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Journal of Clinical Microbiology, October 2000, p. 3555-3560, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Brachyspira aalborgi Infection Diagnosed by Culture and 16S Ribosomal DNA Sequencing Using Human Colonic Biopsy Specimens

Wolfgang Kraaz,1 Bertil Pettersson,2 Ulf Thunberg,3 Lars Engstrand,4 and Claes Fellström5,*

Department of Pathology,1 Department of Oncology,3 and Department of Microbiology,4 University Hospital, S-751 85 Uppsala, Department of Biotechnology, Royal Institute of Technology, S-100 44 Stockholm,2 and Department of Large Animal Clinical Sciences, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, S-750 07 Uppsala,5 Sweden

Received 17 March 2000/Returned for modification 11 June 2000/Accepted 28 July 2000

In this study we report on the isolation and characterization of the intestinal spirochete Brachyspira aalborgi using human mucosal biopsy specimens taken from the colon of a young adult male with intestinal spirochetosis. A selective medium, containing 400 µg of spectinomycin/ml and 5 µg of polymyxin/ml was used for the isolation procedure. A high degree of similarity, in terms of phenotypic properties and 16S ribosomal DNA sequence, was observed between the isolated strain, named W1, and the type strain, 513A, of B. aalborgi. A similarity of 99.7% in the nucleotide sequence was found between W1 and 513AT, based on the almost-complete gene. A short segment of the 16S rRNA gene was amplified by PCR using genetic material enriched from paraffin-embedded biopsy specimens, which were taken from the patient on two occasions. The products showed 16S rRNA gene sequences virtually identical to that of strain 513AT in the actual region. Immunohistochemistry was performed on the colonic biopsy specimens with a polyclonal antibody raised against an intestinal spirochete isolated in a previous case of human intestinal spirochetosis. The antibody reacted strongly with the spirochete on the luminal epithelium. No immune reaction was seen within or below the surface epithelium. Routine histology did not reveal signs of colitis. Electron microscopy showed spirochetes attached end-on to the colonic mucosal surface. The isolate grew poorly on a commonly used selective medium for intestinal spirochetes, which may explain previous failures to isolate B. aalborgi.

* Corresponding author. Mailing address: Department of Large Animal Clinical Sciences, Faculty of Veterinary Medicine, Swedish University of Agricultural Sciences, Box 7018, S-750 07 Uppsala, Sweden. Phone: 46 18 671473. Fax: 46 18 672919. E-mail: Claes.Fellstrom{at}kirmed.slu.se.

Journal of Clinical Microbiology, October 2000, p. 3555-3560, Vol. 38, No. 10
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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