成熟厌氧颗粒污泥的结构及其特征*

厌氧序批式反应器〈AnaerobicSequencingBatchReactorASBR〉在处理啤酒废水过程中 ,能形成厌氧颗粒污泥。文中采用扫描电子显微镜和荧光显微镜技术对成熟厌氧颗粒污泥的结构及微生物群落等进行跟踪观察 ,结果显示 ,颗粒污泥结构复杂 ,细菌以微群落形式分布 ,其中产甲烷菌占一定比例。同时也探讨了厌氧颗粒污泥的形成机制。     

对降解喹啉的厌氧生物反应器中重要功能菌群的鉴定

通过把微生物区系组成的分子水平的动态变化情况与微生物群落的整体功能变化相关联,鉴定重要的功能类群是微生物分子生态学研究的一个重要的策略.应用分子生物学的方法,对一个实验室规模的用于降解喹啉的厌氧反应器生物膜样品的微生物区系组成变化进行解析,找出可能的主要功能菌.通过DGGE对反应器的种子污泥和运行稳定的厌氧生物膜反应器的微生物区系组成进行了对比分析,并对主要的优势条带进行了分子鉴定.同时对以上两个样品构建16S rDNA克隆文库,通过统计学分析对克隆文库的有效性进行验证,并对文库进行测序分析.DGGE条带及克隆文库的序列分析均表明,在驯化过程中,Gamma Proteobacteria亚纲与Desulfobacter postgatei种的微生物显著增加,这种动态变化表明这些细菌可能是在厌氧条件下对喹啉的降解起关键作用的微生物.     

厌氧反应器中颗粒污泥的培育及应用研究进展*

厌氧生物处理技术因其具有有机负荷高、污泥产量低、能耗低等优点被广泛应用于各种废水处理中。厌氧颗粒污泥具有沉降性能好、微生物浓度高、有机负荷高等优点,极大地提高了废水处理效率。尤其在处理含高氨氮废水中,厌氧颗粒污泥的形成对反应器的高效生物脱氮至关重要。但到目前为止,厌氧反应器中的颗粒污泥形成及废水处理效果还缺乏系统的认识。鉴于此,总结了厌氧反应器中颗粒污泥的形成机制,分析了影响厌氧反应器中颗粒污泥形成的因素,论述了厌氧反应器中厌氧颗粒污泥生长的模拟,最后介绍了厌氧颗粒污泥在国内外的主流应用。厌氧反应器中颗粒污泥的形成是综合因素影响的结果,对影响厌氧颗粒污泥形成的每个因素都需要认真对待,可为在厌氧反应器中颗粒污泥的培育和应用提供理论指导和技术支撑。     

河北承德地区两个温泉中细菌的多样性分析

通过构建16S rDNA克隆文库,对承德地区两温泉中的细菌多样性水平及系统发育关系进行了初步研究。研究表明:68°C的A11文库中阳性克隆的16S rDNA序列分属5个细菌类群,分别为Firmicutes(6.25%)、Deinococcus-Thermus(25.0%)、Gammaproteobacteria(12.5%)、Betaproteobacteria(50.0%)、Alphaproteobacteria(6.25%);而74.5°C的A12文库仅属于一个细菌类群:厚壁菌门(Firmicutes)。两温泉中细菌多样性的差异表明,温度是影响温泉中细菌多样性水平的重要因素。此外,A11文库中克隆的16S rDNA序列与许多已知的可产色素的好氧菌相似性很高,而A12文库中的细菌多数为专性厌氧或兼性厌氧型,其中厌氧芽孢杆菌属(Anoxybacillus)中的Anoxybacillus flavithermus可以作为研究泉华形成的理想材料。     

有机污染物对水体真细菌群落结构的影响

为了揭示有机污染物对环境真细菌组成和多样性的影响,应用末端限制性片段长度多态性(tRFLP)和16S rDNA文库技术并结合水质分析方法,比较分析了松花江流域内受不同程度有机污染的4个水体及其沉积物中真细菌的群落结构。tRFLP分析表明各水体及底泥均呈现较为复杂的群落结构模式,不同底泥群落形成的末端限制性片段(TRF)图谱具有很高的相似性,但随着污染程度的加强,部分类群明显富集,而且在水样组和泥样组内,群落结构的相似性同水质相似性是一致的,主成分分析(PCA)显示水样和泥样中的真细菌TRF形成不同的群。16SrDNA文库分析表明松花江哈尔滨段底泥中真细菌分布于10个门,Proteobacteria门占优势,达群落总数的21.92%(β-Proteobacteria亚门占10.96%),而有机染污物严重超标的生活污水排污道底泥中的微生物多样性较低,分布于7个门,Proteobacteria门为优势群,占群落的47.37%(α-Proteobacteria亚门占21.05%,δ/ε-Proteobacteria亚门占15.79%)。该研究表明向水体中长期排放高浓度有机物能使系统中微生物群落多样性降低,与污染物降解相关的功能微生物类群明显富集。     

16S rDNA克隆文库方法分析太岁样品中细菌的多样性

通过构建16S rDNA克隆文库的方法,分析太岁样品中细菌的群落结构及多样性。太岁样品中的细菌归属于4个门9个目,优势类群依次是芽胞杆菌目(Bacillales,33.01%)、柄杆菌目(Caulobacterales,32.04%)和伯克霍尔德氏菌目(Burkholderiales,12.62%);优势属为短波单胞菌属(Brevundimonas,30.10%)、葡萄球菌属(Staphylococcus,29.13%)和食酸菌属(Acidovorax,7.77%)。并且其中的5个目中含有未培养的细菌,红杆菌目(Rhodobacterales)、伯克霍尔德氏菌目和红环菌目(Rhodocyclales)的11个克隆子的细菌16S rDNA序列同源性低于97%。研究表明太岁样品中细菌多样性较丰富,且蕴藏着许多未知的微生物资源。     

颗粒污泥的产甲烷细菌及结构模型初探

本文报道了颗粒污泥中不同形态的产甲烷细菌及其分布特征。电镜观察表明,颗粒表层主要是氢营养型产甲烷菌,如产甲烷短杆菌、产甲烷螺菌等。细菌的分布具有一定的“区位化”。颗粒内层则主要是乙酸营养型产甲烷菌,其中产甲烷丝菌是优势种群,并且与产氢产乙酸细菌之间存在互营共生关系。根据对产甲烷细菌的观察研究结果,作者提出了一个颗粒污泥的初步结构模型,同时对此进行了分析讨论。     

氯苯对EGSB反应器内颗粒污泥性质影响的研究

采用ECSB反应器处理含氯苯有机废水,主要研究了氮苯对颗粒污泥性质的影响。结果表明：氮苯对处理葡萄糖自配水的EGSB反应器内颗粒污泥中的细菌有较强毒害作用,连续投加低浓度氮苯72d后,扫描电镜观察可发现颗粒污泥表面和内部细菌均明显受到损害,停止投加氮苯恢复运行30d和50d后,仍可观察到颗粒污泥内部细菌受损害的现象,且部分颗粒污泥内部存在着明显的空洞;随着运行时间的延长,反应器内颗粒污泥的粒径有较大程度的增大;但长期接触氯苯导致部分颗粒污泥解体,使得小粒径污泥增多,而大粒径污泥相应减少;氮苯对颗粒污泥的损害还表现在使大粒径颗粒的沉速减小,甚至导致部分颗粒污泥内部形成空洞而上浮。     

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。     

厌氧氨氧化颗粒污泥研究进展

厌氧氨氧化(Anaerobic ammonium oxidation,Anammox)工艺是一种新的生物脱氮技术。一经问世即得到人们青睐,现已成为废水脱氮的升级技术。厌氧氨氧化菌(Anaerobicammoniumoxidation bacteria,AnAOB)是Anammox工艺的功能之源。以颗粒污泥形态存在的AnAOB是Anammox颗粒污泥床脱氮系统的重要支柱。由于AnAOB生长缓慢且对环境条件变化敏感,Anammox脱氮系统不仅启动缓慢,而且运行极易失稳甚至崩溃。值得庆幸的是,AnAOB可自主选择、组合和固定功能菌群落而形成Anammox颗粒污泥,并通过其优良的重力沉降性能和高效的基质转化性能保障Anammox脱氮系统的持续工作。本文综述了AnAOB的种类和特性及Anammox颗粒污泥的组成、结构和功能,以期为Anammox工艺的优化和拓展提供参考。     

Cluster Structure of Anaerobic Aggregates of an Expanded Granular Sludge Bed Reactor

The metabolic properties and ultrastructure of mesophilic aggregates from a full-scale expanded granular sludge bed reactor treating brewery wastewater are described. The aggregates had a very high methanogenic activity on acetate (17.19 mmol of CH₄/g of volatile suspended solids [VSS]·day or 1.1 g of CH₄ chemical oxygen demand/g of VSS·day). Fluorescent in situ hybridization using 16S rRNA probes of crushed granules showed that 70 and 30% of the cells belonged to the archaebacterial and eubacterial domains, respectively. The spherical aggregates were black but contained numerous whitish spots on their surfaces. Cross-sectioning these aggregates revealed that the white spots appeared to be white clusters embedded in a black matrix. The white clusters were found to develop simultaneously with the increase in diameter. Energy-dispersed X-ray analysis and back-scattered electron microscopy showed that the whitish clusters contained mainly organic matter and no inorganic calcium precipitates. The white clusters had a higher density than the black matrix, as evidenced by the denser cell arrangement observed by high-magnification electron microscopy and the significantly higher effective diffusion coefficient determined by nuclear magnetic resonance imaging. High-magnification electron microscopy indicated a segregation of acetate-utilizing methanogens (Methanosaeta spp.) in the white clusters from syntrophic species and hydrogenotrophic methanogens (Methanobacterium-like and Methanospirillum-like organisms) in the black matrix. A number of physical and microbial ecology reasons for the observed structure are proposed, including the advantage of segregation for high-rate degradation of syntrophic substrates.     

Analysis of Microbial Community during Biofilm Development in an Anaerobic Wastewater Treatment Reactor

The formation, structure, and biodiversity of a multispecies anaerobic biofilm inside an Upflow Anaerobic Sludge Bed (UASB) reactor fed with brewery wastewater was examined using complementary microbial ecology methods such us fluorescence in situ hybridization (FISH), denaturing gradient gel electrophoresis (DGGE), and cloning. The biofilm development can be roughly divided into three stages: an initial attachment phase (0-36 h) characterized by random adhesion of the cells to the surface; a consolidation phase (from 36 h to 2 weeks) defined by the appearance of microcolonies; and maturation phase (from 2 weeks to 2 months). During the consolidation period, proteobacteria with broad metabolic capabilities, mainly represented by members of alpha-Proteobacteria class (Oleomonas, Azospirillum), predominated. Beta-, gamma-, delta- (both syntrophobacteria and sulfate-reducing bacteria) and epsilon- (Arcobacter sp.) Proteobacteria were also noticeable. Archaea first appeared during the consolidation period. A Methanospirillum-like methanogen was detected after 36 h, and this was followed by the detection of Methanosarcina, after 4 days of biofilm development. The mature biofilm displayed a hill and valley topography with cells embedded in a matrix of exopolymers where the spatial distribution of the microorganisms became well-established. Compared to the earlier phases, the biodiversity had greatly increased. Although alpha-Proteobacteria remained as predominant, members of the phyla Firmicutes, Bacteroidete, and Thermotogae were also detected. Within the domain Archaea, the acetoclastic methanogen Methanosaeta concilii become dominant. This study provides insights on the trophic web and the shifts in population during biofilm development in an UASB reactor.     

厌氧颗粒污泥的超微结构分析

目的：通过透射电镜对厌氧污泥颗粒内部结构进行系统的分析,研究其中微生物的生态情况及分布特征。方法：对颗粒的表面、1／3、1／2等特征部位处进行超薄切片,在透射电镜下观察并分析。结果：厌氧颗粒污泥由多种微生物但主要是丝状菌、杆状菌和球状菌组成。这些菌群整体上以混栖分布的形式存在,菌体密度从外向内看由密变疏,呈递减式的梯度分布。结论：厌氧颗粒污泥是一个微生态系统,不同类型的细菌种群在系统中相互依存,形成互营共生体系,有利于细菌对有机物的降解。透射电镜下对于污泥颗粒内不同层面中的菌群生态及分布情况的观察非常直观,十分有利于了解各菌群之间的生态关系。因此,采用透射电镜技术是研究厌氧污泥颗粒一种好方法。     

Contact Angle Measurement and Cell Hydrophobicity of Granular Sludge from Upflow Anaerobic Sludge Bed Reactors

The contact angle, which is generally used to evaluate the hydrophobicities of pure bacterial strains and solid surfaces, was used to study mixed cell cultures of bacteria involved in anaerobic digestion. Previously published data and data from this study showed that most acidogens are hydrophilic (contact angle, <45(deg)) but most of the acetogens and methanogens isolated from granular sludge are hydrophobic (contact angle, >45(deg)). The hydrophobicities of mixtures of hydrophilic and hydrophobic cells were found to be linearly correlated with the cell mixing ratio. The hydrophobicities of cells present in effluents from upflow anaerobic sludge bed reactors which were treating different types of substrates were different depending on the reactor conditions. When the reactor liquid had a high surface tension, cells sloughing off from sludge granules, as well as cells present on the outer surfaces of the granules, were hydrophobic. Short-term batch enrichment cultures revealed that proteins selected for highly hydrophilic cells. Long-term in-reactor enrichment cultures revealed that sugars selected for hydrophilic acidogens on the surfaces of the granules, while fatty acids tended to enrich for hydrophobic methanogens. When linear alkylbenzenesulfonate was added, the cells on the surfaces of granules became more hydrophilic. Control tests performed with pure cultures revealed that there was no change in the surface properties due to linear alkylbenzenesulfonate; hence, the changes in the wash-out observed probably reflect changes in the species composition of the microbial association. A surface layer with moderate hydrophobicity, a middle layer with extremely high hydrophobicity, and a core with high hydrophobicity could be distinguished in the grey granules which we studied.     

Hydrophobicities and Electrophoretic Mobilities of Anaerobic Bacterial Isolates from Methanogenic Granular Sludge

The hydrophobicities and electrophoretic mobilities of isolates from methanogenic anaerobic granular sludge were measured and compared with those of strains from culture collections. All new isolates were highly hydrophobic, indicating that the upflow anaerobic sludge blanket reactor concept selects for hydrophobic bacteria. Methanothrix soehngenii, a methanogen often observed in methanogenic granular sludge, was highly hydrophobic and showed low electrophoretic mobility at pH 7. The role of this strain in the formation of methanogenic granular sludge is discussed.     

造纸废水活性污泥的微生物群落结构及多样性分析

为探究造纸废水活性污泥中微生物群落结构多样性以及对造纸废水处理效果的影响，利用Illumina MiSeq 高通量测序方法，分析在处理造纸废水过程中，同一运行阶段两个并联氧化沟内活性污泥的微生物群落与多样性组成。结果表明，系统中处理造纸废水的活性污泥在同一废水条件下微生物群落结构总体稳定，优势细菌为绿弯菌门(Chloroflexi)、拟杆菌门（Bacteroidota）、变形菌门（Proteobacteria）、Myxococcota、放线菌门(Actinobacteria）、厚壁菌门(Firmicutes）等。最重要的优势细菌类群为Chloroflexi，相对丰度占比为47.67%~48.22%，远远高于其他废水中Chloroflexi的占比，其中厌氧绳菌纲（Anaerolineae）是其主要成员，占比84.39%~88.34%，可针对性地去除造纸废水中的污染物。造纸废水活性污泥样品中存在大量特殊功能菌群，其在废水中污染物尤其是木质素的去除中发挥着重要作用。     

Phenotypic Properties and Microbial Diversity of Methanogenic Granules from a Full-Scale Upflow Anaerobic Sludge Bed Reactor Treating Brewery Wastewater

Methanogenic granules from an anaerobic bioreactor that treated wastewater of a beer brewery consisted of different morphological types of granules. In this study, the microbial compositions of the different granules were analyzed by molecular microbiological techniques: cloning, denaturing gradient gel electrophoresis and fluorescent in situ hybridization (FISH), and scanning and transmission electron microscopy. We propose here that the different types of granules reflect the different stages in the life cycle of granules. Young granules were small, black, and compact and harbored active cells. Gray granules were the most abundant granules. These granules have a multilayer structure with channels and void areas. The core was composed of dead or starving cells with low activity. The brown granules, which were the largest granules, showed a loose and amorphous structure with big channels that resulted in fractured zones and corresponded to the older granules. Firmicutes (as determined by FISH) and Nitrospira and Deferribacteres (as determined by cloning and sequencing) were the predominant Bacteria. Remarkably, Firmicutes could not be detected in the brown granules. The methanogenic Archaea identified were Methanosaeta concilii (70 to 90% by FISH and cloning), Methanosarcina mazei, and Methanospirillum spp. The phenotypic appearance of the granules reflected the physiological condition of the granules. This may be valuable to easily select appropriate seed sludges to start up other reactors.     

内循环颗粒污泥床硝化反应器流动模型研究

采用脉冲刺激响应技术,对稳态内循环颗粒污泥床硝化反应器进行了示踪试验。根据试验结果,分别运用轴向扩散模型和多釜全混流反应器串联模型,对反应器沉淀区和循环区的流态进行了分析和判断。结果表明,反应器沉淀区的分散数D/uL为0.00148,该区域的流态接近于平推流反应器（PFR）;反应器循环区的串联级数为1.021,该区域的流态接近于全混流反应器（CSTR）。稳态时,反应器的理论水力停留时间为360min,实际水力停留时间为341.2min,反应器中死区所占的体积百分比为5.22％,其中生物体死区为0.75%,水力死区为4.47%,表明反应器结构性能良好。根据试验和分析结果,建立了内循环颗粒污泥床硝化反应器的流动模型,即全混流和平推流的串联组合模型。由流动模型所得的理论停留时间分布曲线与由试验所得的实际停留时间分布曲线吻合良好,两者的平均相对误差为8.56%,表明所建模型具有较高的准确性。