Mesophilic Anaerobic Digestion Research Articles

Evaluation of batch mesophilic anaerobic digestion of raw and trampled llama and dromedary dungs: methane potential and kinetic study

This research was carried out with the aim to evaluate the anaerobic digestion (AD) of llama and dromedary dungs (both untreated and trampled) in batch mode at mesophilic temperature (35 °C). The biochemical methane potential (BMP) tests with an inoculum to substrate ratio of 2:1 (as volatile solids (VS)) were carried out. The methane yield from trampled llama dung (333.0 mL CH4 g−1 VSadded) was considerably higher than for raw llama, raw and trampled dromedary dungs (185.9, 228.4, 222.9 mL CH4 g−1 VSadded, respectively). Therefore, trampled llama dung was found to be the best substrate for methane production due to its high content of volatile solids as well as its high nitrogen content (2.1%) and more appropriate C/N ratio (23.6) for AD. The experimental data was found to be in accordance with both first-order kinetic and transference function mathematical models, when evaluating the experimental methane production against time. By applying the first-order kinetic model, the hydrolysis rate constants, kh, were found to be 19% and 11% higher for trampled dungs in comparison with the raw dung of dromedary and llama, respectively. In addition, the maximum methane production rate (Rm) derived from the transference function model for trampled llama dung (22.0 mL CH4 g−1 VS d−1) was 83.3%, 24.4% and 22.9% higher than those obtained for raw llama manure and for raw and trampled dromedary dungs, respectively.

Mesophilic and Thermophilic Anaerobic Digestion of Wheat Straw in a CSTR System with 'Synthetic Manure': Impact of Nickel and Tungsten on Methane Yields, Cell Count, and Microbiome.

Lignocellulosic residues, such as straw, are currently considered as candidates for biogas production. Therefore, straw fermentations were performed to quantitatively estimate methane yields and cell counts, as well as to qualitatively determine the microbiome. Six fully automated, continuously stirred biogas reactors were used: three mesophilic (41 °C) and three thermophilic (58 °C). They were fed every 8 h with milled wheat straw suspension in a defined, buffered salt solution, called ‘synthetic manure’. Total reflection X-ray fluorescence spectrometry analyses showed nickel and tungsten deficiency in the straw suspension. Supplementation of nickel and subsequently tungsten, or with an increasing combined dosage of both elements, resulted in a final concentration of approximately 0.1 mg/L active, dissolved tungsten ions, which caused an increase of the specific methane production, up to 63% under mesophilic and 31% under thermophilic conditions. That is the same optimal range for pure cultures of methanogens or bacteria found in literature. A simultaneous decrease of volatile fatty acids occurred. The Ni/W effect occurred with all three organic loading rates, being 4.5, 7.5, and 9.0 g volatile solids per litre and day, with a concomitant hydraulic retention time of 18, 10, or 8 days, respectively. A maximum specific methane production of 0.254 m3 CH4, under standard temperature and pressure per kg volatile solids (almost 90% degradation), was obtained. After the final supplementation of tungsten, the cell counts of methanogens increased by 300%, while the total microbial cell counts increased by only 3–62%. The mesophilic methanogenic microflora was shifted from the acetotrophic Methanosaeta to the hydrogenotrophic Methanoculleus (85%) by tungsten, whereas the H2-CO2-converter, Methanothermobacter, always dominated in the thermophilic fermenters.

Impact of Granular Activated Carbon on Anaerobic Process and Microbial Community Structure during Mesophilic and Thermophilic Anaerobic Digestion of Chicken Manure

In this work, the impact of granular activated carbon (GAC) on the mesophilic and thermophilic anaerobic digestion of chicken manure and the structure of microbial communities was investigated. These results demonstrated that GAC supplementation effectively enhanced the consumption of produced organic acids in the mesophilic and thermophilic batch tests, accompanied by faster biomethane production in the presence of GAC than from reactors without GAC. However, since the free ammonia level was 3–6 times higher in the thermophilic reactors, this led to the instability of the anaerobic digestion process of the nitrogen-rich substrate at thermophilic temperatures. Bacteroidia and Clostridia were the two main bacterial classes in the mesophilic reactors, whereas the class Clostridia had a competitive advantage over other groups in the thermophilic systems. The archaeal communities in the mesophilic reactors were mainly represented by representatives of the genera Methanosarcina, Methanobacterium, and Methanotrix, whereas the archaeal communities in the thermophilic reactors were mainly represented by members of the genera Methanosarcina, Methanoculleus, and Methanothermobacter. New data obtained in this research will help control and manage biogas reactors in the presence of GAC at different temperatures.

Optimization of methane production during mesophilic anaerobic digestion using response surface methodology

Optimization of methane production during mesophilic anaerobic digestion using response surface methodology

Influence of volatile fatty acids/alkalinity ratio on methane production during mesophilic anaerobic digestion: stability, efficiency and optimization

Influence of volatile fatty acids/alkalinity ratio on methane production during mesophilic anaerobic digestion: stability, efficiency and optimization

Influence of Thermal Hydrolysis Pretreatment on Mesophilic and Thermophilic Anaerobic Digestion of Sludge

Influence of Thermal Hydrolysis Pretreatment on Mesophilic and Thermophilic Anaerobic Digestion of Sludge

Performance analysis of operating parameters during mesophilic anaerobic digestion of vinasse using principal component analysis

Performance analysis of operating parameters during mesophilic anaerobic digestion of vinasse using principal component analysis

Evaluation of volatile fatty acids (VFAs) production in thermophilic and mesophilic anaerobic digestion of oil palm empty fruit bunch (OPEFB)

Palm oil is one of the leading global commodities, and Indonesia is one of the largest palm oil producers in the world. One of the solid wastes generated from oil palm industrial activities is oil palm empty fruit bunches (OPEFB). The annual accumulation of OPEFB in Indonesia is 126,317.54 tons/year, and it increases with increasing palm oil production. The cellulosic content of OPEFB is a potential substrate for anaerobic digestion to produce volatile fatty acids (VFAs). This study used chemical pretreatment (NaOH 8% w/V) for OPEFB delignification before the anaerobic digestion for VFAs production. Digested cow manure (DCM) was used as the inoculum. Process optimization of producing VFAs is challenging because VFAs act as intermediate products in the metabolic pathway of anaerobic digestion. In general, the temperature can affect the activity of hydrolytic and acidogenic facultative microorganisms. It increases the hydrolysis rate of organic polymers to more soluble forms and improves the performance of anaerobic reactors for the formation of VFAs. This study examines the effect of temperature on the anaerobic digestion process of OPEFB for VFAs production in a batch reactor. The temperature varied in mesophilic (29 °C) and thermophilic (55 °C) conditions. This study showed that anaerobic digestion in the thermophilic conditions produces almost thrice VFAs compared to the mesophilic process. The highest accumulation of VFAs in both reactors occurred on the seventh day, i.e., 3311.57 ± 89 mg.L−1 at ORP -163 ± 16 mV in the thermophilic reactor and 1307 ± 42 mg.L−1 at ORP -224 ± 32 mV in the mesophilic reactor. Acetic acid was the dominant VFA in each reactor. The VFA production in thermophilic reactor (RT) reached 59% of the stoichiometric potential. Although the thermophilic condition improves the VFA yield compared to the mesophilic condition, further study is still needed to increase the VFA production from OPEFB.

Metagenomic Analyses Reveal that Mesophilic Anaerobic Digestion Substantially Reduces the Abundance of Antibiotic Resistance Genes and Mobile Genetic Elements in Dairy Manures

Metagenomic Analyses Reveal that Mesophilic Anaerobic Digestion Substantially Reduces the Abundance of Antibiotic Resistance Genes and Mobile Genetic Elements in Dairy Manures

Anaerobic Mesophilic Digestion of Typha Latifolia Suspensions Using the Fermented Waste of Yeast Production as Fermentation Inoculum

Anaerobic Mesophilic Digestion of Typha Latifolia Suspensions Using the Fermented Waste of Yeast Production as Fermentation Inoculum

A meta-analysis reveals that operational parameters influence levels of antibiotic resistance genes during anaerobic digestion of animal manures

Anaerobic digestion (AD) has shown the potential to reduce the numbers and types of antibiotic-resistance genes (ARG) present in animal manures. However, the variability of the results has limited the ability to draw solid conclusions. To address this issue, we performed a series of meta-analyses to evaluate how AD of pig, cattle, and dairy manures affects ARG levels and how different parameters, such as temperature, pH, digestion times, and the addition of other substances (e.g., solids, antibiotics) influence ARG changes. Twenty studies with enough details on changes in ARG levels during the AD process were identified and used for the meta-analyses. The results suggested that AD could significantly reduce ARG levels regardless of the conditions of the process. Also, thermophilic AD was more effective than mesophilic AD at reducing ARGs, although this difference was only significant for pig manures. The results also suggested that long digestion times (>50 days) yielded better ARG reduction rates, and that the addition of solids from an external source (co-digestion) negatively affected the efficiency of ARG reduction. In general, the results suggested that ARG changes during AD could be linked to the abundance and activity of hydrolytic communities.

Effect of Ammonia Load on Microbial Communities in Mesophilic Anaerobic Digestion of Propionic Acid

Effect of Ammonia Load on Microbial Communities in Mesophilic Anaerobic Digestion of Propionic Acid

Metataxonomics, metagenomics and metabolomics analysis of the influence of temperature modification in full-scale anaerobic digesters

Full-scale anaerobic digesters’ performance is regulated by modifying their operational conditions, but little is known about how these modifications affect their microbiome. In this work, we monitored two originally mesophilic (35 °C) full-scale anaerobic digesters during 476 days. One digester was submitted to sub-mesophilic (25 °C) conditions between days 123 and 373. We characterized the effect of temperature modification using a multi-omics (metataxonomics, metagenomics, and metabolomics) approach. The metataxonomics and metagenomics results revealed that the lower temperature allowed a substantial increase of the sub-dominant bacterial population, destabilizing the microbial community equilibrium and reducing the biogas production. After restoring the initial mesophilic temperature, the bacterial community manifested resilience in terms of microbial structure and functional activity. The metabolomic signature of the sub-mesophilic acclimation was characterized by a rise of amino acids and short peptides, suggesting a protein degradation activity not directed towards biogas production.

Microbial Community Successional Changes in a Full-Scale Mesophilic Anaerobic Digester from the Start-Up to the Steady-State Conditions.

Anaerobic digestion is a widely used technology for sewage sludge stabilization and biogas production. Although the structure and composition of the microbial communities responsible for the process in full-scale anaerobic digesters have been investigated, little is known about the microbial successional dynamics during the start-up phase and the response to variations occurring in such systems under real operating conditions. In this study, bacterial and archaeal population dynamics of a full-scale mesophilic digester treating activated sludge were investigated for the first time from the start-up, performed without adding external inoculum, to steady-state operation. High-throughput 16S rRNA gene sequencing was used to describe the microbiome evolution. The large majority of the reads were affiliated to fermentative bacteria. Bacteroidetes increased over time, reaching 22% of the total sequences. Furthermore, Methanosaeta represented the most abundant methanogenic component. The specific quantitative data generated by real-time PCR indicated an enrichment of bacteria and methanogens once the steady state was reached. The analysis allowed evaluation of the microbial components more susceptible to the shift from aerobic to anaerobic conditions and estimation of the microbial components growing or declining in the system. Additionally, activated sludge was investigated to evaluate the microbial core selected by the WWTP operative conditions.

Can radicals-orientated chemical oxidation improve the reduction of antibiotic resistance genes (ARGs) by mesophilic anaerobic digestion of sludge?

The dissemination of antibiotic resistance genes (ARGs) increases risks towards human health and environmental safety. This work investigates the control of ARGs abundance and bacterial community evolution involved in waste activated sludge (WAS) treatment by chemical conditioning and subsequent mesophilic anaerobic digestion (MAD). The different chemical oxidation processes of ferrous iron-activated oxone and hydrogen peroxide (PMS-Fe2+ and H2O2-Fe2+) and thermal-activated oxone (PMS@80 ℃) were investigated, and the ferric chloride (FeCl3) and inactivated oxone (PMS) were compared. PMS@80 ℃ decreased the absolute abundance of most ARGs by 10.6–99.3% and that of total ARGs by 66.3%. Interestingly, oxidation pretreatment increased rather than decreased the relative abundance of most ARGs. MAD with PMS@80 ℃ pretreatment increased the absolute abundance of total ARGs by 51.6%, and other MAD processes decreased it by 8.6–47.4%. PMS-Fe2+ and PMS@80 ℃ negatively inhibited methane production from 98.3 to 81.7 and 94.4 mL/g VSS in MAD. MAD effluent showed high abundance of Arcobacter genus in the range of 8.1–17.4% upon PMS-based pretreatment, possibly related to sulfur oxidation, nitrate reduction, and blaVEB enrichment. The radicals-orientated chemical oxidation can hardly improve the ARGs elimination by MAD due to the extremely high competitive organics in sludge.

The interaction between lipids and ammoniacal nitrogen mitigates inhibition in mesophilic anaerobic digestion

Ammoniacal nitrogen and long chain fatty acids (LCFA) are common inhibitors of the anaerobic digestion process. However, the interaction between these inhibitors has received little attention. Understanding the interaction between these inhibitors is important to optimise the operation of anaerobic digesters treating slaughterhouse waste or using fat, oil and grease (FOG) as co-substrate among others. To study the interaction between ammoniacal nitrogen and LCFA inhibition, 20 different conditions were trialled in mesophilic batch tests. Experimental conditions included 5 mixtures between slaughterhouse wastewater and LCFA (100:0, 75:25, 50:50, 20:80, 0:100 on a VS basis), each one tested at 4 different ammoniacal nitrogen concentrations (0, 1, 3, 6 gNadded·L-1). Experimental and modelling results showed that ammoniacal nitrogen inhibition was less severe in LCFA-rich mixtures, indicating that LCFA mitigated ammoniacal nitrogen inhibition to a certain extent. However, the positive interaction between inhibitors did not only depend on the LCFA concentration. A protective LCFA coat that limited the diffusion of free ammonia into the cell and/or provided a localised lower pH in the vicinity of the microbial cell could explain the experimental results. However, ammoniacal nitrogen and LCFA inhibition comprise up to 6 different but interrelated inhibitors (i.e. NH3, NH4+, LCFA, VFA, H2 and pH) and therefore the specific mechanism could not be elucidated. Nonetheless, these results suggest that LCFA do not exacerbate TAN-related inhibition and that LCFA-rich substrates can be utilised as co-substrates in mesophilic N-rich digesters.

Anaerobic treatment of swine manure under mesophilic and thermophilic temperatures: Fate of veterinary drugs and resistance genes

The effect of anaerobic treatment of swine manure at 35 °C (mesophilic) and 55 °C (thermophilic) on methane production, microbial community and contaminants of emerging concern was investigated. Pasteurization pretreatment and post treatment was also investigated in combination with anaerobic treatment at 35 °C. Specific methane production (SMP), 26 pharmaceutical compounds (PhACs) and five antibiotic resistance genes (ARGs) (qnrS, tetW, ermB, sul1 and blaTEM) were evaluated. Mesophilic treatment resulted in the highest SMP regardless of whether pasteurization was applied. Marbofloxacin was the most abundant antibiotic in swine manure. In general, all groups of PhACs showed higher removals under thermophilic temperatures as compared to mesophilic. In general, pasteurization pretreatment followed by mesophilic anaerobic digestion provided the highest removals of ARGs. Finally, the genera Streptococcus, Clostridium and Pseudomonas which contain pathogenic species, were present in the swine manure. Streptococcus, which was the most abundant, was decreased during all the treatments, while the others only decreased under certain treatments.

Dynamic changes in anaerobic digester metabolic pathways and microbial populations during acclimatisation to increasing ammonium concentrations

Transitions in microbial community structure in response to increasing ammonia concentrations were determined by monitoring mesophilic anaerobic digesters seeded with a predominantly acetoclastic methanogenic community from a sewage sludge digester. Ammonia concentration was raised by switching the feed to source segregated domestic food waste and applying two organic loading rates (OLR) and hydraulic retention times (HRT) in paired digesters. One of each pair was dosed with trace elements (TE) known to be essential to the transition, with the other unsupplemented digester acting as a control. Samples taken during the trial were used to determine the metabolic pathway to methanogenesis using 14C labelled acetate. Partitioning of 14C between the product gases was interpreted via an equation to indicate the proportion produced by acetoclastic and hydrogenotrophic routes. Archaeal and selected bacterial groups were identified by 16S rRNA sequencing, to determine relative abundance and diversity. Acclimatisation for digesters with TE was relatively smooth, but OLR and HRT influenced both metabolic route and community structure. The 14C ratio could be used quantitatively and, when interpreted alongside archaeal community structure, showed that at longer HRT and lower loading Methanobacteriaceae were dominant and hydrogenotrophic activity accounted for 77% of methane production. At the higher OLR and shorter HRT, Methanosarcinaceae were dominant with the 14C ratio indicating simultaneous production of methane by acetoclastic and hydrogenotrophic pathways: the first reported observation of this in digestion under mesophilic conditions. Digesters without TE supplementation showed similar initial changes but, as expected failed to complete the transition to stable operation.

Quantitative microbial risk assessment associated with ready-to-eat salads following the application of farmyard manure and slurry or anaerobic digestate to arable lands

Farmyard manure and slurry (FYM&S) and anaerobic digestate are potentially valuable soil conditioners providing important nutrients for plant development and growth. However, these organic fertilisers may pose a microbial health risk to humans. A quantitative microbial risk assessment (QMRA) model was developed to investigate the potential human exposure to pathogens following the application of FYM&S and digestate to agricultural land. The farm-to-fork probabilistic model investigated the fate of microbial indicators (total coliforms and enterococci) and foodborne pathogens in the soil with potential contamination of ready-to-eat salads (RTEs) at the point of human consumption. The processes examined included pathogen inactivation during mesophilic anaerobic digestion (M-AD), post-AD pasteurisation, storage, dilution while spreading, decay in soil, post-harvest washing processes, and finally, the potential growth of the pathogen during refrigeration/storage at the retail level in the Irish context. The QMRA highlighted a very low annual probability of risk (Pannual) due to Clostridium perfringens, norovirus, and Salmonella Newport across all scenarios. Mycobacterium avium may result in a very high mean Pannual for the application of raw FYM&S, while Cryptosporidium parvum and pathogenic E. coli showed high Pannual, and Listeria monocytogenes displayed moderate Pannual for raw FYM&S application. The use of AD reduces this risk; however, pasteurisation reduces the Pannual to an even greater extent posing a very low risk. An overall sensitivity analysis revealed that mesophilic-AD's inactivation effect is the most sensitive parameter of the QMRA, followed by storage and the decay on the field (all negatively correlated to risk estimate). The information generated from this model can help to inform guidelines for policymakers on the maximum permissible indicator or pathogen contamination levels in the digestate. The QMRA can also provide the AD industry with a safety assessment of pathogenic organisms resulting from the digestion of FYM&S.

Carbon based conductive materials mediated recalcitrant toxicity mitigation during anaerobic digestion of thermo-chemically pre-treated organic fraction of municipal solid waste

High-temperature thermal pretreatment alone or in conjugation with chemical pretreatment (highly acidic or alkaline) produced recalcitrant compounds, which inhibits the anaerobic digestion (AD) process performance. This study aims to develop a strategy to use carbon-based conductive materials to mitigate the recalcitrant toxicity and enhance the methane generation in the downstream AD. The formation of recalcitrant compounds, mainly the furan derivatives, i.e., furfural and 5-HydroxyMethyl furfurals (5-HMF) during thermo-chemical pretreatment of OFMSW at 150 °C, 175 °C, 200 °C with 3 g/L-NaOH dose, and the alleviation of their inhibitory effects by adding 25 g/L of each of granular activated carbon (GAC) and granular biochar (GBC) during mesophilic AD were studied. The addition of conductive materials resulted in the highest biogas yield of 462 mL/gVSadded (GAC) and 449 mL/gVSadded (GBC) for 175°C-3g/L-NaOH pretreatment, which was >45% higher over control. The highest improvement of >65% in biogas yield was observed for 200°C-3g/L-NaOH pretreatment despite the lower biogas yield. The conductive materials amended digester shows a significant decrease in the 5-HMF and furfurals concertation. The highest reduction in 5-HMF (44%) and furfural (51%) concentrations were observed for 200°C-3g/L-NaOH pretreatment, and 25 g/L GBC amended tests. The score plots from the principal component analysis (PCA) of the characterization of the digestate showed that the data were significant, whereas the loading plots depicted the correlation of different experimental parameters studied (like fate of recalcitrant, biogas yield and other parameters post AD of OFMSW when aided with conductive materials). Application of regression models in all the batch assays depicted that a lag phase of 2–4 days was observed in Modified Gompertz Model (MGM), 4–5 days in Logistic Model (LM) and a rapid hydrolysis was proven with the value of hydrolysis coefficient being between 0.003 and 0.029 from the first-order (FO) model.