Habu Snake Venom Research Articles

Lack of a hemorrhagic principle in Habu snake venom, Trimeresurus flavoviridis, from the Okinawa Islands

Lack of a hemorrhagic principle in Habu snake venom, Trimeresurus flavoviridis, from the Okinawa Islands

Relation between serum creatine phosphokinase activity and the amount of habu ( Trimeresurus flavoviridis) snake venom injected in thigh muscle in rabbits

Relation between serum creatine phosphokinase activity and the amount of habu ( Trimeresurus flavoviridis) snake venom injected in thigh muscle in rabbits

AN ELECTORON MICROSCOPIC OBSERVATION ON THE MICROVASCULAR CHANGES CAUSED BY HABU SNAKE VENOM, WITH SPECIAL REFERENCE TO HEMORRHAGE

Crude Habu snake venom was injected in the femoral muscle of mice and the ultrastructures of the microvessels in the lesions were examined by electoron microscopy. Most of the vessels encountered between the damaged muscle cells were capillaries, partially containing postcapillary venules. The results were as follows.1. There were various injuries of the endothelial cells, such as formation of villous projections and intravascular blebs from cells, degeneration of cellular organelles, especially decrease and disappearance of pinocytotic vesicles, swelling, darkning and thinning of cells, formation of endothelial gaps and celullar necrosis.2. The fluffy changes, discontinuities and deficiency of the basement membrane were observed.3. The hemorrhage by Habu snake venom were occured by discontinuities of both the endothelium and the underlining basement membrane.4. It is considered that the increasing hemorrhage may be caused not only by the destruction of capillary wall, but also by the disturbance of coagulant activities.

Effect of thiol-compound on toxicity of habu snake (Trimeresurus flavoviridis hallowell) venom.

In the previous paper on the effects of various chelating agents on the systemic action of Habu snake venom, we reported that the treatment with EDTA enhanced the toxicity of the venom while thiol-compounds such as BAL and d-penicillarnine markedly reduced it (1). In the present work three thiol-compounds, that is 1-cysteine, thioglycolate-Na and dihydrothioctic acid were examined for their effects on the systemic action of Habu snake venom.

EFFECT OF THIOL-COMPOUND ON TOXICITY OF HABU SNAKE (TRIMERESURUS FLAVOVIRIDIS HALLOWELL) VENOM

In the previous paper on the effects of various chelating agents on the systemic action of Habu snake venom, we reported that the treatment with EDTA enhanced the toxicity of the venom while thiol-compounds such as BAL and d-penicillamine markedly reduced it (1). In the present work three thiol-compounds, that is 1-cysteine, thioglycolate-Na and dihydrothioctic acid were examined for their effects on the systemic action of Habu snake venom.

Immunogenicity in monkeys of a combined toxoid from the main toxic principles separated from Habu snake venom.

Immunogenicity in monkeys of a combined toxoid from the main toxic principles separated from Habu snake venom.

BIOLOGICAL TOXICITY OF MAMUSHI-SNAKE VENOM (AGKISTRODON HALYS) AND MORPHOLOGICAL CHANGES CAUSED BY THE VENOM

In this paper, the biological toxicity of Mamushi- snake venom and morphological changes caused by the venom are described.1. LD50. of the venom to ddN male mice weighing 17to 20g. are 19-23γ intravenously, 27-31γ intraperitoneally and 97-118γ intramuscularly.2. Haemorrhagic action is recognized even in 0.08 γ per 0. 1ml. of the venom, Out of four kinds of land-inhabiting snake-Mamushi -snake venom, Habu-snake venom and Sakishi mahabusnake venom-Mamushi snake venom has the strongest haemorrhagic activity.3. The haemorrhagic and myolytic action are influenced by pH.4. The haemorrhagic and lethal activity are inactivated by heating at 70°C for 5 minutes or at 100°C for 1 minute.5. In mice, and guinea pigs, morphological changes caused by Mamushi-snake venom are as follow.in mice, inoculated with the venom intravenously, intraperitoneally or intramuscularly, haemorrhagic lesions in the injected portion and degenerative and necrotic lesions in the parenchymal organs are common findings.In guinea pigs, inoculated intramuscularly, morphological changes are haemorrhage and coagulations-necrosis at the inoculated portion, and degenerative and necrotic lesions in the parenchymal organs. Further, adrenal glands, liver, thymus, lymphnodes and spleen present reactive changes probably due to stressor.As above montioned, Mamushi snake venom has lethal, haemorrhagic and myonecrotic action. Furthermore it is morphologically considered to have action as stressor.

ハブ蛇毒のアルコール処理による弱毒化とその免疫原性について

Exposure of Habu-snake venom solution to heat at 56°C for 30 minutes causes protein precipitation, which is almost non-toxic. The toxicity of centrifuged supernatant is slightly weakened.When 70% alcohol is added to this supernatants, minute substances which are insoluble in water are precipitated, and the toxicity of the precipitate is remarkably attenuated.By the addition of 0.1%-1% formalin (or Aluminum Potassium Sulfate) to the precipitate, the attenuation of the toxicity tends to increase, and the amount of its protein content is less than 1/2 of that in the non-heated solution. Immunizing effect of formalin-added, alcohol-treated antigen proves effective enough to act as a satisfactory immunogen against the toxicity of Habu-venom-hemorrhagic necrotic damagea t the site.In order to produce antibody in man throng preventive injection against Habu bite, the formalin added, alcohol-treated antigen might be applicable without accompanying any serious by-effects.

Studies on Habu snake venom. VII. Heat-stable myolytic factor and development of its activity by addition of phospholipase A : Kurashige, S., Hara, Y., Kawakami, M. and Mitsuhashi, S. (Department of Microbiology, School of Medicine, Gunma University, Maebashi, Japan). Jap. J. Microbiol.10, 23, 1966

Studies on Habu snake venom. VII. Heat-stable myolytic factor and development of its activity by addition of phospholipase A : Kurashige, S., Hara, Y., Kawakami, M. and Mitsuhashi, S. (Department of Microbiology, School of Medicine, Gunma University, Maebashi, Japan). Jap. J. Microbiol.10, 23, 1966

Studies on Habu snake venom. VII. Heat-stable myolytic factor and development of its activity by addition of phospholipase A.

ABSTRACTIt has been found that there are two factors responsible for myolysis in Habu snake venom; heat‐labile and heat‐stable myolytic factors. As reported previously, the former is a proteinase contained in Habu venom. In this article purification and chemical properties of the heat‐stable myolytic factor are presented. Using the method of fractionation with ammonium sulfate and with column chromatography, a heat‐stable myolytic factor was obtained ultracentrifugally in a homogeneous state. This factor caused severe myolysis with the aid of both Mg++ ion and phospholipase A. It has been found that heat‐stable myolytic factor has no activities such as those of proteinase, esterase or ribonuclease. The relationship between the heat‐stable myolytic factor and phospholipase A in myolytic activity is discussed.

ハブ蛇毒トキソイドの基礎的研究

Mice and rabbits were immunized with Habu Snake Venom treated with equal amount of Freund's incomplete adjuvant, and Cocarboxylase or Dihydrothioctic acid (DHTA). After four weeks, 3 or 5 booster injections of untreated or DHTA treated venom were given weekly. And then, the neutralizing effects of the sera of the animals were tested. They were also challenged intramuscularly into the legs with the venom.It was elucidated that those immunized animals were protected from local hemorrhagic and necrotic action and lethal effect of venom in considerable degree although the protective potency of the sera of the animals was lower than that of antivenin used for treatment of snake bite in man. As it was also confirmed that no side reaction was seen by the injection of DHTA treated venom, an active immunization of men was tried, injecting 5mg of DHTA treated venom divided into two at the interval of two weeks. The immunizing potency of the sera of vaccinated persons was almost the same as that of the animals. Thus, it was suggested that DHTA inactivated toxoid of habu venom was proved to be useful for prophylaxis against the legion caused by Habu snake bite.

ハブ蛇毒トキソイドによるハブ咬症の予防に関する研究

Dihydrothioctic acid inactivated Habu snake toxoid was injected subcutaneously into 2846 people of Kamimotobu Village, Ryukyu Islands. They received shots of 0.5ml. of toxoid, containing 2.5mg. of Habu snake venom and DHTA, at the interval of two weeks. Main local reactions were slight localized pain which continued for a week, and slight swelling. Generalized symptoms such as fever were very few. Effectiveness of the toxoid on Habu snake bite will be further investigated by means of increasing the volunteers who wish the shots.

STUDIES ON THE IMMUNIZATION OF HORSES BY HABU SNAKE VENOM TREATED BY FREUND'S ADJUVANT OR DIHYDROTHIOCTIC ACID

It was elucidated that 5mg. or 100mg. of Habu Snake Venom (Trimeresurus flavoviridis) were available as first immunizing doses in horses when it was treated by Freund's incomplete adjuvant or 100mg. of Dihydrothioctic acid. Four weeks after, 1mg. of plain venom was injected subcutaveously into horses, and the dose was then increased duplicatedly in every week. After ten or fifteen times of booster injection, 0.1ml. of the sera showed to neutralize sufficiently the lethal effect and local lesion due to the venom of 300γ or more. The sera were mixed in vitro with the various amounts of venom and injected intramuscularly into the legs of mice. Thus the total amount of venom used for the immunization and the periods of immunization were reduced to one half in compared with the immunization method without adjuvant adopted until now.

ハブ毒とエラブウミヘビ毒の研究

This paper outlines the natures of Habu snake and Erabu sea snake and properties and actions of their venoms. Moreover was described the effect of tannic acid on the venoms.Habu snake (Trimereserus flavoviridis) is venomous, landinhibiting on the Amami islands and about 150cm long. The victims of Habu snake bite was estimated at 250 to 300 each year. The death rate during recent 7 years was more than 1 per cent. The minimal lethal dosis for mice, weighing 15 to 17g. was about 150γ/0.1ml by intramusculare injection. It was considered that the venom was composed of haemorrhagic, angiotoxic and myolytic factors, which were completly inactivated by heating at 100C for 10 minutes, and heat-stable myolytic factor.Erabu sea snake (Laticauda semifasciata) lives on the coast of Amami Oshima, and has strong fatal venoms. Minimal lethal dosis, in experiments with mice weighing 15 between 17g., was about 6γ/0.1ml by the intramusculare injections. Erabu sea snake venom is considered to have chiefly neurotoxic component which was relatively stable in heating.It was recognized that the toxic activities of the venoms of these different species were inhibited by aqueous solution of tannic acid; a 8.5% solution inactivated lethal and local haemorrhagic activities of 500γ/0.1ml of Habu snake venom, and fatal toxicities of 25γ/0.1ml of Erabu sea snake venom.The above mentioned effect of tannic acid on the venoms may be due to coagulations of the venom and tissue proteins by tannic acid.

EXPERIMENTAL STUDIES ON THE ACTIVE IMMUNIZATION OF GUINÉA PIGS BY HEATED HABU SNAKE VENOM

Immunization with heated Habu Snake venom was attempted on guinea pigs.1. Freund's incomplete adjuvant was added to equivalent volumes of supernatants of heated Habu Snake venom (50mg/1.0ml in 0.85% saline), and 1. 0ml of the mixture was subcutaneously injected into the back of guinea pigs. One week after, 0. 1ml of the heated venom (500γ/0.1ml) was injected one time as additional immunization (booster) 2. When, at two to twelve week after the final immunization, the animals were challenged by injecting 500γ or less of the crude venom into the femoral muscles, the grade and extension of the local damage in them were 1/3 to 1/4 as small as those in the nonimmunized animals.3. When 0. 1ml of the sera obtained from the immunized animals was mixed in vitro with various amounts of the crude and the heated venoms and the mixture were injected intramuscularly into the legs of mice, they depressed the local pathological changes which were visible after injection of 64γ to 34γ of the crude venom or heated venom obtained from 500γ to 250γ of the crude venom.

Studies on Habu snake venom. V. Myolysis caused by phospholipase A in Habu snake venom : Maeno, H., Mitsuhashi, S., Okonogi, T., Hoshi, S. and Homma, M. (Dept. of Microbiology and Pathology, Univ. Gunma, Maebashi, Japan). Jap. J. Exp. Med., 32, 55, 1962

Studies on Habu snake venom. V. Myolysis caused by phospholipase A in Habu snake venom : Maeno, H., Mitsuhashi, S., Okonogi, T., Hoshi, S. and Homma, M. (Dept. of Microbiology and Pathology, Univ. Gunma, Maebashi, Japan). Jap. J. Exp. Med., 32, 55, 1962

Local pathological changes following injection of heat-stable substances in the Habu snake ( Trimeresurus flavoviridis) venom : Okonogi, T., Hoshi, S. Homma, M., Suto, K., Iizuka, H. and Sato, M. (First Dept. of Pathol., Gunma Univ., Maebashi, Japan). Kitakanto Med. J., 12, 199, 1962

Local pathological changes following injection of heat-stable substances in the Habu snake ( Trimeresurus flavoviridis) venom : Okonogi, T., Hoshi, S. Homma, M., Suto, K., Iizuka, H. and Sato, M. (First Dept. of Pathol., Gunma Univ., Maebashi, Japan). Kitakanto Med. J., 12, 199, 1962

Biochemical Analysis of Pathological Lesions Caused by Habu Snake Venom with Special Reference to Hemorrhage

Biochemical Analysis of Pathological Lesions Caused by Habu Snake Venom with Special Reference to Hemorrhage

LOCAL PATHOLOGICAL CHANGES FOLLOWING INJECTION OF HEAT-STABLE SUBSTANCES IN THE HABU-SNAKE (TRIMERESURUS FLAVOV IRIDIS) VENOM

An injection of Habu-snake venom, heated for 10 minutes at 100°C., caused myolysis at the site, histologically. But different from crude venom action, no hemorrhagic changes were recognized. At first, disappearance of mitchondria in the muscular fibers was seen, and then muscular splitting and lysis followed.While the myolytic changes caused by crude venom began immediately af ter the injection, the beginning of the changes were delayed when the heated material were used.Anti-Habu-serum neutralised the toxicity of the heated venom, and also, E.D.T.A.-Ca, Cocarboxylase and α-Thiolactoylglicine-Na repressed the toxicity in vitro.

Studies of H beta-proteinase of Habu snake venom, with special reference to substrate specificity and inhibitory effect of serum.

Studies of H beta-proteinase of Habu snake venom, with special reference to substrate specificity and inhibitory effect of serum.