Section 6
Chapter 5,939

Mouse foot pad test for the determination of anti edema forming and anti hemorrhagic potency of snake anti venom 1. cross neutralization test of habu trimeresurus flavoviridis and sakishima habu trimeresurus elegans venom with its anti venoms

Yamakawa, M.

Snake 13(1): 16-24


Accession: 005938008

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The expression of toxicity of snake venom in terms of lethal activity only was not sufficient; it was the final consequence of the complicated reactions of various toxic components. The toxicity of snake venom should be expressed in local effects such as hemorrhagic, edema-forming and necrotizing activities. Quantitative methods for the determination of edema-forming and hemorrhagic activities of habu and Sakishima-habu venom by injecting the venom into the foot pads of mice were proposed. An attempt was made to apply these methods for the assay of antiedema-forming and antihemorrhagic potencies of snake antivenoms. Lots of antivenom (4) were prepared from plasma of horses hyperimmunized with both crude venoms. Lot 12 and Lot T78 were obtained by immunization with crude habu venom (HC), while Lot 17 and Lot 26 were also prepared by immunization with crude Sakishima-habu venom (SC). These antivenoms were purified with ammonium sulfate, digested with pepsin and lyophilized. Mice in groups of 5 were given injections of 20 .mu.l sample solution into right hind foot pads and saline into left foot pads. After injection (4 h), the mice were killed by inhalation with chloroform. The percentage of the weight of the injected legs to that of the healthy legs was calculated to express the severity of edema. Mouse legs with hemorrhage were used to determine the extent of hemorrhagic activity by measuring Hb content. The cyanmethohemoglobin content of a solution was estimated photometrically at 540 nm with distilled water as a control and was calculated. There was little difference of edema forming activity between crude habu venom and crude Sakishima-habu venom. Crude habu venom showed intensive hemorrhagic effect in the mouse leg, while hemorrhagic activity of crude Sakishima-habu venom was insignificant. The results of cross neutralization tests of habu and Sakishima-habu venom with its antivenoms showed that habu and Sakishima-habu antivenoms effectively neutralized homologous venom. Crude Sakishima-habu venom was fairly neutralized by habu antivenom; Sakishima-habu antivenom had poor neutralizing power against crude habu venom.

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