S Anaswara Krishnan1, R Dileepkumar2, Achuthsankar S Nair3, Oommen V Oommen4. 1. Department of Zoology, University of Kerala, Thiruvananthapuram 695581, Kerala, India. 2. Centre for Venom Informatics, University of Kerala, Thiruvananthapuram 695581, Kerala, India. Electronic address: dileepkamukumpuzha@gmail.com. 3. Department of Computational Biology and Bioinformatics, University of Kerala, Thiruvananthapuram 695581, Kerala, India. 4. Kerala State Biodiversity Board, Pallimukku, Pettah, Thiruvananthapuram 695024, Kerala, India.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: The folklore or traditional therapy in southern India widely utilizes a plethora of local herbs to treat the patients challenged with snake venom. Despite the widespread implementation of antisera therapy, the local population of the country still relies on this century's old medicinal formulas mainly due to the cost effectiveness, lesser side effects and also its cultural acceptability. The present study aims to validate the neutralizing ability of one such traditionally acclaimed antidote Ophiorrhiza mungos root extract against Russell's viper (Daboia russelii) venom in the early developing chick embryos. MATERIALS AND METHODS: The disc impregnated with venom, root extract or the combination of both was placed on the yolk sac membrane preferably over the anterior blood vessel of 6th day chick embryo. The neutralization/inhibition of venom-induced lethality or hemorrhage was achieved by incubating venom and extract before being applied to the embryo. The membrane stabilizing properties of root extract was estimated by HRBC lysis method. The preliminary phytochemical analysis was done to assess the phyto constituents in the root extract. RESULTS: The LD50 of Russell's viper venom in 6th day chick embryo was found to be 3 μg/μl. The neutralising effect of root extract was achieved by pre-incubating venom with various concentrations of extract and at the concentration of 10 μg/μl, 100% recovery of embryos was observed after 6h of incubation. Higher concentration of root extract showed remarkable results by completely abolishing traces of hemorrhagic lesions induced by viper venom. CONCLUSIONS: The above observations confirmed that the root extract of Ophiorrhiza mungos possess potent anti snake venom neutralizing compounds, which inhibit the activity of viper venom. The chick embryo, a new insensate model used in the present study is significant in venom research as it reduces the ruthless suffering of higher mammalian experimental models.
ETHNOPHARMACOLOGICAL RELEVANCE: The folklore or traditional therapy in southern India widely utilizes a plethora of local herbs to treat the patients challenged with snake venom. Despite the widespread implementation of antisera therapy, the local population of the country still relies on this century's old medicinal formulas mainly due to the cost effectiveness, lesser side effects and also its cultural acceptability. The present study aims to validate the neutralizing ability of one such traditionally acclaimed antidote Ophiorrhiza mungos root extract against Russell's viper (Daboia russelii) venom in the early developing chick embryos. MATERIALS AND METHODS: The disc impregnated with venom, root extract or the combination of both was placed on the yolk sac membrane preferably over the anterior blood vessel of 6th day chick embryo. The neutralization/inhibition of venom-induced lethality or hemorrhage was achieved by incubating venom and extract before being applied to the embryo. The membrane stabilizing properties of root extract was estimated by HRBC lysis method. The preliminary phytochemical analysis was done to assess the phyto constituents in the root extract. RESULTS: The LD50 of Russell's viper venom in 6th day chick embryo was found to be 3 μg/μl. The neutralising effect of root extract was achieved by pre-incubating venom with various concentrations of extract and at the concentration of 10 μg/μl, 100% recovery of embryos was observed after 6h of incubation. Higher concentration of root extract showed remarkable results by completely abolishing traces of hemorrhagic lesions induced by viper venom. CONCLUSIONS: The above observations confirmed that the root extract of Ophiorrhiza mungos possess potent anti snake venom neutralizing compounds, which inhibit the activity of viper venom. The chick embryo, a new insensate model used in the present study is significant in venom research as it reduces the ruthless suffering of higher mammalian experimental models.