Mitsuaki Nishibuchi

Characteristics of Vibrio parahaemolyticus O3:K6 from Asia.

ABSTRACT A variety of serovars of the food-borne pathogen Vibrio parahaemolyticus normally cause infection. Since 1996, the O3:K6 strains of this pathogen have caused pandemics in many Asian countries, including Taiwan. For a better understanding of these pandemic strains, the recently isolated clinical O3:K6 strains from India, Japan, Korea, and Taiwan were examined in terms of pulsed-field gel electrophoresis (PFGE) typing and other biological characteristics. After PFGE and cluster analysis, all the O3:K6 strains were grouped into two unrelated groups. The recently isolated O3:K6 strains were all in one group, consisting of eight closely related patterns, with I1(81%) and I5(13%) being the most frequent patterns. Pattern I1 was the major one for strains from Japan, Korea, and Taiwan. All recently isolated O3:K6 strains carried the thermostable direct hemolysin (tdh) gene. No significant difference was observed between recently isolated O3:K6 strains and either non-O3:K6 reference strains or old O3:K6 strains isolated before 1996 with respect to antibiotic susceptibility, the level of thermostable direct hemolysin, and the susceptibility to environmental stresses. Results in this study confirmed that the recently isolated O3:K6 strains of V. parahaemolyticus are genetically close to each other, while the other biological traits examined were usually strain dependent, and no unique trait was found in the recently isolated O3:K6 strains.

Prevalence of the Pandemic Genotype of Vibrio parahaemolyticus in Dhaka, Bangladesh, and Significance of Its Distribution across Different Serotypes

ABSTRACT Sixty-six strains of Vibrio parahaemolyticus belonging to 14 serotypes were isolated from hospitalized patients in Dhaka, Bangladesh, from January 1998 to December 2000. Among these, 48 strains belonging to four serotypes had the pandemic genotype and possessed the tdh gene. A marker (open reading frame ORF8) for a filamentous phage previously thought to correspond to the pandemic genotype was found to have a poor correlation with the pandemic genotype.

Manifestation of the Kanagawa phenomenon, the virulence‐associated phenotype, of Vibrio parahaemolyticus depends on a particular single base change in the promoter of the thermostable direct haemolysin gene

Thermostable direct haemolysin of Vibrio parahaemolyticus has been shown to be a major virulence factor. The Kanagawa phenomenon (KP), haemolysis induced by this haemolysin on a special blood agar medium, is strongly associated with clinical strains. We have been studying the expressions of various tdh genes encoding this haemolysin to elucidate the significance of the tdh genes possessed by KP‐negative strains isolated from patients. We examined the importance of the promoter sequence variation for expression level of the tdh gene in this study. Only the tdh2 gene, one of the two tdh genes (tdh1 and tdh2) present in a KP‐positive strain, was previously shown to be responsible for the haemolytic activity of the KP‐positive strain. The tdh1– and tdh2–lacZ fusions were used to determine and analyse the promoter sequence by primer extension and site‐directed mutagenesis methods. Two bases (positions −24 and −34) within the determined tdh2 promoter sequence were shown to be mostly responsible for the difference in the promoter strength between the tdh2 and tdh1 genes both in Escherichia coli and in V. parahaemolyticus backgrounds. Representative tdh promoters of KP‐negative strains are close to the tdh2 promoter; they differ at position −34 but have the same base at position −24 as the tdh2 promoter. We demonstrated that base substitution of the tdh promoters of KP‐negative strains only at position −34 is sufficient to increase the expression of these genes to the KP‐positive level. Therefore, the tdh genes of KP‐negative strains are considered to be potentially important because they can generate a KP‐positive subclone by a point mutation in their promoters.

Automated ribotyping differentiates vibrio parahaemolyticus O3:K6 strains associated with a Texas outbreak from other clinical strains.

Automated ribotyping with a Qualicon Riboprinter was used to determine whether clinical isolates of Vibrio parahaemolyticus O3:K6 recovered during two U.S. outbreaks of oyster-associated gastroenteritis in 1998 were related to each other and to a previously identified highly virulent Asian clone of this serotype. The patterns produced using the restriction enzymes Eco RI and Pst I suggest that the outbreak in the Northeastern United States was caused by a single strain closely related to the Asian clone. In contrast, it appears that multiple strains were involved in the Texas outbreak and that the predominant type was genetically distinct from the Northeastern and Asian clone.