高效厌氧产甲烷颗粒污泥中微生物多样性及定量化研究

以小试高效厌氧反应器不同运行阶段的颗粒污泥为对象,利用变性梯度凝胶电泳(DGGE)、荧光原位杂交(FISH)和实时定量链式聚合酶反应(RTQ-PCR)等技术研究了其中微生物种群的多样性、特征微生物的空间分布和定量关系,结果表明:随着反应器有机负荷逐渐提高,颗粒污泥中古菌的群落结构的变化较细菌更为明显;细菌多分布在颗粒外层,而古菌则主要分布在颗粒内层;古菌含量略少于细菌,但有逐渐增多的趋势;产甲烷丝菌在古菌中的含量增加明显.     

厌氧颗粒污泥中微生物种群变化的分子生物学解析

荧光定量-聚合酶链式反应(RTQ-PCR)与荧光原位杂交(FISH)技术对厌氧反应器内不同状态下颗粒污泥中微生物种群的数量变化与空间分布进行研究.结果发现,颗粒污泥中真细菌明显多于古细菌,但随着厌氧反应器有机负荷的提高,古细菌明显增加,其中产甲烷丝菌也明显增加;真细菌多分布生长在颗粒污泥的外层,而对环境条件敏感的古细菌多分布生长在内层,且随着反应器有机负荷的提高,这种层状分布的特点更为明显.     

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%.     

20℃ EGSB反应器中颗粒污泥的微生物种群结构分析

利用PCR-DGGE和RTO-PCR等技术,对6个取自20℃下小试EGSB反应器中不同有机负荷下的颗粒污泥进行研究.结果表明,在20℃下反应器有机负荷从1.5 kg/(m3·d)逐渐提升至10.0 ks/(m3·d)的过程中,未对其污泥中占菌和真细菌的种群结构产生明显影响,各污泥样品中古菌种群相似性较高,其多样性指数分别为5.51、5.88、5.47、5.25、5.32和5.11;除接种污泥外,其余5个污泥样品中真细菌种群相似性也较高,其多样性指数分别为2.97、5.07、5.44、6.38、6.66和5.21;6个污泥样品中主要优势古菌为Methanobacterium、Methanocorpusculum、Methanosaeta和Methanospirillum等,随着有机负荷的增加和运行时间的延长,与氧营养型产甲烷菌Methanocorpusculum parvum和Methanospirillum hungatei相似的菌种的优势地位明显增强;各污泥样品中古菌的相对数量先略有降低而后逐渐升高,最终明显超过接种时的水平.