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In situ identification of nocardioform actinomycetes in activated sludge using fluorescent rRNA-targeted oligonucleotide probes

Markus Schuppler^1,,1

Ulf B. Göbel^1,

Institut für Medizinische Mikrobiologie und Hygiene, Klinikum der Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Straße 11, D-79104 Freiburg, Germany
Lehrstuhl für Mikrobiologie, Technische Universität München, ArcisStraße 16, D-80290 München, Germany
Institut für Mikrobiologie der Fakultät für Biologie II, Albert-Ludwigs-Universität Freiburg, SchänzleStraße 1, D-79104 Freiburg, Germany

ABSTRACT

Summary: Hitherto, few environmental samples have been investigated by a ‘full cycle rRNA analysis’. Here the results of in situ hybridization experiments with specific rRNA-targeted oligonucleotide probes developed on the basis of new sequences derived from a previously described comparative 16S rRNA analysis of nocardioform actinomycetes in activated sludge are reported. Application of the specific probes enabled identification and discrimination of the distinct populations of nocardioform actinomycetes in activated sludge. One of the specific probes (DLP) detected rod-shaped bacteria which were found in 13 of the 16 investigated sludge samples from various wastewater treatment plants, suggesting their importance in the wastewater treatment process. Another probe (GLP2) hybridized with typically branched filaments of nocardioforms mainly found in samples from enhanced biological phosphorus removal plants, suggesting that these bacteria are involved in sludge foaming. The combination of in situ hybridization with fluorescently labelled rRNA-targeted oligonucleotide probes and confocal laser scanning microscopy improved the detection of nocardioform actinomycetes, which often showed only weak signals inside the activated-sludge flocs.

Author for correspondence: Markus Schuppler. Tel: +49 351 463 8591. Fax: +49 351 463 8573. e-mail: ms43@irz.inf.tu-dresden.de

Present address: Institut für Medizinische Mikrobiologie und Hygiene, Universitätsklinikum Carl Gustav Carus, Technische Universität Dresden, DürerStraße 24, D-01307 Dresden, Germany.

Present address: Institut für Medizinische Mikrobiologie und Hygiene, Medizinische Fakultät (Charité), Humboldt-Universitat Berlin, DorotheenStraße 96, D-10098 Berlin, Germany.

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