Demonstration of a Temperature-Dependent Inactivating Factor of the Thermostable Direct Hemolysin in
            <i>Vibrio parahaemolyticus</i>

Takeda, Yasuharu; Hori, Yayoi; Miwatani, Toshio

doi:10.1128/iai.10.1.6-10.1974

Cited by 16 publications

(23 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The remarkable heat stability of the Kanagawa hemolysin has been reported previously (Miwatani et al, 1972;Sakurai et al, 1973;Takeda et al, 1974). Our study indicates that heat inactivation rates of this stable hemolysin in buffer, crustacea and shellfish homogenates are linear functions and thus differ sharply from the curvilinear thermal-destruction-rate plots that we obtained for Clostridium sp.…”

Section: Resultssupporting

confidence: 69%

See 1 more Smart Citation

Thermal Inactivation of the Kanagawa Hemolysin of Vibrio parahaemolyticus in Buffer and Shrimp

et al. 1984

View full text Add to dashboard Cite

Growth studiesA semipurified thermostable direct hemolysin (Kanagawa hemolysin) was prepared from a clinical isolate of Vibrio parahnemo(vticus (strain 533-72). Preliminary experiments determined that the hemolytic activity and mouse lethality of the hemolysin were inactivated at the same rate, that the rate was linear, and that the pH range of optimum thermal stability was pH 5.5-6.5. When approximately 256 rat erythrocyte units/g of Kanagawa hemolysin were heated at Il5", l20", l25", and 13O"C, D values ranged from 46.5-13 min (z, = 27°C) in Tris buffer and 48.1-10.4 min (z, = 23.2"C) in shrimp homogenate, both at pH 7.0.V. parahuemof~ticus strains were inoculated into sterile shrimp, crab or oyster meat to study their growth and hemolysin-producing capabilities. Frozen, shelled market shrimp were comminuted in a blender. Live Gulf Coast crabs were heated briefly (121"C, 10 min) in a retort, and the friable meat picked from the shell. Live, wholesale oysters were shucked, and the flesh and liquor comminuted. Each seafood sample was sterilized by autoclauing (l2l"C, 20 min) and recomminuted if necessary. A suitable quantity (400-6OOg) was then transferred asceptically to a sterile beaker.

show abstract

Section: Resultssupporting

confidence: 69%

“…Limited investigations of the time-temperature relationship of Kanagawa hemolysin inactivation were conducted by Miwatani et al (1972) and Takeda et al (1974). Sakurai et al (1973) reported that purified hemolysin in phosphate buffer (pH 7.0) was not inactivated by heating at 100°C for 10 min.…”

Section: Hemolysin Productionmentioning

confidence: 99%

Thermal Inactivation of the Kanagawa Hemolysin of Vibrio parahaemolyticus in Buffer and Shrimp

et al. 1984

View full text Add to dashboard Cite

show abstract

“…The purified preparation of thermostable direct hemolysin contained a single protein with a molecular weight of about 42,000 (7).Assay of hemolytic activity. Hemolytic activity was assayed as described previously (7,13). The standard reaction mixture (2.5 ml) contained 10 mM tris(hydroxymethyl)aminomethane (Tris) -hydrochloride buffer (pH 7.2), 10 mM CaCl12, 0.9% NaCl, the thermostable direct hemolysin, and 1.25 ml of a 1% suspension of human erythrocytes (about 8.3 x 107 cells/ml).…”

mentioning

confidence: 99%

Inactivation of the biological activities of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside Gt1

Takeda¹,

Takeda²,

Honda³

et al. 1976

Infect Immun

View full text Add to dashboard Cite

Biological activities of the thermostable direct hemolysin produced by Vibrio parahaemolyticus, such as its hemolytic activity and lethal activity, were inhibited by neuraminidase-sensitive gangliosides, of which GT1 ganglioside was the most inhibitory. Neuraminidase-resistant gangliosides did not affect the activities of the hemolysin. Results showed that horse erythrocytes, which are resistant to the hemolysin, do not contain the neuraminidase-sensitive gangliosides GT, and GD1a. Therefore, we propose that neuraminidase-sensitive gangliosides, and especially GT1 ganglioside, may be the receptor sites on the membranes for the thermostable direct hemolysin of V. parahaemolyticus.In a previous paper, we reported that the hemolytic activity of the thermostable direct hemolysin of Vibrio parahaemolyticus was inhibited by a ganglioside mixture and that GM, ganglioside was not related to this inhibitory effect (14). To determine the tissue receptor for the thermostable direct hemolysin, we studied the inhibitory effects of various ganglioside components on biological activities of the thermostable direct hemolysin and found that neuraminidase-sensitive gangliosides, and especially GTI ganglioside, inhibited these biological activities. MATERIALS AND METHODSPurification of thermostable direct hemolysin. The thermostable direct hemolysin was isolated from culture filtrates of a Kanagawa phenomenonpositive strain, V. parahaemolyticus WP-1, and purified by successive column chromatographies on diethylaminoethyl-cellulose, hydroxylapatite, and Sephadex G-200, as described previously (7). The purified preparation of thermostable direct hemolysin contained a single protein with a molecular weight of about 42,000 (7).Assay of hemolytic activity. Hemolytic activity was assayed as described previously (7,13). The standard reaction mixture (2.5 ml) contained 10 mM tris(hydroxymethyl)aminomethane (Tris) -hydrochloride buffer (pH 7.2), 10 mM CaCl12, 0.9% NaCl, the thermostable direct hemolysin, and 1.25 ml of a 1% suspension of human erythrocytes (about 8.3 x 107 cells/ml). The reaction mixture was incubated at 37 C for 30 min and then centrifuged at 3,000 rpm for 5 min. The hemolytic activity was determined by measuring the absorbance of the resulting supernatant fluid at 540 nm.Assay of lethal toxicity. Lethal toxicity was assayed by injecting the thermostable direct hemolysin intravenously into 4-to 6-week-old mice (ddO strain) and measuring the survival time of the animals.Gangliosides. The ganglioside mixture used was type II from Sigma Chemical Co. Gangliosides GTI; GDIa, GM,, and GM2 (8). were obtained from Supelco Inc. GM, and GM2 were treated with V. cholerae neuraminidase (Calbiochem, B grade), as described below, before use. Treatment of gangliosides with neuraminidase.Ganglioside was incubated with V. cholerae neuraminidase (Calbiochem, B grade) in 0.2 ml of 0.01 M Tris-hydrochloride buffer (pH 7.2) for 48 h at 37 C. Then, the reaction mixture was dried in a rotary evaporator and the ganglioside was extracted into a mi...

show abstract

“…However, hepatic phosphorylase activity in the poisoned mice was appreciably higher than that in control mice after 2 hr, whereas its activity after 18 hr was not significantly different from that in the control fasted mice (37). Another study (41) in our laboratory has shown that in mice injected intraperitoneally with live Vibrio parahaemolyticus (pathogenic strain), the hepatic phosphorylase and G-6-Pase activities showed a tendency similar to the rise and fall in the blood sugar level although the pathogen exacerbated a heat-stable hemolytic toxin (47) showing lethality to mice, in addition to endotoxin.…”

Section: Discussionmentioning

confidence: 71%

Metabolie Aspects in Mice Administered Live Salmonella typhimurium

Sakaguchi

Hsu

Sakaguchi

1980

Microbiology and Immunology

View full text Add to dashboard Cite

The influence of intraperitoneal inoculation of live Salmonella typhimurium on carbohydrate and lipid metabolisms in mice was investigated at doses of 9.2 × 107 cells, 1.9 × 108 cells, and 3.8 × 108 cells. The hepatic glycogen content in mice at 18 hr after the inoculation decreased in inverse proportion to an increase in the injection dose. The activities of hepatic phosphorylase and G‐6‐Pase increased significantly after 2 hr, but after 18 hr the levels of both enzyme activities, especially G‐6‐Pase, declined in inverse porportion to an increase in dose of viable cells administered to the mice. The levels of serum glucose and free fatty acids (FFA) in mice markedly decreased at doses of 1.9 × 108 and 3.8 × 108 cells after a transient rise at early stage (1 hr) after the injection. Marked hypertriglyceridemia was seen in infected mice. However, the activity in serum lipoprotein lipase (LPL) was reduced by an increase in the injection dose. The effect of intraperitoneal administration of viable cells on the serum triglyceride level was prevented in mice immunized with S. typhimurium endotoxin or administered with the anti‐endotoxin serum. These results indicate that hypertriglyceridemia mainly results from the action of endotoxin in the pathogen. Serum lactate dehydrogenase (LDH) activity markedly increased at the dose of 3.8 × 108 cells within 8–16 hr, and the infected mice exhibited a leakage of isozymes LDH‐3 and 5 in the serum 16 hr post‐inoculation.

show abstract

Demonstration of a Temperature-Dependent Inactivating Factor of the Thermostable Direct Hemolysin in Vibrio parahaemolyticus

Cited by 16 publications

References 2 publications

Thermal Inactivation of the Kanagawa Hemolysin of Vibrio parahaemolyticus in Buffer and Shrimp

Thermal Inactivation of the Kanagawa Hemolysin of Vibrio parahaemolyticus in Buffer and Shrimp

Inactivation of the biological activities of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside Gt1

Metabolie Aspects in Mice Administered Live Salmonella typhimurium

Contact Info

Product

Resources

About