Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia).
Abstract: Naja kaouthia (NK) causes the highest fatality due to snake venom poisoning in Thailand. The specific antivenom produced is of low potency and in short supply. The aim of this study was to improve the antivenom potency. Bentonite and complete Freund's adjuvants (CFA) and various immunogens were compared. Six groups of three to five horses were immunized as follows: Group 1, NK venom adsorbed on bentonite; Group 2, NK venom in CFA; Group 3, NK venom in CFA in multi-emulsion formulation; Group 4, NK venom in 25% CFA; Group 5, NK neurotoxin 3 (NK3) conjugated with tetanus toxoid (NK3-TT) in CFA; Group 6, NK3 conjugated with diphtheria toxoid (NK3-DT) in CFA. Horses in Group 2-6 produced antivenom of very high neutralizing activity, up to four times higher than that of horses of Group 1. CFA (100 or 25%) or as a multi-emulsion formulation, induced comparable neutralizing antibody production with all three immunogens. All horses showed normal weight gain during the course of immunization. Group 1 horses exhibited minimal local reactions while horses in the other five groups had mild and comparable local reactions at the injection sites. No significant differences in the reactions caused by CFA in different formulations or different immunogens were observed. The production of highly potent antivenom against N. kaouthia from these horses should help solve problems associated with the currently available antivenom.
Publication Date: 1997-10-23 PubMed ID: 9330463DOI: 10.1016/s0264-410x(97)00098-4Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article aims to improve the potency of horse antivenom made to combat the venom of the Thai cobra, a snake species causing most fatalities in Thailand due to snakebite. Various methods of immunization were compared before a treatment process was selected that displayed high neutralizing activity without significant side effects.
Objective of the Research
The primary objective of the research was to enhance the effectiveness of the Thai cobra antivenom. This snake’s venom is infamous for causing high death rates in Thailand, but the existing antivenom was found to be low in potency and supply.
- The researchers attempted to create a potent remedy by comparing various immunogens and adjuvants, like Bentonite and Freund’s adjuvant (CFA).
Research Methodology
Six groups of horses were immunized with different venom mixtures to compare their effectiveness. The groupings were as follows:
- Group 1: Horses received the cobra venom absorbed on Bentonite.
- Group 2: Horses were given the cobra venom in CFA.
- Group 3: Horses received the venom in CFA in a multi-emulsion formulation.
- Group 4: Horses were given the venom in 25% CFA.
- Group 5: Horses received the venom conjugated with tetanus toxoid in CFA.
- Group 6: Horses were given the venom conjugated with diphtheria toxoid in CFA.
Findings of the Research
The research revealed that horses from groups 2-6 had substantial neutralization activity. The activity was found to be up to quadruple compared to that of Group 1 horses.
- The proteins in Group 1 appeared to cause minimal local reactions, as observed after analysis.
- Contrarily, horses in groups 2-6 had mild and largely similar local reactions at injection sites.
- Interestingly, no significant differences were observed in the reactions caused by CFA in different formations or with different immunogens.
Significance of the Findings
Due to their comparable neutralizing antibody production across formulations and immunogens, complete Freund’s adjuvants (CFA) at 100% or 25% were concluded to be effective methods for venom neutralization.
- The research paves the way for producing highly potent antivenom against Thai cobra venom.
- The current antivenom problems should be resolved using these horses, which are inducing a strong neutralizing activity with negligible side effects.
- The growth of horses during the immunization period remained normal, suggesting the safety of the process.
Cite This Article
APA
Pratanaphon R, Akesowan S, Khow O, Sriprapat S, Ratanabanangkoon K.
(1997).
Production of highly potent horse antivenom against the Thai cobra (Naja kaouthia).
Vaccine, 15(14), 1523-1528.
https://doi.org/10.1016/s0264-410x(97)00098-4 Publication
Researcher Affiliations
- Department of Microbiology, Faculty of Science, Mahidol University, Bangkok, Thailand.
MeSH Terms
- Animals
- Antivenins / biosynthesis
- Antivenins / immunology
- Elapid Venoms / immunology
- Enzyme-Linked Immunosorbent Assay
- Freund's Adjuvant / immunology
- Horses
- Immunization
- Neutralization Tests
Citations
This article has been cited 9 times.- Song Y, Day CM, Afinjuomo F, Tan JE, Page SW, Garg S. Advanced Strategies of Drug Delivery via Oral, Topical, and Parenteral Administration Routes: Where Do Equine Medications Stand?. Pharmaceutics 2023 Jan 4;15(1).
- Arguedas M, Umaña D, Moscoso E, García A, Pereira C, Sánchez A, Durán G, Cordero D, Sánchez A, Segura Á, Vargas M, Herrera M, Villalta M, Gómez A, Salas C, Díaz C, María Gutiérrez J, León G. Comparison of adjuvant emulsions for their safety and ability to enhance the antibody response in horses immunized with African snake venoms.. Vaccine X 2022 Dec;12:100233.
- Gutiérrez JM, Albulescu LO, Clare RH, Casewell NR, Abd El-Aziz TM, Escalante T, Rucavado A. The Search for Natural and Synthetic Inhibitors That Would Complement Antivenoms as Therapeutics for Snakebite Envenoming.. Toxins (Basel) 2021 Jun 29;13(7).
- Ratanabanangkoon K. A Quest for a Universal Plasma-Derived Antivenom Against All Elapid Neurotoxic Snake Venoms.. Front Immunol 2021;12:668328.
- Lin JH, Lo CM, Chuang SH, Chiang CH, Wang SD, Lin TY, Liao JW, Hung DZ. Collocation of avian and mammal antibodies to develop a rapid and sensitive diagnostic tool for Russell's Vipers Snakebite.. PLoS Negl Trop Dis 2020 Sep;14(9):e0008701.
- Ratanabanangkoon K, Tan KY, Pruksaphon K, Klinpayom C, Gutiérrez JM, Quraishi NH, Tan CH. A pan-specific antiserum produced by a novel immunization strategy shows a high spectrum of neutralization against neurotoxic snake venoms.. Sci Rep 2020 Jul 9;10(1):11261.
- Ratanabanangkoon K, Tan KY, Eursakun S, Tan CH, Simsiriwong P, Pamornsakda T, Wiriyarat W, Klinpayom C, Tan NH. A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia.. PLoS Negl Trop Dis 2016 Apr;10(4):e0004565.
- Sapsutthipas S, Leong PK, Akesowan S, Pratanaphon R, Tan NH, Ratanabanangkoon K. Effective equine immunization protocol for production of potent poly-specific antisera against Calloselasma rhodostoma, Cryptelytrops albolabris and Daboia siamensis.. PLoS Negl Trop Dis 2015 Mar;9(3):e0003609.
- Richard G, Meyers AJ, McLean MD, Arbabi-Ghahroudi M, MacKenzie R, Hall JC. In vivo neutralization of α-cobratoxin with high-affinity llama single-domain antibodies (VHHs) and a VHH-Fc antibody.. PLoS One 2013;8(7):e69495.
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