厌氧污泥降解萘动力学与生物多样性研究

为对工业废水中的萘进行高效的生物处理,采集某石油工业废水处理装置的厌氧活性污泥,以萘和萘与乳酸钠为电子供体进行驯化培养.当一个驯化周期中萘去除率达到90%以上时,研究间歇反应条件下分别以萘和萘与乳酸钠为电子供体时的降解动力学,并提取去除率高的污泥中微生物总DNA,构建16S rDNA基因片段克隆文库分析细菌群落结构.结果表明,以单一萘为电子供体时萘的降解率远远小于萘与乳酸钠共代谢时的降解率;两种驯化模式下萘浓度与时间符合一级反应动力学,动力学常数K分别为3.5×10-3h-1和16×10-3h-1.萘与乳酸钠共代谢污泥中细菌类群主要为Deltaproteobacteria、Thermotogae、Bacteroidetes、Chloroflexi和Unclassified bacteria,其中Deltaproteobacteria类细菌占主导地位.在成熟的厌氧活性污泥中Desulfobulbus sp.所占比例为24.2%;Kosmotoga占21.0%.此外,反应器中还存在Smithella、Syntrophobacter、Levilinea等细菌.对厌氧活性污泥中细菌多样性的研究有利于优化反应条件,从而提高污泥中萘的去除率.     

WSSD 2002, Latin America and Brazil: Biodiversity and Indigenous People

Latin America comprehends notable variations in terms of natural environment, availability of natural resources, living standards, and demographic patterns. Latin America is a mosaic of cultures, post- and pre-Columbian. The rich variety of life forms discovered and described by chroniclers and traveling naturalists in the Neotropics contributed to the proposal, in mid-XVIIIth century, of a new system of classification and a scientific code of nomenclature for all organisms. Biodiversity was, for many centuries, a source of resources to be exploited in natura. In scientific circles, its inventory became the domain of taxonomists. But modern technology showed how important the miriad of life forms really are as sources of chemical molecules to be engineered as drugs and reassembled as novel manufactured products. We are on the brink of a new agricultural and medical revolution, thanks to the techniques of genetic engineering, which will lead eventually to the elimination of hunger and malnutrition.In this essay, the Brazilian environmental and social heterogeneity will serve as an example to illustrate some key points, which have influenced sustainability policies. The Amazon deforestation and indigenous knowledge (IK), subjects often associated with areas of high biodiversity, are usually the focus of environmental debates. The importance of IK in integrating development, reducing poverty and sustainability are considered together with the intellectual property rights of native populations.In the World Summit on Sustainable Development (WSSD) Implementation Plan, a few paragraphs were dedicated to Latin America, because of the pre-existing Action Platform on the Road to Johannesburg 2002, approved in Rio de Janeiro in October 2001. This paper calls attention to the need to draw up specific environmental policies for a region which shows an extremely high cultural and biological diversity, associated with a high availability of forests and water, among other resources.     

农村污水膜生物反应器系统中微生物群落解析

农村污水的随意排放和未加限制的灌溉回用可能会导致水源地环境污染加剧,并且其中潜在的病原微生物对周边淡水资源安全和农村居民的身心健康造成潜在的威胁.为了解农村污水的微生物群落组成,为后续污水灌溉的病原微生物风险评价提供理论依据,采用末端限制性片段长度多态性(T-RFLP)和16S r DNA基因克隆文库技术研究农村污水膜生物反应器(MBR)系统中的微生物群落多样性,并结合实时荧光定量PCR方法监测处理前后典型病原菌——弓形菌(Arcobacter spp.)以及总细菌数量的变化.从未处理的农村污水中获取的73个阳性克隆测序结果表明,其主要细菌类群为变形菌门(91.8%)、厚壁菌门(2.70%)、拟杆菌门(1.40%),以及部分不可培养细菌(4.10%).其中弓形菌属是ε-变形菌门的优势菌,也是农村污水中的优势菌株,占克隆总数的68.5%.TRFLP的结果表明,不同处理工艺阶段的主要细菌类群及丰度明显不同,调节池的物种丰度(S)、Shannon-Wiener(H)和物种均匀度(E)最高,分别为43.0、3.56和0.95.定量PCR结果显示未处理的农村污水中弓形菌数量高达(1.09±0.064 0)×1011copies·L-1,该结果与克隆文库结果均表明弓形菌在污水中确实占较高比例.与未处理的农村污水相比,处理后的出水中所监测的弓形菌及细菌总拷贝数分别减少了2~3个数量级,说明MBR处理系统在一定程度上能够去除微生物.处理后的再生水理化指标及指示菌卫生指标均符合农田灌溉用水标准,但其中残留病原微生物引发的健康风险仍需进一步评价.     

稀有鮈鲫Ras信号通路相关基因的克隆、序列分析及其组织分布

Ras基因是一类在生物进化过程中较为保守的原癌基因,在多种细胞生命活动中起到细胞增殖、分化和细胞骨架构建等重要作用。本文克隆了稀有鮈鲫Ras信号通路相关的k-ras、n-ras、pik3cd、pin1和raf1基因的部分c DNA片段,并分析了其序列。序列分析结果表明,k-ras的氨基酸序列与鲤鱼相对应的氨基酸序列同源性最高,为99%;n-ras、pik3cd、pin1和raf1的氨基酸序列均与斑马鱼相对应的氨基酸序列同源性最高,分别为99%、87%、96%和97%,结果表明稀有鮈鲫与鲤鱼、斑马鱼的亲缘关系较近。组织表达分析表明,k-ras、n-ras、pik3cd、pin1和raf1基因在肝脏、鳃、肾脏、脑和性腺等组织中具有表达差异性。本文为研究环境污染物对鱼类致癌作用机制和安全性评估提供分子生物学基础和科学依据。     

Molecular prenatal diagnosis of 3-hydroxy−3-methylglutaryl coa lyase deficiency

We report the first molecular prenatal diagnosis of 3-hydroxy-3-methylglutaryl CoA lyase (HL) deficiency. The proband had a classic but severe presentation with hypoketotic hypoglycaemia and acidosis, secondary mental retardation, and epilepsy, and HL deficiency was documented in cultured fibroblasts. We found him to be homozygous for the frameshift mutation N46fs (+1), which yields a distinct pattern on single-strand conformation polymorphism (SSCP) analysis. In two subsequent pregnancies, molecular prenatal diagnosis was performed using SSCP. In the first, chorionic villus biopsy was normal. In the second pregnancy, amniocentesis revealed an affected fetus. In both pregnancies, the diagnosis was confirmed enzymatically. HL activity was less than 7 per cent of control values in amniocytes and fetal liver of the affected pregnancy. In the second pregnancy, amniotic fluid metabolite measurements by stable isotope dilution-selected ion monitoring mass spectrometry showed greater than 100-fold increases of 3-hydroxy-3-methylglutaric acid and of 3-methylglutaconic acid levels compared with controls.     

rRNA基因ITS区序列分析在粘帚霉属生防菌株种级分类上的应用

对分离自不同地域的粘帚霉属不同种类的生防菌株rRNA基因转录间区进行了克隆测序,用Clustal X软件进行自动排序,用Njplot程序构建系统进化树.供试26个粘帚霉菌株分在4个组,与传统形态分类结果一致,同一种类的不同来源的菌株差异不明显.研究表明,可以根据ITS区DNA序列差异对粘帚霉属进行种级分类,但不能用于区分种内不同地域的菌株之间的差别.图1表1参17     

高通量测序技术应用于污水处理厂细菌气溶胶群落结构分析

由于污水及其处理设施中细菌气溶胶的逸散作用,导致污水处理厂细菌气溶胶的群落结构较为复杂,因此,选择适宜的采样和分析方法对其多样性的认识和风险评估具有重要意义.本研究以某城市污水厂为研究对象,采用"总悬浮颗粒物采样器+高通量测序技术"分别对污水处理全过程进行细菌气溶胶的收集与分析;作为对照,采用"Andersen六级采样器+克隆文库技术"进行同步采样和分析.结果表明,前者对各采样点气溶胶中细菌的多样性、总体群落组成、优势群落的解析均明显优于后者,分析更简便快捷,数据更全面、更接近其真实的分布状态.本研究结果可为污水处理厂细菌气溶胶的采样和分析方法的选择提供科学依据.     

Eubacteria and Archaea community of simultaneous methanogenesis and denitrification granular sludge

Based on the successful performance of a lab-scale upflow anaerobic sludge blanket (UASB) reactor with the capacity of simultaneous methanogenesis and denitrification (SMD), the specific phylogenetic groups and community structure of microbes in the SMD granule in the UASB reactor were investigated by the construction of the Eubacteria and Archaea 16S rDNA clone libraries, fragment length polymorphism, and sequence blast. Real time quantitative-polymerase chain reaction (RTQ-PCR) technique was used to quantify the contents of Eubacteria and Archaea in the SMD granule. The contents of some special predominant methanogens were also investigated. The results indicated that the Methanosaeta and Methanobacteria were the predominant methanogens in all Archaea in the SMD granule, with contents of 71. 59% and 22. 73% in all 88 random Archaea clones, respectively. The diversity of Eubacteria was much more complex than that of Archaea. The low GC positive gram bacteria and Б-Protebacteria were the main predominant Eubacteria species in SMD granule, their contents were 49. 62% and 12. 03% in all 133 random Eubacteria clones respectively. The results of RTQ-PCR indicated that the content of Archaea was less than Eubacteria, the Archaea content in total microorganisms in SMD granule was about 27. 6%.     

贫营养条件下EPS、SMP和微生物多样性的研究

为了考察活性污泥在营养缺乏的条件下,胞外聚合物（EPS）、溶解性微生物产物（SMP）和微生物种群结构自身的变化情况,为优化MBR系统运行、延缓膜污染等提供理论依据,对天津大学游泳馆MBR中的污泥混合液进行贫营养实验,测定了污泥混合液中EPS和SMP的含量,通过聚合酶链式反应-变性梯度凝胶电泳（PCR-DGGE）技术和克隆测序技术对微生物多样性进行分析,根据序列数据进行同源性分析并构建系统进化树.实验初期,EPS和SMP的浓度由15.04 mg/g和0 mg/g分别上升到17.99 mg/g和3.29 mg/g.随着实验的进行,EPS有很大的降低,最终只有2.40 mg/g;SMP则一直在3.5 mg/g左右变化.实验表明,EPS和SMP对外界环境变化具有一定的缓冲作用,并且在营养缺乏的条件下微生物能够以降解EPS和SMP来维持自身生命活动.由于对EPS和SMP的利用,污泥的Shannon多样性指数由最初的0.81上升到最高时的1.09,随后开始降低,并最终稳定在0.95.克隆测序的结果表明,污泥中微生物的种类比较丰富,并且优势菌种大部分为未经培养菌种.部分菌种能够通过产生蛋白质和多糖水解酶来实现对EPS和SMP的降解,主要属于拟杆菌(Bacteroidetes)、黄杆菌(Flavobacterium）、腐螺旋菌(Saprospiraceae)和厚壁门菌(Firmicutes)等.     

多功能细菌复合系NSC-7的菌种组成多样性

NSC-7是一组具有降解纤维素和林丹双重功能的细菌复合系.为系统了解复合系的菌种组成,在有氧条件下,利用传统的平板画线法分离到11株单菌,将11株单菌按体积比1∶1重新组合并不具备分解纤维素的能力,利用单层和双层平板滤纸法检测NSC-7的分解能力,发现只有双层平板上的滤纸变黄且降解,说明复合系内纤维素降解的关键菌是厌氧或微好氧菌.利用现代分子生物学技术对NSC-7构建克隆文库,获得了195个16S rDNA片断,经DGGE筛选获得25个代表克隆,其序列数据库比对结果中有60％的近缘种为已知菌,分别归属于Clostridium、Petrobacter、Bacteria、Paenibacillus、Proteobacterium 5个属,其余40％的近缘种为难培养菌株.