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Int J Syst Evol Microbiol 53 (2003), 1609-1614; DOI  10.1099/ijs.0.02541-0
© 2003 International Union of Microbiological Societies

Flow cytometric sorting, phylogenetic analysis and in situ detection of Oscillospira guillermondii, a large, morphologically conspicuous but uncultured ruminal bacterium

Kazuhiro Yanagita1,2, Akira Manome1,2, Xian-Ying Meng1, Satoshi Hanada1, Takahiro Kanagawa1, Takayasu Tsuchida1,2, Roderick I. Mackie3 and Yoichi Kamagata1

1 Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Central 6, Higashi 1-1-1, Tsukuba, Ibaraki 305-8566, Japan
2 Center Research Laboratories, Ajinomoto Co., Kawasaki 210-8681, Japan
3 Department of Animal Sciences, University of Illinois Urbana–Champaign, Urbana IL 61801, USA

Correspondence
Yoichi Kamagata
y.kamagata{at}aist.go.jp

Flow cytometric sorting based on its large cell size enabled an enriched fraction of Oscillospira guillermondii cells to be obtained from the rumen contents of a sheep. Phylogenetic analysis based on cloned 16S rDNA sequences indicated that the bacterium is a member of the low-G+C Gram-positive bacterial cluster. Sporobacter termitidis and Papillibacter cinnamivorans were the most closely related known species, with sequence similarities of only 86·3–88·1 %. Fluorescently labelled 16S rRNA-targeted oligonucleotide probes specific for Oscillospira were designed and applied to the rumen sample from which the enriched fraction was obtained. The probes hybridized specifically with the large, morphologically conspicuous Oscillospira cells.

Abbreviations: FCM, flow cytometry; FISH, fluorescent in situ hybridization

Published online ahead of print on 9 May 2003 as DOI 10.1099/ijs.0.02541-0.

The GenBank/EMBL/DDBJ accession numbers for the O. guillermondii 16S rDNA sequences OSC1–OSC5 are AB040495–AB040499.




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