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Bacterial Classification of Fish-Pathogenic Mycobacterium Species by Multigene Phylogenetic Analyses and MALDI Biotyper Identification System
Authors:Satoru Kurokawa  Jun Kabayama  Tsuguaki Fukuyasu  Seong Don Hwang  Chan-Il Park  Seong-Bin Park  Carmelo S del Castillo  Jun-ichi Hikima  Tae-Sung Jung  Hidehiro Kondo  Ikuo Hirono  Haruko Takeyama  Takashi Aoki
Institution:1. Animal Health Department of Research and Development Agricultural and Veterinary Division, Meiji Seika Pharma, 2-4-16, Kyobashi, Chuo-ku, Tokyo, 104-8002, Japan
2. Institute of Marine Industry, Department of Marine Biology and Aquaculture, College of Marine Science, Gyeongsang National University, 38 Cheondaegukchi-Gil, Tongyeong, 650-160, South Korea
3. Aquatic Biotechnology Center of WCU Project, College of Veterinary Medicine, Gyeongsang National University, 900 Gajwa-dong, Jinju, Gyeongnam, 660-710, South Korea
4. Laboratory of Genome Science, Tokyo University of Marine Science and Technology, Konan 4-5-7, Minato-ku, Tokyo, 108-8477, Japan
5. Department of Life Science and Medical Bioscience, Waseda University, 2-2 Wakamatsu-cho, Shinjuku, Tokyo, 162-8480, Japan
6. Consolidated Research Institute for Advanced Science and Medical Care, Waseda University, 513, Wasedatsurumaki-cho, Shinjuku-ku, Tokyo, 162-0041, Japan
Abstract:Mycobacterium marinum is difficult to distinguish from other species of Mycobacterium isolated from fish using biochemical methods. Here, we used genetic and proteomic analyses to distinguish three Mycobacterium strains: M. marinum strains MB2 and Europe were isolated from tropical and marine fish in Thailand and Europe, and Mycobacterium sp. 012931 strain was isolated from yellowtail in Japan. In phylogenetic trees based on gyrB, rpoB, and Ag85B genes, Mycobacterium sp. 012931 clustered with M. marinum strains MB2 and Europe, but in trees based on 16S rRNA, hsp65, and Ag85A genes Mycobacterium sp. 012931 did not cluster with the other strains. In proteomic analyses using a Bruker matrix-assisted laser desorption ionization Biotyper, the mass profile of Mycobacterium sp. 012931 differed from the mass profiles of the other two fish M. marinum strains. Therefore, Mycobacterium sp. 012931 is similar to M. marinum but is not the same, suggesting that it could be a subspecies of M. marinum.
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