Production of an effective anti-Bothrops-tetanus mixed hyperimmune serum of equine origin

The present investigation reveals the possibility of simultaneous immunization of horses with Bothrops or Crotalus snake venoms and Tetanus antigens for the production of anti-Bothrops-Tetanus or anti-Crotalus-Tetanus mixed serum, with high titers of the respective specific antibodies. Bothrops antivenoms with an average neutralizing titer of 4.16 mg venom/ml were obtained from plasma of horses with titers lower than 0.5 mg venom/ml when Tetanus antigens were not used. This suggests the existence of a synergism between Bothrops venoms and Tetanus antigens in the stimulation of the antibody response. The pooled plasma of the animal had a neutralizing titer of 21.0 mg/ml reference Bothrops venoms and 3,300 IU/ml to Tetanus antigens after purification by enzymatic digestion and ammonium sulphate precipitation. These experiments lead us to conclude that Bothrops envenomation therapy can be successfully performed using Anti-Bothrops-Tetanus serum also serving as Tetanus prophylaxis. anti-Crotalus-Tetanus serum can also be produced, although it is not of medical interest as Crotalus envenomation rarely results in local necrotizing lesions.

Neutralization of the effect of Crotalus durissus terrificus venom by gangliosides

We determined the ability of a mixture of gangliosides (16 percent) GDlb, 19 percent GT1b, 21 percent GM1, 40 percent GD1a) to neutralize the effect of Crotalus durissus terrificus (Cdt) venom in vitro and in vivo. Protection was indicated by the absence of muscular contractions, hind limb paralysis or death of BLB/c mice (16-18g) after receiving Cdt venom (1µgCdt venom containing 0.6 µg protein) at the doses indicated. A dose of Cdt venom above 0.9µg (ip) or 1 µg (im) induced muscular contraction and above 1.2 µg (ip) or 5.5 µg (im) the venom induced muscular contraction and hind limb paralysis. Cdt venom BOVE 2.5 µG (IP) OR 9 µg (im) induced all these symptoms and 95 to 100 percent death in experimental animals. The lethal dose 50 percent of the Cdt venom used was 8µ (im) and 1.5 µg (ip). In vitro studies, 4 mg gangliosides neutralized the effect of up to 1.5 µg Cdt venom. Quantities as low as 0.2 mg gangliosides were capable neutralizing 0.9 µg of Cdt venom in vitro. Intramuscular treatment with 1 mg gangliosides performed 60 min after the intramuscular injection of 5 µg Cdt venom protected 100 percent of the animals. In contrast, no protection was achieved with intraperitoneal treatment with gangliosides. The data show that gangliosides were effective in neutralizing the toxic effect induced by Crotalus durissus terrificus venom both in vitro and in vivo and that post-exposure intramuscular treatment with gangliosides could protect animals experimentally inoculated with the venom