Robustness of archaeal populations in anaerobic co-digestion of dairy and poultry wastes

The objective of this study is to investigate the responses of methanogen populations to poultry waste addition by comparing the archaeal microbial populations in continuous anaerobic digesters with or without the addition of poultry waste as a co-substrate. Poultry waste was characterized as an organic/nitrogen-rich substrate for anaerobic digestion. Supplementing dilute dairy waste with poultry waste for anaerobic co-digestion to increase organic loading rate by 50% resulted in improved biogas production. Elevated ammonia derived from poultry waste did not lead to process inhibition at the organic loadings tested, demonstrating the feasibility of the anaerobic co-digestion of dairy and poultry wastes for improved treatment efficiency. The stability of the anaerobic co-digestion process was linked to the robust archaeal microbial community, which remained mostly unchanged in community structure following increases in organic loading and ammonia levels. Surprisingly, Crenarchaeota archaeal populations, instead of the Euryarchaeota methanogens, dominated the archaeal communities in the anaerobic digesters. The ecological functions of these abundant non-methanogen archaeal populations in anaerobic digestion remain to be identified.     

典型有机固废厌氧消化微生物研究现状与发展方向

经过人工富集和驯化的兼性和严格厌氧微生物是厌氧消化工艺的核心。不同厌氧消化体系中存在的问题大多可以通过改变微生物群落的代谢活性来得到有效改善。得益于微生物组学检测技术的快速发展,对厌氧消化系统中微生物多样性的认识获得了极大的拓展,同时在微生物类群间、微生物与环境的互作关系研究方面也取得了一系列新的进展。然而,有机固废厌氧消化系统中,各种微生物以及微生物和物质的相互作用构成了更为复杂的代谢网络,所以目前对这些互作关系的解析尚不完善。本文重点关注了厌氧消化过程中的典型菌群互作关系,阐述了典型有机固废厌氧消化系统中存在的问题及微生物在其中发挥的作用,最后,立足于现有组学技术推动的微生物组研究进展,对未来有机固废厌氧消化系统微生物组的研究提出展望。     

Production of biogas from solid organic wastes through anaerobic digestion: a review

Anaerobic digestion treatments have often been used for biological stabilization of solid wastes. These treatment processes generate biogas which can be used as a renewable energy sources. Recently, anaerobic digestion of solid wastes has attracted more interest because of current environmental problems, most especially those concerned with global warming. Thus, laboratory-scale research on this area has increased significantly. In this review paper, the summary of the most recent research activities covering production of biogas from solid wastes according to its origin via various anaerobic technologies was presented.     

A multi-criteria ranking of different technologies for the anaerobic digestion for energy recovery of the organic fraction of municipal solid wastes

This paper describes a conceptual framework and methodological tool developed for the evaluation of different anaerobic digestion technologies suitable for treating the organic fraction of municipal solid waste, by introducing the multi-criteria decision support method Electre III and demonstrating its related applicability via a test application. Several anaerobic digestion technologies have been proposed over the last years; when compared to biogas recovery from landfills, their advantage is the stability in biogas production and the stabilization of waste prior to final disposal. Anaerobic digestion technologies also show great adaptability to a broad spectrum of different input material beside the organic fraction of municipal solid waste (e.g. agricultural and animal wastes, sewage sludge) and can also be used in remote and isolated communities, either stand-alone or in conjunction to other renewable energy sources. Main driver for this work was the preliminary screening of such methods for potential application in Hellenic islands in the municipal solid waste management sector. Anaerobic digestion technologies follow different approaches to the anaerobic digestion process and also can include production of compost. In the presented multi-criteria analysis exercise, Electre III is implemented for comparing and ranking 5 selected alternative anaerobic digestion technologies. The results of a performed sensitivity analysis are then discussed. In conclusion, the performed multi-criteria approach was found to be a practical and feasible method for the integrated assessment and ranking of anaerobic digestion technologies by also considering different viewpoints and other uncertainties of the decision-making process.     

Inhibition of anaerobic digestion process: a review

Anaerobic digestion is an attractive waste treatment practice in which both pollution control and energy recovery can be achieved. Many agricultural and industrial wastes are ideal candidates for anaerobic digestion because they contain high levels of easily biodegradable materials. Problems such as low methane yield and process instability are often encountered in anaerobic digestion, preventing this technique from being widely applied. A wide variety of inhibitory substances are the primary cause of anaerobic digester upset or failure since they are present in substantial concentrations in wastes. Considerable research efforts have been made to identify the mechanism and the controlling factors of inhibition. This review provides a detailed summary of the research conducted on the inhibition of anaerobic processes. The inhibitors commonly present in anaerobic digesters include ammonia, sulfide, light metal ions, heavy metals, and organics. Due to the difference in anaerobic inocula, waste composition, and experimental methods and conditions, literature results on inhibition caused by specific toxicants vary widely. Co-digestion with other waste, adaptation of microorganisms to inhibitory substances, and incorporation of methods to remove or counteract toxicants before anaerobic digestion can significantly improve the waste treatment efficiency.     

Simple and rapid methods to evaluate methane potential and biomass yield for a range of mixed solid wastes

This paper describes rapid techniques to evaluate the methane potential and biomass yield of solid wastes. A number of solid wastes were mixed to provide a range of C:N ratios. Empirical formulae were calculated for each waste based on the results of chemical analysis and these formulae were used to estimate the COD equivalent and stoichiometric methane potential (SMP). The actual COD and biochemical methane potential (BMP) were determined experimentally for each waste and for both parameters there was a good agreement between the empirical and experimental values. The potential of adenosine triphosphate (ATP) to act as an indicator of biomass yield (mg VSS mg(-1) COD removed) was determined during the anaerobic digestion process. The biomass yield determined from ATP analysis was in the range 0.01-0.25mg VSS mg(-1) COD removed which corroborated well with previously reported studies. Empirical formula based SMP together with ATP measurement were shown to provide rapid methods to replace or augment the traditional BMP and VSS measurements and are useful for evaluating the bioenergy and biomass potential of solid wastes for anaerobic digestion.     

Effect of anaerobic digestion on oocysts of the protozoan Eimeria tenella.

The effect of anaerobic digestion of poultry waste on oocysts of the protozoan Eimeria tenella, a common enteric pathogen that causes coccidiosis in poultry, was investigated in this study. Thermophilic (50 degrees C) and mesophilic (35 degrees C) anaerobic digestors, with poultry manure as the substrate, were inoculated with the oocysts. The oocysts were damaged during anaerobic digestion, as determined by morphological change and loss of their ability to sporulate. The recovered oocysts were tested for their infectivity in young chicks, as measured by body weight gain, mortality, and cecal lesions. Oocysts lost all their infectivity during thermophilic digestion, while oocysts subjected to mesophilic digestion remained moderately infective in comparison with untreated oocysts, which produced severe coccidiosis, high mortality, and low body weight gain in chicks. Oocysts were inactivated at 50 degrees C when they were suspended in digestor fluid or saline. Inactivation at 35 degrees C was significantly stronger in the digestor fluid than in the saline, which implied that factors other than temperature were involved in the lethal effect of anaerobic digestion on protozoan oocysts. In this study we demonstrated that the treatment of animal waste by anaerobic digestion, especially at a thermophilic temperature, has the benefits of pathogen control and protection of human and animal health in a farm environment.     

Effect of anaerobic digestion on oocysts of the protozoan Eimeria tenella

The effect of anaerobic digestion of poultry waste on oocysts of the protozoan Eimeria tenella, a common enteric pathogen that causes coccidiosis in poultry, was investigated in this study. Thermophilic (50 degrees C) and mesophilic (35 degrees C) anaerobic digestors, with poultry manure as the substrate, were inoculated with the oocysts. The oocysts were damaged during anaerobic digestion, as determined by morphological change and loss of their ability to sporulate. The recovered oocysts were tested for their infectivity in young chicks, as measured by body weight gain, mortality, and cecal lesions. Oocysts lost all their infectivity during thermophilic digestion, while oocysts subjected to mesophilic digestion remained moderately infective in comparison with untreated oocysts, which produced severe coccidiosis, high mortality, and low body weight gain in chicks. Oocysts were inactivated at 50 degrees C when they were suspended in digestor fluid or saline. Inactivation at 35 degrees C was significantly stronger in the digestor fluid than in the saline, which implied that factors other than temperature were involved in the lethal effect of anaerobic digestion on protozoan oocysts. In this study we demonstrated that the treatment of animal waste by anaerobic digestion, especially at a thermophilic temperature, has the benefits of pathogen control and protection of human and animal health in a farm environment.     

Survival of bacterial pathogens during the thermophilic anaerobic digestion of biowaste: laboratory experiments and in situ validation

Anaerobic digestion is continually gaining importance for the processing of the organic fraction of municipal solid wastes. Although methods for studying the survival of pathogen exist, these methods often need adaptations, are expensive, time consuming or generally not well suited for the harsh conditions within an anaerobic digestion system. In the present study we investigated the applicability of commercially available, mechanically stable and inexpensive pathogen carriers to validate in situ pathogen inhibition within a 750,000l thermophilic, bio-waste treating anaerobic digester. None of the pathogens investigated (Listeria monocytogenes, Salmonella enterica, Escherichia coli, and Campylobacter jejuni) was capable of survival under the conditions of the biogas reactor for more than 24 h indicating that the temperature and physico-chemical properties of the sludge of the fermenter were effective in inhibiting the survival of these microorganisms.     

Alternative methods for determining anaerobic biodegradability: A review

The characterization of solid wastes is a necessary step before they can be used in anaerobic digestion. The quantities of different compounds (carbohydrates, proteins, lipids and fibers) and anaerobic biodegradability (capacity to produce methane) are important information required to characterize waste. The Biochemical Methane Potential (BMP) test is one of the most relevant tests for assessing the biodegradability of waste materials. The BMP test is run under anaerobic conditions, using bacteria populations, which makes it very time consuming, i.e., about 30 days. This paper presents alternative methods for determining the anaerobic biodegradability of solid waste. First, we describe the already existing tests for characterizing organic matter. Then we correlate an aerobic test with an anaerobic test in order to estimate anaerobic biodegradability and biogas production. This shortens the analysis time to 5 days. Models using physico-chemical characteristics as input data (total carbohydrate, total nitrogen, fiber, etc.) can predict the amount of methane produced by correlation. Pyrolysis is a very fast analytical test that can be used to characterize solid waste. Lastly, spectroscopy techniques seem to be useful for determining biodegradability, in particular by taking into account the interaction between different molecules in the organic matter.     

An Analysis of the Energy Potential of Anaerobic Digestion of Agricultural By-Products and Organic Waste

Anaerobic digestion is a promising option for recycling agricultural by-products and some organic wastes. While both agricultural by-products and wastes have no direct commercial value, their management is both complicated and costly. One option to simplify by-product management and reduce the costs associated with biogas plant feedstock is to substitute dedicated crops with vegetal by-products. Given that the chemical composition of some of these by-products can differ considerably from more typical biogas plant feedstock (such as maize silage), more complete knowledge of these alternatives to produce environmentally friendly energy is warranted. To this end, batch trials under mesophilic conditions were conducted to evaluate the potential biogas yield of many agricultural by-products: maize stalks, rice chaff, wheat straw, kiwi fruit, onions, and two expired organic waste products (dairy and dry bread) from the retail mass-market. Among the considered biomasses, the highest methane producer was the expired dairy product mixture, which yielded 554 l_NCH₄ kg⁻¹ volatile solids (VS). Maize stalks and wheat straw produced the lowest yields of 214 and 285 l_NCH₄ kg⁻¹VS, respectively. An assessment of the biogas and methane yields of each biomass was also undertaken to account for the specific chemical composition of each biomass as it can affect the anaerobic digestion operating system. Finally, the total Italian green energy production that might be derived from feeding all these biomasses to a biogas digester was estimated, in order to understand its potential impact.     

Anaerobic digestion of municipal solid waste and agricultural waste and the effect of co-digestion with dairy cow manure

Anaerobic digestion of dairy cow manure (CM), the organic fraction of municipal solid waste (OFMSW), and cotton gin waste (CGW) was investigated with a two-phase pilot-scale anaerobic digestion (AD) system. The OFMSW and CM were digested as single wastes and as combined wastes. The single waste digestion of CM resulted in 62m(3) methane/ton of CM on dry weight basis. The single waste digestion of OFMSW produced 37m(3) methane/ton of dry waste. Co-digestion of OFMSW and CM resulted in 172m(3) methane/ton of dry waste. Co-digestion of CGW and CM produced 87m(3) methane/ton of dry waste. Comparing the single waste digestions with co-digestion of combined wastes, it was shown that co-digestion resulted in higher methane gas yields. In addition, co-digestion of OFMSW and CM promotes synergistic effects resulting in higher mass conversion and lower weight and volume of digested residual.     

Production and characterization of β-glucosidase from Aspergillus niger fermentation: Application for organic fraction of municipal solid waste hydrolysis and methane enhancement

The anaerobic digestion of the organic fraction of municipal solid waste (OFMSW) is currently an attractive treatment process with energy production in the form of biogas. Hydrolysis is the rate-limiting step for the anaerobic digestion of solid wastes. Thus, in the present study fungal enzymatic pretreatment of OFMSW was applied to enhance biogas production. Two enzyme cocktails rich on β-glucosidase were produced from submerged fermentation of Aspergillus niger on basal medium using OFMSW as carbon source and urea (Urea cocktail) and Ulva rigida as nitrogen source (Ulva cocktail). Ulva cocktail displayed an important effect on OFMSW solubilization. Therefore, an increase of reducing sugar concentration about 60% was obtained which was in correlation with chemical oxygen demand (COD) increase. The performance of enzymatic pretreatment on anaerobic digestion of OFMSW was studied by conducting biochemical methane potential tests. Results showed that the enzymatic pretreatment improved methane yield of OFMSW even at high solid concentration. High methane yield about 500 ml/g total volatile solid was obtained, which corresponds up to 68% enhancement over the control.     

Continuous biomethanization of agrifood industry waste: A case study in Spain

The anaerobic digestion technology is a biological treatment widely used to reduce the pollution load of wet waste biomass. In this work we present the results obtained by performing extensive experiments of anaerobic digestion of slaughterhouse waste, tomato industry waste and olive oil industry waste in continuous mode, which were designed to demonstrate that anaerobic digestion is an effective technology from an environmental and economic point of view.Biogas yields obtained are between 35.22 and 5.45 Nm³ biogas/m³ olive oil industry waste and tomato industry waste respectively and the slaughterhouse wastes achieve intermediate production, 30.86 Nm³ biogas/m³ municipal slaughterhouse waste and 22.53 Nm³ biogas/m³ Iberian pig slaughterhouse waste. Moreover, it possible to degrade between 63.46 and 75.3% of the initial organic matter.If these results are analyzed, the environmental, energetic economic benefits of anaerobic digestion can be quantified. Biomethanation of all these wastes generated annually in Extremadura could prevent the emission of 134,772 t of equivalent carbon dioxide, generate an energy similar to that provided by 2826 toe and reach payback times from 3.29 to 3.75 years for anaerobic digestion plant designed to treat the wastes generated by a medium-sized industry. So, we have fulfilled all the planned aims.     

Fungal fermentation on anaerobic digestate for lipid-based biofuel production

Background

Anaerobic digestate is the effluent from anaerobic digestion of organic wastes. It contains a significant amount of nutrients and lignocellulosic materials, even though anaerobic digestion consumed a large portion of organic matters in the wastes. Utilizing the nutrients and lignocellulosic materials in the digestate is critical to significantly improve efficiency of anaerobic digestion technology and generate value-added chemical and fuel products from the organic wastes. Therefore, this study focused on developing an integrated process that uses biogas energy to power fungal fermentation and converts remaining carbon sources, nutrients, and water in the digestate into biofuel precursor-lipid.

Results

The process contains two unit operations of anaerobic digestion and digestate utilization. The digestate utilization includes alkali treatment of the mixture feed of solid and liquid digestates, enzymatic hydrolysis for mono-sugar release, overliming detoxification, and fungal fermentation for lipid accumulation. The experimental results conclude that 5 h and 30 °C were the preferred conditions for the overliming detoxification regarding lipid accumulation of the following fungal cultivation. The repeated-batch fungal fermentation enhanced lipid accumulation, which led to a final lipid concentration of 3.16 g/L on the digestate with 10% dry matter. The mass and energy balance analysis further indicates that the digestate had enough water for the process uses and the biogas energy was able to balance the needs of individual unit operations.

Conclusions

A fresh-water-free and energy-positive process of lipid production from anaerobic digestate was achieved by integrating anaerobic digestion and fungal fermentation. The integration addresses the issues that both biofuel industry and waste management encounter—high water and energy demand of biofuel precursor production and few digestate utilization approaches of organic waste treatment.

     

Enzyme research and applications in biotechnological intensification of biogas production

Biogas technology provides an alternative source of energy to fossil fuels in many parts of the world. Using local resources such as agricultural crop remains, municipal solid wastes, market wastes and animal waste, energy (biogas), and manure are derived by anaerobic digestion. The hydrolysis process, where the complex insoluble organic materials are hydrolysed by extracellular enzymes, is a rate-limiting step for anaerobic digestion of high-solid organic solid wastes. Biomass pretreatment and hydrolysis are areas in need of drastic improvement for economic production of biogas from complex organic matter such as lignocellulosic material and sewage sludge. Despite development of pretreatment techniques, sugar release from complex biomass still remains an expensive and slow step, perhaps the most critical in the overall process. This paper gives an updated review of the biotechnological advances to improve biogas production by microbial enzymatic hydrolysis of different complex organic matter for converting them into fermentable structures. A number of authors have reported significant improvement in biogas production when crude and commercial enzymes are used in the pretreatment of complex organic matter. There have been studies on the improvement of biogas production from lignocellulolytic materials, one of the largest and renewable sources of energy on earth, after pretreatment with cellulases and cellulase-producing microorganisms. Lipids (characterised as oil, grease, fat, and free long chain fatty acids, LCFA) are a major organic compound in wastewater generated from the food processing industries and have been considered very difficult to convert into biogas. Improved methane yield has been reported in the literature when these lipid-rich wastewaters are pretreated with lipases and lipase-producing microorganisms. The enzymatic treatment of mixed sludge by added enzymes prior to anaerobic digestion has been shown to result in improved degradation of the sludge and an increase in methane production. Strategies for enzyme dosing to enhance anaerobic digestion of the different complex organic rich materials have been investigated. This review also highlights the various challenges and opportunities that exist to improve enzymatic hydrolysis of complex organic matter for biogas production. The arguments in favor of enzymes to pretreat complex biomass are compelling. The high cost of commercial enzyme production, however, still limits application of enzymatic hydrolysis in full-scale biogas production plants, although production of low-cost enzymes and genetic engineering are addressing this issue.     

Acidogenic fermentation of proteinaceous solid waste and characterization of different bioconversion stages and extracellular products

The purpose of this study was to investigate hydrolysis of animal fleshing (ANFL), a predominant tannery solid waste and to characterize the acetogenic fermentation products of anaerobic digestion. The acidogenic digestibility of the tannery solid wastes were evaluated up to 120 h using batch anaerobic digestion tests performed under mesophilic condition at 37 degrees C. The degradation of ANFL starts with non-fibrillar proteins and proceeds with fibrillar proteins. The release of aliphatic amino acid in the early stages of hydrolysis (24 h) and followed by aromatic amino acids (24-72 h) were evidenced by HPLC analysis. The maximum production of propionic and valeric acid were observed in 72 h followed by rapid increase in acetic acid in 96 h using GC-MS. Breakdown of ANFL and formations of other metabolites were evidenced by FT-IR and (1)H-NMR spectroscopy.     

Enzyme research and applications in biotechnological intensification of biogas production

Biogas technology provides an alternative source of energy to fossil fuels in many parts of the world. Using local resources such as agricultural crop remains, municipal solid wastes, market wastes and animal waste, energy (biogas), and manure are derived by anaerobic digestion. The hydrolysis process, where the complex insoluble organic materials are hydrolysed by extracellular enzymes, is a rate-limiting step for anaerobic digestion of high-solid organic solid wastes. Biomass pretreatment and hydrolysis are areas in need of drastic improvement for economic production of biogas from complex organic matter such as lignocellulosic material and sewage sludge. Despite development of pretreatment techniques, sugar release from complex biomass still remains an expensive and slow step, perhaps the most critical in the overall process. This paper gives an updated review of the biotechnological advances to improve biogas production by microbial enzymatic hydrolysis of different complex organic matter for converting them into fermentable structures. A number of authors have reported significant improvement in biogas production when crude and commercial enzymes are used in the pretreatment of complex organic matter. There have been studies on the improvement of biogas production from lignocellulolytic materials, one of the largest and renewable sources of energy on earth, after pretreatment with cellulases and cellulase-producing microorganisms. Lipids (characterised as oil, grease, fat, and free long chain fatty acids, LCFA) are a major organic compound in wastewater generated from the food processing industries and have been considered very difficult to convert into biogas. Improved methane yield has been reported in the literature when these lipid-rich wastewaters are pretreated with lipases and lipase-producing microorganisms. The enzymatic treatment of mixed sludge by added enzymes prior to anaerobic digestion has been shown to result in improved degradation of the sludge and an increase in methane production. Strategies for enzyme dosing to enhance anaerobic digestion of the different complex organic rich materials have been investigated. This review also highlights the various challenges and opportunities that exist to improve enzymatic hydrolysis of complex organic matter for biogas production. The arguments in favor of enzymes to pretreat complex biomass are compelling. The high cost of commercial enzyme production, however, still limits application of enzymatic hydrolysis in full-scale biogas production plants, although production of low-cost enzymes and genetic engineering are addressing this issue.     

Modeling solid waste decomposition

The hydrolysis rate coefficients of sorted municipal waste were evaluated from the biochemical methane potential tests using non-linear regression. A distributed mathematical model of anaerobic digestion of rich (food) and lean (non-food) solid wastes with greatly different rates of polymer hydrolysis/acidogenesis was developed to describe the balance between the rates of hydrolysis/acidogenesis and methanogenesis. The model was calibrated using previously published experimental data [Biores. Technol. 52 (1995) 245] obtained upon various initial food waste loadings. Simulations of one- and two-stage digestion systems were carried out. The results showed that initial spatial separation of food waste and inoculum enhances methane production and waste degradation in a one-stage solid-bed digester at high waste loading. A negative effect of vigorously mixing at high waste loading reported in some papers was discussed. It was hypothesized that the initiation methanogenic centers developing in time and expanding in space under minimal mixing conditions might be a key factor for efficient anaerobic conversion of solid waste into methane.     

大型奶牛场粪便厌氧消化工程的研究

本文报导了大型牛场粪便厌氧消化工程研究的概况。该工程针对牛粪草多、浮渣多的特点设计了除长草、除浮渣装置。以保证了厌氧消化的稳定运行。工程每天可处理奶牛粪便８０余吨、鸡粪及工业发酵废渣２０余吨,生产沼气供应２８６８户职工家庭作燃气使用,平均沼气率为１．２ｍ＾３／ｍ＾３．ｄ。经后处理的发酵残留物被充分利用,制成肥料、绿色营养土和鱼饵,这样既免除了二次污染,又提高了工程的经济效益。该工程为我国大中型沼气