Short-term effect of temperature variation on the competition between PAOs and GAOs during acclimation period of an EBPR system

Sequencing batch reactor (SBR) for enhanced biological phosphorus removal (EBPR) processes was used to investigate the impact of the temperature shock on the competition between phosphorus-accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs) in start-up stage. During the 34 days operation, SBR was set with temperature variation(0–5 d, 22±1°C; 6–13 d, 29±1°C; 14–34 d, 14±1°C). PAOs and GAOs were analyzed by fluorescent in situ hybridization (FISH), and intracellular polyphosphate granules were stained by Neisser-stain. The results showed that the influence of temperature shock on PAOs’ abundance was more serious than that on GAOs in the enriching process. Under sudden and substantially temperature variation, from 22±1°C to 29±1°C and then to 14±1°C, the domination of PAOs was deteriorated. After temperature shock, PAOs’ competitive advantages at low temperature that concluded in other study did not appear in our study. As mesophilic, GAOs (indicated by Alphaproteobacteria and Gammaproteobacteria) were more temperature adaptive and better grew and took the domination at 14±1°C in the end. In the competition process, organisms of tetrad forming organisms (TFOs)-like shape which were considered as typical GAOs, were observed. With the evidence of poly-P granules containing by Neisser-straining and result of FISH, these organisms of TFOs-like shape were better to be assumed as adaption state or a special self-protecting shape of PAOs.     

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。     

应用FISH法对污水生物处理系统中细菌数量的计数

近年来,FISH(Fluorescence in Situ Hybridization,荧光原位杂交)法作为一种应用分子生物学技术对细胞及细菌等微生物进行定性以及定量分析的研究方法,在国际上得到广泛的应用。本文介绍了应用FISH法对污水处理工艺中细菌数量的计数方法。同时介绍了活性污泥试样制作、FISH法观测试样的制作以及各种试剂的配制、作用以及使用方法,基因探针的选择原则、杂交方法等实际操作方法与过程。     

Chromosome 18 analysis by fluorescence in situ hybridization (fish) in human blastomeres of abnormal embryos after in vitro fertilization (ivf) attempt

We performed fluorescence in situ hybridization (FISH) with a chromosome 18-specific probe on human abnormal cleaved embryos, fertilized either by two spermatozoa and exhibiting three pronuclei (3 PN) or normally fertilized and exhibiting two pronuclei (2 PN) with subsequent severe fragmentation and/or blocking. The aim of the study was to evaluate the incidence of chromosome 18 anomalies among these embryos, in order to evaluate the FISH efficiency on such material and to obtain more precise and complete data than those obtained with classical cytogenetic analysis. For the 3 PN cleaved embryos, FISH confirmed the frequent regulation towards diploidy (25 per cent) and the high frequency of mosaics (53 per cent). For the 2 PN blocked or damaged embryos, FISH permitted chromosome evaluation, which was otherwise impossible with classical cytogenetic techniques: we also found a high mosaic frequency (45 per cent) with these embryos. If this frequency were the same for normally developing embryos, it would be a major obstacle to the reliability of either chromosomal or genetic preimplantation diagnosis.     

荧光原位杂交法在活性污泥硝化细菌检测中的应用

采用荧光原位杂交(FISH)法对活性污泥样品中的硝化细菌进行了检测研究。与传统方法相比,FISH法具有快速便捷的特点,可原位检测出活性污泥菌胶团上的硝化细菌的分布情况,结合稀释的方法,可对环境样品中的硝化细菌进行定量分析。此法既可定性,又可对环境中的硝化细菌进行直接计数,为研究硝化细菌类群的空间和数量分布提供有效的检测工具。     

A/O工艺膜生物反应器处理生活污水的脱氮特性及硝化菌群的分子检测

采用A/O工艺膜生物反应器(MBR),以生活污水为处理对象,考察了系统的脱氮特性,并采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)与荧光原位杂交(FISH)技术对系统中硝化菌群进行了分子检测.结果表明,A/O工艺MBR处理生活污水,TN去除率在85%左右,NH3-N去除率在95%以上;DGGE图谱显示,随着系统运行时间的延长,硝化菌群数量逐渐增加,并且不同菌种的丰度也发生了变化;FISH检测显示,系统中硝化菌的优势菌种为氨氧化菌和亚硝酸氧化菌.应用Motic Fluo 1.0软件对FISH结果进行分析,结果显示,系统运行初期到末期,氨氧化菌占硝化菌的比例一直保持在25%左右;亚硝酸氧化菌占硝化菌的比例由系统运行初期的35%逐渐增加到系统运行末期的55%左右.     

序批式生物膜反应器不同填料挂膜及短程硝化特性研究

以实际生活污水为研究对象,采用序批式生物膜反应器(SBBR),填充不同种类的填料,针对其各自的挂膜特征和短程硝化的实现与稳定的特性进行了研究.结果表明,与立方体海绵填料相比,炭纤维填料的SBBR能够更快地实现挂膜启动,硝化效果稳定、高效(NH_4~+-N去除率高达99.3%);立方体海绵填料更易在常温下,实现NO_2~--N大量积累的短程硝化,但是相比而言,硝化效率不高.升高温度至30 ℃左右时,能够在30 d内实现炭纤维填料的短程硝化,通过过程控制可以实现短程硝化的稳定.荧光原位杂交技术(FISH)检测结果证实,SBBR中短程硝化的实现与稳定是因为菌群得到了优化,氨氧化细菌成为优势菌种.