Study On The Preparation Of Antivenin Against The Common Sea Snake Venom

It is suggested by the study on the marine poisonous pest along the coast that sea snake injury is most commonly seen and most severe marine life injury. For lack of specific antidote, sea snake injury has become a common problem for the health safeguard of sea exploration, marine work and transoceanic fight. There are 9 genus 16 kinds of sea snake mainly in the South Sea, Northern Gulf and the coast along Hai Nan, Tai Wan, Guang Xi, Guang Dong, and Fu Jian. The snakes are predominantly Laticaudinae and Hydrophiinae. In the pairing-season and when attacked, the sea snake is apt to attack human being actively. When lack of specific antivenom, about 80% of the patients will die from pulmonary failure in 48h after being injured. Now the management of sea snake bite is just for the symptoms. And there is no antivenom specific for snakebite. Furthermore, the antivenom for land snake is of no use for sea snake. So there are reports of death from sea snake bite every year.Objectives:To prepare the antivenom for Hydrophis cynocinctus and to study its practical usage in emergency. To provide the effective safeguard for sea creature injury.Methods:1. The Hydrophis cynocinctus were beheaded before the venom was got and cryodesiccated. The venom was attenuated with 2% methanal and sterilized. And then it was used to immunize the rabbit with the adjuvant. The potency was assessed with immune-double diffusion and ELISA. After the standardization of the potency, blood was taken from the animal and the serum was isolated.2. The serum was salted out with ammonium sulfate and purified withimmunoaffinity chromatography. After purified, the serum was used in the in vivo and ex vivo experiments on mouse to evaluate its neutralizing capacity.3. In the establish of snake bite animal model, 40 New Zealand rabbits were randomized into 4 groups: the control one, intravenous antivenom at 0.5h after poisoned, intravenous antivenom at 2h after poisoned, intravenous antivenom at 4h after poisoned. In all the four groups, the physio-biochemical indicators (including HR, SBP, DBP, RR, RI, ALT, LDH, CK, UREA, CR, PaO₂, PaCO₂, PH, SaO₂) and ECG and electrolyte (K⁺, Na⁺) were determined before being poisoned and at 0.5h, 1h, 2h, 3h, 4h, 5h, 6h, 7h, 8h, 10h after being poisoned to evaluate the early emergency management of antivenom of Hydrophis cynocinctus and its outcome.Results:1. After 19 weeks of immunization, the immunopotency was 1:8 by immune-double diffusion with very clear line of sedimentation and the potency reaches 1,280,000 by ELISA. When the potency was stable, blood was taken from the experimental animal.2. After twice of ammonium sulfate salt-out, SDS-PAGE electrophoresis of the serum was adopted to get the better purified globulin with higher recovery rate. By immunoaffinity chromatography, highly purified antibody can be collected but with low recovery rate (only 16.7mg of antibody from 10ml serum). It was indicated by the trial blood test that 1ml crudely extracted serum and 1ml serum get by immunoaffinity chromatography can neutralize 0.45mg (24.5 LD₅₀) and 0.80mg(46.2 LD₅₀)of the venom of Hydrophis cynocinctus. In the cross trial blood test, lml of highly purified serum can neutralize 0.34mg (18.8LD₅₀) venom of Lapemis hardwickii.3. Administration of adequate antivenom at various time points for sea snake bite shows that in all the animals the indicators had been ameliorated by different degree. In the 0.5h group, 1-2 hours after the administration of antivenom, the indicators had significant difference with that of the group of being poisoned and were in the normal range. In the 2h group, 2⁴h after the administration of antivenom, some indicators were in the normal range and some were higher than normal. In all the groups, the ECG shows...