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Journal of Clinical Microbiology, March 2001, p. 1057-1066, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.1057-1066.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Molecular Typing and Epidemiological Study of Salmonella enterica Serotype Typhimurium Isolates from Cattle by Fluorescent Amplified-Fragment Length Polymorphism Fingerprinting and Pulsed-Field Gel Electrophoresis

Yukihiro Tamada,1 Yuji Nakaoka,2 Kei Nishimori,3 Akira Doi,4 Takahiro Kumaki,5 Nobuko Uemura,6 Kiyoshi Tanaka,3 Sou-Ichi Makino,7 Toshiya Sameshima,8 Masato Akiba,9 Muneo Nakazawa,8 and Ikuo Uchida3,*

Nemuro Livestock Hygiene Service Center, Betsukaimidorimachi, Betsukai, Notsukegun, 086-0214,1 Kamikawa Livestock Hygiene Service Center, 4-15 Higashitakasu, Asahikawa 071-8154,2 Hokkaido Research Station, National Institute of Animal Health, 4 Hitsujigaoka,3 and Ishikari Livestock Hygiene Service Center, 3 Hitsujigaoka,5 Toyohira, Sapporo 062-0045, Kushiro Livestock Hygiene Service Center, 127-1 Otanoshike, Kushiro, 084-0917,4 Soya Livestock Hygiene Service Center, Hamatonbetu, Esashigun 098-5736,6 Obihiro University of Agriculture and Veterinary Medicine, Inada, Obihiro 080-8555,7 National Institute of Animal Health, Tsukuba Science City 305-0856,8 and Kyushu Research Station, National Institute of Animal Health, 2702 Chuzan, Kagoshima 891-0105,9 Japan

Received 14 September 2000/Returned for modification 16 December 2000/Accepted 6 January 2001

One hundred twenty Salmonella enterica serotype Typhimurium strains, including 103 isolates from cattle gathered between 1977 and 1999 in the prefecture located on the northern-most island of Japan, were analyzed by using fluorescent amplified-fragment length polymorphism (FAFLP) and pulsed-field gel electrophoresis (PFGE) to examine the genotypic basis of the epidemic. Among these strains, there were 17 FAFLP profiles that formed four distinct clusters (A, B, C, and D). Isolates that belonged to cluster A have become increasingly common since 1992 with the increase of bovine salmonellosis caused by serotype Typhimurium. PFGE resolved 25 banding patterns that formed three distinct clusters (I, II, and III). All the isolates that belonged to FAFLP cluster A, in which all the strains of definitive phage type 104 examined were included, were grouped into PFGE cluster I. Taken together, these results indicate that clonal exchange of serotype Typhimurium has taken place since 1992, and they show a remarkable degree of homogeneity at a molecular level among contemporary isolates from cattle in this region. Moreover, we have sequenced two kinds of FAFLP markers, 142-bp and 132-bp fragments, which were identified as a polymorphic marker of strains that belonged to clusters A and C, respectively. The sequence of the 142-bp fragment shows homology with a segment of P22 phage, and that of the 132-bp fragment shows homology with a segment of traG, which is an F plasmid conjugation gene. FAFLP is apparently as well suited for epidemiological typing of serotype Typhimurium as is PFGE, and FAFLP can provide a source of molecular markers useful for studies of genetic variation in natural populations of serotype Typhimurium.

* Corresponding author. Mailing address: Hokkaido Research Station, National Institute of Animal Health, 4 Hitsujigaoka, Toyohira, Sapporo, Hokkaido 062-0045, Japan. Phone: (81) 11-851-5226. Fax: (81) 11-853-0767. E-mail: ikuouchi{at}affrc.go.jp.

Journal of Clinical Microbiology, March 2001, p. 1057-1066, Vol. 39, No. 3
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.3.1057-1066.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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