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Journal of Clinical Microbiology, March 2009, p. 727-733, Vol. 47, No. 3
0095-1137/09/$08.00+0 doi:10.1128/JCM.01574-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Genetic Heterogeneity among Strains of Treponema phagedenis-Like Spirochetes Isolated from Dairy Cattle with Papillomatous Digital Dermatitis in Japan 
Takahisa Yano,1
Ryoko Yamagami,1
Kazuhiro Misumi,2
Chikara Kubota,3
Kyaw Kyaw Moe,1
Tetsuya Hayashi,4,5,8
Kazunori Yoshitani,6
Osamu Ohtake,7 and
Naoaki Misawa1,8*
Laboratory of Veterinary Public Health, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan,1 Laboratory of Veterinary Surgery, Department of Veterinary Science, Faculty of Agriculture, Kagoshima University, 1-21-24 Kourimoto, Kagoshima 890-0065, Japan,2 Laboratory of Veterinary Reproduction, Department of Veterinary Science, Faculty of Agriculture, Kagoshima University, 1-21-24 Kourimoto, Kagoshima 890-0065, Japan,3 Division of Microbiology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, 5200 Kiyotake, Miyazaki 889-1692, Japan,4 Division of Bioenvironmental Science, Frontier Science Research Center, University of Miyazaki, 5200 Kiyotake, Miyazaki 889-1692, Japan,5 Hokubu Veterinary Clinical Center, NOSAI Chiba, 99-1 Katori, Chiba 289-0407, Japan,6 Tsuyama Veterinary Clinical Center, NOSAI Okayama, 8-9 Kokubunji, Tsuyama, Okayama 708-0843, Japan,7 Project for Zoonoses Education and Research, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan8
Received 14 August 2008/ Returned for modification 11 December 2008/ Accepted 3 January 2009
Papillomatous digital dermatitis (PDD) is an infectious foot disease of cattle that is prevalent throughout the world. Although it has been prevalent in Japan since the first case was reported in 1992, full epidemiological and bacteriological examinations have not been conducted. We collected 91 lesions of PDD from 80 dairy cattle on 12 farms in eight regions of Japan to isolate the spirochetes that are frequently detected in lesions. We isolated 40 strains of spirochetes from 24 cattle (30.0%) by a simple two-step culture technique, in which the biopsy samples were incubated at 4°C for 48 to 72 h in an enrichment broth supplemented with antibiotics, which improved the rate of isolation, and then inoculated on selective agar plates. All spirochetes examined were catalase positive and oxidase negative and showed weak beta-hemolytic activity. Enzyme activities were identical to those of Treponema phagedenis ATCC 27087. Sequencing of the 16S rRNA gene showed that all strains isolated had >99% identity to those of the T. phagedenis type strain and of T. phagedenis-like strains isolated from PDD lesions in the United States and Europe. Pulsed-field gel electrophoresis and PCR-based random amplified polymorphism DNA methods revealed considerable diversity among strains isolated not only from different cattle but also from the same individuals. These findings may provide further evidence for the role of these treponemes in the pathogenesis of persistent PDD.
* Corresponding author. Mailing address: Laboratory of Veterinary Public Health, Department of Veterinary Science, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan. Phone and fax: 81-985-58-7284. E-mail: a0d901u{at}cc.miyazaki-u.ac.jp
Published ahead of print on 14 January 2009.
Journal of Clinical Microbiology, March 2009, p. 727-733, Vol. 47, No. 3
0095-1137/09/$08.00+0 doi:10.1128/JCM.01574-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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