527T). In addition, an emended description of Clostridium colinum is provided.

Review on solid-state anaerobic digestion of lignocellulosic biomass and organic solid waste.

Sustainable management of organic solid wastes especially the municipal solid waste (MSW) is essential for the realization of various sustainable development goals (SDGs). Resource recovery centric waste processing technologies generate valorizable products to meet the operations and maintenance (O&M) costs while reducing the GHG emissions. Solid-state anaerobic digestion (SSAD) of organic solid wastes is a biomethanation process performed at a relatively higher total solids (TS) loading in the range of 10-45%. SSAD overcomes various limitations posed by conventional anaerobic slurry digesters such as higher degradable matter per unit volume of the bioreactor resulting in a smaller footprint, low freshwater consumption, low wastewater generation, simple upstream and downstream processes, relatively lower operation, and maintenance costs. This review elucidates the recent developments and critical assessment of different aspects of SSAD, such as bioreactor design, operational strategy, process performances, mass balance, microbial ecology, applications, and mathematical models. A critical assessment revealed that the operating scale of SSAD varies between 1000 and 100,000 ts/year at organic loading rate (OLR) of 2-15 g volatile solids (VS)/L·day. The SSAD experiences process failures due to the formation of volatile fatty acids (VFAs), biogas pockets and clogging of the digestate outlet. Acclimatization of microbes accelerates the startup phase, steady-state performances, and the enrichment of syntrophic microbes with 10-50 times greater population of cellulolytic and xylanolytic microbes in thermophilic SSAD over mesophilic SSAD. Experimental limitations in the accurate determination of rate constants and the oversimplification of biochemical reactions result in an inaccurate prediction by the models.

Metagenomic approaches: effective tools for monitoring the structure and functionality of microbiomes in anaerobic digestion systems.

Microbial metagenome analysis has proven its usefulness to investigate the microbiomes present in technical engineered ecosystems such as anaerobic digestion systems. The analysis of the total microbial genomic DNA allows the detailed determination of both the microbial community structure and its functionality. In addition, it enables to study the response of the microbiome to alterations in technical process parameters. Strategies of functional microbial networks to face abiotic stressors, e.g., resistance, resilience, and reorganization, can be evaluated with respect to overall process optimization. The objective of this paper is to review the main metagenomic tools used for effective studies on anaerobic digestion systems in monitoring the dynamic of the microbiomes, as well as the factors that have been identified so far as limiting the metagenomic studies in this ecosystems.

Proteiniborus indolifex sp. nov., isolated from a thermophilic industrial-scale biogas plant.

A novel strictly anaerobic bacterium, designated strain BA2-13T, was isolated from a thermophilic industrial-scale biogas plant. Cells were rod-shaped and Gram-stain-positive. Growth occurred at temperatures of 25 to 50 °C and between pH 6.3 and 9.5. Strain BA2-13T produced indole. Cell growth was stimulated by yeast extract, peptone, meat extract, a mixture of 20 amino acids, glucose, pyruvate and ribose. When grown on peptone and yeast extract, the main fermentation products were acetic acid, H2 and CO2. The predominant cellular fatty acids were iso-C15 : 0 and iso-C14 : 0 3-OH. Major polar lipids were diphosphatidylglycerol, glycolipids, phospholipids and phosphatidylgycerol. Phylogenetic analysis based on 16S rRNA gene nucleotide sequence analysis placed strain BA2-13T within the order Clostridiales showing closest affiliation with Proteiniborusethanoligenes with 95.9 % sequence identity. Physiological, genotypic and chemotaxonomic differences of strain BA2-13T from P. ethanoligenes support the description of a new species within the genus Proteiniborus for which we suggest the name Proteiniborusindolifex sp. nov. (type strain BA2-13T=DSM 103060T=LMG 29818T).

Bioenergy production from diluted poultry manure and microbial consortium inside Anaerobic Sludge Bed Reactor at sub-mesophilic conditions.

In this study, anaerobic treatability of diluted chicken manure (with an influent feed ratio of 1 kg of fresh chicken manure to 6 L of tap water) was investigated in a lab-scale anaerobic sludge bed (ASB) reactor inoculated with granular seed sludge. The ASB reactor was operated at ambient temperature (17-25°C) in order to avoid the need of external heating up to higher operating temperatures (e.g., up to 35°C for mesophilic digestion). Since heat requirement for raising the temperature of incoming feed for digestion is eliminated, energy recovery from anaerobic treatment of chicken manure could be realized with less operating costs. Average biogas production rates were calculated ca. 210 and 242 L per kg of organic matter removed from the ASB reactor at average hydraulic retention times (HRTs) of 13 and 8.6 days, respectively. Moreover, average chemical oxygen demand (COD) removal of ca. 89% was observed with suspended solids removal more than 97% from the effluent of the ASB reactor. Influent ammonia, on the other hand, did not indicate any free ammonia inhibition due to dilution of the raw manure while pH and alkalinity results showed stability during the study. Microbial quantification results indicated that as the number of bacterial community decreased, the amount of Archaea increased through the effective digestion volume of the ASB reactor. Moreover, the number of methanogens displayed an uptrend like archaeal community and a strong correlation (-0.645) was found between methanogenic community and volatile fatty acid (VFA) concentration especially acetate.

Ecobiotechnological strategy to enhance efficiency of bioconversion of wastes into hydrogen and methane.

Vegetable wastes (VW) and food wastes (FW) are generated in large quantities by municipal markets, restaurants and hotels. Waste slurries (250 ml) in 300 ml BOD bottles, containing 3, 5 and 7 % total solids (TS) were hydrolyzed with bacterial mixtures composed of: Bacillus, Acinetobacter, Exiguobacterium, Pseudomonas, Stenotrophomonas and Sphingobacterium species. Each of these bacteria had high activities for the hydrolytic enzymes: amylase, protease and lipase. Hydrolysate of biowaste slurries were subjected to defined mixture of H2 producers and culture enriched for methanogens. The impact of hydrolysis of VW and FW was observed as 2.6- and 2.8-fold enhancement in H2 yield, respectively. Direct biomethanation of hydrolysates of VW and FW resulted in 3.0- and 1.15-fold improvement in CH4 yield, respectively. A positive effect of hydrolysis was also observed with biomethanation of effluent of H2 production stage, to the extent of 1.2- and 3.5-fold with FW and VW, respectively. The effective H2 yields were 17 and 85 l/kg TS fed, whereas effective CH4 yields were 61.7 and 63.3 l/kg TS fed, from VW and FW, respectively. This ecobiotechnological strategy can help to improve the conversion efficiency of biowastes to biofuels.

Bioaugmentation by enriched hydrogenotrophic methanogens into trickle bed reactors for H2/CO2 conversion.

Biomethanation represents a promising approach for biomethane production, with biofilm-based processes like trickle bed reactors (TBRs) being among the most efficient solutions. However, maintaining stable performance can be challenging, and both pure and mixed culture approaches have been applied to address this. In this study, inocula enriched with hydrogenotrophic methanogens were introduced to to TBRs as bioaugmentation strategy to assess their impacts on the process performance and microbial community dynamics. Metagenomic analysis revealed a metagenome-assembled genome belonging to the hydrogenotrophic genus Methanobacterium, which became dominant during enrichment and successfully colonized the TBR biofilm after bioaugmentation. The TBRs achieved a biogas production with > 96 % methane. The bioaugmented reactor consumed additional H2. This may be due to microbial species utilizing CO2 and H2 via various CO2 reduction pathways. Overall, implementing bioaugmentation in TBRs showed potential for establishing targeted species, although challenges remain in managing H2 consumption and optimizing microbial interactions.

An overview of biomethanation and the use of membrane technologies as a candidate to overcome H2 mass transfer limitations.

Energy produced from renewable sources such as sun or wind are intermittent, depending on circumstantial factors. This fact explains why energy supply and demand do not match. In this context, the interest in biomethanation has increased as an interesting contribution to the Power-to-gas concept (P2G), transforming the extra amount of produced electricity into methane (CH4). The reaction between green hydrogen (H2) (produced by electrolysis) and CO2 (pollutant present in biogas) can be catalysed by different microorganisms to produce biomethane, that can be injected into existing natural gas grid if reaching the standards. Thus, energy storage for both hydrogen and electricity, as well as transportation problems would be solved. However, H2 diffusion to the liquid phase for its further biological conversion is the main bottleneck due to the low solubility of H2. This review includes the state-of-the-art in biological hydrogenotrophic methanation (BHM) and membrane-based technologies. Specifically, the use of hollow-fiber membrane bioreactors as a technology to overcome H2 diffusion limitations is reviewed. Furthermore, the influence of operating conditions, microbiology, H2 diffusion and H2 injection methods are critically discussed before setting the main recommendations about BHM.

External ceramic membrane contactor for in-situ H2 assisted biogas upgrading.

Biological H2-assisted biogas upgrading has gained significant attention as an environmentally friendly substitute to common physico-chemical upgrading techniques, but is largely limited by the low solubility of H2. This study evaluated the design of a ceramic membrane contactor module for H2 injection. H2 dissolution was maintained at high efficiency by controlling gas supply and sludge recirculation rate, achieving a biogas quality of average 98.8% CH4 during the stable operation phase with a 108% increase in the CH4 production rate. This also outperforms conventional H2 injection using diffuser sparging which could only achieve a biogas quality of 84% CH4 content. Microbial community analysis found high Methanobacterium spp. abundance within the archaea at 95.2% at the end of the operation, allowing the dominance of the hydrogenotrophic methanogenesis pathway for high upgrading efficiencies. The system is a high-performance external membrane connector module coupled to common anaerobic digestion systems for biogas upgrading.

[Research advances of microbial transformation of CO2 in geological sequestration].

Carbon capture, utilization and storage is the vital technology for China to achieve the goals of carbon peaking and carbon neutrality. Microbial activities in situ are an indispensable part in the process of geological CO2 sequestration. Some microorganisms can convert CO2 into methane and organics as the resource for utilization or into carbonate to achieve long-term sequestration. These activities contribute to the stable storage of CO2 and even negative carbon emission. This paper focuses on the processes of bio-methanation, bio-liquefaction, and bio-precipitation that may be involved in CO2 sequestration in deep stratum and discusses the research progress in the bio-transformation pathways. Bio-methanation and bio-liquefaction can convert CO2 into methane or high-value organic compounds to realize resource reuse. The two technologies can be used alone or coupled to expand the application range of CO2 biotransformation. Bio-mineralization can convert CO2 into calcite by microorganism-induced carbonate precipitation, being a technology of great potential in fixing CO2 and limiting CO2 escape. At present, this field is still in the infancy stage, and there is an urgent need to establish and improve the theoretical and technical systems of CO2 in-situ biotransformation from transformation principle, influencing factors, conversion efficiency, economy, environmental protection, and technological conditions. Moreover, it can be combined with CCUS to establish a technical system integrating capture, transport, displace, storage, transfer, and exploit, so as to promote the value-added application of CCUS and the achievement of carbon peaking and carbon neutrality.

Making waves: Power-to-X for the Water Resource Recovery Facilities of the future.

The wastewater industry and the energy system are undergoing significant transformations to address climate change and environmental pollution. Green hydrogen, which will be mainly obtained from renewable electricity water electrolysis (Power-to-Hydrogen, PtH), has been considered as an essential energy carrier to neutralize the fluctuations of renewable energy sources. PtH, or Power-to-X (PtX), has been allocated to multiple sectors, including industry, transport and power generation. However, considering its large potential for implementation in the wastewater sector, represented by Water Resource Recovery Facilities (WRRFs), the PtX concept has been largely overlooked in terms of planning and policymaking. This paper proposes a concept to implement PtX at WRRFs, where sourcing of water, utilization of the oxygen by-product, and PtX itself can be sustainable and diversified strategies. Potential value chains of PtX are presented and illustrated in the frame of a WWRF benchmark simulation model, highlighting the applications of oxygen from PtX through pure oxygen aeration and ozone disinfection. Opportunities and challenges are highlighted briefly, and so is the prospective outlook to the future. Ultimately, it is concluded that 'coupling PtX to WRRFs' is a promising solution, which will potentially bring sustainable opportunities for both WRRFs and the energy system. Apart from regulatory and economic challenges, the limitations in coupling PtX to WRRFs mainly come from energy efficiency concerns and the complexity of the integration of the water framework and the energy system.

Unraveling prevalence of homoacetogenesis and methanogenesis pathways due to inhibitors addition.

Three inhibitors targeting different microorganisms, both from Archaea and Bacteria domains, were evaluated for their effect on CO2 biomethanation: sodium ionophore III (ETH2120), carbon monoxide (CO), and sodium 2-bromoethanesulfonate (BES). This study examines how these compounds affect the anaerobic digestion microbiome in a biogas upgrading process. While archaea were observed in all experiments, methane was produced only when adding ETH2120 or CO, not when adding BES, suggesting archaea were in an inactivated state. Methane was produced mainly via methylotrophic methanogenesis from methylamines. Acetate was produced at all conditions, but a slight reduction on acetate production (along with an enhancement on CH4 production) was observed when applying 20 kPa of CO. Effects on CO2 biomethanation were difficult to observe since the inoculum used was from a real biogas upgrading reactor, being this a complex environmental sample. Nevertheless, it must be mentioned that all compounds had effects on the microbial community composition.

Influence of applied voltage on bioelectrochemical enhancement of biomethanation for low-rank coal and microbial community distribution.

The performance of peat biomethanation was investigated in bioelectrochemical anaerobic digestion at different applied voltages, and compared to conventional anaerobic digestion. The methane yield was stabilized at 16 mL/g peat in the conventional anaerobic digestion. However, in the bioelectrochemical anaerobic digestion, the methane yield was significantly increased to 264 mL/g peat at the applied voltage of 4 V, followed by 1 V, 2 V, 0.5 V and 0 V. The bioelectrochemical system could enrich more electroactive microorganisms on the electrode, as well as in the bulk solution, and further improve the direct interspecies electron transfer for methane production. The 16S rRNA analysis showed a significant increase in the abundance of specific microorganisms in the bulk solution, including Firmicutes phylum and Proteobacteria phylum, in addition to a gradual increase in acetoclastic methanogenesis with an increase in applied voltage. These results provide a solution to turn low-rank coal into a new alternative energy.

Combined sodium citrate and ultrasonic pretreatment of waste activated sludge for cost effective production of biogas.

This study aimed to pretreat the waste activated sludge (WAS) by ultrasonication in an energy efficient way by combining sodium citrate with ultrasonic pretreatment at 0.03 g/g suspended solids (SS) of dosage. The ultrasonic pretreatment was done at various (20-200 W) power levels, sludge concentration (7 to 30 g/L), sodium citrate dosages (0.01 to 0.2 g/g SS). An elevated COD solubilization of 26.07 ± 0.6 % was achieved by combined pretreatment at a treatment time of 10 min, ultrasonic power level of 160 W when compared to individual ultrasonic pretreatment (18.6 ± 0.5 %). A higher biomethane yield of 0.26 ± 0.009 L/g COD was achieved in sodium citrate combined ultrasonic pretreatment (SCUP) than ultrasonic pretreatment (UP) 0.145 ± 0.006 L/g COD. Almost 50% of the energy can be saved through SCUP when compared to UP. Future study evaluating SCUP in continuous mode anaerobic digestion is vital.

Syngas biomethanation: In a transfer limited process, is CO inhibition an issue?

Syngas biomethanation is a promising technology in the process chain converting wastes to methane. However, gas-liquid mass transfer is a limiting factor of the biomethanation process. To reach high methane productivity, increasing the pressure is an interesting strategy to improve mass transfer. However, the CO content in the syngas raises concerns about a potential inhibition of the microorganisms. Therefore, the aim of the research was to assess the ability to work at high CO partial pressures. In this regard, a pressurized continuous stirred column with a working volume of 10L was implemented and a consortium adapted for syngas-biomethanation for 22 months was submitted to 100% CO and increasing pressure. No inhibition phenomenon was observed for logarithmic PCO as high as 1.8 bar (inlet pressure 5.0 bar), which was the first time that such a high CO partial pressure was tested in continuous mode. Mass transfer limitations allowed for the carboxydotrophic microorganisms to consume CO faster than it was transferred, allowing for the dissolved CO concentration to remain under inhibitory concentrations. These results question the habitual consensus that CO inhibition is a limiting factor of syngas biomethanation.

Perspective on the use of methanogens in lithium recovery from brines.

Methanogenic archaea stand out as multipurpose biocatalysts for different applications in wide-ranging industrial sectors due to their crucial role in the methane (CH4) cycle and ubiquity in natural environments. The increasing demand for raw materials required by the manufacturing sector (i.e., metals-, concrete-, chemicals-, plastic- and lubricants-based industries) represents a milestone for the global economy and one of the main sources of CO2 emissions. Recovery of critical raw materials (CRMs) from byproducts generated along their supply chain, rather than massive mining operations for mineral extraction and metal smelting, represents a sustainable choice. Demand for lithium (Li), included among CRMs in 2023, grew by 17.1% in the last decades, mostly due to its application in rechargeable lithium-ion batteries. In addition to mineral deposits, the natural resources of Li comprise water, ranging from low Li concentrations (seawater and freshwater) to higher ones (salt lakes and artificial brines). Brines from water desalination can be high in Li content which can be recovered. However, biological brine treatment is not a popular methodology. The methanogenic community has already demonstrated its ability to recover several CRMs which are not essential to their metabolism. Here, we attempt to interconnect the well-established biomethanation process with Li recovery from brines, by analyzing the methanogenic species which may be suitable to grow in brine-like environments and the corresponding mechanism of recovery. Moreover, key factors which should be considered to establish the techno-economic feasibility of this process are here discussed.