厌氧颗粒污泥微生物群落结构与功能的研究进展

厌氧颗粒污泥(anaerobicgranularsludge,AnGS)是由多种功能微生物组成的自固定化聚集体,具有容积负荷高、工艺简单、剩余污泥产量低等优点,在废水处理领域中显示出巨大的技术和经济潜力,被认为是一种很有前景的低碳废水处理工艺。本文系统总结了近年来厌氧颗粒污泥微生物结构和功能的研究成果,从微生物学角度讨论了厌氧颗粒污泥形成及稳定的影响因素,并对今后厌氧颗粒污泥的研究进行了展望,以期为后续厌氧颗粒污泥技术的深入研究和实际工程应用提供参考。     

Preservation characteristics of anaerobic granular sludge

Summary The preservation characteristics of granular sludges were investigated in terms of storing age and environmental condition. As a preservative index, physicochemical, microbiological, and morphological parameters were measured in order to quantify the change in granular sludge. Results from this research indicated that, as the anaerobic granular sludge had an outstanding preservation characteristic, UASB system could be effectively applied to the industries operating seasonally or intermittently.     

Inorganic composition and microbial characteristics of methanogenic granular sludge grown in a thermophilic upflow anaerobic sludge blanket reactor

An upflow anaerobic sludge blanket reactor was operated under thermophilic conditions (55° C) for 160 days by feeding a wastewater containing sucrose as the major carbon source. The reactor exhibited a satisfactory performance due to the formation of well-settling granulated sludge, achieving a total organic carbon (TOC) removal of above 80% at an organic loading rate of 30 kg total organic C m^–3 day^–1. Structural and microbial properties of the methanogenic granular sludge were examined using scanning electron microscope X-ray analyses and serum vial activity tests. All the thermophilic granules developed showed a double-layered structure, comprised of a black core portion and a yellowish exterior portion. The interior cope portion contained abundant crystalline precipitates of calcium carbonate. Calcium-bound phosphorus was also present more prominently in the core portion than in the exterior portion. Methanogenic activities of the thermophilic granules both from acetate and from H₂ increased with increasing vial-test temperature in the range of 55–65° C [from 1.43 to 2.36 kg CH₄ chemical oxygen demand (COD) kg volatile suspended solids (VSS)^–1 day^–1 for acetate and from 0.85 to 1.11 kg CH₄ COD kg VSS^–1 day^–1 for H₂]. On the other hand, propionate-utilizing methanogenic activity was independent of vial-test temperature, and was much lower (0.1–0.12 kg CH₄ COD kg VSS^–1 day^–1) than that from either acetate or H₂. Acetate consumption during vial tests was considerably inhibited by the presence of H₂ in the headspace, indicating that a syntrophic association between acetate oxidizers and H₂-utilizing methane-producing bacteria was responsible for some portion of the overall acetate elimination by the theromophilically grown sludge.     

Acidification of methanol-fed anaerobic granular sludge bioreactors by cobalt deprivation: Induction and microbial community dynamics

The acidification of mesophilic (30 degrees C) methanol-fed upflow anaerobic sludge bed (UASB) reactors induced by cobalt deprivation from the influent was investigated by coupling the reactor performance (pH 7.0; organic loading rate 4.5 g COD . L(-1) . d(-1)) to the microbial ecology of the bioreactor sludge. The latter was investigated by specific methanogenic activity (SMA) measurements and fluorescence in situ hybridization (FISH) to quantify the abundance of key organisms over time. This study hypothesized that under cobalt limiting conditions, the SMA on methanol of the sludge gradually decreases, which ultimately results in methanol accumulation in the reactor effluent. Once the methanol accumulation surpasses a threshold value (about 8.5 mM for the sludge investigated), reactor acidification occurs because acetogens outcompete methylothrophic methanogens at these elevated methanol concentrations. Methanogens present in granular sludge at the time of the acidification do not use methanol as the direct substrate and are unable to degrade acetate. Methylotrophic/acetoclastic methanogenic activity was found to be lost within 10 days of reactor operation, coinciding with the disappearance of the Methanosarcina population. The loss of SMA on methanol can thus be used as an accurate parameter to predict reactor acidification of methanol-fed UASB reactors operating under cobalt limiting conditions.     

The effect of calcium on the treatment of fresh leachate in an expanded granular sludge bed bioreactor

This research investigated the calcium effect on the anaerobic treatment of fresh leachate in an expanded granular sludge bed (EGSB) bioreactor under mesophilic conditions. The observations show that the bioreactor, inoculated with anaerobic granular sludge, can be started up only in about 40 days for the treatment of calcium-containing fresh leachate with chemical oxygen demand (COD) removal efficiency above 90% and organic loading rate up to 72.84 kg COD/m³ day. The calcium accumulation onto the granules was monotonically related to the calcium concentration, accounting for 17-18 wt.% of Ca in the suspended solid in the form of calcium carbonate, phosphates/phosphonates and carboxylates. The mineral formation significantly increased the granule settling velocity (by ∼50%) and the suspended solid concentration (by ∼100%). However, the effect of calcium precipitation on the specific methanogenic activity and the CH₄ production rate was complex, first positive during the start-up but later on negative.     

Physicochemical characteristics of anaerobic H2-producing granular sludge

Granule-based biological H₂ production processes are gaining great popularity in recent years. An efficient and stable operating of such systems relies heavily on the performance of the H₂-producing granules (HPGs), which possess many unique properties compared with floc sludge and methanogenic granules. Hence, a full understanding of the sludge characteristics is essential. Especially, the physicochemical properties of HPGs may provide useful information for effective evaluation of system status. This review offers a systematical introduction of the physicochemical properties of HPGs, including size, morphology, settling velocity, permeability, rheology, surface charge, hydrophobicity and extracellular polymeric substances (EPS). We also analyze the relationships between these physicochemical factors and the system performance, and discuss the remaining challenges and future implications for sludge characterization and process monitoring. This work may facilitate a better understanding of granule-based biological H₂ production processes and offer a basis for timely process monitoring and manipulation.     

Treatment of fresh leachate with high-strength organics and calcium from municipal solid waste incineration plant using UASB reactor

Treatment of a fresh leachate with high-strength organics and calcium from municipal solid waste (MSW) incineration plant by an up-flow anaerobic sludge blanket (UASB) reactor was investigated under mesophilic conditions, emphasizing the influence of organic loading rate (OLR). When the reactor was fed with the raw leachate (COD as high as 70,390-75,480 mg/L) at an OLR of 12.5 kg COD/(m³ d), up to ∼82.4% of COD was removed suggesting the feasibility of UASB process for treating fresh leachates from incineration plants. The ratio of volatile solids/total solids (VS/TS) of the anaerobic sludge in the UASB decreased significantly after a long-term operation due to the precipitation of calcium carbonate in the granules. Scanning electron microscopy (SEM) observation shows that Methanosaeta-like species were in abundance, accompanied by a variety of other species. The result was further confirmed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and sequencing.     

Temporal microbial diversity changes in solvent-degrading anaerobic granular sludge from low-temperature (15 degrees C) wastewater treatment bioreactors

Anaerobic sludge granules were obtained from laboratory-scale anaerobic bioreactors used to treat pharmaceutical-like (methanol-, acetone- and propanol-contaminated) wastewater under low-temperature conditions (15 degrees C). The microbial diversity and diversity changes of the sludge samples were ascertained by applying 16S rRNA gene cloning and terminal restriction fragment length polymorphism (TRFLP) analyses, respectively, and using sludge samples from the inoculum, throughout and at the conclusion of the bioreactor trial. Data from genetic fingerprinting correlated well with those from physiological activity assays of the reactor biomass. Specifically, for example, TRFLP profiles indicated the dominance of hydrogenotrophic methanogens within the archaeal community, thus supporting the findings of specific methanogenic activity measurements. TRFLP data supported the hypothesis that the deviation between the replicated reactors, in terms of treatment efficiency, was associated with succession within the microbial communities present, and indicated that community development was linked to both operating temperature and wastewater composition. Fluorescence in situ hybridization (FISH) was also applied, to quantitatively assess the abundance of selected microbial groups, and revealed the underestimation of the abundance Methanosarcina by gene cloning analysis and demonstrated the spatial arrangement of these organisms within the architecture of the low-temperature solvent-degrading anaerobic biofilms.     

Long-term effects of multi-walled carbon nanotubes on the performance and microbial community structures of an anaerobic granular sludge system

Multi-walled carbon nanotubes (MWCNTs) released into the sewage may cause negative and/or positive effects on the treatment system. The objective of this study was to explore over 110 days’ effect of MWCNTs on the performance of anaerobic granular sludge and microbial community structures in an upflow anaerobic sludge blanket (UASB) reactor. The results showed that MWCNTs had no significant effect on the removal of chemical oxidation demand (COD) and ammonia in UASB reactor, but the total phosphorus (TP) removal efficiency increased by 29.34%. The biogas production of the reactor did not change. The anaerobic granular sludge tended to excrete more EPS to resist the effects of MWCNTs during the long-term impact. Illumina MiSeq sequencing of 16S rRNA gene revealed that MWCNTs did not affect the microbial diversity, but altered the composition and structure of microbial community in the reactor. In this process, Saccharibacteria replaced Proteobacteria as the highest abundant bacterial phylum. MWCNTs promoted the differentiation of methanogen structure, resulting in increase of Methanomassiliicoccus, Methanoculleus, and the uncultured WCHA1–57. These results indicated that MWCNTs impacted the performance of UASB reactor and the structures of the microbial community in anaerobic granular sludge.

     

Quantitative analysis of a high-rate hydrogen-producing microbial community in anaerobic agitated granular sludge bed bioreactors using glucose as substrate

Fermentative H₂ production microbial structure in an agitated granular sludge bed bioreactor was analyzed using fluorescence in situ hybridization (FISH) and polymerase chain reaction-denatured gradient gel electrophoresis (PCR-DGGE). This hydrogen-producing system was operated at four different hydraulic retention times (HRTs) of 4, 2, 1, and 0.5 h and with an influent glucose concentration of 20 g chemical oxygen demand/l. According to the PCR-DGGE analysis, bacterial community structures were mainly composed of Clostridium sp. (possibly Clostridium pasteurianum), Klebsiella oxytoca, and Streptococcus sp. Significant increase of Clostridium/total cell ratio (68%) was observed when the reactor was operated under higher influent flow rate. The existence of Streptococcus sp. in the reactor became more important when operated under a short HRT as indicated by the ratio of Streptococcus probe-positive cells to Clostridium probe-positive cells changing from 21% (HRT 4 h) to 38% (HRT 0.5 h). FISH images suggested that Streptococcus cells probably acted as seeds for self-flocculated granule formation. Furthermore, combining the inspections with hydrogen production under different HRTs and their corresponding FISH analysis indicated that K. oxytoca did not directly contribute to H₂ production but possibly played a role in consuming O₂ to create an anaerobic environment for the hydrogen-producing Clostridium.     

Effect of sulfate and iron on physico-chemical characteristics of anaerobic granular sludge

This research investigated the effect of the substrate composition (no substrate, glucose, glucose + sulfate or glucose + sulfate + iron) on the physico-chemical characteristics of two different anaerobic granular sludges as a function of time. The sludges were fed batch wise (pH 7, 30 °C) at an organic loading rate of 1.2 g COD l⁻¹ d⁻¹ (0.04 g COD g VSS⁻¹ day⁻¹) for 30 days. The presence of sulfate (COD/sulfate ratio = 1) in the feed of glucose fed anaerobic sludges did not change the physico-chemical characteristics throughout the incubation. In contrast, the presence of iron in the feed (in addition to glucose and sulfate, COD/iron ratio = 1) reduced the protein and carbohydrate content in the SMP and EPS with about 50% after 30 days incubation compared to the other feeding conditions. The sludge grown on glucose + sulfate + iron contained much more iron (+300–500%) and sulfur (+200–350%) than the other incubated sludges both after 14 and 30 days. The higher mineral content (lower VSS content) and the decrease of the EPS content contributed to the disintegration of iron fed granules, as shown by their lower size particles. However, the iron fed sludge displayed a higher granule strength than the other incubated sludges. Although an appreciable variation in the granule strength was noticed between the sludges investigated, it was not possible to relate these differences to their inorganic composition, the chemical composition of the extracted polymers or to the physical characteristics investigated.     

好氧-厌氧混合污泥启动微生物燃料电池产电性能及微生物群落动态特征

【目的】为探讨好氧-厌氧混合污泥启动微生物燃料电池(Microbial fuel cell,MFC)产电性能以及MFC对微生物群落的选择作用,【方法】以乳酸为底物,应用不依赖于培养的微生物分子生物学技术解析单室MFC启动过程中微生物群落的组成和结构动态学特征。【结果】结果表明,MFC经过3个周期启动成功,最高输出电压230 m V。当MFC外电阻为1656Ω时,最大功率密度11.15 W/m3,电池运行稳定。混合污泥启动MFC以后,阳极生物膜微生物群落结构同种泥差异较大,且多样性降低。生物膜中微生物类群按丰度依次为β-变形菌纲(Betaproteobacteria)24.90%、拟杆菌门(Bacteroidetes)21.30%、厚壁菌门(Firmicutes)9.70%、γ-变形菌纲(Gammaproteobacteria)8.50%、δ-变形菌纲(Deltaproteobacteria)7.90%、绿弯菌门(Chloroflexi)4.20%以及α-变形菌纲(Alphaproteobacteria)3.60%。有利于生物膜形成与稳定的动胶菌属(Zoogloea)和不动杆菌属(Acinetobacter)序列丰度分别占生物膜群落的5.00%和3.90%,与MFC产电能力直接相关的地杆菌属(Geobacter)序列由混合污泥中的0.60%上升至阳极生物膜中的2.60%。【结论】本研究表明,MFC阳极生物膜在驯化过程中对污泥中的微生物进行淘汰和选择,最终驯化形成了有利于生物膜形成与稳定、有机物厌氧发酵与产电的微生物菌群。     

Hydrophobic molecular features of EPS extracted from anaerobic granular sludge treating wastewater from a paper recycling plant

1. Download : Download high-res image (244KB)
2. Download : Download full-size image

     

Kinetics and mass-transfer phenomena in anaerobic granular sludge

The kinetic properties of acetate-degrading methanogenic granular sludge of different mean diameters were assessed at different up-flow velocities (V(up)). Using this approach, the influence of internal and external mass transfer could be estimated. First, the apparent Monod constant (K(S)) for each data set was calculated by means of a curve-fitting procedure. The experimental results revealed that variations in the V(up) did not affect the apparent K(S)-value, indicating that external mass-transport resistance normally can be neglected. With regard to the granule size, a clear increase in K(S) was found at increasing granule diameters. The experimental data were further used to validate a dynamic mathematical biofilm model. The biofilm model was able to describe reaction-diffusion kinetics in anaerobic granules, using a single value for the effective diffusion coefficient in the granules. This suggests that biogas formation did not influence the diffusion-rates in the granular biomass.     

Performance of an anaerobic membrane bioreactor in which granular sludge and dynamic filtration are integrated

To alleviate the fouling of a filter, simple substrates, dynamic filtration, and granular sludge were applied in an anaerobic membrane bioreactor (AnMBR). The results showed that under a transmembrane pressure < 20 kPa, the filter flux ranged between 15 and 20 l (m^?2 h)^?1 for a period of 30 days. The flux was higher than the typical flux of AnMBRs with conventional membranes and most current dynamic filters. In addition, the low cost of the filter avoided the need for a higher flux. Moreover, a stable granular sludge bed, which consumed all volatile fatty acids, was maintained. A compact fouling/filtration layer formed on the filter, which contributed to low effluent chemical oxygen demand concentrations and turbidity. In addition, substrate scarcity in the filtration zone resulted in the evolution of diverse bacteria on the filter.     

Methanogenic and perchloroethylene-dechlorinating activity of anaerobic granular sludge

The biodegradation and toxicity of tetrachloroethylene (C₂Cl₄) and trichloroethylene (C₂HCl₃) were studied with different anaerobic enrichment cultures using the following electron donors: acetate, propionate, butyrate, methanol, formate and hydrogen. All of them sustained dechlorination except propionate, for which C₂Cl₄ biodegradation rates were not significant. The best results were obtained with butyrate. Hydrogen appeared to be a relevant electron donor for dechlorination with the present cultures. In the presence of specific inhibitors such as bromoethanesulphonate or molybdate, a slight inhibition of dechlorination was observed. According to dechlorination kinetics, Monod-type behaviour was observed up to 120 μM C₂Cl₄ or 200 μM C₂HCl₃ with K _s values around 7 μM for both compounds. Dechlorination was partially inhibited at higher concentrations. In contrast, methanogens, or at least methane production, were more sensitive to the presence of chlorinated ethylenes and inhibition of methanogenesis was observed to different extents over all the C₂Cl₄/C₂HCl₃ concentration range tested, even at the lowest concentrations. Received: 17 April 1998 / Received revision: 18 June 1998 / Accepted: 19 June 1998     

厌氧处理低浓度污水颗粒污泥的形成和生物特性研究

用UASB反应器在室温条件下处理模拟低浓度污水,研究了颗粒污泥的形成过程和特性。UASB反应器进水浓度为100～200mgCOD／L,水温为25℃～9℃,60天内形成了成熟的颗粒污泥。颗粒污泥中以丝状菌为主,存在少量球状菌。     

Isolation of hydrogen-producing bacteria from granular sludge of an upflow anaerobic sludge blanket reactor

H₂-producing bacteria were isolated from anaerobic granular sludge. Out of 72 colonies (36 grown under aerobic conditions and 36 under anaerobic conditions) arbitrarily chosen from the agar plate cultures of a suspended sludge, 34 colonies (15 under aerobic conditions and 19 under anaerobic conditions) produced H₂ under anaerobic conditions. Based on various biochemical tests and microscopic observations, they were classified into 13 groups and tentatively identified as follows: From aerobic isolates,Aeromonas spp. (7 strains),Pseudomonas spp. (3 strains), andVibrio spp. (5 strains); from anaerobic isolates,Actinomyces spp. (11 strains),Clostridium spp. (7 strains), andPorphyromonas sp. When glucose was used as the carbon substrate, all isolates showed a similar cell density and a H₂ production yield in the batch cultivations after 12h (2.24–2.74 OD at 600 nm and 1.02–1.22 mol H₂/mol glucose, respectively). The major fermentation by-products were ethanol and acetate for the aerobic isolates, and ethanol, acetate and propionate for the anaerobic isolates. This study demonstrated that several H₂ producers in an anaerobic granular sludge exist in large proportions and their performance in terms of H₂ production is quite similar.     

Anaerobic ammonium oxidation process in an upflow anaerobic sludge blanket reactor with granular sludge selected from an anaerobic digestor

The purpose of this work was to evaluate the development of the anammox process by the use of granular sludge selected from a digestion reactor as a potential seed source in a lab-scale UASB (upflow anaerobic sludge blanket) reactor system. The reactor was operated for approximately 11 months and was fed by synthetic wastewater. After 200 days of feeding with NH₄ ⁺ and NO₂ ⁻ as the main substrates, the biomass showed steady signs of ammonium consumption, resulting in over 60% of ammonium nitrogen removal. This report aims to present the results and to more closely examine what occurs after the onset of anammox activity, while the previous work described the start-up experiment and the presence of anammox bacteria in the enriched community using the fluorescencein situ hybridization (FISH) technique. By the last month of operation, the consumed NO₂ ⁻N/NH₄ ⁺-N ratio in the UASB reactor was close to 1.32, the stoichiometric ratio of the anammox reaction. The obtained results from the influentshutdown test suggested that nitrite concentration would be one key parameter that promotes the anammox reaction during the start-up enrichment of anammox bacteria from granular sludge. During the study period, the sludge color gradually changed from black to red-brownish.