Sea snake venoms contain less protein than those of land snakes (Toom et al., 1969). Sea snake venoms lack arginine ester hydrolyzing activity, whereas those of Crotalidae and Viperidae have such activity (Tu et al., 1966). Sea snakes live in salty water, and their venoms may be different from those of land snakes. Because of the difficulty in obtaining sea snake venoms, information about sea snake venoms is quite incomplete. NGF is commonly present in the venoms of land snakes such as Elapidae, Viperidae, and Crotalidae (Cohen and Levi-Montalcini, 1956; Lipps, 2002). It is therefore of interest to investigate the presence or absence of NGF in sea snake venoms. In order to investigate the presence or absence of NGF, five sea snake venoms were selected. Lapemis hardwickii (Hardwick's sea snake) and Acalyptophis peronii venom were obtained from the Gulf of Thailand. Hydrophis cyanocinctus (common sea snake) and Enhydrina schistosa (beaked sea snake) venom were obtained from the Strait of Malacca. Laticauda semifasciata (broad band blue sea snake) venom was also examined and the venom was obtained from Gato Island in the Philippines.
Immunization with Bungarus candidus venom was performed in four rabbits at high dose (initial dose, 75 microg/kg) and low dose (initial dose, 50 microg/kg). Each dose group consisted of two rabbits; one rabbit received the venom subcutaneously (s.c.) and the other intradermally (i.d.). The venom was injected as emulsified solutions with the same volume of Freund's complete adjuvant until the 4th immunization, thereafter as plain solutions. By stepwise increments of the immunizing dose, the higher dose group received a dose of 200 microg/kg and the lower dose group 150 microg/kg after the 5th immunization, respectively. Thereafter, seven additional immunizations were performed within six months. All rabbits were sacrificed two weeks after the last immunization (12th). Antilethal activity of the immunized antisera thus obtained was determined not only with the homologous venom but also with two heterologous venoms from Bungarus fasciatus and Bungarus flaviceps. Immunodiffusion analysis was also performed with these venoms. The results obtained in this pilot trial provided useful information for production of Malayan krait antivenom at Queen Saovabha Memorial Institute.