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Abstract
Echis pyramidum (Epy) is a venomous snake belongs to Viperidae family; it causes fetal coagulopathy systemic effects and death. Searching for more effective and safe antivenin is mandatory for viper bites treatment. Proteases are the most lethal components in viper venom inducing hemorrhage, edema and coagulation problems. Thus, the study aims to evaluate the potency of the prepared antivenin and their neutralizing activities against the biological, biochemical and histopathological activities induced by whole Epy venom individually. Echis pyramidum metalloprotease enzyme was purified using gel filtration chromatography followed by DEAE-ion exchange chromatography. The purified Epy metalloprotease enzyme (SVMP) was detoxified with 1.5 kGy gamma rays from cobalt60 gamma cell and used for immunization. 1.5 kGy irradiated Epy metalloprotease (SVMPi) showed less lethal activity (LD50) compared to the corresponding native immunogen. The prepared antivenins boosted against whole Epy venom (WV), 1.5 kGy irradiated whole Epy venom (WVi), SVMP and SVMPi were tested for neutralization of lethality and biological, biochemical and histopathological activities induced by Epy venom. The ELISA titers of the sera pool from rabbits immunized by WVi and SVMPi were higher than their corresponding native ones. The anti-SVMPi serum showed the highest neutralization of lethality (ED50) compared to the other prepared antivenins. In addition, it prolonged the clotting time from 49.0 ± 2.5 to 176.2 ± 1.4 s. Furthermore, it demonstrated a highly neutralizing activity against edema induction and hemorrhage of Epy venom by 66.8% and 94.3%, respectively compared with the other prepared antivenins. Native Epy venom was very toxic to the kidney as it produced after 4h significant increase in urea, creatinine levels, MDA, TNF-Ü, VCAM-1, IL-18, renal caspase-1, NGAL and Kim-1 contents and significant decrease in albumin level, GSH and SOD contents compared to the normal group