间歇好氧硫酸盐废水处理系统微生物区系解析

应用PCR-DGGE(polymerase chain reaction-denaturing gradient gel electrophoresis)技术和16S rDNA序列测定对间歇好氧硫酸盐废水处理工艺的微生物群落结构进行了研究.采集味精厂好氧池原始污泥以及实验室内间歇好氧工艺驯化后不同条件下的活性污泥样品,通过基因组DNA的提取、PCR扩增和DGGE分离,初步分析了各污泥样品的微生物群落多样性,结果表明,PCR-DGGE方法可以在一定程度上反映工艺以及操作条件对微生物群落结构的影响.通过DGGE反复分离纯化及割胶回收,DGGE检验为单一条带后进行测序并提交到GenBank数据库比对,结果表明,间歇好氧硫酸盐系统中优势菌株大多数为未培养细菌,来源于不同的污染环境,具有重要污染物降解的生态功能,其中包括与硫酸盐还原菌(Desulfobulbus propionicus)在系统发育上非常接近的菌株.     

环境微生态研究中的分子生物学新技术

随着分子生物学的发展,可用于环境微生态研究的新方法新手段不断出现.利用这些分子生物学技术,可直接对环境样品的总DNA进行分析,绕开菌株分离和培养瓶颈,可以最大限度地获得相关微生物的遗传信息,全面地分析环境中微生物的多样性.对目前广泛应用于环境微生态研究的分子生物学新技术,包括rDNA基因序列分析、变性梯度凝胶电泳、温度梯度凝胶电泳、单链构象多态性、限制性片段长度多态性、随机扩增多态性DNA、核酸杂交和DNA芯片等技术进行了综述.     

河流沉积物中微生物DNA的提取方法比较研究

提取河流沉积物中DNA是研究河流沉积物微生物多样性的难点之一,实验开发了一种酚与SDS的DNA提取方法,与OMEGA和BioDee 2种DNA提取试剂盒方法进行了比较,并通过DNA纯度、PCR扩增结合16S rDNA克隆文库检验对几种方法进行了评价.结果表明,OMEGA DNA提取试剂盒法方法简单,提取的DNA纯度较高...     

醌指纹法及其在环境微生物领域的应用

微生物在环境中的丰度和多样性受到广泛关注.系统介绍了一种通过分析微生物菌群中电子传递体的种类和数量来表征其群落丰度和多样性的方法——醌指纹法.综述了醌指纹法的原理和检测分析手段,归纳了醌指纹法在废水处理、水体、土壤和堆肥环境场景中的应用,此外还分析了醌指纹法的优缺点.最后,对醌指纹法在未来微生物治理领域的应用进行了展望...     

PCR-DGGE技术在水解酸化-缺氧法处理采油废水的微生物研究中的应用

采用PCR-DGGE技术直接从水解酸化和缺氧反应器中的污泥样品提取DNA,测定部分菌种的16S rDNA V3区片段序列,通过NCBI基因库比对,初步确定不同生物反应器内优势菌种,并进行了多样性指数分析.结果表明,水解酸化反应器中的生物膜与缺氧反应器中悬浮污泥微生物种群结构存在较大的差异,显示了在不同环境条件下,微生物群落结构的连续动态变化过程.     

荧光原位杂交技术在环境微生物生态学解析中的应用研究

近年来 ,诸多国家在环境微生物领域先后开展了分子生物学研究方法的建立和生物学评价工作。一些不依靠纯培养的微生物群落的分析方法已得到广泛应用和发展。荧光原位杂交 (FISH)技术 ,具有细胞在测定过程中不被破坏、形状不改变、特异性强、能够真实反映在自然环境下微生物的情况及分布等特点 ,在环境微生物群落探测分析中已逐渐被广泛应用。该技术利用带有荧光标记的特异性寡核苷酸探针 ,与细胞内相应的靶核糖体结合 ,能将微生物探测、鉴定到属和种的水平。运用于硝化细菌、除磷细菌和丝状微生物等废水处理中常见的特征性微生物种群和群落生态学研究中 ,颇为高效。该技术的应用避免了传统培养方法进行鉴定和计数的局限性 ,在环境微生物生态学解析中具有较高应用价值。     

荧光原位杂交技术在环境微生物生态学解析中的应用研究

近年来，诸多国家在环境微生物领域先后开展了分子生物学研究方法的建立和生物学评价工作。一些不依靠纯培养的微生物群落的分析方法已得到广泛应用和发展。荧光原位杂交(FISH)技术，具有细胞在测定过程中不被破坏、形状不改变、特异性强、能够真实反映在自然环境下微生物的情况及分布等特点，在环境微生物群落探测分析中已逐渐被广泛应用。该技术利用带有荧光标记的特异性寡核苷酸探针，与细胞内相应的靶核糖体结合，能将微生物探测、鉴定到属和种的水平。运用于硝化细菌、除磷细菌和丝状微生物等废水处理中常见的特征性微生物种群和群落生态学研究中，颇为高效。该技术的应用避免了传统培养方法进行鉴定和计数的局限性，在环境微生物生态学解析中具有较高应用价值。     

真菌胞外聚合物及其与重金属作用机制研究进展

真菌胞外聚合物(EPS)是近年来真菌微生物研究的热点,其在食品行业、医药工业和环境重金属污染治理中起着非常重要的作用.综述了国内外关于真菌EPS提取方法、产量影响因素、组成和结构的研究进展;介绍了真菌EPS在环境中与重金属作用机制的研究进展,主要包括胞外吸附和络合机制、草酸盐微沉淀机制和胞外重金属还原机制.在此基础上,展望了真菌EPS在环境重金属污染治理特别是重金属废水处理上的应用前景和亟待解决的研究内容,以便更好地为国内真菌EPS和重金属污染治理等方面的研究提供参考.     

废水处理污泥中邻苯二甲酸酯降解菌的多样性研究

系统探究了常规废水处理污泥对典型邻苯二甲酸酯(PAEs)的微生物群落响应特征和污泥中PAEs降解菌的多样性。结果表明:(1)废水处理污泥经过10d培养后,对1 000mg/L的邻苯二甲酸二丁酯(DBP)和邻苯二甲酸二异辛酯(DEHP)的降解率分别可达99.9%和83.6%。DEHP的降解速率低于DBP。(2)DBP和DEHP的添加显著降低了废水处理污泥的微生物多样性;DBP、DEHP降解菌富集培养后污泥物种组成相似性较高,且均与废水处理污泥差异较大。(3)废水处理污泥中存在多种类型PAEs降解菌。高浓度DBP和DEHP的存在促使具有DBP/DEHP降解能力的微生物群落不断增长,进而成为优势微生物,最终达到DBP和DEHP的有效降解。     

活性污泥DNA样品的保存条件对聚磷菌FQ-PCR结果的影响

对目标微生物的准确定量是研究微生物群落的组成与功能的重要前提。荧光定量PCR技术作为目前流行的核酸定量分析技术,具有操作时间短,敏感性高,特异性强等特点,但也存在由客观因素导致实验结果出现显著误差的缺点。针对目前环境微生物定量方法中容易忽略的污泥样品DNA保存步骤与上机时间等环节,详细探讨了保存条件、上机批次对聚磷菌（PAOs）荧光定量PCR结果的影响。结果显示保存温度、保存时间均对标准质粒和污泥DNA样品中PAOs定量结果产生影响：模板DNA的浓度及保存时间会影响标准曲线Ct值的重复性;同一样品在相同保存温度（4℃）、不同保存时间、不同批次条件下的定量结果存在显著性差异（PPP>0.05）。因此,在对活性污泥样品进行定量分析时,DNA样品应平行保存于-20℃条件,尽量避免反复冻融,并确保对比的生物样品保存及上机条件完全一致,从而减少定量结果误差。     

一种活性污泥总DNA提取方法的优化

对3种活性污泥DNA提取方法———传统蛋白酶K-SDS-氯仿异戊醇法(CPSCI法)、液氮研磨法和脱腐处理法进行了对比,并针对CPSCI法从污泥量、保温时间、裂解方式及沉淀时间等4个方面进行了优化。结果表明,优化的蛋白酶K-氯仿-异戊醇法(OPSCI法)采用污泥量0.10 g、37℃静置10 min,加SDS常温旋涡振荡及异丙醇直接离心等条件可获得长度在23.1 kb左右的DNA,OD260nm/OD280nm为1.86,稀释10倍后即可进行16S rDNA PCR。该方法重复性好,提取得率高,纯度好,操作简便,为常规实验室开展活性污泥微生物多样性研究提供了帮助。     

应用于PCR-DGGE分析的活性污泥微生物总DNA提取方法比较

探讨适用于PCR-DGGE分析研究的活性污泥细菌和真菌的DNA提取方法。采用5种方法提取活性污泥微生物基因组DNA,以DNA纯度、含量、片段大小及DGGE条带多样性作为考察指标评价提取方法的优劣,以确定最佳实验方案。紫外吸收法和琼脂糖凝胶电泳结果显示,试剂盒法提取的DNA含量最低,其余4种方法获得的DNA含量无显著差异,就DNA纯度而言,试剂盒法最优;除高温裂解法对真菌细胞壁裂解效果较差外,其他4种方法均能不同程度地裂解细菌和真菌细胞;DGGE结果表明,高温裂解法获得的细菌条带最多,基于SDS的细胞裂解法得到的真菌条带最多。综合分析,高温裂解法更适合于活性污泥中细菌的PCR-DGGE分析,基于SDS的细胞裂解法则更适合于污泥中真菌的PCR-DGGE分析。     

活性污泥胞外聚合物提取方法研究

考察4种方法(蒸气法、EDTA法、NaOH法和搅拌法)对3种活性污泥(可乐厌氧活性污泥、可乐好氧活性污泥和市政污水污泥)胞外聚合物的提取效率,对比分析了各提取方法对不同样品提取所得胞外聚合物的组成影响,结果表明,EDTA法提取多糖的效率最高,提取市政污水污泥、可乐好氧活性污泥和可乐厌氧活性污泥胞外聚合物糖分别在8.3、16.7和9.7 mg/g VSS左右;NaOH提取蛋白质的效率最高,提取市政污水污泥、可乐好氧活性污泥和可乐厌氧活性污泥胞外聚合物蛋白质分别为:3.15、0.55和5.57 mg/g VSS.对于3种不同种类污泥,EDTA法提取EPS总量效率最高,分别为市政污水污泥10.7 mg/g VSS、可乐好氧活性污泥16.8 mg/g VSS和可乐厌氧活性污泥9.97 mg/g VSS,分别比效率最低的搅拌法高出5倍、5倍和3倍.     

不同萃取方法对长江口边滩多环芳烃萃取效果的影响

选取索氏提取法和加速溶剂萃取仪(ASE)提取法,对长江口边滩样品进行多环芳烃萃取实验,并利用气相色谱/质谱仪对萃取出来的多环芳烃进行定量分析,研究了不同萃取方法以及不同溶剂条件下多环芳烃的总量、分布特征.结果表明,长江口边滩多环芳烃最高值出现在吴淞口;索氏提取法在多环芳烃萃取效果上优于加速溶剂萃取仪提取法;混合溶剂萃取...     

Lead removal from foundry waste by solvent extraction

Solvent extraction is used to reduce lead concentrations from millpond wastewater solids, a type of foundry process waste. Toluene and toluene mixed with di-(2-ethyl-hexyl) phosphoric acid (HDEHP) have been tried as leaching solvents. Toluene is ineffective as a solvent in extracting lead, but the toluene-HDEHP mixture effectively removes lead from solid foundry waste. The effects of the HDEHP concentration, the contact time, and the amount of solvent used on lead extraction have been investigated. The mass transfer process is rapid: contact time of 1/2 hour has been found to be sufficient to accomplish the leaching process. The concentration of HDEHP significantly impacts lead removal. The optimum concentration of HDEHP is determined to range from 0.05 to 0.1 mol/l. The Toxicity Characteristic Leaching Procedure (TCLP) test of the treated samples gives leachable lead in much lower quantities than those found in the untreated samples. Thus the solvent extraction process appears to be an effective method to significantly reduce the lead content of millpond wastewater solids.     

Extraction of arsenic in a synthetic arsenic-contaminated soil using phosphate

An environment-friendly and cost-effective extraction method has been studied for the removal of arsenic from contaminated soil. A yellow-brown forest soil was contaminated with arsenic(V) and used as a model soil. Among various potassium and sodium salts, potassium phosphate was most effective in extracting arsenic, attaining more than 40% extraction in the pH range of 6–8 with minimum damage to the soil properties. Exchange mechanism is proposed for the extraction of arsenic from soil by phosphate. Sequential extraction shows that phosphate is effective in extracting arsenic of Al- and Fe-bound forms. Arsenic of residual form was not extracted. Arsenic was efficiently extracted by phosphate solution of pH 6.0 at 300 mM phosphate concentration and at 40°C.     

砷形态分析中的样品前处理方法

本文对近 10多年来砷形态分析中样品的前处理技术及其在不同样品中的应用进行了总结和评述 ,指出 ,断续低功率微波辅助提取、经典索氏萃取和纯乙醇低温冷冻离心提纯这 3种砷化物前处理技术较其他方法更具优势     

Application of ultrasonic assisted extraction of chemically diverse organic compounds from soils and sediments

The objective of this research was the development, optimization, and demonstration of an ultrasonic assisted extraction (UAE) based method for organic anthropogenic waste indicators (AWIs) with a range of physicochemical properties from soil and sediment samples. The optimized method was designed to be cost effective compared to existing extraction methods, which may require large quantities of consumables, produce substantial volumes of organic waste, or require costly instrumentation or equipment. Reagent grade sand, soil collected from the native grassland in proximity to Eastern Washington University (EWU), and sediment samples collected from the Spokane river were used as sample matrices during method development. These matrices were fortified with eight AWIs of varying chemical properties that are representative of a variety of household, industrial, and agricultural sources. The recoveries of the AWIs spiked onto these matrices were determined in the extracts using gas chromatography/mass spectrometry (GC/MS). These values reflect the efficiency of the method for extraction of these analytes from representative environmental matrices. Recoveries ranged from 46.1% to 110% in the fortified soil and from 49.2% to 118.6% in the fortified sediment samples, which is comparable with existing methods for the study analytes. The optimized method was then used to quantify AWIs in a biosolid-amended soil. Indole and p-cresol were detected in the biosolid-amended soil.     

Extraction of organic contaminants from marine sediments and tissues using microwave energy.

In this study, we compared microwave solvent extraction (MSE) to conventional methods for extracting organic contaminants from marine sediments and tissues with high and varying moisture content. The organic contaminants measured were polychlorinated biphenyl (PCB) congeners, chlorinated pesticides, and polycyclic aromatic hydrocarbons (PAHs). Initial experiments were conducted on dry standard reference materials (SRMs) and field collected marine sediments. Moisture content in samples greatly influenced the recovery of the analytes of interest. When wet sediments were included in a sample batch, low recoveries were often encountered in other samples in the batch, including the dry SRM. Experiments were conducted to test the effect of standardizing the moisture content in all samples in a batch prior to extraction. SRM1941a (marine sediment). SRM1974a (mussel tissue), as well as QA96SED6 (marine sediment), and QA96TIS7 (marine tissue), both from 1996 NIST Intercalibration Exercise were extracted using microwave and conventional methods. Moisture levels were adjusted in SRMs to match those of marine sediment and tissue samples before microwave extraction. The results demonstrated that it is crucial to standardize the moisture content in all samples, including dry reference material to ensure good recovery of organic contaminants. MSE yielded equivalent or superior recoveries compared to conventional methods for the majority of the compounds evaluated. The advantages of MSE over conventional methods are reduced solvent usage, higher sample throughput and the elimination of halogenated solvent usage.     

Comparative study of extraction methods for the determination of PAHs from contaminated soils and sediments

The following four methods were compared on the extraction efficiency of 16 EPA (US Environmental Protection Agency) polycyclic aromatic hydrocarbons (PAHs): German method of the Verband Deutscher Landwirtschaftlicher Untersuchungs und Forschungsanstalten (VDLUFA), two methods of the International Organization for Standardization using shaking (ISO A) and Soxhlet extraction (ISO B) and an ultrasonic method. Recovery rates of 16 PAHs were determined in two soils. Extraction efficiency was evaluated in five soils and three sediments. Effect of drying soils and sediments on extraction efficiency was tested using the VDLUFA and the ultrasonic methods. Our study shows that the number of aromatic rings, rather than extraction procedures, significantly influenced recovery rates of individual PAHs. No significant differences in extraction efficiency of the four methods were observed for less polluted samples. For highly polluted soils, extraction efficiency decreased in the following order: VDLUFA method > ISO A > ultrasonic method > ISO B. Influence of soil moisture on extraction efficiency depended to some extent on both solvent used and content of PAHs in samples. A mixture of dichloromethane/acetone (5:1) is recommended for PAH extraction from moist samples when the ultrasonic method is used.