PCR-DGGE研究处理垃圾渗滤液序批式生物膜反应器(SBBR)中的细菌多样性

为了研究序批式生物膜反应器中的细菌多样性及其脱氮的微生物学机理,为工艺改进提供依据,从同步高效去除垃圾渗滤液中高氨氮和高COD的SBBR生物膜和渗滤液原水中采集微生物样品并提取微生物总DNA,使用细菌通用引物对（GC341F/907R）从总DNA中成功扩增出目标16S rDNA片段,然后对扩增的16S rDNA进行DGGE,对凝胶染色并进行条带统计分析和切胶测序,使用序列数据进行同源性分析并建立了系统发育树.结果表明,该驯化后的SBBR生物膜和渗滤液原水中都有比较丰富的细菌多样性,驯化的生物膜细菌主要来自渗滤液原水,而且生物膜细菌在反应器正常运行时不会出现明显的群落结构变化;在该SBBR中有多种硝化细菌与反硝化细菌、好氧反硝化细菌和厌氧氨氧化细菌共存,说明该反应器中可能同时存在全程硝化反硝化、同步硝化反硝化和厌氧氨氧化3种脱氮方式.研究结果为SBBR脱氮微生物机理研究提供了一些有价值的参考依据.     

二苯并噻吩降解菌筛选及16S rDNA序列分析

以DBT(dibenzothiophene,二苯并噻吩)为模型化合物,分离得到了一株能降解DBT的微生物HB2,应用HPLC对其脱硫特性进行了检测。应用PCR技术克隆到16SrDNA片段,核苷酸序列分析结果表明,该菌的16SrDNA全序列与假单胞菌存在98%的同源性,初步确定该菌在微生物系统发育学上的地位。     

一株反硝化菌的分离鉴定和系统发育分析

从反硝化反应器中分离到一株异养型兼氧反硝化细菌,命名为菌株JIN1,分离菌株为革兰氏染色阴性杆状。茵落颜色为乳白色。该菌株能以硝酸钠为唯一氮源进行反硝化作用并产生亚硝酸。部分长度的16SrDNA序列分析表明,分离菌株JIN1与Aquaspirillum菌具有99%相似,与Microvirgulaaerodenitrificans菌具有99%相似。其生化性状与Aquaspirillum菌不相符。系统发育学分析表明菌株JIN1与Microvirgulaaerodenitrificans菌关系最近。故确认菌株JIN1属Microvirgulaaerodenitrificans菌。     

Isolation, characterization and phylogenetic analysis of a bacterial strain capable of degrading acetamiprid

An aerobic bacterium, capable of degrading the new chloronicotine pesticide acetamiprid, was isolated from the sludge of pesticide factory after successive enrichment cultures and named strain FH2 which is a Gram-negative, rod-shaped, obligate aerobic org     

浮游动物DNA宏条形码标志基因比较研究

DNA宏条形码技术作为一种新型生物监测方法,在未来生态环境监测中有巨大的应用潜力。目前,浮游动物DNA宏条形码监测仍在发展阶段,需要首先对其(采样方法、引物选择和数据分析等)进行标准化和调整,然后才能用于常规流域生态监测。其中,如何选择合适的PCR扩增引物是DNA宏条形码生物监测标准化的关键问题之一。本研究比较了COI、18SV9和16S通用引物在浮游动物DNA宏条形码监测中的差异,为初步建立规范化的浮游动物DNA宏条形码监测方法提供技术支撑。结果表明,16S引物对浮游动物具有更好的特异性,其产生的操作分类单元(operational taxonomic unit, OTU)有88.1%属于浮游动物。虽然18SV9引物具有更高的物种覆盖度,不仅能扩增出浮游动物,还能扩增出大量藻类和真菌,但其物种识别敏感性较差,不适合浮游动物物种水平多样性监测。COI引物的浮游动物物种特异性、物种覆盖度和物种识别敏感性都适中,检出的浮游动物物种数量高于18SV9引物和16S引物,更加适合浮游动物DNA宏条形码监测。     

Validation of Bacteroidales-based microbial source tracking markers for pig fecal pollution and their application in two rivers of North China

• Pig feces is the predominant excrement produced by animal husbandry in China. • The PF, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan MST assays showed better performance. • The pig-specific MST assays can contribute to managing the pig fecal pollution. In China, pig feces is the predominant source of excrement produced by animal husbandry. Improper use or direct discharge of pig feces can result in contamination of natural water systems. Microbial source tracking (MST) technology can identify the sources of fecal pollution in environmental water, and contribute to the management of pig fecal pollution by local environmental protection agencies. However, the accuracy of such assays can be context-dependent, and they have not been comprehensively evaluated under Chinese conditions. We aimed to compare the performance of five previously reported pig-specific MST assays (PF, Pig-Bac1^SYBR, Pig-Bac2^SYBR, Pig-1-Bac^TaqMan, and Pig-2-Bac^TaqMan, which are based on Bacteroidales 16S rRNA gene markers) and apply them in two rivers of North China. We collected a total of 173 fecal samples from pigs, cows, goats, chickens, humans, and horses across China. The PF assay optimized in this study showed outstanding qualitative performance and achieved 100% specificity and sensitivity. However, the two SYBR green qPCR assays (Pig-Bac1^SYBR and Pig-Bac2^SYBR) cross-reacted with most non-pig fecal samples. In contrast, both the Pig-1-Bac^TaqMan and Pig-2-Bac^TaqMan assays gave 100% specificity and sensitivity. Of these, the Pig-2-Bac^TaqMan assay showed higher reproducibility. Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand, Japan, and America. Using the PF and Pig-2-Bac^TaqMan assays, a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.     

Effects of temperature and fertilization on total vs. active bacterial communities exposed to crude and diesel oil pollution in NW Mediterranean Sea

The dynamics of total and active microbial communities were studied in seawater microcosms amended with crude or diesel oil at different temperatures (25, 10 and 4 °C) in the presence/absence of organic fertilization (Inipol EAP 22). Total and hydrocarbon-degrading microbes were enumerated by fluorescence microscopy and Most Probable Number (MPN) method, respectively. Total (16S rDNA-based) vs. active (16S rRNA) bacterial community structure was monitored by Capillary-Electrophoresis Single Strand Conformation Polymorphism (CE-SSCP) fingerprinting. Hydrocarbons were analyzed after 12 weeks of incubation by gas chromatography-mass spectrometry. Total and hydrocarbon-degrading microbial counts were highly influenced by fertilization while no important differences were observed between temperatures. Higher biodegradation levels were observed in fertilized microcosms. Temperature and fertilization induced changes in structure of total bacterial communities. However, fertilization showed a more important effect on active bacterial structure. The calculation of Simpson's diversity index showed similar trends among temperatures whereas fertilization reduced diversity index of both total and active bacterial communities.     

The evolution of the Canadian mining industry: The role of regulatory punctuation

In this study, we analyze the evolution of Canada's mining industry from 1929 to 2006, focussing on the determinants of the number of firms in the industry and why this number changed over that period. Most empirical studies of industry evolution have focused on manufacturing industries that share similar structural characteristics. Perhaps because of this, extant models of industry evolution tend to ignore industry-specific and national-specific factors that can cause atypical trajectories, that is, heterogeneous industry evolution. Initial inspection of the Canadian mining industry shows that it is atypical in that it exhibits “negative skew” over time in the number of firms rather than the typical “positive skew.” We review two dominant theoretical approaches to industry evolution: the density-dependence theory and variants of industrial organization economics. We also consider possible sources of industry evolution heterogeneity, focussing particularly on “regulatory punctuation”. Using Canadian mining data, we find that the traditional models do not fully explain the changes in population size in Canada's mining industry. As a result, we introduce a number of hybrid models. The results from these hybrid models suggest that Canadian-specific regulatory punctuations, particularly the introduction of significant new taxes, environmental legislation, and incentives have shaped the trajectory of mining firm participation.     

降解SO2功能菌的16S rRAN基因测序及降解特性

用低浓度SO2诱导驯化方法获得高效脱硫菌群,并用分离培养与16S rRNA基因测序技术相结合的方法鉴定菌群种属,分析驯化过程中种群结构的动态变化,同时研究分离纯菌种的脱硫性能。结果表明,从诱导驯化7 d和14 d菌液中分别分离出23株菌和22株菌,16S rRNA序列分析发现这些菌归属于13个种,其中有6个种(Rhodococcus erythropolis、Pseudomonas putida、Microbacterium oxydans、Sphingomonas koreensis、Acinetobacter junii、Acinetobacter johnsonii)对SO2-3有较强的降解能力,并在持续驯化过程中稳定的生长传代,降解产物以硫酸根为主,还有极少量的单质硫。与含混合菌的驯化菌液降解SO2-3的能力相比,单一脱硫菌的脱硫性能较弱。脱硫功能菌株及其基本特性的研究为微生物处理SO2烟气提供了丰富的菌源信息和理论基础。     

Reduction of selenite to red elemental selenium by moderately halotolerant Bacillus megaterium strains isolated from Bhitarkanika mangrove soil and characterization of reduced product

Two Gram (+) bacterial strains, BSB6 and BSB12, showing resistance and potential for Se(IV) reduction among 26 moderately halotolerant isolates from the Bhitarkanika mangrove soil were characterized by biochemical and 16S rDNA sequence analyses. Both of them were strictly aerobic and able to grow in a wide range of pH (4-11), temperature (4-40 °C) and salt concentration (4-12%) having an optimum growth at 37 °C, pH ∼7.5 and 7% salt (NaCl). The biochemical characteristics and 16S rDNA sequence analysis of BSB6 and BSB12 showed the closest phylogenetic similarity with the species Bacillus megaterium. Both the strains effectively reduced Se(IV) and complete reduction of selenite (up to 0.25 mM) was achieved within 40 h. SEM with energy dispersive X-ray and TEM analyses revealed the formation of nano size spherical selenium particles in and around the bacterial cells which were also supported by the confocal micrograph study. The UV-Vis diffuse reflectance spectra and XRD of selenium precipitates revealed that the selenium particles are in the nanometric range and crystalline in nature. These bacterial strains may be exploited further for bioremediation process of Se(IV) at relatively high salt concentrations and green synthesis of selenium nanoparticles.