Studies on the Quantitative Method for Determination of Hemorrhagic Activity of Habu Snake Venom

Abstract

Hemorrhage is one of the most prominent manifestations produced by the parenteral administration of various snake venoms especially of Habu (Mitsuhashi et al., 1959; Ohsaka et al., 1961) and other species of Crotalidae (Fidler et al., 1940; Taube et al., 1937). In order to study the principle(s) responsible for hemorrhage caused by snake venom, a quantitative method for the determination of hemorrhagic activity is required. Up to date, however, few attempts have been made to establish the quantitative estima tion of hemorrhagic activity of snake venom. Minton (1956) and Mitsuhashi et al. (1959) tried to estimate the hemorrhagic activity but the results they obtained were not repro ducible in our hands. The present authors proposed a new method, which consists of the following procedures; (1) intracutaneous injection of venom into the depilated back skin of rabbits; (2) accurate measurement of the size of hemorrhagic spot carried out from the inside of the removed skin; (3) application of the parallel line assay method for the estimation. By this method it was possible to study the relationships of hemorrhagic activity to proteolytic and other pathological activities of Habu venom (Ohsaka et al., 1960, 1961). In this paper, details of the new method will be presented. MATERIALS AND METHODS Snake venom: The work to be reported here dealt with the venom from a species of Trimeresurus flavoviridis (Habu).* Each pool of the crude venom was dried, powdered and then stored at room temperature until used. These preparations, dissolved in M/30 phosphate buffered saline (pH

Análise das atividades biológicas dos venenos de Philodryas olfersii (Lichtenstein) e P. patagoniensis (Girard) (Serpentes, Colubridae)

“…Na atividade hemorrágica utilizou-se o método descrito por KONDO et al (1960), modificado por THEAKSTON & REID (1983). Para a determinação do tempo de ação (cinética) dos venenos quanto ao desenvolvimento da atividade hemorrágica, alíquotas de 0,1 ml de uma solução de 20 µg dos venenos de P. olfersii e P. patagoniensis foram injetadas, i.d., na pele previamente depilada do dorso de ratos machos adultos, pesando de 230 a 250 g, sob anestesia leve de CO 2 .…”

“…Foi utilizado o método descrito por KONDO et al (1960), onde estipula-se a Dose Mínima Necrosante, que é definida como a menor quantidade de veneno, em microgramas que, quando injetada i.d. na pele do dorso de ratos, produz, em 24 h, uma lesão necrótica de 5 mm de diâmetro.…”

Caracterización biológica y acción de inhibidores de una fosfolipasa A2 del veneno de Lachesis muta

“…Se realizó según el Método de Kondo et al (1960); ratones (18 -22 g) fueron inyectados intraperitonealmente con 0,1 mL de la muestra, luego de 2 horas se sacrificaron. Retirándose la piel y evaluando el área hemorrágica formada.…”

Neutralization of edema, hemorrhage and myonecrosis induced by North American crotalid venoms in simulated first-aid treatments

“…Edema was based on the relative increase in diameter of the envenomated lower hindlimb, similar to the measurements of the footpad, as described by Lomonte et al (1993a). Usually, hemorrhage is measured using Kondo's method (Kondo et al, 1960) which requires an i.d. injection, not an Lm.…”