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In situ identification of bacteria in drinking water and adjoining biofilms by hybridization with 16S and 23S rRNA-directed fluorescent oligonucleotide probes.

机译:通过与16S和23S rRNA定向的荧光寡核苷酸探针杂交在饮用水和相邻生物膜中原位鉴定细菌。

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摘要

Free-water-phase and surface-associated microorganisms from drinking water were detected and roughly identified by hybridization with fluorescence-labeled oligonucleotide probes complementary to regions of 16S and 23S rRNA characteristic for the domains Bacteria, Archaea, and Eucarya and the beta and gamma subclasses of Proteobacteria. Samples of glass-attached biofilms and plankton were taken from a Robbins device installed in a water distribution system. More than 70% of the surface-associated cells and less than 40% of the planktonic cells visualized by 4',6-diamidino-2-phenylindole staining bound detectable amounts of rRNA-targeted probes. These findings are an indication for higher average rRNA content and consequently higher physiological activity of the attached microbial cells compared with the free-living cells. All detectable cells hybridized with the bacterial probe, whereas no Archaea and no Eucarya cells could be detected. Simultaneous hybridization with probes specific for the beta and gamma subclasses of Proteobacteria revealed that microcolonies already consisted of mixed populations in early stages with fewer than 50 cells. These observations provide further evidence that the coexistence and interaction of bacteria in drinking water biofilms may be an integral part of their growth and survival strategies.
机译:通过与荧光标记的寡核苷酸探针杂交检测并粗略鉴定了饮用水中的自由水相和与表面相关的微生物,这些探针与16S和23S rRNA区域互补,具有细菌,古细菌和Eucarya结构域以及β和γ亚类的特征的细菌。玻璃附着生物膜和浮游生物的样品是从安装在水分配系统中的Robbins装置中获取的。通过4',6-diamidino-2-phenylindole染色可见,超过70%的表面相关细胞和不到40%的浮游细胞结合了可检测量的rRNA靶向探针。这些发现表明,与游离细胞相比,附着的微生物细胞具有更高的平均rRNA含量,因此具有更高的生理活性。所有可检测的细胞均与细菌探针杂交,而未检测到古生菌和真核细胞。与针对变形杆菌β和γ亚型的特异性探针同时杂交表明,小克隆已经由早期混合种群组成,少于50个细胞。这些观察结果提供了进一步的证据,证明饮用水生物膜中细菌的共存和相互作用可能是其生长和生存策略不可或缺的一部分。

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