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      Characterization of Daboia russelii and Naja naja venom neutralizing ability of an undocumented indigenous medication in Sri Lanka

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          Abstract

          Background

          Indigenous medicinal practice in Sri Lanka talks about powerful compounds extracted from native plants for treating venomous snake bites which are hardly documented in literature but are used by the indigenous doctors for thousand years.

          Objective

          We screened the neutralizing ability of a herbal preparation practiced in indigenous medicine of Sri Lanka, consisting of Sansevieria cylindrica, Jatropha podagrica and Citrus aurantiifolia, for its ability to neutralize venom toxins of Naja naja (Common Cobra) and Daboia russelii (Russell's viper).

          Materials and methods

          The venom toxicity was evaluated using a 5-day old chicken embryo model observing the pathophysiology and the mortality for six hours, in the presence or absence of the herbal preparation. The known toxin families to exist in snake venom, such as Phospholipase A 2, Snake venom Metalloprotease, were evaluated to understand the mechanism of venom neutralizing ability of the herbal preparation.

          Results

          The LD 50 of D. russelii venom, as measured using the 5-day old chicken embryo model, was 4.8 ± 0.865 ug (R 2 = 84.8%, P = 0.079). The pre-incubation of venom with the herbal preparation increased the LD 50 of D. russelii venom to 17.64 ± 1.35 μg (R 2 = 81.0%, P = 0.100), showing a clear neutralizing action of D. russelii venom toxicity by the herbal medicine. Whereas the pre-incubation of venom with the 1× venom neutralizing dose of commercially available polyvalent anti-venom serum shifted the LD 50 venom only up to 5.5 ± 1.35 μg (R 2 = 98.8%, P = 0.069). In the presence of the herbal preparation, Phospholipase A 2 activity of D. russelii venom was significantly reduced from 9.2 × 10 −3 mM min −1 to 8.0 × 10 −3 mM min −1 and that of N. naja from 2.92 × 10 −2 mM min −1 to 0.188 × 10 −2 mM min −1. Further, the pre-incubation of N. naja venom with the herbal preparation significantly reduced its Metalloprotease activity from 0.069 units min −1 to 0.019 units min −1.

          Conclusion

          The herbal preparation shows a clear neutralizing action against the toxicities of D. russelii and N. naja venoms demonstrating the potential to be used as a plant based antidote for snake envenomation.

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          Most cited references13

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          Snakebite mortality in the world.

          In examining the relative importance of snakebite mortality in different parts of the world, the authors review the information collected concerning both snakebite mortality and the species of snake incriminated. Available statistical data are known to be unreliable and at best can serve to provide only an approximate and highly conservative estimate of the relative magnitude of the snakebite problem. The sources of error inherent in the data are discussed, and estimates are made of the probable mortality from snakebite in various areas of the world.
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            An alternative in vitro method for testing the potency of the polyvalent antivenom produced in Costa Rica.

            The ability of several batches of polyvalent antivenom to neutralize indirect hemolytic activity of Bothrops asper venom was studied using a sensitive plate test. All samples of antivenom tested effectively neutralized this activity. A highly significant correlation was observed between neutralization of indirect hemolysis and neutralization of lethal activity. This simple and sensitive in vitro test could be used to monitor antibody levels in horses immunized to produce polyvalent antivenom.
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              An open, randomized comparative trial of two antivenoms for the treatment of envenoming by Sri Lankan Russell's viper (Daboia russelii russelii).

              Russell's viper (Daboia russelii russelii) is an important cause of morbidity and mortality in Sri Lanka. In a study in 1985, Haffkine equine polyspecific antivenom in doses up to 20 g proved ineffective in clearing antigenaemia and caused a high incidence of anaphylactoid reactions. A new, monospecific ovine Fabantivenom (Polonga TAb) has been developed against the venom of Sri Lankan Russell's viper and, to assess its safety and efficacy, we carried out (in 1997) an open, randomized comparison of this with the Haffkine antivenom currently in use in the country. Patients with systemic envenoming following Russell's viper bite were randomized to receive an initial intravenous dose of either 1 g of Polonga TAb (n = 23) or 10 g of Haffkine antivenom (n = 20). One dose of Polonga TAb permanently restored blood coagulability in only 9 (41%) of 22 patients and 13 needed repeated doses, whereas the majority (14/20; 70%) had restored coagulability after 1 dose of Haffkine antivenom. There was a tendency towards more rapid resolution of local swelling and systemic manifestations in the Haffkine group. Venom antigenaemia was eliminated more quickly in the Haffkine group and ovine Fab was cleared from the circulation more rapidly than equine F(ab')2. To evaluate safety, patients were closely observed for adverse reactions. Following a severe reaction with Haffkine antivenom all subsequent patients in this group were treated prophylactically with hydrocortisone and chlorpheniramine. Despite this, the incidence of adverse reactions was significantly higher in the Haffkine group compared with the PolongaTAb group (81% compared with 48%) and 4 patients had a severe anaphylactic reaction in the former group. In conclusion, the new antivenom is safer than Haffkine antivenom but, to avoid repeated doses, an initial dose higher than 1 g is needed in the treatment of Sri Lankan Russell's viper envenoming. The safety of this larger dose is the subject of further studies.
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                Author and article information

                Contributors
                Journal
                J Ayurveda Integr Med
                J Ayurveda Integr Med
                Journal of Ayurveda and Integrative Medicine
                Elsevier
                0975-9476
                0976-2809
                13 March 2017
                Jan-Mar 2017
                13 March 2017
                : 8
                : 1
                : 20-26
                Affiliations
                [a ]Faulty of Medicine, General Sir John Kotelawala Defence University, Ratmalana, 10390, Sri Lanka
                [b ]Department of Zoology, Faculty of Science, University of Colombo, Colombo, 03, Sri Lanka
                Author notes
                []Corresponding author. charithalg@ 123456kdu.ac.lk
                Article
                S0975-9476(16)30315-1
                10.1016/j.jaim.2016.10.001
                5377483
                28302413
                dc0e9df2-d851-4701-b293-3057f77f3533
                © 2016 Transdisciplinary University, Bangalore and World Ayurveda Foundation. Publishing Services by Elsevier B.V.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 28 September 2016
                : 4 October 2016
                : 5 October 2016
                Categories
                Original Research Article (Experimental)

                Complementary & Alternative medicine
                daboia russelii,naja naja,venom,neutralization,herbal preparation

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