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Applied and Environmental Microbiology, July 2003, p. 3883-3891, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3883-3891.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Prevalence of Pandemic Thermostable Direct Hemolysin-Producing Vibrio parahaemolyticus O3:K6 in Seafood and the Coastal Environment in Japan

Yukiko Hara-Kudo,1,{dagger} Kanji Sugiyama,2 Mitsuaki Nishibuchi,3 Ashrafuzzaman Chowdhury,4 Jun Yatsuyanagi,5 Yoshimitsu Ohtomo,6 Akinobu Saito,7 Hidetoshi Nagano,8 Tokuhiro Nishina,9 Hiroshi Nakagawa,10 Hirotaka Konuma,11 Michiko Miyahara,11 and Susumu Kumagai12*

Department of Biomedical Food Research, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640,1 Shizuoka Institute of Environment and Hygiene, Kitaandoh, Shizuoka 420-8637,2 Center for Southeast Asian Studies,3 Graduate School of Medicine, Kyoto University, Kyoto 606-8501,4 Akita Prefectural Institute of Public Health, Akita 010-0874,5 Aomori Prefectural Institute of Public Health and Environment, Aomori 030-8566,6 Saitama Institute of Public Health, Saitama 338-0824,7 Fukuoka Institute of Health and Environmental Sciences, Dazaifu, Fukuoka 818-0135,8 Tokai University Junior College, Shizuoka 420-8511,9 Tokyo Kenbikyoin Foundation, Chuo-ku, Tokyo 103-0015,10 Division of Microbiology, National Institute of Health Sciences, Setagaya-ku, Tokyo 158-8501,11 Department of Veterinary Public Health, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan,12

Received 25 October 2002/ Accepted 28 April 2003

Although thermostable direct hemolysin (TDH)-producing Vibrio parahaemolyticus has caused many infections in Asian countries, the United States, and other countries, it has been difficult to detect the same pathogen in seafoods and other environmental samples. In this study, we detected and enumerated tdh gene-positive V. parahaemolyticus in Japanese seafoods with a tdh-specific PCR method, a chromogenic agar medium, and a most-probable-number method. The tdh gene was detected in 33 of 329 seafood samples (10.0%). The number of tdh-positive V. parahaemolyticus ranged from <3 to 93/10 g. The incidence of tdh-positive V. parahaemolyticus tended to be high in samples contaminated with relatively high levels of total V. parahaemolyticus. TDH-producing strains of V. parahaemolyticus were isolated from 11 of 33 tdh-positive samples (short-necked clam, hen clam, and rock oyster). TDH-producing strains of V. parahaemolyticus were also isolated from the sediments of rivers near the coast in Japan. Representative strains of the seafood and sediment isolates were examined for the O:K serovar and by the PCR method specific to the pandemic clone and arbitrarily primed PCR and pulsed-field gel electrophoresis techniques. The results indicated that most O3:K6 tdh-positive strains belonged to the pandemic O3:K6 clone and suggested that serovariation took place in the Japanese environment.

* Corresponding author. Mailing address: Department of Veterinary Public Health, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan. Phone and fax: 81 3 5841 5469. E-mail: askuma{at}mail.ecc.u-tokyo.ac.jp.

{dagger} Present address: Division of Microbiology, National Institute of Health Sciences, Setagaya-ku, Tokyo 158-8501, Japan.

Applied and Environmental Microbiology, July 2003, p. 3883-3891, Vol. 69, No. 7
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.7.3883-3891.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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