管网生物膜菌株胞外聚合物的提取方法比较

以饮用水管网生物膜样品中一株强成膜能力的菌株Pleomorphomonas oryzae作为研究对象,考察了8种方法(高速离心法、超声法、加热法、EDTA法、H2SO4法、NaOH法、SDS法、甲醛法)对菌株胞外聚合物(EPS)的提取效果,并结合三维荧光光谱(EEM)和傅立叶红外光谱(FTIR)对提取的EPS进行成分分析.结果表明,EDTA法和H2SO4法既能提高EPS的提取效率,提取量分别为64.77 mg.g-1SS和74.43 mg.g-1SS,是离心方法提取量的1.62倍和1.86倍,又不会在提取过程中对菌株细胞造成破坏,是较为理想的EPS提取方法.EEM分析进一步证实,NaOH法对菌株细胞破坏严重,造成EPS成分变化较大.FTIR分析则说明,化学提取方法相较于物理提取方法会引入杂质对组分测定造成干扰.     

阳离子交换树脂法提取一株除铬(Ⅵ)优势菌(Bacillus sp.)的胞外聚合物

采用阳离子交换树脂法对一株除铬优势菌(Bacillussp.)的胞外聚合物(EPS)进行提取,综合考察树脂量、振摇频率、提取时间三个主要因素对EPS提取效率的影响,确定阳离子交换树脂法提取优势菌Bacillussp.的最佳EPS提取条件.试验结果表明:当菌悬液OD为0.833,树脂量20g,振摇频率120r.min-1,提取时间8h时,EPS的提取效果最佳,提取量高达132.15μg.ml-1菌悬液,而DNA提取量仅1.48μg.ml-1菌悬液.     

三维荧光光谱结合化学分析评价胞外多聚物的提取方法

采用高速离心法、EDTA法、阳离子交换树脂法(CER)、甲醛法和甲醛 NaOH法提取普通活性污泥和好氧颗粒污泥的胞外多聚物(EPS),结合三维荧光光谱对五种方法进行评价,并对EPS中的成分进行分析.结果表明,5个EPS的三维荧光光谱峰值中,2个是类蛋白峰,2个是类富里酸峰,1个是类腐殖酸峰.匀浆对好氧颗粒污泥EPS的提取必不可少.甲醛严重干扰核酸的化学测定.透析前后的EPS提取液均应作为分析对象.单纯高速离心法无法有效提取EPS.CER法不容易造成细胞破壁,是一种比较有效的方法.虽然甲醛 NaOH法对EPS的提取量最大,但对细胞的破壁程度也最大.EDTA会导致溶胞,并严重污染EPS.甲醛 NaOH法和EDTA法都会严重干扰胞外蛋白的三维荧光光谱分析.     

莱茵衣藻胞外聚合物的提取和红外光谱表征

胞外聚合物(EPS)是微生物细胞代谢过程中产生的高分子物质,其中的多种官能团通过络合、转化等反应改变周围环境的元素形态和活性,在污染生物修复中具有应用前景.以模式微藻——莱茵衣藻为研究材料,采用加热法、NaOH法、EDTA法、阳离子交换树脂(CER)法、高速离心法提取莱茵衣藻的EPS,比较分析每种方法得到的EPS的含量及其组成成分,并结合LIVE/DEAD BacLight染色和激光共聚焦显微镜观察细胞活性,以确定莱茵衣藻EPS最适宜的提取方法.结果表明,不同提取方法得到的EPS各组分含量有显著差异,5种方法的提取总量依次为NaOH法加热法CER法EDTA法离心法.荧光染色和激光共聚焦显微镜检测结果表明,相对于NaOH法,加热法对微藻细胞的破坏程度较小,且傅里叶红外光谱分析结果表明,加热法提取的EPS出现N-H、C=O或C-N(蛋白质)和C-H、C-O-C、RHC(OH)(OR)(糖类)等吸收峰.本研究表明加热法是莱茵衣藻EPS的最适宜提取方法,EPS的官能团信息sk可为后续研究莱茵衣藻EPS与重金属之间的相互作用奠定基础.     

超声波提取活性污泥胞外聚合物的研究

采用超声波提取活性污泥的胞外聚合物(EPS),以EPS中葡萄糖-6-磷酸脱氢酶(G6PD)活性衡量细胞破损程度.结果表明:超声波的最佳作用频率为21kHz,最佳作用功率范围为32W-40W,提取时间不宜超过3min.低泥龄污泥的EPS易被提取,并且超声波法容易引起低泥龄污泥中细菌细胞的破损.     

微生物胞外聚合物对水体重金属和富营养元素的环境生化效应研究展望

综述了胞外聚合物(Extracellular polymeric substance,EPS)环境生物化学效应已有的研究成果,阐述了EPS对重金属和富营养元素环境生物化学效应的影响过程,指出了研究存在的问题和未来方向.EPS作为微生物分泌的产物,不仅对微生物的生存起到至关重要的作用,还对有毒重金属物质和富营养元素的环境生物化学效应影响显著.目前有关EPS环境效应的研究主要集中在实验室EPS提取、EPS凝絮性和EPS吸附动力学3个方面,而针对EPS与环境重金属和富营养元素的结合状态、EPS对重金属行为归宿的影响以及EPS与污染元素之间的微界面反应过程研究较少.     

活性污泥胞外聚合物的分层组分及溶解性微生物代谢产物的特性

提取了活性污泥胞外聚合物(EPS)和溶解性微生物代谢产物(SMP),并按EPS的紧密程度对松散EPS、紧密EPS的分层组分以及SMP属性进行了分析鉴定.结果表明,以葡萄糖为基质时,松散EPS的主要组成是多糖,紧密EPS的主要组成是蛋白质,而SMP主要由腐殖质类物质构成.SMP、松散EPS、紧密EPS和总样EPS的三维荧光图谱显示其只有类色氨酸峰和类腐殖酸峰,其类腐殖酸峰主要是由生物代谢产生的腐殖质组成.大分子量腐殖质类物质在几种组分中均出现,但是具有腐殖质特性的分子量为8500 Da的组分会在SMP中大量生成,而具有腐殖质特性的分子量为400 Da的组分只存在于EPS的紧密层中,这与微生物对这3种物质的生物可利用性有关.     

废水处理系统中生物聚集体胞外多聚物研究进展

胞外多聚物(Extracellular polymeric substances,EPS)是废水生物处理系统中生物聚集体(包括絮体污泥、生物膜、颗粒污泥等)的重要组成部分,直接包裹于微生物细胞壁外,其理化性质及所处的特殊位置决定了它在生物聚集体中的重要作用.综述了EPS对废水生物处理系统污泥沉降性能、脱水性以及膜生物反应器膜污染影响的相关研究,分析认为EPS组成与结构特性改变污泥表面电位、疏水性等,进而影响污泥沉降与脱水性能、膜污染程度;以好氧颗粒污泥为典型的生物聚集体代表,总结了EPS组分含量与分布对颗粒污泥的形成与结构稳定性的影响,并在EPS提取方法标准化、现代理化技术与分子生物学技术综合分析等方面进行了展望,进一步的研究有望揭示生物絮凝体形成过程EPS的产生与调控机制.     

一种基于SDS溶液高效提取土壤残留Bt蛋白的方法

通过对SDS浓度、孵育温度和孵育时间3种提取条件的研究,建立了一种基于SDS溶液的提取土壤中残留Bt蛋白的方法(简称SDS法).在ρ(SDS)为2 g·L-1、孵育温度为50℃、孵育时间≥4 h条件下,采用SDS法可有效地从不同类型土壤样品中提取残留Bt蛋白,提取效果明显好于碳酸盐法、人造蠕虫肠道蛋白提取液法和PBST(phosphate buffer solution with Tween-20)法.     

MBR中胞外聚合物累积对污泥性质的影响

以一体式膜生物反应器(MBR)处理模拟生活污水为研究体系,探讨累积胞外聚合物(EPS)的组成、含量及其分布对污泥特性的影响.结果表明,EPS经历一个低谷后,在35 d时出现明显累积,污泥比基质降解速率和污泥体积指数(SVI)也出现相应变化.体系中EPS总量及污泥中蛋白质与多糖比值的增加有利于污泥活性及其沉降性能的改善,EPS累积期大部分EPS存在于污泥中,从另一个角度证实了EPS与污泥活性及其降解性能之间的相关关系.     

pH调节对活性污泥混合液膜过滤性的影响

探讨了用NaOH调节pH值对膜-生物反应器混合液膜过滤性能的影响,并通过分析污泥混合液性质的变化研究其作用机理.试验表明,适当调节pH到碱性,可以使膜过滤性能得到改善.投加碱液使污泥胞外多聚物(EPS)减少,污泥容积指数(SVI)降低,但上清液中总有机碳(TOC)浓度增加.混合液膜过滤性能的改善与污泥SVI的降低密切相关.在碱性条件下,从细胞表面脱落下来的EPS对絮体颗粒的絮凝性有一定促进作用,但效果有限.推测碱液处理使细胞表面的LB-EPS脱落,从而使细胞表面疏水性增加,有利于絮体颗粒之间的絮凝,从而有利于混合液膜过滤性的改善.     

膜生物反应器投加PAC处理生活污水效能的试验

在一体式MBR系统中投加少量的粉末活性炭，运行效果良好，并且可以很好地降低膜污染。粉末活性炭在形成生物活性炭后，对难降解有机物具有很好的降解能力；NH4^ -N的去除率得到进一步提高，NO3^-的含量升高；但反硝化作用不明显，致使总氮去除率不高；生物活性炭很好地吸附并降解了易引起膜污染的有机物，改变了污泥的性质，对膜组件起到了很好的保护作用。     

A new polystyrene-latex-based and EPS-containing synthetic sludge

Since the living microorganisms in activated sludge continuously change, it is difficult to conduct controlled experiments and achieve reproducible results for evaluating sludge characteristics. Synthetic sludge, as a chemical surrogate to activated sludge, could be used to investigate the sludge physicochemical properties, and it is desirable to prepare synthetic sludge with similar structure and properties to real activated sludge to explore the flocculation and settlement processes in activated sludge systems. In this work, a high-strength synthetic sludge was prepared with functional polystyrene latex particles as the framework and extracellular polymeric substances (EPS) to modify its surface. The flocculation and settling characteristics of the microspheres and the prepared synthetic sludge were tested. Compared with other three functional polystyrene latex microspheres, the synthetic sludge prepared with EPS-modified polystyrene latex microspheres showed good settling characteristics and a significantly higher strength. They could be used for studying the physicochemical properties of activated sludge.     

活性污泥的絮体结构模型

采用Motic数码显微镜观察SRT为3 d和15 d的活性污泥絮体以及它们的LEPS和TEPS絮凝污泥悬浮液形成的生物絮体的结构。结果表明,高SRT（15 d）活性污泥絮体较低SRT（3 d）活性污泥絮体的颜色深且密实;同一活性污泥中,内层的絮体结构较外层的絮体结构密实,细菌细胞与菌胶团之间的结合更为紧密。最后,结合絮体结构图片,从大分子作用力的角度,提出了活性污泥絮体结构模型,以形象地描述不同SRT活性污泥EPS及其表面性质变化对活性污泥絮凝沉降性能的影响。     

不同泥龄下活性污泥絮体性状的研究

采用序批式反应器对不同泥龄下污泥絮体的化学性状(胞外聚合物成分及含量)、物理性状(表面电荷)、形态性状(粒度分布、分形维数)等进行了对比研究.结果表明:泥龄对胞外聚合物总量及各组分含量的影响规律并不明显;多糖/蛋白质愈大,污泥絮体表面电负性愈强;污泥絮体的平均粒径随泥龄的延长呈逐渐减小的趋势,且粒度分布愈来愈均匀;不同泥龄下,污泥絮体形态结构亦不相同,泥龄短时,絮体表面粗糙,结构开放疏松;泥龄长时,絮体表面平滑,结构紧凑;随着泥龄的增大,絮体分形维数逐渐增加.由于不同泥龄下所表现出的污泥絮体性状的差异,直接影响了污泥的絮凝和沉降性能.     

Extracellular polymeric substances (EPS) from cyanobacterial mats: characterisation and isolation method optimisation

Extracellular polymeric substances (EPS) play an important role in bacterial mat formation and sediment stabilisation of coastal zones. The analysis of these secretion products on a molecular level is a prerequisite to understand their formation mechanisms and ecological function in microbial consortia. The present study focuses on the optimisation of EPS isolation and characterisation from cohesive cyanobacterial mats. Extracted EPS were analysed for quantity, content of total organic carbon and nitrogen, and bulk contents of neutral sugars, uronic acids, and proteins. These criteria are strongly influenced by the extraction conditions applied and therefore, the effects of different extraction media (NaCl or artificial seawater), addition of EDTA, centrifugal force, and purification via repeated ethanol precipitation on extraction success were determined. From this an optimised extraction procedure for EPS resulted. When using fresh mat samples, the yield of EPS amounted to 3.3 ± 0.8 mg g⁻¹ mat (dw). The isolated EPS were composed of nearly equal amounts of proteins and uronic acids (12.7 ± 1.5 and 11.8 ± 1.1%, respectively) and the bulk content of neutral sugars was 30.5 ± 2.6%. High contents of TOC and TN indicated relatively pure EPS and only a low contribution of inorganic compounds. Furthermore, low contents of 2-keto-3-deoxyoctonate and the small protein/polysaccharide-ratio in the EPS extracted by our method, signified low contaminations by intracellular polymers and hence a low rupture of cells. Our method provides an useful tool to evaluate further investigations of the composition, characteristics and properties of EPS on a sound basis.     

不同基质条件对生物膜细胞外聚合物组成和含量的影响

对比研究了处于胶体和溶解状态的有机物对生物膜胞外聚合物组成和含量的影响，研究发现，不同基质条件对胞外聚合物的总量、多糖和蛋白质含量产生影响，另外，基质条件与会对生物膜的表面结构造成一定的影响。     

Influence of aeration intensity on the performance of A/O-type sequencing batch MBR system treating azo dye wastewater

Among the numerous parameters affecting the membrane bioreactor (MBR) performance, the aeration intensity is one of the most important factors. In the present investigation, an anoxic/aerobic-type (A/O-type) sequencing batch MBR system, added anoxic process as a pretreatment to improve the biodegradability of azo dye wastewater, was investigated under different aeration intensities and the impact of the aeration intensity on effluent quantity, sludge properties, extracellular polymeric substances (EPS) amount generated as well as the change of permeation flux were examined. Neither lower nor higher aeration intensities could improve A/O-type sequencing batch MBR performances. The results showed 0.15 m³·h^-1 aeration intensity was promising for treatment of azo dye wastewater under the conditions examined. Under this aeration intensity, chemical oxygen demand (COD), ammonium nitrogen and color removal as well as membrane flux amounted to 97.8%, 96.5%, 98.7% and 6.21 L·m^-2·h^-1, respectively. The effluent quality, with 25.0 mg·L^-1COD, 0.84 mg·L^-1 ammonium nitrogen and 8 chroma, could directly meet the reuse standard in China. In the meantime, the sludge relative hydrophobicity, the bound EPS, soluble EPS and EPS amounts contained in the membrane fouling layer were 70.3%, 52.0 mg·g^-1VSS, 38.8 mg·g^-1VSS and 90.8 mg·g^-1VSS, respectively, which showed close relationships to both pollutant removals and membrane flux.     

Co-application of energy uncoupling and ultrafiltration in sludge treatment: Evaluations of sludge reduction,supernatant recovery and membrane fouling control

• Effects of metabolic uncouplers addition on sludge reduction were carried out. • TCS addition effectively inhibited ATP synthesis and reduced sludge yield. • The effluent quality such as TOC and ammonia deteriorated but not significantly. • Suitable dosage retarded biofouling during sludge water recovery by UF membrane. Energy uncoupling is often used for sludge reduction because it is easy to operate and does not require a significant amount of extra equipments (i.e. no additional tank required). However, over time the supernatant extracted using this method can deteriorate, ultimately requiring further treatment. The purpose of this study was to determine the effect of using a low-pressure ultrafiltration membrane process for sludge water recovery after the sludge had undergone an energy uncoupling treatment (using 3,3′,4′,5-tetrachlorosalicylanilide (TCS)). Energy uncoupling was found to break apart sludge floc by reducing extracellular polymeric substances (EPS) and adenosine triphosphate (ATP) content. Analysis of supernatant indicated that when energy uncoupling and membrane filtration were co-applied and the TCS dosage was below 30 mg/L, there was no significant deterioration in organic component removal. However, ammonia and phosphate concentrations were found to increase as the concentration of TCS added increased. Additionally, due to low sludge concentrations and EPS contents, addition of 30–60 mg/L TCS during sludge reduction increased the permeate flux (two times higher than the control) and decreased the hydraulic reversible and cake layer resistances. In contrast, high dosage of TCS aggravated membrane fouling by forming compact fouling layers. In general, this study found that the co-application of energy uncoupling and membrane filtration processes represents an effective alternative method for simultaneous sludge reduction and sludge supernatant recovery.