膜生物反应器中不同阶段微生物群落结构演变的研究

为了研究膜生物反应器（membrane bioreactor, 简称MBR）中微生物群落结构的演变情况,对不同阶段运行状况下(包括培养驯化阶段、初期增长阶段、污泥流失阶段和恢复成熟阶段4个阶段)污泥中的微生物进行了考察。从MBR污泥中提取细菌总基因组DNA,进行聚合酶链式反应变性梯度凝胶电泳(PCR-DGGE)图谱的直观分析,以及总细菌的Shannon多样性指数、各污泥样品的相似性分析和聚类分析。研究表明,反应器中的微生物群落比较丰富。不同的阶段状况下,微生物的群落结构变化比较明显,能够比较好地反映MBR的运行状况和系统处理效能的关系,并且发现微生物的群落结构能够随着反应器的运行状况的变化做出调整恢复。     

复合生物活性填料对活性污泥生物特性的影响

为了考察生物活性填料对活性污泥生物特性的影响,采用电子扫描显微镜、激光粒度仪和Miseq高通量测序技术,分别对SBR中污泥的形态、粒径和微生物群落分布进行了研究。结果表明,投加生物填料可以降低污泥的产率,促进污泥沉降,增强脱氮除磷效能;投加生物活性填料的SBR相比对照组SBR的污泥结构紧密,污泥颗粒分布均匀,且颗粒粒径大;内置填料和外置填料的活性污泥粒径在28.988~124.573μm的颗粒占据了大部分的比例,而对照组的活性污泥粒径在0.554~23.538μm的颗粒占据了大部分的比例;3个微生物群落主要分布在Sphingobacteriales、Xanthomonadales、Burkholderiales、Rhodocyclales、Flavobacteriales和Spirochaetales等6个目内;内置填料活性污泥微生物群落(17.12%)和外置填料活性污泥微生物群落(23.17%)中的黄色单胞菌目,要比对照组活性污泥微生物群落(5.51%)的比例高,说明通过复合生物活性填料的作用,能够改善活性污泥微生物群落的除磷能力,这与反应器实际运行效果和污泥粒径分析研究内容一致。     

膨胀颗粒污泥床反应器启动过程中颗粒污泥性质变化研究

采用小试规模的膨胀颗粒污泥床(EGSB)反应器,考察其启动规律及运行特点.在60 d内EGSB反应器的COD容积负荷达到12 kg/(m3·d),COD去除率保持在95%以上.试验中对不同水力负荷下污泥床状态进行了分析.结果表明,将水力负荷控制在1.0 m3/(m2·h)以上,可以确保污泥颗粒化的进行及EGSB反应器的稳定,否则易发生沟流、活塞式漂浮等污泥床异常现象.同时,对颗粒污泥性质及微生物相变化进行了跟踪分析.随着运行条件的改变,由上流式厌氧污泥床(UASB)反应器接种的污泥结构、性能和微生物群落都在不断发生变化.最终得到的颗粒污泥结构密实,沉降速率为38.8～64.6 m/h,比产甲烷活性达到314.25 mL/(g·d),内部微生物相丰富,各菌种呈混杂分布.     

高负荷条件下好氧颗粒污泥同步脱氮除碳特性及微生物群落结构分析

为研究高负荷条件下好氧颗粒污泥的形成过程、同步脱氮除碳效果和微生物群落结构特点,构建了一个序批式反应器(sequencing batch reactor,SBR)。结果表明,C/N=40进水条件下能够完成颗粒化,成熟后的好氧颗粒污泥呈表面光滑结构紧实的椭球体。随着颗粒粒径增大,其比好氧速率提高、含水率下降、沉降性能变好、生物量增加。颗粒形成过程产生的胞外聚合物(extracellular polymeric substances,EPS)先增加后受水质冲击减少,之后又明显提高,整个过程中多糖与蛋白质之比(PS/PN)持续下降,EPS中的蛋白质对颗粒的形成影响较大。SBR中的好氧颗粒污泥能够同时高效去除进水中的COD、N H_4~+-N和TN,去除率分别为94%、96%和93%,反应器的反硝化性能良好。C/N=40时,采用MiSeq高通量测序方法对成熟好氧颗粒污泥中的群落结构进行研究,发现存在促进颗粒化的优势菌门(包括Saccharibacteria、Proteobacteria、Bacteroidetes、Actinobacteria、Firmicutes和Chloroflexi)。同时,在颗粒污泥中,异养硝化、好氧/缺氧反硝化菌属丰度较高,表明异养硝化-好氧/缺氧反硝化菌属可能存在于好氧颗粒污泥中。     

不同污泥负荷下SBR处理游泳馆污水的微生物群落的演变

污泥负荷直接影响微生物的生长模式,当污泥负荷发生变化时,短时间内微生物群落结构将发生明显变化。为了研究污泥负荷冲击对SBR系统内活性污泥微生物群落结构的影响,应用聚合酶链式反应-变性梯度凝胶电泳(PCRDGGE)技术,对不同污泥负荷冲击时,SBR处理游泳馆污水中的活性污泥微生物进行了考查。研究表明,在不同污泥负荷冲击的条件下,以MBR污泥为接种污泥,SBR工艺处理游泳馆污水系统内活性污泥微生物群落结构变化明显,多样性指数随着污泥负荷升高而逐渐增加并趋于稳定,但污泥冲击负荷过高多样性指数反而下降,SBR系统内微生物菌种大部分为未经培养菌种,肠杆菌属、甲苯单胞菌属以及γ-变形菌纲细菌等。微生物通过对不同负荷阶段环境条件的适应及演变,逐渐形成了适应相应污泥负荷的微生物种群。     

膜污染状态下的胞外聚合物、细菌数量及微生物群落结构研究

研究了膜污染状态下膜生物反应器(MBR)中混合液污泥和膜丝表面污泥的胞外聚合物(EPS)含量、细菌数量以及微生物种群结构的情况。结果表明:EPS主要由蛋白质组成,膜丝表面污泥中的EPS含量较高,高达68.3mg/g;MBR污泥中细菌的数量级基本达109个/g,其中膜丝表面污泥细菌数量较高;MBR混合液污泥和膜丝表面污泥微生物群落的Shannon-Weaver指数分别为2.639和2.303,两者微生物群落结构的相似性系数达到0.833,两者的优势菌均为黄色单胞菌,变形杆菌门和拟杆菌门是膜丝表面种类较多的菌群,可能与膜污染有密切的联系。深入分析膜污染形成原因和过程,对膜污染的防治具有重要的意义。     

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。     

垃圾渗滤液厌氧处理过程颗粒污泥生长特性

以上流式厌氧反应器为例,研究了渗滤液厌氧处理过程中颗粒污泥的生长特性。结果表明,反应器运行阶段,颗粒污泥粒径由0.96 mm增长至1.06 mm,MLVSS/MLSS大于0.76,污泥能够保持较高的活性。高污泥负荷有利于0.90 mm颗粒污泥生长,0.60~0.90 mm污泥颗粒生长过程中结构不稳定,易发生解体。厌氧颗粒污泥生长过程中,粒径较大的颗粒污泥的孔隙结构较为疏松,它对污泥的活性产生影响。随着孔隙率增大,污泥沉降速度由0.88 cm/s升至0.97cm/s。颗粒污泥表层与内部微生物分布不同,表层微生物以球菌为主,各微生物种群间紧密联系,颗粒污泥内部以短杆菌为主。     

污泥好氧颗粒化过程中氨氧化菌群结构的演替与分析

为了揭示颗粒污泥形成过程中氨氧化菌(AOB)群落结构的演替规律,利用变性梯度凝胶电泳(DGGE)、克隆测序和实时定量聚合酶链式反应(real-time PCR)等分子生物学技术对AOB群落的演替进行了研究。DGGE结果表明,污泥接种驯化期,AOB群结构的变化较为剧烈,在外加选择压的作用下,种群多样性迅速下降;但随着污泥颗粒化的完成而趋于稳定。测序结果表明,接种污泥中的大多数亚硝化单胞菌属因可快速适应工艺的淘洗过程而被保留在系统内,而亚硝化螺菌属逐渐被淘汰。real-time PCR结果表明,在经历了运行初期的淘洗后,AOB含量随着污泥浓度的提高而逐渐增长;但污泥的氨氧化活性随着污泥浓度的增长而降低。     

微生物在水处理技术研究中的新进展

微生物水处理法具有投资少、操作简单、处理时间短等优点,广泛应用于水处理工程之中.2000年3月,在东京大学召开了名为"运用复杂微生物群落机能的高级水处理技术体系的建立与评估"国际研讨会.本文主要就该国际会议从以下五个方面--微生物群落分析技术、水净化微生物技术、生物营养物去除、污泥减量与资源化技术和亚洲热带与亚热带地区的水处理技术等进行综述,并指出微生物在水处理技术中进一步发展的方向和趋势.     

生物膜水解-好氧循环系统处理活性艳蓝RB-19的微生物群落动态变化

为了研究生物膜水解-好氧循环系统处理蒽醌类染料活性艳蓝RB-19效果及其中微生物群落动态变化,利用基于16S rDNA的PCR-DGGE技术获得了微生物群落的DNA特征指纹图谱。结果表明,该系统能有效地降解活性艳蓝RB-19模拟废水,在RB-19浓度≤400 mg/L时,RB-19去除率维持在82%～96%之间,COD去除率维持在95%左右,但当RB-19浓度提高到500 mg/L时,RB-19去除率降低到58%,COD去除率降低到85%。DGGE分析表明,生物膜上的微生物群落结构随着RB-19浓度递增有显著变化,好氧、水解反应器内的细菌Shannon指数分别从1.32和1.20降低到1.11和1.19。UPGMA聚类分析和NMDS散点分析表明,水解、好氧反应器内的微生物并没有因为同处一个系统内而使得其菌群落结构产生明显的趋同倾向。系统内的多种优势菌群为兼性细菌,克隆测序的结果发现,在水解反应器存在一类具有很强还原能力的古细菌——Methanobacterium sp.MB4。     

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.     

Influence of benomyl and prometryn on the soil microbial activities and community structures in pasture grasslands of Slovakia

The effects of pesticides (a herbicide and a fungicide) on the microbial community structure and their activity were analyzed in soil from four alpine pasture grasslands in Slovakia. Specifically, the effects of the herbicide, Gesagard (prometryn active ingredient), and fungicide, Fundazol 50 WP (benomyl active ingredient), on the microbial respiration activity (CO₂ production), the numbers of selective microbial physiological groups (CFU.g^?1) and the structure (relative abundance) of soil microbial communities [(phospholipid fatty acid (PLFA)] were analyzed under controlled laboratory conditions. All treatments including the treatments with pesticides increased (statistically significantly) the production of CO₂ in all fields during 21 days of incubation and posed a statistically insignificant negative influence on the numbers of the observed physiological groups of microorganisms. The significantly negative influence was evaluated only in the numbers of two physiological groups; spores of bacteria utilizing organic nitrogen and bacteria, and their spores utilizing inorganic nitrogen. A shift in the microbial composition was evident when the PLFA patterns of samples from different sites and treatments were compared by the Principal Component Analysis (PCA). According to the second component PCA 2 (15.95 %) the locations were grouped into two clusters. The first one involved the Donovaly and Dubakovo sites and the second one contained the Velka Fatra and Mala Fatra locations. The PLFA composition of the soils showed important changes after the treatment with pesticides according to PCA 1 (66.06 %). Other treatments had not had a significant effect on the soil microbial community with the exception of the population of fungi. The lower relative abundance (significant effect) of Gram-positive bacteria, actinomycetes and general group of bacteria were determined in samples treated by the herbicide Gesagard. The application of fungicide Fundazol decreased (statistically significantly) the relative abundance of actinomycetes and general group of bacteria and paradoxically increased the population of fungi.     

A+OSA污泥减量工艺的微生态特性

采用16S rDNA序列与PCR-DGGE(polymerase chain reaction-denaturing gradient gel electrophoresis)分析技术相结合的方法,研究了A+OSA(the anoxic+oxic-settling-anaerobic)污泥减量工艺在不同工况下的减量效果及其微生态特性。结果显示,在自然条件下,A+OSA工艺可有效减少剩余污泥27%左右。分子生物研究表明,解耦联池的插入可以明显改变系统微生物的群落结构,且随着解耦联池水力停留时间的延长,系统中部分微生物被"淘洗",微生物丰富度和多样性指数均有所降低。相似性分析表明,参照系统和A+OSA工艺分属于2个不同的集群,但在A+OSA工艺内部各反应池样品间具有较高的相似性,且各反应池在HRT为5.16 h和7.14 h时,表现为显著相似。通过上述研究可为该工艺优化及调控提供理论指导。     

Impact of long‐term pesticide applications on some soil biological parameters

Abstract

The soil oxidative and anaerobic processes, as well as, the microbial biomass were followed during three years in a cotton farm (Tatuí) where the recommended pesticides have been used for several years, and in an experimental field (São Paulo) treated first time with the same pesticides. The oxidative process was monitored by the dehydrogenase (DHA)‐activity using triphenyltetrazolium chloride (TTC) as substrate. The anaerobic process was followed by the iron‐oxide reduction, and the microbial biomass was estimated by the substrate (glucose)‐indiced respiration. Increases in DHA‐activity and in the microbial biomass occurred only in the farm soil, with concomitant decreases in iron‐reduction. In the experimental field soil, the increases in DHA‐activity were followed only by decreases in iron‐reduction. Soil characteristics were the determining factor for different biological parameters after pesticide inputs. All the pesticides produced at least one clear but transient effect.     

抗生素菌渣堆肥进程中微生物群落的变化

将青霉素菌渣、林可霉素菌渣与牛粪等原料分别进行好氧堆肥实验,以考察堆肥过程中不同菌渣对微生物群落的影响。在堆制的41d里,根据温度变化分阶段采集堆肥样品,采用稀释倒平板法测定细菌、放线菌和真菌的数量。结果表明,菌渣不同,其堆肥中的微生物群落变化趋势不同。青霉素菌渣堆肥中细菌数量变化趋势为高一低,真菌数量变化趋势为高一低．高,放线菌数量为逐渐增加;林可霉素菌渣堆肥过程中细菌数量变化趋势为低一高一低,放线菌和真菌数量变化趋势为高．低．高。依据真菌菌落形态观察,菌渣堆肥中的真菌种类比对照牛粪堆肥单一,表明两种菌渣对堆肥中的微生物多样性均产生了不利影响。林可霉素菌渣堆肥初始时的细菌数量比对照低1个数量级,放线菌数量在整个堆肥进程中都明显低于对照,堆肥结束时,随着菌渣含量的增加,放线菌数量逐渐下降,高温期真菌数量下降幅度随着菌渣含量增加而加大,表明林可霉素菌渣对细菌、放线菌和真菌均有不同程度的抑制。堆肥化后菌渣中林可霉素残留量的减少表明,在一定条件下堆肥处理可以将抗生素菌渣无害化和资源化。     

Characterization of microbial consortia that reductively dechlorinate 4-chlorophenol and transform phenol to benzoate enriched from estuarine sediment of Lake Shinji

Monochlorophenols were degraded to benzoate via phenol by the initial dechlorination and the subsequent conversion of phenol to benzoate in anaerobic sediment samples of estuarine Lake Shinji under methanogenic conditions. To characterize bacteria that dechlorinate 4-chlorophenol and transform phenol to benzoate, we analyzed the microbial community structure of the enrichment culture with each 4-chlorophenol and phenol by the limiting dilution method with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rRNA gene. After serial dilution of the culture, the 4-chlorophenol-dechlorinating culture consisted of two dominant bacteria, one of which was most homologous with Dehalobacter sp. In the enriched culture with phenol, minor band homologous with Cryptanaerobacter phenolicass corresponded to the transformation activity.     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

Effects of glyphosate and foliar amendments on activity of microorganisms in the soybean rhizosphere

A field study was conducted to determine the effects of glyphosate on microbial activity in the rhizosphere of glyphosate-resistant (GR) soybean and to evaluate interactions with foliar amendments. Glyphosate at 0.84 kg ae ha^{? 1} was applied GR soybean at the V4–V5 development stages. Check treatments included a conventional herbicide tank mix (2003 study only) and no herbicides (hand-weeded). Ten days after herbicide application, a commercially available biostimulant and a urea solution (21.0% N) were applied to soybean foliage at 33.5 mL ha^{? 1} and 9.2 kg ha^{? 1}, respectively. Soil and plant samples were taken 0, 5, 10, 15, 20 and 25 days after herbicide application then assayed for enzyme and respiration activities. Soil respiration and enzyme activity increased with glyphosate and foliar amendment applications during the 2002 growing season; however, similar increases were not observed in 2003. Contrasting cumulative rainfall between 2002 and 2003 likely accounted for differences in soil microbial activities. Increases in soil microbial activity in 2002 suggest that adequate soil water and glyphosate application acted together to increase microbial activity. Our study suggests that general soil microbial properties including those involving C and N transformations are not sensitive enough to detect effects of glyphosate on rhizosphere microbial activity. Measurements of soil-plant-microbe relationships including specific microbial groups (i.e., root-associated Fusarium spp.) are likely better indicators of impacts of glyphosate on soil microbial ecology.     

接种菌剂和外加能源对污泥生物干化效果的影响

采用自主设计的试验装置,研究了接种菌剂和外加能源对城市污水处理厂脱水污泥生物干化效果的影响。结果表明:(1)试验7d,添加接种菌剂的物料升温累积值为66.0℃.d,比不添加接种菌剂(18.3℃.d)大261%;添加接种菌剂物料的水分去除率(27.33%)比不添加接种菌剂(18.56%)提高了8.77百分点;添加接种菌剂物料的挥发性固体(VS)降解率(20.90%)比不添加接种菌剂(12.31%)高8.59百分点;添加接种菌剂物料的减重率(18.67%)比不添加接种菌剂(10.80%)高7.87百分点。(2)试验8d,添加外加能源的升温累积值(69.5℃.d)比不加外加能源(46.2℃.d)大50.43%;添加外加能源物料的水分去除率(33.50%)比不加外加能源(28.56%)高4.94百分点;添加外加能源的物料VS降解率(22.62%)比不加外加能源(19.67%)高2.95百分点;添加外加能源物料的减重率(19.56%)比不加外加能源(17.87%)高1.69百分点。