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Int J Syst Evol Microbiol 59 (2009), 1336-1341; DOI  10.1099/ijs.0.000760-0
© 2009 International Union of Microbiological Societies

Mycobacterium kyorinense sp. nov., a novel, slow-growing species, related to Mycobacterium celatum, isolated from human clinical specimens

Mitsuhiro Okazaki1, Kiyofumi Ohkusu2, Hiroyuki Hata2, Hiroaki Ohnishi1, Keiko Sugahara3, Chizuko Kawamura4, Nagatoshi Fujiwara5, Sohkichi Matsumoto5, Yukiko Nishiuchi6, Kouichi Toyoda7, Hajime Saito8, Shota Yonetani1, Yoko Fukugawa1, Masayuki Yamamoto9, Hiroo Wada9, Akiko Sejimo3, Akio Ebina4, Hajime Goto9, Takayuki Ezaki2 and Takashi Watanabe1

1 Department of Laboratory Medicine, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka-shi, Tokyo 181-8611, Japan
2 Department of Microbiology, Regeneration and Advanced Medical Science, Gifu University Graduate School of Medicine, Gifu, 1-1 Yanagido, Gifu City, Gifu 501-1194, Japan
3 National Hospital Organization Tokyo National Hospital, 3-1-1 Takeoka, Kiyose-shi, Tokyo 204-0023, Japan
4 Central Laboratory, Aomori Prefectural Hospital, 2-1-1 Higashizoudou, Aomori-shi, Aomori 030-8553, Japan
5 Department of Host Defense, Osaka City University, Graduate School of Medicine, 1-4-3 Asahi-machi, Abeno-ku, Osaka-shi 545-8585, Japan
6 Toneyama Institute for Tuberculosis Research, Osaka City University Medical School, Osaka, Japan
7 Kyokuto Pharmaceutical Industrial Co. Ltd, 7-8, Nihonbashi-kobunachou, Chuuou-ku, Tokyo, 103-0024, Japan
8 Hiroshima Environment & Health Association, Health Science Center, 9-1 Hirosekita-machi, Naka-ku, Hiroshima 730-8631, Japan
9 Department of Respiratory Medicine, Kyorin University School of Medicine, 6-20-2 Shinkawa, Mitaka-shi, Tokyo 181-8611, Japan

Correspondence
Hiroaki Ohnishi
onishi{at}ks.kyorin-u.ac.jp

A novel, non-pigmented, slow-growing mycobacterium was identified on the basis of biochemical and nucleic acid analyses, as well as growth characteristics. Three isolates were cultured from clinical samples (two from sputum and one from pus in lymph nodes) obtained from three immunocompetent patients with infections. Bacterial growth occurred at 28–42 °C on Middlebrook 7H11-OADC agar. The isolates showed negative results for Tween hydrolysis, nitrate reductase, semiquantitative catalase, urease activity, 3 day arylsulfatase activity, pyrazinamidase, tellurite reduction and niacin accumulation tests, but positive results for 14 day arylsulfatase activity and heat-stable catalase tests. The isolates contained {alpha}-, keto-, and dicarboxymycolates in their cell walls. Sequence analysis revealed that all isolates had identical, unique 16S rRNA sequences. Phylogenetic analysis of the 16S rRNA, rpoB, hsp65 and sodA gene sequences confirmed that these isolates are unique but closely related to Mycobacterium celatum. DNA–DNA hybridization of the isolates demonstrated less than 50 % reassociation with M. celatum and Mycobacterium branderi. On the basis of these findings, a novel species designated Mycobacterium kyorinense sp. nov. is proposed. The type strain is KUM 060204T (=JCM 15038T=DSM 45166T).

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA, hsp65, proB and sodA gene sequences of strains KUM 060204T, NTH 512-121 and AHM 060905 are, respectively: AB370111, AB370169 and AB370170 (16S rRNA); AB370171, AB370176, and AB370177 (hsp65); AB370178, AB370182, and AB370183 (rpoB); AB370184, AB370188, and AB370189 (sodA).






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