Influence of fermentation metabolites on redox potential in anaerobic digestion of proteinaceous solid wastes by Synergistes sp.

The anaerobic digestion of animal fleshing from tannery solid waste was investigated with regard to hydrolytic enzymes, protease and lipase, fermentative enzyme deaminase, soluble protein and amino acids, redox potential (Eh), volatile fatty acids, ammonia and carbon dioxide up to 120 h of retention time. The release of these fermentation metabolites at various retention times greatly influenced the Eh. In the hydrolytic phase, the maximum value of Eh was ?50 mV and it reached the minimum of ?350 mV in 24 h in the fermentative phase. The minimum and maximum values of Eh were ?387 and ?452 mV at 80 h of anaerobic digestion. The release of extracellular metabolites was confirmed by HPLC and GC‐MS. In this study, we have found that the ammonia and pH had a substantial influence on the Eh during the anaerobic digestion of animal fleshing.     

粪便收集时间和饲料中肉骨粉含量对异育银鲫(Carassius auratus gibelio) 消化率的影响

本研究探讨了通过收集器不同粪便收集时间和饲料中肉骨粉(MBM)含量对异育银鲫干物质、蛋白、能量、磷的表观消化率 (ADC) 影响.不同梯度(0, 20, 40, 60, 80 ,100%)的肉骨粉替代鱼粉(FM) 蛋白配制成六种等氮 (粗蛋白: 410 g/kg) 等能 (总能: 18 kJ/g) 的饲料,通过11周的饲养实验,实验开始后2周开始收集粪便,收集时间分别是:排粪后 1 min, 投喂后4h和16h. 结果表明,干物质、蛋白、能量、磷的消化率明显随粪便收集时间增加而升高(p<0.05),但在高肉骨粉替代饲料中差异不显著(p>0.05). 干物质、蛋白、能量、磷的消化率随饲料中肉骨粉含量的增加呈线性或近线性下降.因此,在消化率测定中应该尽快收集排出的粪便以保证消化率的真实性.消化率是影响肉骨粉利用一个因素.     

Methane recovery from the anaerobic codigestion of municipal sludge and FOG

The anaerobic biodegradability of a mix of municipal primary sludge (PS), thickened waste activated sludge (TWAS) and fat, oil, and grease (FOG) was assessed using semi-continuous feed, laboratory-scale anaerobic digesters operated at mesophilic (35 °C) and thermophilic (52 °C) temperature. Addition of a large FOG fraction (48% of the total VS load) to a PS + TWAS mix, resulted in 2.95 times larger methane yield, 152 vs. 449 mL methane @ STP/g VS added at 35 °C and 2.6 times larger methane yield, 197 vs. 512 mL methane @ STP/g VS added at 52 °C. The high FOG organic load fraction was not inhibitory to the process. The results of this study demonstrate the benefit of sludge and FOG codigestion.     

On-field study of anaerobic digestion full-scale plants (Part II): new approaches in monitoring and evaluating process efficiency

Biogas plants need easy and practical tools for monitoring and evaluating their biological process efficiency. As soon as, in many cases, biomass supply present considerable costs, full-scale anaerobic digestion (AD) processes must approach, as much as possible, the potential biogas yield of the organic mixture fed to the biodigesters. In this paper, a new indicator is proposed (the bio-methane yield, BMY), for measuring the efficiency in full-scale AD processes, based on a balance between the biochemical methane potential (BMP) of the input biomass and the residual BMP of the output materials (digestate). For this purpose, a one-year survey was performed on three different full-scale biogas plants, in the Italian agro-industrial context, and the bio-chemical processes were fully described in order to calculate their efficiencies (BMY = 87-93%) and to validate the new indicator proposed, as useful and easily applicable tool for full-scale AD plants operators.     

Anaerobic digestion of secondary residuals from an anaerobic bioreactor at a brewery to enhance bioenergy generation

Many beer breweries use high-rate anaerobic digestion (AD) systems to treat their soluble high-strength wastewater. Biogas from these AD systems is used to offset nonrenewable energy utilization in the brewery. With increasing nonrenewable energy costs, interest has mounted to also digest secondary residuals from the high-rate digester effluent, which consists of yeast cells, bacteria, methanogens, and small (hemi)cellulosic particles. Mesophilic (37 °C) and thermophilic (55 °C) lab-scale, low-rate continuously-stirred anaerobic digestion (CSAD) bioreactors were operated for 258 days by feeding secondary residuals at a volatile solids (VS) concentration of ∼40 g l⁻¹. At a hydraulic retention time (HRT) of 15 days and a VS loading rate of 2.7 g VS l⁻¹ day⁻¹, the mesophilic bioreactor showed an average specific volumetric biogas production rate of 0.88 l CH₄ l⁻¹ day⁻¹ and an effluent VS concentration of 22.2 g VS l⁻¹ (43.0% VS removal efficiency) while the thermophilic bioreactor displayed similar performances. The overall methane yield for both systems was 0.21 l CH₄ g⁻¹ VS fed and 0.47–0.48 l CH₄ g⁻¹ VS removed. A primary limitation of thermophilic digestion of this protein-rich waste is the inhibition of methanogens due to higher nondissociated (free) ammonia (NH₃) concentrations under similar total ammonium (NH₄ ⁺) concentrations at equilibrium. Since thermophilic AD did not result in advantageous methane production rates or yields, mesophilic AD was, therefore, superior in treating secondary residuals from high-rate AD effluent. An additional digester to convert secondary residuals to methane may increase the total biogas generation at the brewery by 8% compared to just conventional high-rate digestion of brewery wastewater alone. JIMB-2008: BioEnergy—Special issue.     

Potential errors in the quantitative evaluation of biogas production in anaerobic digestion processes

Errors that are commonly made in the quantification of biogas from anaerobic digestion experiments were investigated.For liquid displacement gasometers where a barrier solution separates the biogas and the atmosphere, inaccuracy due to gas diffusion was examined experimentally. Acidified saturated saline solution was the most suitable barrier solution, as biogas characteristics changed least with time. Using acidified or tap water caused considerable biogas losses and should therefore be avoided where biogas is stored before measurement.Errors associated with volume calculation from three common liquid displacement gasometer types were investigated theoretically. Corrections that must be made to obtain gas volumes at standard temperature and pressure when using this equipment are discussed. Regarding experimental errors, gasometer designs where displaced liquid is weighed to determine the volume are the most versatile since errors depend mainly upon balance sensitivity. Using liquid heights to calculate volume requires appropriate sizing of the gasometer relative to the volume of gas measured.The calibration of a low flow gas meter was investigated and an approximately linear variation with flow rate was found; hence in situ calibration is advised for this type of instrument. Correction for atmospheric conditions should be performed in real time to reduce errors.     

Two-stage anaerobic digestion of biodegradable municipal solid waste using a rotating drum mesh filter bioreactor and anaerobic filter

A rotating drum mesh filter bioreactor (RDMFBR) with a 100 μm mesh coupled to an anaerobic filter was used for the anaerobic digestion of biodegradable municipal solid waste (BMW). Duplicate systems were operated for 72 days at an organic loading rate (OLR) of 7.5 gVS l⁻¹ d⁻¹. Early in the experiment most of the methane was produced in the 2nd stage. This situation gradually reversed as methanogenesis became established in the 1st stage digester, which eventually produced 86–87% of the total system methane. The total methane production was 0.2 l g⁻¹ VS_added with 60–62% volatile solids destruction. No fouling was experienced during the experiment at a transmembrane flux rate of 3.5 l m⁻² h⁻¹. The system proved to be robust and stably adjusted to a shock loading increase to 15 gVS l⁻¹ d⁻¹, although this reduced the overall methane production to 0.15 l g⁻¹ VS_added.     

Effects of metal oxide nanoparticles (TiO2, Al2O3, SiO2 and ZnO) on waste activated sludge anaerobic digestion

The effect of metal oxide nanoparticles (nano-TiO2, nano-Al2O3, nano-SiO2 and nano-ZnO) on anaerobic digestion was investigated by fermentation experiments using waste activated sludge as the substrates. Nano-TiO2, nano-Al2O3 and nano-SiO2 in doses up to 150 milligram per gram total suspended solids (mg/g-TSS) showed no inhibitory effect, whereas nano-ZnO showed inhibitory effect with its dosages increased. The methane generation was the same as that in the control when in the presence of 6 mg/g-TSS of nano-ZnO, however, which decreased respectively to 77.2% and 18.9% of the control at 30 and 150 mg/g-TSS. The released Zn2+ from nano-ZnO was an important reason for its inhibitory effect on methane generation. It was found that higher dosages of nano-ZnO inhibited the steps of sludge hydrolysis, acidification and methanation. Also, the activities of protease, acetate kinase (AK) and coenzyme F420 were inhibited by higher dosages of nano-ZnO during WAS anaerobic digestion.     

Cellulolytic activity in leachate during leach-bed anaerobic digestion of municipal solid waste

The degradation of municipal solid waste (MSW) under mesophilic conditions can be enhanced by exchanging leachate between fresh waste and stabilised waste. The optimum point in time when leachate from an anaerobically digesting waste bed can be used to initiate degradation of another waste bed might occur when the leachate of the digesting waste bed is highly active with cellulolytic and methanogenic bacteria. In this study, the cellulolytic activity of the leachate was measured using the cellulose-azure assay. As products of hydrolysis are soluble compounds, the rate of generation of these compounds was estimated based on a soluble chemical oxygen demand (SCOD) balance around the fresh waste bed. It was found that once the readily soluble material present in MSW was washed out there was very little generation of SCOD without the production of methane, indicating that flushing leachate from a stabilised waste bed resulted in a balanced inoculation of the fresh waste bed. With the onset of sustained methanogenesis, the rate of SCOD generation equalled the SCOD released from the digester as methane. The experimental findings also showed that cellulolytic activities of the leachate samples closely followed the trend of SCOD generation.     

Biogas production from potato-juice, a by-product from potato-starch processing, in upflow anaerobic sludge blanket (UASB) and expanded granular sludge bed (EGSB) reactors

In this study, the utilization of potato-juice, the organic by-product from potato-starch processing, for biogas production was investigated in batch assay and in high rate anaerobic reactors. The maximum methane potential of the potato-juice determined by batch assay was 470 mL-CH₄/gVS-added. Anaerobic digestion of potato-juice in an EGSB reactor could obtain a methane yield of 380 mL-CH₄/gVS-added at the organic loading rate of 3.2 gCOD/(L-reactor.d). In a UASB reactor, higher organic loading rate of 5.1 gCOD/(L-reactor.d) could be tolerated, however, it resulted in a lower methane yield of 240 mL-CH₄/gVS-added. The treatment of reactor effluent was also investigated. By acidification with sulfuric acid to pH lower than 5, almost 100% of the ammonia content in the effluent could be retained during the successive up-concentration process step. The reactor effluent could be up-concentrated by evaporation to minimize its volume, and later be utilized as fertilizer.     

Effect of increasing the surface area of primary sludge on anaerobic digestion at low temperature

Two sets of reactors were operated at 15 °C and at sludge retention times (SRTs) of 65 days and 30 days, respectively. Each set was operated at six different mixing velocities. Shear forces provided by mixing affected particle size distribution and resulted in different sludge surface areas. The aim was to investigate the effect of increasing primary sludge surface area on anaerobic digestion at low temperature. The maximum surface areas at the applied mixing velocities were 5926 cm²/cm³of sludge and 4672 cm²/cm³ of sludge at SRTs of 65 days and 30 days, respectively. The corresponding calculated methanogenesis were 49% and 15% at SRTs of 65 days and 30 days, respectively. Maximum total solids (TS) reductions were 26% and 11% at 65 days and 30 days SRTs, respectively. Lipase activity increased with increasing SRT and sludge surface area. Results clearly showed that increasing sludge surface area improved sludge digestion at 15 °C.     

The Survival of antibiotic resistant and sensitiveEscherichia coli strains during anaerobic digestion

Summary Total counts ofEscherichia coli were followed during anaerobic digestion of pig slurry laboratory scale digesters at 37° C. Counts decreased rapidly during anaerobic digestion. Antibiotic resistant strains in most cases appeared to be more persistent in anaerobic digesters than sensitiveE. coli strains as calculated from the decimal decay rates.     

A meta-analysis of the microbial diversity observed in anaerobic digesters

In this study, the collective microbial diversity in anaerobic digesters was examined using a meta-analysis approach. All 16S rRNA gene sequences recovered from anaerobic digesters available in public databases were retrieved and subjected to phylogenetic and statistical analyses. As of May 2010, 16,519 bacterial and 2869 archaeal sequences were found in GenBank. The bacterial sequences were assigned to 5926 operational taxonomic units (OTUs, based on ?97% sequence identity) representing 28 known bacterial phyla, with Proteobacteria (1590 OTUs), Firmicutes (1352 OTUs), Bacteroidetes (705 OTUs), and Chloroflexi (693 OTUs) being predominant. Archaeal sequences were assigned to 296 OTUs, primarily Methanosaeta and the uncharacterized WSA2 group. Nearly 60% of all sequences could not be classified to any established genus. Rarefaction analysis indicates that approximately 60% of bacterial and 90% of archaeal diversity in anaerobic digesters has been sampled. This analysis of the global bacterial and archaeal diversity in AD systems can guide future studies to further examine the microbial diversity involved in AD and development of comprehensive analytical tools.     

Laboratory scale examination of the effects of overloading on the anaerobic digestion by glycerol

The anaerobic digestion of pure glycerol, which produces a baseline acetic acid to propionic acid ratio of 0.2, was studied in laboratory scale reactors (3 l working volume) at mesophilic temperature (37 °C) with 3000 mg chemical oxygen demand (COD) l⁻¹d⁻¹. During the experiment tVFA and C2-C6 VFA analysis and daily biogas yield measurement were carried out. Following 10 days of a 15% d⁻¹ increase in the organic loading rate (OLR) of 3.0-10.5 g COD l⁻¹d⁻¹, the concentration of propionic acid increased to 6200-8000 mg l⁻¹. Then the inoculum was divided into three parts feeding with 100% glycerol, 50% glycerol + 50% acetic acid, and 50% glycerol + 50% thick stillage, (presented in % of 2.60 g COD l⁻¹d⁻¹ OLR), respectively. The application of co-substrates reduced the recovery period by 5 days compared to feeding with pure glycerol. When the reactors were loaded with glycerol again (10% OLR raise per day) the previously applied co-substrates had a positive effect on the VFA composition and the biogas yield as well.     

Enhancement of bioenergy production from organic wastes by two-stage anaerobic hydrogen and methane production process

The present study investigated a two-stage anaerobic hydrogen and methane process for increasing bioenergy production from organic wastes. A two-stage process with hydraulic retention time (HRT) 3 d for hydrogen reactor and 12 d for methane reactor, obtained 11% higher energy compared to a single-stage methanogenic process (HRT 15 d) under organic loading rate (OLR) 3 gVS/(L d). The two-stage process was still stable when the OLR was increased to 4.5 gVS/(L d), while the single-stage process failed. The study further revealed that by changing the HRT_hydrogen:HRT_methane ratio of the two-stage process from 3:12 to 1:14, 6.7%, more energy could be obtained. Microbial community analysis indicated that the dominant bacterial species were different in the hydrogen reactors (Thermoanaerobacterium thermosaccharolyticum-like species) and methane reactors (Clostridiumthermocellum-like species). The changes of substrates and HRT did not change the dominant species. The archaeal community structures in methane reactors were similar both in single- and two- stage reactors, with acetoclastic methanogens Methanosarcina acetivorans-like organisms as the dominant species.     

The future of anaerobic digestion and biogas utilization

One of the common tendencies of animal production activities in Europe and in developed countries in general is to intensify the animal production and to increase the size of the animal production units. High livestock density is always accompanied by production of a surplus of animal manure, representing a considerable pollution threat for the environment in these areas. Avoiding over-fertilization is not only important for environmental protection reasons but also for economical reasons. Intensive animal production areas need therefore suitable manure management, aiming to export and to redistribute the excess of nutrients from manure and to optimize their recycling.     

Integration of on-farm biodiesel production with anaerobic digestion to maximise energy yield and greenhouse gas savings from process and farm residues

Anaerobic co-digestion of residues from the cold pressing and trans-esterification of oilseed rape (OSR) with other farm wastes was considered as a means of enhancing the sustainability of on-farm biodiesel production. The study verified the process energy yields using biochemical methane potential (BMP) tests and semi-continuous digestion trials. The results indicated that high proportions of OSR cake in the feedstock led to a decrease in volatile solids destruction and instability of the digestion process. Co-digestion with cattle slurry or with vegetable waste led to acceptable specific and volumetric methane productions, and a digestate low in potentially toxic elements (PTE). The results were used to evaluate energy balances and greenhouse gas emissions of the integrated process compared with biodiesel production alone. Co-digestion was shown to provide energy self-sufficiency and security of supply to farms, with sufficient surplus for export as fuel and electricity.     

Specific detection of prion antigenic determinants retained in bovine meat and bone meal by flow microbead immunoassay

AIMS: The purpose of this study was to develop an effective method for detecting prion (PrP) antigenic determinants remaining in bovine meat and bone meal (MBM) using pressurized fluid extraction (PSE) equipment and flow microbead immunoassay (FMI). METHODS AND RESULTS: Using the FMI, bovine recombinant PrP could be determined quantitatively in the 7 pmol-7 nmol range using anti-PrP peptide polyclonal antibody-coupled microbeads and anti-PrP monoclonal antibody (SAF61) as a detection antibody. PSE extraction at 120 degrees C for 5 min under high pressure was most effective for eluting PrP determinants from bovine MBMs. The FMI was capable of detecting PrP determinants in bovine MBM extracts with high specificity and indicated that the MBMs contained high levels of PrP determinants. This assay was also applied to the detection of PrP(Sc) determinants in bovine MBM spiked with a scrapie-infected brain at a weight ratio of 50 : 1. CONCLUSIONS: These data indicate that this assay was effective for the specific detection of PrP determinants contained in bovine MBM extracts. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first report detailing the detection of PrP determinants in bovine MBM. The assay could be applied to securing the safety of bovine MBM.