Wagner M, Amann R, Lemmer H, Schleifer K H
Lehrstuhl für Mikrobiologie, Technische Universität München, Germany.
Appl Environ Microbiol. 1993 May;59(5):1520-5. doi: 10.1128/aem.59.5.1520-1525.1993.
Bacterial community structures in activated sludge samples from aeration tanks of a two-stage system with a high-load first stage and a low-load second stage were analyzed with oligonucleotide probes. The probes were complementary to conserved regions of the rRNA of the alpha, beta, and gamma subclasses of proteobacteria and of all bacteria. Group-specific cell counts were determined by in situ hybridization with fluorescent probe derivatives. Contributions of the proteobacterial subclasses to total bacterial rRNA were quantified by dot blot hybridization with digoxigenin-labeled oligonucleotides. The activated sludge samples were dominated by proteobacteria from the alpha, beta, or gamma subclass. These proteobacteria account for about 80% of all active bacteria found in the activated sludge. For both samples the community structures determined with molecular techniques were compared with the composition of the heterotrophic saprophyte flora isolated on nutrient-rich medium. Probes were used to rapidly classify the isolates and to directly monitor population shifts in nutrient-amended, activated sludge samples. The rich medium favored growth of gamma-subclass proteobacteria (e.g., enterobacteria) and selected against beta-subclass proteobacteria. The culture-dependent community structure analysis of activated sludge produced partial and heavily biased results. A more realistic view will be obtained by using in situ techniques.
采用寡核苷酸探针分析了一个两级系统曝气池活性污泥样品中的细菌群落结构，该系统第一级为高负荷，第二级为低负荷。这些探针与变形菌门α、β和γ亚类以及所有细菌的rRNA保守区域互补。通过与荧光探针衍生物进行原位杂交来确定特定菌群的细胞计数。通过与地高辛标记的寡核苷酸进行斑点杂交，对变形菌门亚类在总细菌rRNA中的贡献进行了定量。活性污泥样品中主要是α、β或γ亚类的变形菌。这些变形菌约占活性污泥中所有活性细菌的80%。对于这两个样品，将用分子技术确定的群落结构与在富营养培养基上分离的异养腐生菌群的组成进行了比较。使用探针快速对分离株进行分类，并直接监测营养改良活性污泥样品中的种群变化。富营养培养基有利于γ亚类变形菌（如肠杆菌）的生长，而不利于β亚类变形菌。活性污泥的基于培养的群落结构分析产生了不完整且严重有偏差的结果。通过使用原位技术将获得更真实的情况。
