Low-temperature anaerobic digestion for wastewater treatment

Methanogenesis is an important biogeochemical process for the degradation of organic matter within cold environments, and is associated with the release of the potent greenhouse gas, methane. Cold methanogenesis has been harnessed, in engineered systems, as low-temperature anaerobic digestion (LTAD) for wastewater treatment and bioenergy generation. LTAD represents a nascent wastewater treatment biotechnology, which offers an attractive alternative to conventional aerobic and anaerobic processes. Successful, high-rate, LTAD of sewage and industrial wastewaters (e.g. from the brewery, food-processing and pharmaceutical sectors), with concomitant biogas generation, has been demonstrated at laboratory-scale and pilot-scale. A holistic, polyphasic approach, which integrates bioprocess, physiological and molecular biological datasets has been critical to the development of the LTAD concept.     

餐厨垃圾厌氧消化处理主要过程的微生物群落结构分析

【背景】厌氧消化是我国餐厨垃圾处理的主要方法,微生物在其处理过程中起到关键作用,但是目前对其不同工艺单元微生物群落结构的研究较少。【目的】通过分析各工艺单元的微生物多样性与群落结构,为改进餐厨垃圾资源化处理技术、提高资源利用效率提供科学依据。【方法】采集某餐厨垃圾处理厂油水分离、厌氧发酵、沼渣脱水等3个工艺单元产生的废液样品,采用16S rRNA基因高通量测序技术,研究其菌群组成、丰度、优势菌群及其与环境因子的相关性。【结果】初始油水分离样品中的微生物群落种类相对较少,而经厌氧发酵和沼渣脱水处理后样品中的微生物群落种类较丰富。在门水平上,厚壁菌门(Firmicutes)在各单元样品中所占平均比例最高,为81.1%,其次为拟杆菌门(Bacteroidetes)和绿弯菌门(Chloroflexi),分别占15.81%和4.59%;在属水平上,相对丰度较高的菌属为乳酸菌属(Lactobacillus)、互营单胞菌属(Syntrophomonas)等。餐厨垃圾处理过程中的部分菌属可能具有资源-环境双重属性,例如在沼渣脱水单元相对丰度高达32.67%的假单胞菌属(Pseudomonas),该菌属中既存在少部分致病菌或条件致病菌,也具有生产聚羟基脂肪酸酯的功能菌。影响各组样品微生物群落组成结构最显著的因子是p H值,其次是总糖的含量。【结论】研究明确了典型餐厨垃圾厌氧消化处理工艺单元的微生物群落结构和多样性,并提出了优化处理工艺、强化资源利用效率的建议。     

Microbial diversity of methanogenic communities in the systems for anaerobic treatment of organic waste

Methane production via anaerobic degradation of organic-contaminated wastewater, semiliquid, or solid municipal waste of complex composition by methanogenic microbial communities is a multistage process involving at least four groups of microorganisms. These are hydrolytic bacteria (polysaccharolytic, proteolytic, and lipolytic), fermentative bacteria, acetogenic bacteria (syntrophic, proton-reducing), and methanogenic archaea; complex trophic interactions exist between these groups. The review provides information concerning the diversity of the major microbial groups identified in the systems for wastewater and concentrated waste treatment, solid-phase anaerobic fermentation, and landfills for disposal of municipal solid waste, and also specifies the sources of isolation of the type strains. The research demonstrates that both new microorganisms and those previously isolated from natural habitats may be found in waste treatment systems. High microbial diversity in the systems for organic waste treatment provides for stable methanogenesis under fluctuating environmental conditions.     

Microbial processes and bacterial populations associated to anaerobic treatment of sulfate-rich wastewater

A pilot-scale (1.2 m³) anaerobic sequencing batch biofilm reactor (ASBBR) containing mineral coal for biomass attachment was fed with sulfate-rich wastewater at increasing sulfate concentrations. Ethanol was used as the main organic source. Tested COD/sulfate ratios were of 1.8 and 1.5 for sulfate loading rates of 0.65–1.90 kgSO₄²⁻/cycle (48 h-cycle) or of 1.0 in the trial with 3.0 gSO₄²⁻ l⁻¹. Sulfate removal efficiencies observed in all trials were as high as 99%. Molecular inventories indicated a shift on the microbial composition and a decrease on species diversity with the increase of sulfate concentration. Beta-proteobacteria species affiliated with Aminomonas spp. and Thermanaerovibrio spp. predominated at 1.0 gSO₄²⁻ l⁻¹. At higher sulfate concentrations the predominant bacterial group was Delta-proteobacteria mainly Desulfovibrio spp. and Desulfomicrobium spp. at 2.0 gSO₄²⁻ l⁻¹, whereas Desulfurella spp. and Coprothermobacter spp. predominated at 3.0 gSO₄²⁻ l⁻¹. These organisms have been commonly associated with sulfate reduction producing acetate, sulfide and sulfur. Methanogenic archaea (Methanosaeta spp.) was found at 1.0 and 2.0 gSO₄²⁻ l⁻¹. Additionally, a simplified mathematical model was used to infer on metabolic pathways of the biomass involved in sulfate reduction.     

Microbial and enzymatic improvement of anaerobic digestion of waste biomass

Metabolic activities of different microorganisms (Bacillus subtilis, B. licheniformis and Aspergillus niger) and hydrolytic enzymes (concentrations: 1 to 200 mg enzyme solids g^–1 feed) were studied individually and in combinations with respect to H₂ and methane production from damaged wheat grains. Bacillus subtilis, B. licheniformis and pre-existing hydrogen producers (control) produced 45 to 64 l H₂ kg^–1 total solids and subsequently, with the help of added methanogens, 155 to 220 l methane kg^–1 total solids could be produced. H₂ production from damaged wheat grains could be decreased to 28% or enhanced up to 152% with respect to control, by employing various microbial and enzymatic treatments. Similarly, it has been made possible to vary methane production capacities from as low as 17% to as high as 110% with respect to control.     

Thermophilic anaerobic digestion: the best option for waste treatment

After introducing thermophilic anaerobic digestion (AD), characteristics of thermophilic methanogens are provided. Accordingly, (a) site of occurrence, (b) morphological characteristics (shape and motility), (c) biochemical characteristics (Gram character and % G+C profile), (d) nutritional characteristics (NaCl requirement and substrate specificity), and (e) growth characteristics (pH and temperature) of thermophilic methanogens are described. Some studies of the thermophilic AD are cited with their operational management problems. Subsequently, strategies to maximize net energy production are given, including mode of heating the bioreactors, role of agitation to promote AD performance and mode/intensity of mixing. Finally, advantages as well as drawbacks of AD under thermophilic conditions are given, concluding with its applications.     

Mesophilic anaerobic digestion: first option for waste treatment in tropical regions

Rural India derives its energy needs for cooking and heating through the use of fuel wood and for lighting and agricultural operations through kerosene and diesel. Use of fuel wood has aggravated the problem of de-forestation, while availability of kerosene and diesel cannot be guaranteed due to corrupt practices in the public distribution systems. In contrast, urban India derives its energy needs through LPG cylinders, petrol, and electricity. However, their cost and uncertainty rendered them beyond the reach of lower income population. This scenario is more or less true with many developing countries. To meet these objectives, biogas generation from biodegradable waste using anaerobic digestion (AD) appears to be a sustainable avenue as it could be used for (a) water and space heating of farmhouses, animal shelters, (b) generating steam for food processing plants, and (c) electricity generation, in addition to reducing the pollution/hazard potential of these wastes. Many of the underdeveloped and developing countries are in the temperate zone and thus mesophilic AD could provide a desired pathway to achieve a long delayed need of energy for comfortable living, farming, and industrial operations. Efforts made in this direction are reviewed in the present article.     

Microbial communities and their enzymes facilitate degradation of recalcitrant polymers in anaerobic digestion

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Dynamic simulator for anaerobic digestion processes

A transient, two-culture model simulating methane production from biomass has been developed. The simulator, based partially on the work by Andrews and McCarty, is capable of calculating the hydrolysis products of several common organic materials, accommodating various substrate feeding modes, and simulating the transient physico-biochemical transport and conversion processes occurring in the biological, liquid, and gaseous phases of a well-mixed reactor. The mathematical representation of this bioconversion system consists of a set of 11 coupled, nonlinear first-order rate equations based on the principles of mass conservation and biochemical reaction kinetics. The model can be used in conjunction with laboratory investigations and as a simulator for evaluating process control strategies and cost developments.     

Microbial community structure and dynamics during anaerobic digestion of various agricultural waste materials

The influence of the feedstock type on the microbial communities involved in anaerobic digestion was investigated in laboratory-scale biogas reactors fed with different agricultural waste materials. Community composition and dynamics over 2 months of reactors’ operation were investigated by amplicon sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Major bacterial taxa belonged to the Clostridia and Bacteroidetes, whereas the archaeal community was dominated by methanogenic archaea of the orders Methanomicrobiales and Methanosarcinales. Correlation analysis revealed that the community composition was mainly influenced by the feedstock type with the exception of a temperature shift from 38 to 55 °C which caused the most pronounced community shifts. Bacterial communities involved in the anaerobic digestion of conventional substrates such as maize silage combined with cattle manure were relatively stable and similar to each other. In contrast, special waste materials such as chicken manure or Jatropha press cake were digested by very distinct and less diverse communities, indicating partial ammonia inhibition or the influence of other inhibiting factors. Anaerobic digestion of chicken manure relied on syntrophic acetate oxidation as the dominant acetate-consuming process due to the inhibition of aceticlastic methanogenesis. Jatropha as substrate led to the enrichment of fiber-degrading specialists belonging to the genera Actinomyces and Fibrobacter.     

Recent advances in anaerobic biological processes for textile printing and dyeing wastewater treatment: a mini-review

Textile printing and dyeing wastewater is usually characterized by high pH, high turbidity, poor bio-degradability, complex composition, and high chrominance, and is discharged in large amounts. It has been regarded as one of the hardest to treat forms of industrial wastewater. Conventional physicochemical technologies can remove these contaminants from water bodies, but at the expense of high energy consumption and high cost. Alternatively, biological processes with limited energy consumption, low cost and high efficiency are considered as promising technologies. Among them, the anaerobic biological processes have been proven to be effective for the treatment of high-concentration textile printing and dyeing wastewater. In this mini-review, recent advances on high-rate anaerobic technologies for such purposes are reviewed. Current limitations of these technologies are summarized, and future research directions are indicated.

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Microbial activity and community structure in a lake sediment used for psychrophilic anaerobic wastewater treatment

Aims: The aim of the study was to investigate the feasibility of a continuous reactor for psychrophilic anaerobic wastewater treatment by using the sludge from cold natural environment. Methods and Results: Six sludge samples (S1–S6) were collected from different cold natural locations to select sludge with high anaerobic microbial activity under low temperatures. After a 225‐day incubation, the maximum specific methane production rate of a waterfowl lake sediment (S1) at 15°C (70·5 mLCH₄ gVSS^?1 day^?1) was much higher than all other samples. S1 was thus chosen as the seed sludge for the reactor treating synthetic brewery wastewater at 15°C, by immobilizing the micro‐organisms on polyurethane foam carriers. The chemical oxygen demand (COD) removal efficiency reached over 80% after 240‐day operation at an organic loading rate of 5·3 kg m^?3 day^?1, and significant enrichment of biomass was observed. Clone libraries of the microbial communities in the inoculum had high diversities for both archaea and bacteria. Along with a decrease in microbial community diversities, the dominant bacteria (79·5%) at the end of the operation represented the phylum Firmicutes, while the dominant archaeon (41·5%) showed a similarity of 98% with the psychrotolerant methanogen Methanosarcina lacustris. Conclusions: The possibility of using anaerobic micro‐organisms from cold environments in anaerobic wastewater treatment under psychrophilic conditions is supported by these findings. Significance and Impact of the Study: This study enriches the theory on microbial community and the application on anaerobic treatment of sludge from cold natural environments.     

Microbial community in a pilot-scale bioreactor promoting anaerobic digestion and sulfur-driven denitrification for domestic sewage treatment

A pilot-scale reactor treating domestic sewage was operated to promote anaerobic digestion and denitrification using endogenous electron donors. While 55 % of organic matter was removed, nitrogen and sulfur showed a different dynamics during the operation. Pyrosequencing analysis clarified this behavior revealing that specific microbial communities inhabited the anaerobic (47.05 % of OTUs) and anoxic (31.39 % of OTUs) chambers. Analysis of 16S rRNA gene partial sequences obtained through pyrosequencing revealed a total of 1727 OTUs clustered at a 3 % distance cutoff. In the anaerobic chamber, microbial community was comprised of fermentative, syntrophic and sulfate-reducing bacteria. The majority of sequences were related to Aminobacterium and Syntrophorhabdus. In the anoxic chamber, the majority of sequences were related to mixotrophic and strictly autotrophic denitrifiers Arcobacter and Sulfuricurvum, respectively, both involved in sulfur-driven denitrification. These results show that pyrosequencing was a powerful tool to investigate the microbial panorama of a complex system, providing new insights to the improvement of the system.     

Determining specific biomass activity in anaerobic wastewater treatment processes

An experimental method for the measurement of specific gas production rate was developed and tested with biomass samples taken from anaerobic fluidized bed reactors, operating with a variety of carriers with molasses, condensate from cellulose production and brewery wastewater as feeds. The method is based on reactor sampling and offline gas volume measurement during a known time interval. Important factors are biomass and liquid sampling under oxygen-free conditions, using the liquid from the reactor as substrate, providing sufficient mixing and maintaining the physical integrity of the biomass. The method was developed in such a way that small samples (20 ml) were taken under anaerobic conditions (poising agent) for short-term (2–3 min.) gas rate measurements in a small fluidized bed (25 ml) batch reactor with U-tube. Biomass content was measured by an instrumental nitrogen method (Dumas), followed by weight determination of the carrier. The gas rates measured with the test system, and their dependence on substrate concentration, were in good agreement with those directly measured from the continuous fluidized bed reactor. Additions of molasses and acetate to the sample proved that the influence of concentration on the biomass activity can be obtained only by operating the continuous reactor at the concentration levels of interest. Comparison between the reactors showed large differences in the specific activity and the total reactor activity. It was found when comparing two reactors, that the values of the specific and the total activities permitted the calculation of the relative biomass quantities. In this way the influence of the carrier-type could be evaluated.     

Semi-continuous anaerobic digestion of a food industry wastewater in an anaerobic filter

The experimental results of semi-continuous tests of anaerobic digestion of confectionery industry wastewater, carried out at different residence times and organic loads in an upflow anaerobic filter, are presented and discussed. Giving COD removals higher than 80% under the whole range of conditions tested, the anaerobic filter demonstrated not only a great ability of biomass to adapt itself to a new carbon source but also an excellent capability to deal with organic load fluctuations and to utilise dilute feeds. Sampling at different filter heights demonstrated that the biogas development ensured mixing within the filter and that most of the organic substances were utilised at the bottom of the reactor, especially when very dilute wastewater was fed. The results of this work could be taken as a starting basis for scaling-up the process to the industrial scale.     

Transition of microbial communities during the adaption to anaerobic digestion of carrot waste

In this study a microbial community suitable for anaerobic digestion of carrot pomace was developed from inocula obtained from natural environmental sources. The changes along the process were monitored using pyrosequencing of the 16S rRNA gene. As the community adapted from a diverse natural community to a community with a definite function, diversity decreased drastically. Major bacterial groups remaining after enrichment were Bacilli (31-45.3%), Porphyromonadaceae (12.1-24.8%) and Spirochaetes (12.5-18.5%). The archaeal population was even less diverse and mainly represented by a single OTU that was 99.7% similar to Methanosarcina mazei. One enrichment which failed to produce large amounts of methane had shifts in the bacterial populations and loss of methanogenic archaea.     

Microbial communities in an anaerobic dynamic membrane bioreactor (AnDMBR) for municipal wastewater treatment: Comparison of bulk sludge and cake layer

454 high-throughput pyrosequencing was employed to investigate microbial communities in bulk sludge (BS) and cake layer (CL) in an anaerobic dynamic membrane bioreactor (AnDMBR). Pyrosequencing produced 7402 and 7026 high-quality V1–V3 tags of the 16S rRNA-gene for the BS and CL samples, and the number of total OTUs estimated by Chao1 was 2906 (BS) and 2532 (CL) at a 3% distance. Phylogenetic spectrum and Venn diagrams revealed that the bacterial phyla in the two samples were nearly identical, and that the predominant microorganisms of them exhibited a high homology. Nevertheless, population distribution was different between them. Proteobacteria were the most dominant in the BS, while Bacteroidetes, Firmicutes and Spirochaetes were highly enriched in the CL, accounting for 57.6% of the total microbes. The main microbial groups in the CL were highly functionally organized. Bacteroides, Clostridia and Delta-proteobacteria were likely the pioneers for surface colonization on membranes in the AnDMBR.