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Applied and Environmental Microbiology, November 2000, p. 5043-5052, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Phylogenetic Analysis of and Oligonucleotide Probe Development for Eikelboom Type 021N Filamentous Bacteria Isolated from Bulking Activated Sludge

Takahiro Kanagawa,1,* Yoichi Kamagata,1 Shinobu Aruga,1 Tetsuro Kohno,2 Matthias Horn,3 and Michael Wagner3

National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, Tsukuba 305-8566,1 and Department of Civil and Environmental Engineering, Yamanashi University, Kofu 400-0016,2 Japan and Lehrstuhl für Mikrobiologie, Technische Universität München, D-85350 Freising, Germany3

Received 8 May 2000/Accepted 21 August 2000

Fifteen filamentous strains, morphologically classified as Eikelboom type 021N bacteria, were isolated from bulking activated sludges. Based on comparative 16S ribosomal DNA (rDNA) sequence analysis, all strains form a monophyletic cluster together with all recognized Thiothrix species (88.3 to 98.7% 16S rDNA sequence similarity) within the gamma-subclass of Proteobacteria. The investigated Eikelboom type 021N isolates were subdivided into three distinct groups (I to III) demonstrating a previously unrecognized genetic diversity hidden behind the uniform morphology of the filaments. For in situ detection of these bacteria, 16S rRNA-targeted oligonucleotide probes specific for the entire Eikelboom type 021N-Thiothrix cluster and the Eikelboom type 021N groups I, II, and III, respectively, were designed, evaluated, and successfully applied in activated sludge.

* Corresponding author. Mailing address: National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, 1-1 Higashi, Tsukuba 305-8566, Japan. Phone: 81-298-61-6026. Fax: 81-298-61-6009. E-mail: kanagawa{at}nibh.go.jp.

Applied and Environmental Microbiology, November 2000, p. 5043-5052, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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