Evaluation of health status of horses immunized with snake venom and montanide adjuvants, IMS 3012 (nanoparticle), ISA 206 and ISA 35 (emulsion based) during polyvalent snake antivenom production: hematological and biochemical assessment.
Abstract: Several biochemical and hematological changes in horses are observed during production of snake antivenom. Although conventional adjuvants like Freund's (Complete and Incomplete) are good immunopotentiators, they produce considerable local reactions in animals. Variety of commercial adjuvants, like montanide adjuvants, having high immunopotentiation and showing lesser side effects are available. The prime objective during antivenom production is to strike a balance between safety of immunized horses and efficacy of the product. In our earlier work, efficacy of montanide group of adjuvants in antivenom production has already been established. The aim of the present work was to assess the safety parameters in horses, viz.: biochemical and hematological, during production of snake antivenom. In the present study, 33 new horses were randomly divided into four groups and hyperimmunized using mixture of snake venoms, viz.: Cobra venom, Russell's viper venom, Krait venom and Echis venom along with montanide adjuvants, IMS 3012, ISA 206, ISA 35 and Incomplete Freund's adjuvant as a control adjuvant; through subcutaneous route at intervals of two weeks. During the immunization period, biochemical and hematological parameters were monitored at 0th, 14th, 21st, 30th and 42nd weeks. The mean hemoglobin values dropped slightly during initial immunization but subsequently regained to normal levels. The mean serum total protein values and globulin levels showed an increment in all the four groups, compared to day zero, vice-versa a slight drop was observed in albumin levels. No significant changes were observed in serum creatinine, bilirubin, alkaline phosphatase and blood urea nitrogen values. Finally, we conclude that montanide adjuvants could be a safer alternative to the conventional adjuvants for primary phase of immunization in antivenom production.
Copyright © 2014 Elsevier Ltd. All rights reserved.
Publication Date: 2014-02-28 PubMed ID: 24582735DOI: 10.1016/j.toxicon.2014.02.012Google Scholar: Lookup
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- Journal Article
Summary
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The study examined the health of horses immunized with snake venom and various montanide adjuvants during the production of snake antivenom. Results indicate that montanide adjuvants are a safer alternative for primary immunization in antivenom production.
Montanide Adjuvants and Immunization
- The study was centered around the production of snake antivenom where horses are typically immunized with snake venom and various immunostimulants (adjuvants).
- Different adjuvants, such as Freund’s Complete and Incomplete, are often used but are known to produce significant side effects. An alternative, montanide adjuvants, which come in different variations such as IMS 3012, ISA 206, and ISA 35, show promise for providing the necessary immune response with fewer side effects.
- A previous study confirmed the efficacy of montanide adjuvants for antivenom production, laying the groundwork for this study to investigate the health effects on the horses.
Research Methodology
- 33 horses were assigned into four groups and were immunized with a mix of snake venoms – Cobra, Russell’s viper, Krait, and Echis venom – and montanide adjuvants or Freund’s adjuvant (serving as the control group). Immunizations were given subcutaneously every two weeks.
- The researchers assessed biochemical and hematological parameters on week 0, 14, 21, 30, and 42.
Findings and Conclusion
- While there was a small initial decrease in hemoglobin levels, they returned to normal during the course of the study. Total protein and globulin levels in the blood serum of the horses increased over the study period compared to baseline measurements, but albumin levels saw a slight decline.
- Key kidney and liver function markers, including creatinine, bilirubin, alkaline phosphatase, and blood urea nitrogen values, did not show significant changes during the immunization period. This suggests that the immunization process, particularly with the montanide adjuvants, does not adversely impact these biologically significant areas.
- The study concludes that montanide adjuvants could be safer alternatives to conventional adjuvants for the primary phase of immunization during snake antivenom production, as it does not negatively affect the horses’ health. However, further study is needed for additional validation.
Cite This Article
APA
Waghmare AB, Salvi NC, Deopurkar RL, Shenoy PA, Sonpetkar JM.
(2014).
Evaluation of health status of horses immunized with snake venom and montanide adjuvants, IMS 3012 (nanoparticle), ISA 206 and ISA 35 (emulsion based) during polyvalent snake antivenom production: hematological and biochemical assessment.
Toxicon, 82, 83-92.
https://doi.org/10.1016/j.toxicon.2014.02.012 Publication
Researcher Affiliations
- Antitoxins & Sera Department, Haffkine Biopharmaceutical Corporation Limited, Pune, Maharashtra, India. Electronic address: arunw2000@yahoo.com.
- Antitoxins & Sera Department, Haffkine Biopharmaceutical Corporation Limited, Pune, Maharashtra, India.
- Department of Microbiology, University of Pune, Pune, Maharashtra, India. Electronic address: writetodeopurkar@gmail.com.
- Department of Pharmacology, Progressive Education Society's Modern College of Pharmacy, Nigdi, Pune, Maharashtra, India. Electronic address: priyank.shenoy@rediffmail.com.
- Indian Institute of Toxicology (IIT), Pune, Maharashtra, India.
MeSH Terms
- Adjuvants, Immunologic / administration & dosage
- Adjuvants, Immunologic / chemistry
- Adjuvants, Immunologic / pharmacology
- Animals
- Antivenins / biosynthesis
- Antivenins / immunology
- Blood Cell Count
- Blood Chemical Analysis
- Emulsions
- Female
- Freund's Adjuvant
- Health Status
- Horses / immunology
- Male
- Nanoparticles
- Snake Venoms / immunology
Citations
This article has been cited 13 times.- Mirzaei Z, Sari S, Moghaddam Pour M, Hassanzadeh SM, Damizadeh B, Taghizadeh M, Mahdavi M. Yavar-70A, a novel water-in-oil adjuvant: A potency study in HPV-16E7d vaccine model. Iran J Basic Med Sci 2025;28(2):224-229.
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- Miranda ALS, Antunes BC, Minozzo JC, Lima SA, Botelho AFM, Campos MTG, Chávez-Olórtegui C, Soto-Blanco B. The Health Status of Horses Used for at Least Six Complete Cycles of Loxoscelic Antivenom Production. Toxins (Basel) 2023 Sep 26;15(10).
- Ratanabanangkoon K. Polyvalent Snake Antivenoms: Production Strategy and Their Therapeutic Benefits. Toxins (Basel) 2023 Aug 24;15(9).
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- Cardoso-Arenas S, Clement H, Arenas I, Olvera F, Zamudio F, Caliskan F, Corrales-García LL, Corzo G. Recombinant expression and antigenicity of two peptide families of neurotoxins from Androctonus sp. J Venom Anim Toxins Incl Trop Dis 2022;28:e20220026.
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- Ramos MGDSN, Campos SDE, Strauch MA, Ott LC, Macieira DB, de Alencar NX, Lessa DAB. Serum proteins and electrophoretic profile in horses undergoing crotalid venom hyperimmunization. J Equine Sci 2022 Sep;33(3):31-35.
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- Clement H, Corrales-García LL, Bolaños D, Corzo G, Villegas E. Immunogenic Properties of Recombinant Enzymes from Bothrops Ammodytoides Towards the Generation of Neutralizing Antibodies against Its Own Venom. Toxins (Basel) 2019 Dec 2;11(12).
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