Purification of the thermostable direct hemolysin of Vibrio parahaemolyticus by immunoaffinity column chromatography

1986 ◽  
Vol 32 (1) ◽  
pp. 71-73 ◽  
Author(s):  
Takeshi Honda ◽  
Myonsun Yoh ◽  
Ikuyo Narita ◽  
Toshio Miwatani ◽  
Huilan Sima ◽  
...  
Keyword(s):  
Column Chromatography ◽  
Vibrio Parahaemolyticus ◽  
Culture Supernatant ◽  
Immunoaffinity Column ◽  
Simple Method ◽  
Thermostable Direct Hemolysin

A simple method involving immunoaffinity column chromatography to purify the thermostable direct hemolysin of Vibrio parahaemolyticus was developed. The thermostable direct hemolysin purified from the culture supernatant of a strain isolated from the first reported case of V. parahaemolyticus infection in China in 1985 was indistinguishable from the hemolysins purified from strains isolated in Japan.

scholarly journals Contribution of Vibrio parahaemolyticus Virulence Factors to Cytotoxicity, Enterotoxicity, and Lethality in Mice

2010 ◽  
Vol 78 (4) ◽  
pp. 1772-1780 ◽  
Author(s):  
Hirotaka Hiyoshi ◽  
Toshio Kodama ◽  
Tetsuya Iida ◽  
Takeshi Honda
Keyword(s):  
Cell Lines ◽  
Virulence Factors ◽  
Vibrio Parahaemolyticus ◽  
Biological Activities ◽  
Infection Model ◽  
Cytotoxic Activities ◽  
Mutant Strains ◽  
Thermostable Direct Hemolysin ◽  
Clinical Illness ◽  
Pathogenic Vibrios

ABSTRACT Vibrio parahaemolyticus, one of the human-pathogenic vibrios, causes three major types of clinical illness: gastroenteritis, wound infections, and septicemia. Thermostable direct hemolysin (TDH) secreted by this bacterium has been considered a major virulence factor of gastroenteritis because it has biological activities, including cytotoxic and enterotoxic activities. Previous reports revealed that V. parahaemolyticus strain RIMD2210633, which contains tdh, has two sets of type III secretion system (T3SS) genes on chromosomes 1 and 2 (T3SS1 and T3SS2, respectively) and that T3SS1 is responsible for cytotoxicity and T3SS2 is involved in enterotoxicity, as well as in cytotoxic activity. However, the relative importance and contributions of TDH and the two T3SSs to V. parahaemolyticus pathogenicity are not well understood. In this study, we constructed mutant strains with nonfunctional T3SSs from the V. parahaemolyticus strain containing tdh, and then the pathogenicities of the wild-type and mutant strains were evaluated by assessing their cytotoxic activities against HeLa, Caco-2, and RAW 264 cells, their enterotoxic activities in rabbit ileal loops, and their lethality in a murine infection model. We demonstrated that T3SS1 was involved in cytotoxic activities against all cell lines used in this study, while T3SS2 and TDH had cytotoxic effects on a limited number of cell lines. T3SS2 was the major contributor to V. parahaemolyticus-induced enterotoxicity. Interestingly, we found that both T3SS1 and TDH played a significant role in lethal activity in a murine infection model. Our findings provide new indications that these virulence factors contribute to and orchestrate each distinct aspect of the pathogenicity of V. parahaemolyticus.

Determination of Benzo (a) pyrene in Foods

1978 ◽  
Vol 61 (1) ◽  
pp. 129-135
Author(s):  
Yukio Saito ◽  
Hiroshi Sekita ◽  
Mitsuharu Takeda ◽  
Mitsuru Uchiyama
Keyword(s):  
Sulfuric Acid ◽  
Silica Gel ◽  
Column Chromatography ◽  
Analytical Method ◽  
Selective Extraction ◽  
Acid Extraction ◽  
Simple Method ◽  
Extraction Step ◽  
Concentrated Sulfuric Acid

Abstract An analytical method was developed for determining benzo (a) pyrene in foods, suitable for routine use. The method consists of 4 cleanup steps: (1) alkali cleavage of sample, (2) preliminary silica gel column chromatography, (3) selective extraction with concentrated sulfuric acid, and (4) further silica gel column chromatography. Recoveries of benzo- (a) pyrene added to 50 g (or 10 g) food at levels of 0.4 ppb (or 2 ppb) ranged from 70% for short-necked clam and mackerel to 85% for chicken meat. The sulfuric acid extraction step affords a simple method for isolating benzo (a)- pyrene from various kinds of interfering substances which could not be separated by existing methods.

scholarly journals An Estuarine Neritid Gastropod, Clithon corona, a Potential Reservoir of Thermostable Direct Hemolysin-Producing Vibrio parahaemolyticus

2005 ◽  
Vol 67 (8) ◽  
pp. 833-835 ◽  
Author(s):  
Aiyamperumal VELAMMAL ◽  
Mina KATO ◽  
Sayaka MIYAGI ◽  
Megumi TOYOZATO ◽  
Norichika H. KUMAZAWA
Keyword(s):  
Vibrio Parahaemolyticus ◽  
Potential Reservoir ◽  
Thermostable Direct Hemolysin
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