Jpn. J. Infect. Dis., 52, 1999

Laboratory and Epidemiology Communications
Emergence and Prevalence of a Novel Ⅵbrio parahaemolyticus

03:K6 Clone in Japan
Eiji Arakawa, Toshiyuki Murasel , Toshio Shimada, Tadayuki Okitsul,

Shiro Yamailand Haruo Watanabe*
DepartmenE of Bacteriology, National Institute of lnjTectious Diseases,
7Toyatna 1‑23‑1, Shinjuku, To砂0 162‑8640 andJDepartment of Bacteriology and Pathology, Kanagawa PrejTectural Public Health Laboratory,
Asahi, Yokoha〝は, Kanagawa 241 ‑0815

Communicated by Hiroshi Yoshikura(Accepted January 5, 2000)

Ⅵあrio parahaemolyticus was atthe top of the list of path0‑

from patients, food or the environment in 1997‑1998 ingens isolated from patients with food‑bom diseases in 1998
in Japan･ One of the dominant serotypes amongthe isolates

AomoriPrefecture; 61 isolates from patients, food or theenvironment in 198 I I 1998 in Kanagawa Prefecture; 6 isolates

was 03:K6･ This serotypehas been increasing since 1996

from patients in 1997‑1998 at Narita Airport Quarantine(1)･ Here we examined clonality of the 03:K6 isolates byusing pulsed‑field gel electrophoresis (PFGE) as previously

Station; and 3 isolates from patients in 1999 in Thailand

descritxxl (2). We used a total of 86 V. parahaeTWlyticus 03:K6(kindly provided from the National Institute of Health inisolates discovered between 1962 and 1999 Crable). TheseThailand). Fifteen isolates withother serotypes, including
03:K48, 03:K56, 04:K8, 04:K12, and 04:K68, were included

consisted of one isolate obtained in 1962; 5 isolates obtained
from patients in 1998 in the USA (kindly provided from CDC,of tdh (thermostable direct hemolysin) and trh (tdh‑relatedUSA); 2 isolates from patients in 1998 in Tokyo; 8 isolateshemolysin) genes and urease test were pe血med･in the analysis. In addition to PFGE analysis, PCR detection

Table･ PFGE and other markers of V･ parahaemolyticus 03:K6 isolates
Prw〇一〇en○&ｨｸ

tdhtrJI+

偖ﾂﾂ+

7DﾗG

PFqBEpatternS

ｲﾈ

ｶﾈ耳

烹

I$○tdedy○▲r(N○.dLS○lat○叫儂otlSWIﾂ+十ﾈ痔

AA田R

8澱冂c途

1996(5).1997(15),1998(42),1999(3)1981(6)

1981(2).1984(2),1995(1),1996(2)DDTotal1982(1)

EE

1986(1)FF

1985(1),1988(1)GG

1983(1)

HH

1962(1)

Il1987(1)JJ

1984(1)

剴ッ*Corresponding author: E‑mail : haruwata@nih･ goJP

246

Jpn. J. Infect. Dis., 52, 1999
03

K5648

K81268isolates in the Thai and US isolates. The same PFGE pattem

03:K604

A8C

D

E

ドwas repohed in India, SoutheastAsia, and the USA (3‑6).

GTaken together, these data may suggest the emergence of anew V. parahaemolyticus 03:K6 clonethat is spreading world‑
wide and already affecting Japan.

kb582

‑

485‑388‑

We thank the AomoriPrefectural Institute of Public Healthand Environment,the Tokyo Metropolitan Research Labora‑
tory and Public Health and the Narita Airport Quarantine

291 ̀‑

Station for providing strains.
194‑

145.5‑

97

This study was supported by a grant from the Ministry ofHealth and Welfare of Japan.
REFERENCES‑‑

I. National Institute of Infectious Diseases and Infectious

Diseases Division, Ministry of Health and Welfare (1999):
48.5‑

Vibrio parahaemolyticus, Japan, 19961 1998. Infect.
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2. Arakawa, E., Murase, T., Matsushita, S., Shimada, T.,

Yamai, S., Ito, T. and Watanabe, H. (2000): Pulsed‑field
gel electrophoresis‑based molecular comparison of Ⅵbriocholerae 0 1 isolates tNBtWeen domesticand imported cases

in Japan in 1997. J･Clin･ Microbiol･, 38, 424‑426･

Figure･ Representahve NotI‑cleaved PFGE pattems of V･ fWahaemolyticus

isolates･ hes from left toright indicate PFGEs of serotypes 03 :K56,

3. Okuda, ∫., Ishibashi, M., Hayakawa, E･, Nishino, T･,

03:K48, 04:K8, 04:K12, 04:K68, 03:K6, and 03:K6, designatedas groups A to G, respectively･ The position of each DNAISize (kb)

Takeda, Y., Mukhopadhyay, A. K., Garg, S. , Bhattacharya,

marker is shown onthe left.

of a unique 03:K6 clone of Vibrio parahaemolyticus ln

S･ K･, Nair, G･ B･ and Nishibuchi, M･ (1997): Emergen?らCalcutta, India, and isolation of strains from the same

clonalgroup from southeastAsiantravelers amvlng ln
PFGE pattems of NotI‑ or SPI‑digests of DNAsderived

Japan. ∫. Clin. Microbio1., 35, 3150‑3155･

from the 03:K6 isolates could be classified into 10 distinct
trh‑, and urease negative isolates obtained later than 1996

4. Bag, P. K., Nandi, S., Bhadra, R･ K･, Ramamurthy, T･,
Bhattacharya, S. K., Nishibuchi, M･, Hamabata, T･,
Yamasaki, S., Takeda, Y. and Nair, G. B. (1999): Clonal(65 strains including isolates from Thailand and the USA)

diversity among recently emerged strains of Vibrio

constituted group A, while strains with the same phenotype

parahaemolyticus 03:K6 associated with pandemic

groups, designated as A to J (Fig. and Table). All of the tdh',

Obtained in 198 1 constituted group B (Table). Thoughdiffer‑spread. ∫. Clin. Microbio1., 37, 2354‑2357･

ences were found between the PFGE pattem of each group ln

5. Centers for Disease Control and Prevention (1998):

03:K6 and other serotypes, recent isolates of the 04:K68
serotype were very similar to recent isolates of the 03:K6
serotype (compare the lane of 04:K68 andthat of groupA of

Outbreak of Ⅵあrio parahaemolyticus infections associ‑ated witheating oysters ‑ Pacific Northwest, 1 997･ Morbid.
Mortal. Wkly. Rep., 47, 457‑462･03:K6 in Fig.).
We showed here that recent V. parahaemolyticus 03:K6

6. Centers for Disease Control and Prevention (1999):
Outbreak of Vibrio parahaemolyticus infections associ‑isolates in Japanhad a PFGE pattem distinct from that

ated with eating raw oysters and clams haⅣested fromLeng Island Sound ‑ Connecticut, New Jersey,and NewYork, 1998. Morbid. Mortal. Wkly. Rep., 48, 48‑51.

prevalent in previous isolates. In addition, We identified aPFGE pattem indistinguishable from those of recent Japanese247