A cyclic shift-temperature operation method to train microbial communities of mesophilic anaerobic digestion.

Temperature is the critical factor affecting the efficiency and cost of anaerobic digestion (AD). The current work develops a shift-temperature AD (STAD) between 35 °C and 55 °C, intending to optimise microbial community and promote substrate conversion. The experimental results showed that severe inhibition of biogas production occurred when the temperature was firstly increased stepwise from 35 °C to 50 °C, whereas no inhibition was observed at the second warming cycle. When the organic load rate was increased to 6.37 g VS/L/d, the biogas yield of the STAD reached about 400 mL/g VS, nearly double that of the constant-temperature AD (CTAD). STAD promoted the proliferation of Methanosarcina (up to 57.32 %), while severely suppressed hydrogenophilic methanogens. However, when the temperature was shifted to 35 °C, most suppressed species recovered quickly and the excess propionic acid was quickly consumed. Metagenomic analysis showed that STAD also promoted gene enrichment related to pathways metabolism, membrane functions, and methyl-based methanogenesis.

Enhancing proton-coupled electron transfer drives efficient methanogenesis in anaerobic digestion.

The enhancement of electron or proton transfer between syntrophic microbes has been widely recognised as a means for improving methane generation. However, the uncoupled supplementation of electrons and protons in multiphase anaerobic environment hinders the balanced uptake of electrons and protons in the cytoplasm of methanogens, limiting methanogenesis efficiency. Herein, the cooperative effect of a proton-conductive material (PM) and an electron-conductive material (EM) in enhancing proton-coupled electron transfer (PCET) and driving efficient methanogenesis in anaerobic digestion was investigated. The cooperation of the PM and EM significantly increased methane production and the maximum methane generation rate by 78.9 % and 103.5 %, respectively, indicating enhanced methanogenesis efficiency. Analysis of the physicochemical properties, biochemical components, and microbial dynamics revealed that the cooperation of the PM and EM improved the metabolism of syntrophic microbes, which was critically dependent on electron and proton transfer. This enhancement was primarily due to the improvement in PCET, as mainly supported by hydrogen/deuterium kinetic isotope effect measurements, multi-omics integration analyses and reaction thermodynamics and kinetics analyses. Our findings suggest that the PCET enhancement stimulated efficient membrane-bound enzymatic reactions related to electron-driven proton translocation and facilitated electron and proton supply for CO2 reduction to realise highly efficient methane generation. These findings are expected to provide a new insight into effective electron and proton coupling transfer for methanogenic metabolism in multiphase anaerobic environments.

The Sulfur Conversion Functional Microbial Communities in Biogas Liquid Can Participate in Coal Degradation.

The addition of biogas liquid is a practical way to improve the yield of biological coalbed methane. The microbial composition in biogas liquid is complex, and whether it could participate in the sulfur conversion of coal remains unknown. In this study, sulfur conversion-related microbial communities were enriched from biogas liquid, which was dominated by genera Anaerosolibacter, Bacillus, Hydrogenispora, and Oxobacter. The co-culture of these groups with coal significantly changed the coal microbial community composition but did not increase the content of CH4 and H2S. The changed microbial communities mainly belonged to phyla Firmicutes, Proteobacteria, and Actinobacteriota, and increased the relative abundance of genera Bacillus, Thermicanus, Hydrogenispora, Oxobacter, Lutispora, Anaerovorax, Desulfurispora, Ruminiclostridium, and Fonticella. From the microscopic structure of coal, an increase in the number of holes and roughness on the surface of the coal was found but the change of surface functional groups was weak. In addition, the addition of S-related microbial communities increased the contents of phoxim, methylthiobenzoylglycine and glibornuride M5 in aromatic compounds, as well as the content of lauryl hydrogen sulfate in alkyl compounds. Furthermore, the dibenzothiophene degradation-related microbial communities included Bacillus, Brevibacillus, Brevundimonas, Burkholderia-Caballeronia-Paraburkholderia, and Thermicanus, which can break C-S bonds or disrupt benzene rings to degrade dibenzothiophene. In conclusion, the S-related microbial communities in biogas liquid could rebuild the coal microbial community and be involved in the conversion process of organic sulfur in coal.

Generic mitigating and promoting effect of zeolite on anaerobic digestion: Physicochemical and metataxonomic data.

This article provides comprehensive data on degradation performance and microbial dynamics derived from a set of 24 lab-scale batch anaerobic digesters involving various types of inhibitors and the addition of zeolite as a support material. In the first series of 12 digesters, three inhibitors were investigated at the following concentrations: 20 g/L of sodium chloride, 400 mg/L of erythromycin, and 5 mg/L of S-metolachlor. Each inhibitor was tested in triplicate, along with a control condition without inhibition. A parallel series was set up identically, except that 15 g/L of zeolite was introduced into each digester to mitigate the inhibition and promote the degradation process. The provided data comprises information regarding the experimental setup, monitoring measurements that assess the degradation performance (production, composition, and apparent isotopic factor of biogas, pH, dissolved inorganic and organic carbon and volatile fatty acids concentrations), microbial samples information, and 16S rRNA gene sequencing data that decipher changes in microbial structure. This datapaper is associated with research article [1] and presents both the sequencing data and the associated physicochemical data in a structured table format. The sequencing data were generated using the Ion Torrent PGM sequencer and have been deposited in the European Nucleotide Archive (ENA) database at EMBL-EBI under accession number PRJEB65129 (https://www.ebi.ac.uk/ena/browser/view/PRJEB65129), with sample accession numbers ranging from ERS16257742 to ERS16257691 [2]. The data serves as a valuable resource for comparisons with data from other studies on lab-scale batch anaerobic digesters, particularly those utilizing zeolite as a support material or involving inhibition caused by similar types of inhibitors (salts, antibiotics, or pesticides).

Nitrogen-doped activated carbon-based steel slag composite material as an accelerant for enhancing the resilience of flexible biogas production process against shock loads: Performance, mechanism and modified ADM1 modeling.

Anaerobic digestion for flexible biogas production can lead to digestion inhibition under high shock loads. While steel slag addition has shown promise in enhancing system buffering, its limitations necessitate innovation. This study synthesized the nitrogen-doped activated carbon composite from steel slag to mitigate intermediate product accumulation during flexible biogas production. Material characterization preceded experiments introducing the composite into anaerobic digestion systems, evaluating its impact on methane production efficiency under hydraulic and concentration sudden shocks. Mechanistic insights were derived from microbial community and metagenomic analyses, facilitating the construction of the modified Anaerobic Digestion Model No. 1 (ADM1) to quantitatively assess the material's effects. Results indicate superior resistance to concentration shocks with substantial increment of methane production rate up to 33.45% compared with control group, which is mediated by direct interspecies electron transfer, though diminishing with increasing shock intensity. This study contributes theoretical foundations for stable flexible biogas production and offers an effective predictive tool for conductor material reinforcement processes.

Sustainable valorization of slaughterhouse waste through anaerobic digestion: A circular economy perspective.

Slaughterhouse waste (SHW) poses significant environmental challenges due to its complex composition. In response, a novel review exploration of anaerobic digestion (AD) as a means of valorising SHW within the context of the circular economy (CE) is presented. The physicochemical properties of individual SHW, representing key parameters for the correct management of the AD process, are scrutinized. These parameters are further connected with identifying suitable pretreatment methods to enhance biogas production. Subsequently, the review examines the diverse technologies employed in the AD of SHW, considering the complexities of mono- or co-digestion. Various AD systems are evaluated for their effectiveness in harnessing the substantial biogas production potential from SHW, encompassing key parameters, reactor configurations, and operational conditions that influence the AD process. Moreover, the review interestingly extends its scope to the recovery and management of digestate, the by-product of AD. Along with the digestate composition, strategies for various utilization of this by-product are discussed. This investigation thus underscores, within the principles of the CE, the dual sustainable benefits of SHW processing via AD in biogas production and utilization of the resultant nutrient-rich digestate in various sectors.

Hydrodynamic disintegration effects assessment by CFD modelling integrated with bench tests.

The hydrodynamic disintegration process depends, among others, on operational parameters like rotational speed or introduced energy. The study presents an interdisciplinary approach to the hydrodynamic disintegration parameters impact assessment on the internal processes and disintegration effects on the example of sewage sludge treatment. Three rotational speeds were considered, including fluid properties change at selected disintegration stages. Disintegration effects were measured in the bench tests. Soluble chemical oxygen demand (SCOD) and volatile fatty acids (VFA) were measured before and after disintegration process. The assessment of the effects of disintegration employed the disintegration degree and the assessment of the course of methane production employed biochemical methane potential (BMP) tests. Fluid properties change during the disintegration stages does not cause a significant change in the flow structure. Due to the mathematical modelling results, at 1500 rpm no cavitation phenomenon was observed. Although, the bench tests results indicates, for the rotational speed 1500 rpm, organic compounds released to the liquid were characterised by higher susceptibility to biological decomposition than those released for 2500 and 3000 rpm (as suggested by the low SCOD/VFA values for 1500 rpm). Obtained results have confirmed, that the main phenomenon responsible for the disintegration effect is mechanical shredding not cavitation.

Ultrasonic disintegration of municipal sludge: fundamental mechanisms, process intensification and industrial sono-reactors.

Sludge disintegration is an environmental and industrial challenge that requires intensive research and technological development. Sludge contains a complex structure with a high yield of various chemical and biological compounds. Anaerobic digestion is the most used process for sludge disintegration to produce biogas, detoxify the sludge, and generate biosolids that can be used in agricultural. Biological cell lysis is the rate-limiting cell lysis. This review discusses the application of sonolysis as a sludge pretreatment for enhanced anaerobic digestion via three combined processes: thermal destruction, hydrochemical shear forces, and radical oxidation. The mechanistic pathways of sono-pretreatment to enhance biogas, sludge-enhanced dewatering, activation of filamentous bacteria, oxidation of organic pollutants, release of heavy metals, reduction of bulking and foaming sludge, and boosting ammonia-oxidizing bacteria activity are discussed in this report. The combination of ultrasound with other chemical processes, such as Fenton and cation binding agents for enhanced sludge disintegration, is discussed. Finally, we reviewed the most common large-scale sono-reactors available on the market for sludge disintegration.

Techno-economic and environmental analysis of organic municipal solid waste for energy production.

Addressing the critical conundrum of escalating municipal solid waste (MSW) and shrinking landfill spaces in urban areas, this research pioneers a sustainable approach for Bangladesh by exploring the potential of biogas production from MSW. Distinctly, it fills the research gap by providing a detailed techno-economic and environmental analysis of decentralized fixed-dome anaerobic digestion facilities in the urban context of Chittagong, Bangladesh, a domain previously underexplored. Our findings demonstrate the feasibility of converting MSW into a renewable energy source, offering an innovative solution that simultaneously tackles waste management and energy generation challenges. Each proposed plant showcases the capability to generate 536 m³ of biogas daily, sufficient to power a 50 kW gas engine and supply 44 households, thereby contributing significantly to urban waste reduction and CO2 emissions mitigation by approximately 500 tons monthly. The economic analysis reveals an attractive investment payback period of two years, underscoring the model's viability and its potential as a replicable framework for similar urban settings grappling with waste management crises. This study not only bridges a critical knowledge gap but also introduces a novel, sustainable waste-to-energy model, marking a pivotal step towards achieving energy security and environmental sustainability in developing nations.

Catabolism and interactions of syntrophic propionate- and acetate oxidizing microorganisms under mesophilic, high-ammonia conditions.

Microbial inhibition by high ammonia concentrations is a recurring problem that significantly restricts methane formation from intermediate acids, i.e., propionate and acetate, during anaerobic digestion of protein-rich waste material. Studying the syntrophic communities that perform acid conversion is challenging, due to their relatively low abundance within the microbial communities typically found in biogas processes and disruption of their cooperative behavior in pure cultures. To overcome these limitations, this study examined growth parameters and microbial community dynamics of highly enriched mesophilic and ammonia-tolerant syntrophic propionate and acetate-oxidizing communities and analyzed their metabolic activity and cooperative behavior using metagenomic and metatranscriptomic approaches. Cultivation in batch set-up demonstrated biphasic utilization of propionate, wherein acetate accumulated and underwent oxidation before complete degradation of propionate. Three key species for syntrophic acid degradation were inferred from genomic sequence information and gene expression: a syntrophic propionate-oxidizing bacterium (SPOB) "Candidatus Syntrophopropionicum ammoniitolerans", a syntrophic acetate-oxidizing bacterium (SAOB) Syntrophaceticus schinkii and a novel hydrogenotrophic methanogen, for which we propose the provisional name "Candidatus Methanoculleus ammoniitolerans". The results revealed consistent transcriptional profiles of the SAOB and the methanogen both during propionate and acetate oxidation, regardless of the presence of an active propionate oxidizer. Gene expression indicated versatile capabilities of the two syntrophic bacteria, utilizing both molecular hydrogen and formate as an outlet for reducing equivalents formed during acid oxidation, while conserving energy through build-up of sodium/proton motive force. The methanogen used hydrogen and formate as electron sources. Furthermore, results of the present study provided a framework for future research into ammonia tolerance, mobility, aggregate formation and interspecies cooperation.

Mild alkaline conditions affect digester performance and community dynamics during long-term exposure.

This paper examines the adaptive responses of microbial communities to gradual shifts in pH toward the mild alkaline range in anaerobic digestion (AD) systems. The results indicate that a pH of 8.0 serves as a critical upper limit for stable AD operation, beyond which microbial efficiency declines, underscoring the importance of microbial resilience against elevated pH stress. Specifically, hydrolysis genera, e.g. Eubacterium and Anaerobacterium, and syntrophic bacteria were crucial for reactor stability. Fibrobacter had also been shown to play a key role in the accumulation of propionate, thus leading to its dominance in the volatile fatty acid profile throughout the experimental phases. Overall, this investigation revealed the potential adaptability of microbial communities in AD systems to mild alkaline pH shifts, emphasizing the hydrolysis bacteria and syntrophic bacteria as key factors for maintaining metabolic function in elevated pH conditions.

Gasification chars and activated carbon: Systematic physico-chemical characterization and effect on biogas production.

Gasification residues/chars (GR) and activated carbon (AC) are added to wastewater treatment processes mainly as a fourth purification stage, e.g., to adsorb heavy metals or pharmaceutical residues. However, the effects of GR or AC, which are transferred to the anaerobic digestion (AD) via the sludge, are not yet fully understood. Although, the positive effect of char addition on AD has been demonstrated in several investigations, systematic studies with chemically well described chars are still missing. Therefore, in this study, different chars were characterized in detail, subjected to AD in different concentrations, and their effect on methane production investigated. GR of a gasification plant with a floating fixed bed technology, carbon made by chemical impregnation with ZnCl2 from waste-wood, carbon produced by thermochemical activation with CO2 from GR and commercial powdered AC were used for the experiments. Among others, thermogravimetric analysis, physisorption, pH, and conductivity analysis were used to characterize the chars. Mesophilic AD batch tests with different concentrations (0.025, 0.05, 0.5, 1.0, 7.0, 14.0 gL-1) of all chars (GR and ACs, respectively) were performed with digester sludge from a wastewater treatment plant for a period of 47 d. Volatile fatty acids (VFA) as well as biogas production and CH4 concentrations were monitored. It could be shown, that concentrations below 1.0 g char L-1 did not result in significant effects on CH4 and/or VFA production, whereas high concentrations of GR and AC influenced both, the CH4 yield and kinetics. Depending on the production process and the characteristics of the chars, the effect on AD varied, whereby both, positive and negative effects on biogas yield and methane production were observed. This study provides the first systematic evaluation of char application to AD processes, and therefore allows for better predictions of char applicability and effect.

Construction of a Synthetic Microbial Community for Enzymatic Pretreatment of Wheat Straw for Biogas Production via Anaerobic Digestion.

Biological pretreatment is a viable method for enhancing biogas production from straw crops, with the improvement in lignocellulose degradation efficiency being a crucial factor in this process. Herein, a metagenomic approach was used to screen core microorganisms (Bacillus subtilis, Acinetobacter johnsonii, Trichoderma viride, and Aspergillus niger) possessing lignocellulose-degrading abilities among samples from three environments: pile retting wheat straw (WS), WS returned to soil, and forest soil. Subsequently, synthetic microbial communities were constructed for fermentation-enzyme production. The crude enzyme solution obtained was used to pretreat WS and was compared with two commercial enzymes. The synthetic microbial community enzyme-producing pretreatment (SMCEP) yielded the highest enzymatic digestion efficacy for WS, yielding cellulose, hemicellulose, and lignin degradation rates of 39.85, 36.99, and 19.21%, respectively. Furthermore, pretreatment of WS with an enzyme solution, followed by anaerobic digestion achieved satisfactory results. SMCEP displayed the highest cumulative biogas production at 801.16 mL/g TS, which was 38.79% higher than that observed for WS, 22.15% higher than that of solid-state commercial enzyme pretreatment and 25.41% higher than that of liquid commercial enzyme pretreatment. These results indicate that enzyme-pretreated WS can significantly enhance biogas production. This study represents a solution to the environmental burden and energy use of crop residues.

Effects of nanobubble water on digestate soaking hydrolysis of rice straw.

This study investigated the performance of combined nanobubble water (NW) and digestate in the soaking hydrolysis process. Two types of NW (CO2NW and O2NW) with digestate were used to soak rice straw for 1, 2, 3, 5, and 7 days. During soaking process, the volatile fatty acids (VFA) concentration in the treatment with O2NW and digestate for 3 days (O2NW-3 d) reached 7179.5 mg-HAc/L. Moreover, the highest specific methane yield (SMY) obtained in this treatment could reach 336.7 NmL/gVS. Although the addition of NW did not significantly increase SMY from digestate soaking, NW could accelerate the rate of methane production and reduce digestion time of T80. The enrichment of Enterobacter in the soaking process was observed when using CO2NW and O2NW as soaking solutions which played important roles in VFA production. This study provides a new insight into environment-friendly enhanced crop straw pretreatment, combining NW and digestate soaking hydrolysis.

Anaerobic digestion integrated with microbial electrolysis cell to enhance biogas production and upgrading in situ.

Anaerobic digestion (AD) is an effective and applicable technology for treating organic wastes to recover bioenergy, but it is limited by various drawbacks, such as long start-up time for establishing a stable process, the toxicity of accumulated volatile fatty acids and ammonia nitrogen to methanogens resulting in extremely low biogas productivities, and a large amount of impurities in biogas for upgrading thereafter with high cost. Microbial electrolysis cell (MEC) is a device developed for electrosynthesis from organic wastes by electroactive microorganisms, but MEC alone is not practical for production at large scales. When AD is integrated with MEC, not only can biogas production be enhanced substantially, but also upgrading of the biogas product performed in situ. In this critical review, the state-of-the-art progress in developing AD-MEC systems is commented, and fundamentals underlying methanogenesis and bioelectrochemical reactions, technological innovations with electrode materials and configurations, designs and applications of AD-MEC systems, and strategies for their enhancement, such as driving the MEC device by electricity that is generated by burning the biogas to improve their energy efficiencies, are specifically addressed. Moreover, perspectives and challenges for the scale up of AD-MEC systems are highlighted for in-depth studies in the future to further improve their performance.

Machine learning for high solid anaerobic digestion: Performance prediction and optimization.

Biogas production through anaerobic digestion (AD) is one of the complex non-linear biological processes, wherein understanding its dynamics plays a crucial role towards process control and optimization. In this work, a machine learning based biogas predictive model was developed for high solid systems using algorithms, including SVM, ET, DT, GPR, and KNN and two different datasets (Dataset-1:10, Dataset-2:5 inputs). Support Vector Machine had the highest accuracy (R2) of all the algorithms at 91 % (Dataset-1) and 87 % (Dataset-2), respectively. The statistical analysis showed that there was no significant difference (p = 0.377) across the datasets, wherein with less inputs, accurate results could be predicted. In case of biogas yield, the critical factors which affect the model predictions include loading rate and retention time. The developed high solid machine learning model shows the possibility of integrating Artificial Intelligence to optimize and control AD process, thus contributing to a generic model for enhancing the overall performance of the biogas plant.

Microwave pretreatment of wastewater sludge technology-a scientometric-based review.

This manuscript presents a scientometric review of recent advances in microwave pretreatment processes for sewage sludge, systematically identifying existing gaps and prospects. For this purpose, 1763 papers on the application of microwave technology to sludge pretreatment were retrieved from the Web of Science (WoS) using relevant keywords. These publications were then analyzed using diverse scientometric indices. The results show that research in this field encompasses applications based on the non-thermal effects of microwaves, enhanced effectiveness of anaerobic digestion (AD), and the energy balance of this pretreatment system. Overcoming existing technical challenges, such as the cleavage of extracellular polymers, reducing microwave energy consumption, understanding the non-thermal effects of microwaves, promoting AD of sludge in combination with other chemical and physical methods, and expanding the application of the technology, are the main scientific focuses. Additionally, this paper thoroughly examines both the constraints and potential of microwave pretreatment technology for wastewater treatment.

Predictive modelling of methane yield in biochar-amended cheese whey and septage co-digestion: Exploring synergistic effects using Gompertz and neural networks.

This study performed bench scale studies on anaerobic co-digestion of cheese whey and septage mixed with biochar (BC) as additive at various dosages (0.5 g, 1 g, 2 g and 4 g) and total solids (TS) concentrations (5%, 7.5%, 10%,12.5% and 15%). The experimental results revealed 29.58% increase in methane yield (486 ± 11.32 mL/gVS) with 27% reduction in lag phase time at 10% TS concentration and 50 g/L of BC loading. The mechanistic investigations revealed that BC improved process stability by virtue of its robust buffering capacity and mitigated ammonia inhibition. Statistical analysis indicates BC dosage had a more pronounced effect (P < 0.0001) compared to the impact of TS concentrations. Additionally, the results were modelled using Gompertz model (GM) and artificial neural network (ANN) algorithm, which revealed the outperformance of ANN over GM with MSE 17.96, R2 value 0.9942 and error 0.27%. These findings validated the practicality of utilizing a high dosage of BC in semi-solid anaerobic digestion conditions.

Life-cycle and economic assessments of microalgae biogas production in suspension and biofilm cultivation systems.

Biogas production via anaerobic digestion is highly attractive for microalgae. The technology of microalgae cultivation has profound impacts on biogas production system as it is the most energy-consuming process. However, a comprehensive evaluation of the environmental and economic benefits of different cultivation systems has yet to be sufficiently conducted. Here, life-cycle and economic assessments of open raceway ponds, photobioreactors and biofilm systems were investigated. Results showed greenhouse gas emissions of all systems were positive because more than two-thirds of carbon in fuel gas was lost and the fixed carbon in product gas and solid fertilizer was less than the emitted carbon during energy input. Particularly, biofilm system achieved the least greenhouse gas emissions (9.3 g CO2-eq/MJ), net energy ratio (0.7) and levelized cost of energy (0.9 $/kWh), indicating the optimum cultivation system. Open raceway ponds and photobioreactors failed to achieve positive benefits because of low harvesting efficiency and biomass concentration.

Biodegradability of PBAT/PLA coated paper and bioplastic bags under anaerobic digestion.

Poly (lactic acid) (PLA) and Poly(butylene adipate-co-terephthalate) (PBAT) are two of biodegradable plastics with the highest production capacities in 2021. Bioplastic waste management can be easily integrated with organic waste management, especially when bioplastics are used as food packaging material, since they are potentially biodegradable. The aim of this study was to assess the biodegradability of biodegradable polymer-coated paper (BPCP) and bioplastic bags made from PBAT/PLA blend during mesophilic and thermophilic anaerobic digestion (AD) and to reveal the changes in the physicochemical properties of the bioplastics. BPCP obtained 155 NmL-CH4/g VS and 307.3 NmL-CH4/g VS under mesophilic and thermophilic conditions, respectively, but left bioplastic film residues. The bioplastic bags did not exhibit significant biodegradation during the AD processes. 1H NMR results indicated that the ratio of PLA to PBAT decreased significantly after AD of the BPCP film and that PLA monomers were formed from the bioplastic bags, leading to a decrease in the hydrophobicity on the surfaces of the materials. Methanoculleus was found to be enriched on the bioplastic surface after mesophilic AD. From the perspective of coupling bioplastic waste management with the food waste management, the incorporation of BPCP into the AD reactor not only enhances system stability and methane production to a greater extent than biodegradable plastic bags but also raises concerns regarding the residual biofilm when utilizing the digestate for direct land applications.