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Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY)

Received: 17 October 2014     Accepted: 28 October 2014     Published: 2 February 2015
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Abstract

Infusion of mammalian antivenom is considered to be the best available treatment for snake bites; but, production of cost-effective IgG in pure form is challenging. Purification of egg yolk immunoglobulins (IgY) raised against various toxins has been found to be relatively easier. But to use IgY for therapeutic purpose its efficacy and safety need to be experimentally proven which is hardly done due to lack of an appropriate model. In this study, pure IgY against viper venom was isolated and its efficacy and safety for intravenous infusion was tested in rabbits. Rabbit snake bite model was created by subcutaneous injection of 2x lethal dose50 (LD50) venom. Animals were given intravenous infusion of pure anti-viper IgY and recovery was monitored. Isolated chicken immunoglobulin (IgY) was >90% homogenous and showed 1:32 titre in immunodiffusion experiment. The minimum hemorrhagic dose (MHD) of viper venom was 0.2mg and antihaemorragic dose (AHD) of IgY was 4x concentration (0.8 mg) of native venom. Subcutaneously injected venom at LD50 resulted in severe local reaction, coagulation abnormality and mortality in rabbits. When anti viper IgY was infused within 2 h of envenomation, the animals survived, clotting parameters were reversed to normal and animals showed steady weight gain like healthy animals. No adverse effect of IgY was noticed on renal or hepatic function. The efficacy of commercially available mammalian IgG was lower than that of anti-viper IgY. Long term stability of the purified and lyophilized IgY was demonstrated. The effective IgY dose required to prevent mortality in the envenomed rabbits was found to be 4x of the injected venom estimated by Lowry’s protein assay. It has been demonstrated that rabbit model of snake bite is successfully cured by anti-snake IgY infusion at a specific dose.

Published in American Journal of Clinical and Experimental Medicine (Volume 3, Issue 1)
DOI 10.11648/j.ajcem.20150301.14
Page(s) 32-38
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2015. Published by Science Publishing Group

Keywords

Antivenom, Chicken IgY, Russell’s Viper, Effective Dose, Animal Model

References
[1] Chippaux JP. Snake-bites: appraisal of the global situation. Bull World Health Org. 1998; 76:515.
[2] Gutiérrez JM, Lomonte B, Leon G, Rucavado A, Chaves F, et al. Trends in snakebite envenomation therapy: scientific, technological and public health considerations. Curr Pharm Des 2007; 13: 2935.
[3] Maya DC, Mary VB, Lal AV, Umashankar, PR and Krishnan LK.An Improved Method for Isolation of Anti- Viper Venom Antibodies from Chicken Egg Yolk . Journal of Biochemical and Biophysical Methods. 2001;51:129.
[4] Paul K, Manjula ., Deepa EP, Selvanayagam ZE, Ganesh KA and Subba Rao PV. Anti-Echis carinatus venom antibodies from chicken egg yolk: isolation, purification and neutralization efficacy. Toxicon. 2007;50: 893.
[5] Almeida CMC, Da Silva CL, Couto HP, Escocard Rde C, DA, ROCHA DG, Sentinelli Lde P, Kipnis TL, da Silva WD. Development of process to produce polyvalent IgY antibodies anti-African snake venom. 2008; Toxicon. 52:293.
[6] Maya DC, Mary VB, Krishnan LK. Development of anti viper venom antibodies in chicken egg yolk and assay of its antigen binding capacity. Toxicon, 2001;40: 857.
[7] Simpson ID. The “worldwide shortage” of antisnake venom: is the only right answer “produce more” or is it also “use it smarter?”. Wilderness Environ Med 2008;19: 99.
[8] Theakston RDG, Warrell DA and Griffiths E. Erratum to report of WHO workshop on the standardization and control on antivenoms. Toxicon. 2003;41:541
[9] Seneviratne SL, Opanayaka CJ, Ratnayake NS, Kumara KE, Sugathadasa AM, et al. Use of antivenom serum in snake bite: a prospective study of hospital practice in the Gampaha district. Ceylon Med J. 2000;45:65.
[10] Premawardhena AP, de Silva CE, Fonseka MM, Gunatilake SB, de Silva HJ. Low dose subcutaneous adrenaline to prevent acute adverse reactions to antivenom serum in people bitten by snakes: randomised, placebo controlled trial. BMJ. 1999;318:1041.
[11] Phillips RE, Theakston RD, Warrell DA, Galigedara Y, Abeysekera DT, et al.. Paralysis, rhabdomyolysis and haemolysis caused by bites of Russell's viper (Vipera russelli pulchella) in Sri Lanka: failure of Indian (Haffkine) antivenom. Q J Med. 1988;68:691.
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    Lissy Kalliyana Krishnan, Juliet Baby Saroja, Madhumati Rajalingam, Vimala John, Mohanan Parayathala Valappil, et al. (2015). Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY). American Journal of Clinical and Experimental Medicine, 3(1), 32-38. https://doi.org/10.11648/j.ajcem.20150301.14

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    ACS Style

    Lissy Kalliyana Krishnan; Juliet Baby Saroja; Madhumati Rajalingam; Vimala John; Mohanan Parayathala Valappil, et al. Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY). Am. J. Clin. Exp. Med. 2015, 3(1), 32-38. doi: 10.11648/j.ajcem.20150301.14

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    AMA Style

    Lissy Kalliyana Krishnan, Juliet Baby Saroja, Madhumati Rajalingam, Vimala John, Mohanan Parayathala Valappil, et al. Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY). Am J Clin Exp Med. 2015;3(1):32-38. doi: 10.11648/j.ajcem.20150301.14

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  • @article{10.11648/j.ajcem.20150301.14,
      author = {Lissy Kalliyana Krishnan and Juliet Baby Saroja and Madhumati Rajalingam and Vimala John and Mohanan Parayathala Valappil and Harikrishnan Vijayakumar Sreelatha},
      title = {Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY)},
      journal = {American Journal of Clinical and Experimental Medicine},
      volume = {3},
      number = {1},
      pages = {32-38},
      doi = {10.11648/j.ajcem.20150301.14},
      url = {https://doi.org/10.11648/j.ajcem.20150301.14},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ajcem.20150301.14},
      abstract = {Infusion of mammalian antivenom is considered to be the best available treatment for snake bites; but, production of cost-effective IgG in pure form is challenging. Purification of egg yolk immunoglobulins (IgY) raised against various toxins has been found to be relatively easier. But to use IgY for therapeutic purpose its efficacy and safety need to be experimentally proven which is hardly done due to lack of an appropriate model. In this study, pure IgY against viper venom was isolated and its efficacy and safety for intravenous infusion was tested in rabbits. Rabbit snake bite model was created by subcutaneous injection of 2x lethal dose50 (LD50) venom. Animals were given intravenous infusion of pure anti-viper IgY and recovery was monitored. Isolated chicken immunoglobulin (IgY) was >90% homogenous and showed 1:32 titre in immunodiffusion experiment. The minimum hemorrhagic dose (MHD) of viper venom was 0.2mg and antihaemorragic dose (AHD) of IgY was 4x concentration (0.8 mg) of native venom. Subcutaneously injected venom at LD50 resulted in severe local reaction, coagulation abnormality and mortality in rabbits. When anti viper IgY was infused within 2 h of envenomation, the animals survived, clotting parameters were reversed to normal and animals showed steady weight gain like healthy animals. No adverse effect of IgY was noticed on renal or hepatic function. The efficacy of commercially available mammalian IgG was lower than that of anti-viper IgY. Long term stability of the purified and lyophilized IgY was demonstrated. The effective IgY dose required to prevent mortality in the envenomed rabbits was found to be 4x of the injected venom estimated by Lowry’s protein assay. It has been demonstrated that rabbit model of snake bite is successfully cured by anti-snake IgY infusion at a specific dose.},
     year = {2015}
    }
    

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    T1  - Rabbit Snake-Bite Model to Assess Safety and Efficacy of Anti Viper Chicken Antibodies (IgY)
    AU  - Lissy Kalliyana Krishnan
    AU  - Juliet Baby Saroja
    AU  - Madhumati Rajalingam
    AU  - Vimala John
    AU  - Mohanan Parayathala Valappil
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    DO  - 10.11648/j.ajcem.20150301.14
    T2  - American Journal of Clinical and Experimental Medicine
    JF  - American Journal of Clinical and Experimental Medicine
    JO  - American Journal of Clinical and Experimental Medicine
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    EP  - 38
    PB  - Science Publishing Group
    SN  - 2330-8133
    UR  - https://doi.org/10.11648/j.ajcem.20150301.14
    AB  - Infusion of mammalian antivenom is considered to be the best available treatment for snake bites; but, production of cost-effective IgG in pure form is challenging. Purification of egg yolk immunoglobulins (IgY) raised against various toxins has been found to be relatively easier. But to use IgY for therapeutic purpose its efficacy and safety need to be experimentally proven which is hardly done due to lack of an appropriate model. In this study, pure IgY against viper venom was isolated and its efficacy and safety for intravenous infusion was tested in rabbits. Rabbit snake bite model was created by subcutaneous injection of 2x lethal dose50 (LD50) venom. Animals were given intravenous infusion of pure anti-viper IgY and recovery was monitored. Isolated chicken immunoglobulin (IgY) was >90% homogenous and showed 1:32 titre in immunodiffusion experiment. The minimum hemorrhagic dose (MHD) of viper venom was 0.2mg and antihaemorragic dose (AHD) of IgY was 4x concentration (0.8 mg) of native venom. Subcutaneously injected venom at LD50 resulted in severe local reaction, coagulation abnormality and mortality in rabbits. When anti viper IgY was infused within 2 h of envenomation, the animals survived, clotting parameters were reversed to normal and animals showed steady weight gain like healthy animals. No adverse effect of IgY was noticed on renal or hepatic function. The efficacy of commercially available mammalian IgG was lower than that of anti-viper IgY. Long term stability of the purified and lyophilized IgY was demonstrated. The effective IgY dose required to prevent mortality in the envenomed rabbits was found to be 4x of the injected venom estimated by Lowry’s protein assay. It has been demonstrated that rabbit model of snake bite is successfully cured by anti-snake IgY infusion at a specific dose.
    VL  - 3
    IS  - 1
    ER  - 

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Author Information
  • Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum-695012, India

  • Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum-695012, India

  • Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum-695012, India

  • Thrombosis Research Unit, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum-695012, India

  • Division of Toxicology, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum-695012, India

  • Division Laboratory Animal Science, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Trivandrum, INDIA-695012

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