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 pathogenic bacteria during mesophilic anaerobic digestion of animal waste

The survival of pathogenic bacteria was investigated during the operation of a full-scale anaerobic digester which was fed daily and operated at 28°C. The digester had a mean hydraulic retention time of 24 d. The viable numbers of Escherichia coli, Salmonella typhimurium, Yersinia enterocolitica, Listeria monocytogenes and Campylobacter jejuni were reduced during mesophilic anaerobic digestion. Echerichia coli had the smallest mean viable numbers at each stage of the digestion process. Its mean T₉₀ value was 76.9 d. Yersinia enterocolitica was the least resistant to the anaerobic digester environment; its mean T₉₀ value was 18.2 d. Campylobacter jejuni was the most resistant bacterium; its mean T₉₀ value was 438.6 d. Regression analysis showed that there were no direct relationships between the slurry input and performance of the digester and the decline of pathogen numbers during the 140 d experimental period.     

Survey of wastewater indicators and human pathogen genomes in biosolids produced by class a and class B stabilization treatments

Accurate modeling of the infectious aerosol risk associated with the land application of biosolids requires an in-depth knowledge of the magnitudes and changes in pathogen concentrations for a variety of class A and class B stabilization methods. The following survey used quantitative PCR (qPCR) and culture assays to detect environmentally resistant bacterial and viral pathogens and biosolid indicator organisms for 36 biosolid grab samples. Biosolids were collected from 14 U.S. states and included 16 class B mesophilic anaerobic digestion (MAD) samples and 20 class A biosolid samples from temperature-phased anaerobic digestion (TPAD), MAD plus composting (COM), and MAD plus heat pelletization processes. The indicator concentrations of fecal coliforms and male-specific coliphages as well as pathogen genome concentrations for human adenovirus species, Legionella pneumophila, Staphylococcus aureus, and Clostridium difficile were significantly lower in the class A samples, and a multivariate analysis of variance ranked the stabilization processes from the lowest pathogen/indicator load to the highest as (i) class A COM, (ii) class A TPAD, and (iii) class B MAD. Human adenovirus genomes were found in 88% of the class B samples and 70 to 100% of the class A samples. L. pneumophila, S. aureus, and C. difficile genomes were detected at the qPCR assay detection limits in 19 to 50% of the class B and class A anaerobic digestion samples, while L. pneumophila was detected in 50% of the class A compost samples. When considering all the stabilization methods, both the fecal coliform and the male-specific coliphage concentrations show a significant linear correlation with the pathogen genome concentrations. This survey provides the necessary pathogen concentrations to add to biosolid aerosol risk and pathogen exposure analyses and clarifies the effectiveness of class A stabilization methods with the pathogen and indicator loads in biosolids.     

Bacterial survival and association with sludge flocs during aerobic and anaerobic digestion of wastewater sludge under laboratory conditions.

The fate of indicator bacteria, a bacterial pathogen, and total aerobic bacteria during aerobic and anaerobic digestion of wastewater sludge under laboratory conditions was determined. Correlation coefficients were calculated between physical and chemical parameters (temperature, dissolved oxygen, pH, total solids, and volatile solids) and either the daily change in bacterial numbers or the percentage of bacteria in the supernatant. The major factor influencing survival of Salmonella typhimurium and indicator bacteria during aerobic digestion was the temperature of sludge digestion. At 28 degrees C with greater than 4 mg of dissolved oxygen per liter, the daily change in numbers of these bacteria was approximately -1.0 log10/ml. At 6 degrees C, the daily change was less than -0.3 log10/ml. Most of the bacteria were associated with the sludge flocs during aerobic digestion of sludge at 28 degrees C with greater than 2.4 mg of dissolved oxygen per liter. Lowering the temperature or the amount of dissolved oxygen decreased the fraction of bacteria associated with the flocs and increased the fraction found in the supernatant.     

Comparison of Methods to Identify Pathogens and Associated Virulence Functional Genes in Biosolids from Two Different Wastewater Treatment Facilities in Canada

The use of treated municipal wastewater residues (biosolids) as fertilizers is an attractive, inexpensive option for growers and farmers. Various regulatory bodies typically employ indicator organisms (fecal coliforms, E. coli and Salmonella) to assess the adequacy and efficiency of the wastewater treatment process in reducing pathogen loads in the final product. Molecular detection approaches can offer some advantages over culture-based methods as they can simultaneously detect a wider microbial species range, including non-cultivable microorganisms. However, they cannot directly assess the viability of the pathogens. Here, we used bacterial enumeration methods together with molecular methods including qPCR, 16S rRNA and cpn60 gene amplicon sequencing and shotgun metagenomic sequencing to compare pre- and post-treatment biosolids from two Canadian wastewater treatment plants (WWTPs). Our results show that an anaerobic digestion WWTP was unsuccessful at reducing the live indicator organism load (coliforms, generic E. coli and Salmonella) below acceptable regulatory criteria, while biosolids from a dewatering/pelletization WWTP met these criteria. DNA from other pathogens was detected by the molecular methods, but these species were considered less abundant. Clostridium DNA increased significantly following anaerobic digestion treatments. In addition to pathogen DNA, genes related to virulence and antibiotic resistance were identified in treated biosolids. Shotgun metagenomics revealed the widest range of pathogen DNA and, among the approaches used here, was the only approach that could access functional gene information in treated biosolids. Overall, our results highlight the potential usefulness of amplicon sequencing and shotgun metagenomics as complementary screening methods that could be used in parallel with culture-based methods, although more detailed comparisons across a wider range of sites would be needed.     

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.     

Thermophilic anaerobic digestion of solid waste for fuel gas production.

Anaerobic digestion offers a potential means of converting organic solid waste into fuel gas and thereby provide a supplemental and readily utilizable source of energy. We are particularly interested in the use of thermophilic digestion over a mesophilic operation for it can achieve higher rates of digestion, greater conversion of waste organics to gas, faster solid-liquid separation, and minimization of bacterial and viral pathogen accumulation. Our results comparing mesophilic (37 degree C) and thermophilic (65 degree C) anaerobic digestion of domestic solid waste confirm the increased rate and conversion of waste to methane. In addition, utilizing radioactive labeling of glucose and acetic acid, we have measured the volumetric rates of volatile acid production and disappearance under both mesophilic and thermophilic conditions.     

Enhancement on biodegradation and anaerobic digestion efficiency of activated sludge using a dual irradiation process

A dual irradiation process involving aerobic thermophilic irradiation pretreatment (ATIP) and intermittent irradiation anaerobic digestion was developed to improve the digestion of waste-activated sludge. First, the effect of ATIP on further anaerobic digestion of activated sludge in batch mode was investigated. When exposed to ATIP for 24 h, the digestion reactor gave the highest methane yield, removed the most dissolved organic carbon (DOC) and showed the most effective reduction of VS compared to other irradiation times. This process was further enhanced by using an anaerobic fluidised-bed reactor packed with carbon felt in semi-continuous mode for digesting the pretreated activated sludge under intermittent irradiation conditions. Dual irradiation for 24 h followed by 60 min of anaerobic irradiation processing per day turned out to be optimal. This resulted in 65.3% of VS reduction, 83.9% of DOC removal ratio and 538 ml/g-VS of methane yield.     

Optimisation of the anaerobic digestion of agricultural resources

It is in the interest of operators of anaerobic digestion plants to maximise methane production whilst concomitantly reducing the chemical oxygen demand of the digested material. Although the production of biogas through anaerobic digestion is not a new idea, commercial anaerobic digestion processes are often operated at well below their optimal performance due to a variety of factors. This paper reviews current optimisation techniques associated with anaerobic digestion and suggests possible areas where improvements could be made, including the basic design considerations of a single or multi-stage reactor configuration, the type, power and duration of the mixing regime and the retention of active microbial biomass within the reactor. Optimisation of environmental conditions within the digester such as temperature, pH, buffering capacity and fatty acid concentrations is also discussed. The methane-producing potential of various agriculturally sourced feedstocks has been examined, as has the advantages of co-digestion to improve carbon-to-nitrogen ratios and the use of pre-treatments and additives to improve hydrolysis rates or supplement essential nutrients which may be limiting. However, perhaps the greatest shortfall in biogas production is the lack of reliable sensory equipment to monitor key parameters and suitable, parallelised control systems to ensure that the process continually operates at optimal performance. Modern techniques such as software sensors and powerful, flexible controllers are capable of solving these problems. A direct comparison can be made here with, for instance, oil refineries where a more mature technology uses continuous in situ monitoring and associated feedback procedures to routinely deliver continuous, optimal performance.     

Comparison of static,in-vessel composting of MSW with thermophilic anaerobic digestion and combinations of the two processes

The biological stabilisation of the organic fraction of municipal solid waste (OFMSW) into a form stable enough for land application can be achieved via aerobic or anaerobic treatments. To investigate the rates of degradation (e.g. via electron equivalents removed, or via carbon emitted) of aerobic and anaerobic treatment, OFMSW samples were exposed to computer controlled laboratory-scale aerobic (static in-vessel composting), and anaerobic (thermophilic anaerobic digestion with liquor recycle) treatment individually and in combination. A comparison of the degradation rates, based on electron flow revealed that provided a suitable inoculum was used, anaerobic digestion was the faster of the two waste conversion process. In addition to faster maximum substrate oxidation rates, anaerobic digestion (followed by post-treatment aerobic maturation), when compared to static composting alone, converted a larger fraction of the organics to gaseous end-products (CO₂ and CH₄), leading to improved end-product stability and maturity, as measured by compost self-heating and root elongation tests, respectively. While not comparable to windrow and other mixed, highly aerated compost systems, our results show that in the thermophilic, in-vessel treatment investigated here, the inclusion of a anaerobic phase, rather than using composting alone, improved hydrolysis rates as well as oxidation rates and product stability. The combination of the two methods, as used in the DiCOM^® process, was also tested allowing heat generation to thermophilic operating temperature, biogas recovery and a low odour stable end-product within 19 days of operation.     

Strain differentiation of epidemic typhus rickettsiae (Rickettsia prowazekii) by DNA restriction endonuclease analysis

We have investigated the number of methanogenic bacteria in different samples of fresh feces and effluents from the anaerobic digestion of cattle and swine waste and municipal sludges. We have further compared conventional anaerobic techniques with the rigorous, oxygen-free, anaerobic, experimental conditions obtained in a controlled glove cabinet atmosphere. It was found that the total number of oxygen-intolerant methanogenic bacteria was about 10–100 times higher than that of anaerobic oxygen-tolerant methanogenic bacteria. We have also developed a simple technique for the growth of methanogenic bacteria that permits the realization of rigorous anaerobic conditions without the use of a glove cabinet.     

Energy Metabolism and Survival of the Infective Juveniles of Steinernema carpocapsae under Oxygen-Deficient Conditions

Energy metabolism and its relation to survival of the infective juveniles (IJ) of S. carpocapsae under anaerobic and oxygen-deficient conditions were studied by monitoring changes in survival rate, levels of key energy reserve materials, oxygen consumption, and respiratory quotient (RQ). The effects of various factors on the survival of IJ under anaerobic conditions were also investigated. Under anaerobic conditions, the IJ were inactivated but could survive for several days in an immobile state, using the carbohydrate reserves glycogen and trehalose for energy supply. The survival time of IJ was mainly dependent on the availability of energy supply, which, in turn, was influenced by factors such as temperature and metabolic by-products. Surviving, anaerobically incubated IJ fully recovered upon return to aerobic conditions. Recovering IJ were characterized by regaining mobility and restoration of carbohydrate reserves consumed during the anaerobic period. Carbohydrate reserves were restored by conversion from lipid reserves and possibly from anaerobic metabolic by-products. The infectivity of IJ recovered from the anaerobic state was not affected. At 1% oxygen level, IJ were also immobile and mainly depended on carbohydrate reserves for energy supply and the RQ was greater than 1. However, some oxygen was consumed; the survival time of these IJ was shorter than those kept in natural air but longer than those under anaerobic conditions. When IJ were incubated at oxygen levels of 3% to 21%, the RQs were maintained at 0.7 to 0.8. Oxygen consumption rates and the reduction in both mean dry weight and lipid levels were proportional to oxygen levels while the survival time of IJ was inversely proportional to oxygen levels.     

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。     

Role of anaerobiosis in virulence of Salmonella typhimuirium

Intestinal pathogens are exposed to various stress conditions during their infectious cycle. Anaerobiosis, one of such hostile condition, is offered by the host within gut and intestinal lumen, where survival, multiplication and entry into intestinal epithelial cells is priority for the invading pathogen. In the present study, a virulent strain of S. typhimurium (1402/84) was grown under anaerobic conditions and its virulence characteristics such as host cell binding, penetration and intracellular survival were compared with aerobic S. typhimurium. Anaerobically grown S. typhimurium showed significantly higher binding to immobilized mice enterocytes and intestinal mucus as compared to bacteria grown aerobically. Anaerobic bacteria also showed an early penetration of mucus and subsequent binding to underlying immobilized enterocytes, in vitro. Anaerobic S. typhimurium exhibited increased intracellular survival within spleen macrophages of mice and caused significantly higher fluid accumulation in ligated rabbit ileal loops as compared to aerobic bacteria. LD₅₀ of anaerobic S. typhimurium was also observed to be 2 fold lower when compared to aerobic bacteria. Cell surface hydrophobicity of anaerobic S. typhimurium was also found to be significantly higher than aerobic bacteria. Thus, it appears that exposure of S. typhimurium to anaerobiosis results in its enhanced virulence, adhesion and penetration of host cells.     

Cultivation of Ulva lactuca with manure for simultaneous bioremediation and biomass production

The potential of liquid manure as sole nutrient source for cultivation of Ulva lactuca was investigated with the perspective of utilizing the produced biomass for feed and/or energy. Algae grown with manure demonstrated equal growth rates to algae grown with standard f/2-medium. The optimum manure concentration, expressed as ammonium concentration, was 25?μM. At these conditions, the biomass produced was potentially suitable for anaerobic digestion, due to a relative high carbon/nitrogen ratio (approximately 19). At higher manure concentrations, tissue concentrations of nitrogen, phosphorus, proteins, and amino acids increased, making the biomass less suitable for anaerobic digestion but potentially interesting as a feed supplement. Cultivating U. lactuca with manure as nutrient source has potential in terms of bioremediation as well as production of bioenergy and protein-feed.     

Viability of Plant–Pathogenic Fungi Reduced by Anaerobic Digestion

Feedstock of anaerobic digestion infected with phytopathogens could enhance the risk of spreading those pathogens to uninfested field through digestate. The viability of Fusarium proliferatum, Fusarium verticillioides, Sclerotinia sclerotiorum, and Rhizoctonia solani was investigated in anaerobic digestion experiments using infected plant material of sorghum (Sorghum bicolor), sugar beet (Beta vulgaris subsp. vulgaris var. altissima), and potato (Solanum tuberosum L.). Results from lab-scale reactors were confirmed in full-scale biogas plants. Anaerobic digestion under mesophilic conditions (35–42 °C) reduced most of the phytopathogens of feedstocks investigated. Thus, S. sclerotiorum and R. solani lost their viability within 6 h. In the case of sorghum, however, Fusarium spp. infected feedstock required a maximum of 138 h for sanitation. Thus, the risk of spreading plant pathogens with the digestate can only be decreased when the feedstock would undergo an additional treatment before anaerobic digestion or of the resulting digestate.     

Anaerobic co-digestion of mechanically biologically treated municipal waste with primary sewage sludge - a feasibility study

This bench scale study investigated the suitability of MBT material for treatment by anaerobic digestion and the impacts of co-digestion of these wastes with sewage sludge. The results suggest that MBT material is amenable to anaerobic digestion with sewage sludge. The main problems for scale-up are related to the physical composition of the MBT material, the accumulation of heavy metals and other inert contaminants and the impact of both of these factors on final sludge quality. Full-scale trials would be required to assess the long-term impacts of MBT waste on anaerobic digestion, if this form of co-digestion were to be pursued. The material contamination that presents a barrier to the direct recycling of MBT material in land-applications is also a major hurdle in commercial co-digestion. Better quality input material would be likely to result in higher methane yields and fewer restrictions on the utilisation of the product in recycling.     

微生物组学及其在厌氧消化中的研究进展

我国每年产生大量的有机废弃物,如果处置不当将会对生态、气候以及人类健康造成重大影响。厌氧消化是一种可靠的、绿色的、可持续的有机废弃物处理方式,但由于缺乏准确有效的监测手段,厌氧消化微观过程常常被视为“黑盒”。随着微生物组学的发展,学者们在菌群与运行参数关联性分析、代谢途径分析等方面有了更深入的认识。本文从“三阶段、四菌群”的厌氧消化过程出发,介绍了常用微生物组学的类型,包括:16S rRNA基因组、宏基因组、宏转录组和宏蛋白组;详细阐述了物种组成分析、α多样性分析、OTU相似性分析以及多元统计学分析等6种常用的微生物群落生物信息学分析方法;系统回顾了厌氧消化过程的微生物学研究进展,以期能为分析厌氧消化的微生物群落结构和功能、开发新的厌氧消化工艺和技术提供支持。     

厌氧消化酸抑制研究进展

厌氧消化工艺目前已广泛应用于各类废水的处理处置过程中,但在实际运行中,受消化条件和物料性质的影响,消化系统经常遭受由挥发性脂肪酸积累过多导致的酸抑制问题,引发产气量下降、产甲烷率降低等问题。近年来,有研究者发现,挥发性脂肪酸的种类和浓度及pH、温度是影响酸抑制的主要因素。基于此,相关研究者分别尝试了添加碱性化学药剂和微量元素及利用生物强化技术与微生物电化学技术来解除酸抑制的尝试,并都取得了不错的效果。本文综述了厌氧消化过程中酸抑制的产生过程、抑制机理及恢复方法,以期为解决厌氧消化酸抑制问题提供参考。