International Journal of Tropical Insect Science

, Volume 11, Issue 6, pp 907–916 | Cite as

Rhipicephalus Appendiculatus: Immunochemical Isolation, Identification and Characterization of Tick Gut Antigens Recognized by IgGs Derived from Rabbits Repeatedly Infested with Ticks

  • Abdul O. Mongi
  • C. A. Aganyo
Research Article


Immune resistance to infestation by the brown ear tick, Rhipicephalus appendiculatus Neumann, was induced in rabbits by repeated tick feeding. Antibodies from the rabbits resistant to ticks were purified and used to isolate tick midgut antigens. Purified immunoglobulin G (IgGs) from rabbits immune to tick infestations demonstrated, by immunoblotting, nine protein subunits with molecular weights ranging from 12,000 to 92,000 daltons. Immuno-adsorption studies revealed an aggregated native protein with a molecular weight greater than 500,000 daltons as resolved by Polyacrylamide gel electrophoresis (PAGE). Sodium dodecyl sulphate (SDS)-PAGE of this protein molecule revealed the presence of 14 protein subunits similar to some of those detected by the immunoblots. Their molecular weights ranged from 14,000 to 165,000 daltons. Sera from rabbits immunized with the 500,000 dalton molecule formed a weak precipitating arc in the immunodiffusion test, confirming its immunogenicity. The reproductive potential of ticks fed on rabbits immunized with the purified protein molecule was slightly affected compared to controls. The protein molecule also had the capacity to induce immediate and delayed type skin reactions when injected into rabbits resistant to adult R. appendiculatus, as opposed to control tick-naive rabbits.

Key Words

Rhipicephalus appendiculatus midgut antigens resistant rabbits immunoglobulins immunoblotting immunoaffinity immunization immunodiffusion ion exchange native protein protein subunits immunization skin tests 


La resistance immunitaire à l’infestation des tiques Rhipicephalus appendiculatus Neumann, a été induite sur des lapins après alimination repetée. Les anticorps des lapins resistants aux tiques furent purifies et utilisés pour isoler les antigenes de l’intestin des tiques. L’immunoglobuline G (IgGs) purifié des lapins immunisés à l’infestion des tiques a demontré à l’immunoblotting 9 sous unités de proteines avec un poids moleculaire allant de 12,000 à 92,000 daltons. L’immunoabsorption a relèvé une proteine avec un poids moleculaire plus grand que 500,000 daltons grace à l’électrophorese du gel Polyacrylamide (PAGE). Le SDS-PAGE de cette molecule a relèvé la presence de 14 sous unités de proteines. Ces 14 proteines furent detectées à l’immunoblots. Le poids moleculaire de ces proteines se situait entre 14,000 et 165,000 daltons. Les serums des lapins immunizés avec la proteines au poids moleculaire 500,000 daltons forma un arc de precipitation au test de (’immunodiffusion pour confirme l’immunogenecité de cattée proteine. Le potentiel reproductive des tiques nourris sur des lapin immunizés avec la molecule de proteine purifiée a été legèrement affectée par rapport aux lapins utilisés comme contrôle. La molecule de proteine aussi la capacité d’induire des reactions cutanées immediates ou retardées lorsqu’elle est injectée aux lapins resistants aux adultes du R. appendiculatus a l’opposé des contrôles qui sont des lapins qui ne pas exposés aux tiques.


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  1. Agbede R. I. S. and Kemp D. H. (1986) Immunization of cattle against Boophilus microplus using extracts derived from adult female ticks: Histopathology of ticks feeding on vaccinated cattle. Int. J. Parasitol. 16, 35–41.CrossRefGoogle Scholar
  2. Allen J. R. and Humphreys S. J. (1979) Immunization of guinea pigs and cattle against ticks. Nature (London) 280, 491–493.CrossRefGoogle Scholar
  3. Bailey K. P. (1960) Notes on the rearing of Rhipicephalus appendiculatus and their infection with Theileria parva for experimental transmission. Bull. Epizoot. Dis. Afr. 8, 33–43.Google Scholar
  4. Binnington K. C and Kemp D. H. (1980) Role of tick salivary glands in feeding and disease transmission. Adv. Parasitol. 18, 315–339.CrossRefGoogle Scholar
  5. Brown S. J., Shapiro S. Z. and Askenase P. W. (1984) Characterization of tick antigens inducing host immune resistance. I. Immunization of guinea pigs with Amblyomma americanum-deriwed salivary gland extract and identification of an important salivary gland protein antigen with guinea pig anti-tick antibodies. J. Immunol. 133, 3319–3325.Google Scholar
  6. Irvin A. D. and Broadesky D. W. (1970) Rearing and maintaining Rhipicephalus appendiculatus in the laboratory. J. Inst. Anim. Technol. 21, 106–112.Google Scholar
  7. Johnston L. A. Y., Kemp D. H. and Pearson R. D. (1986) Immunization of cattle against Boophilus microplus using extracts derived from adult female ticks: Effects of induced immunity to tick populations. Int. J. Parasitol. 16, 27–34.CrossRefGoogle Scholar
  8. Kemp D. H., Stone B. F. and Binnington K. C. (1982) Tick attachment and feeding: Role of the mouthparts, feeding apparatus, salivary gland secretions and the host response. In Physiology of Ticks (Edited by Obenchain F. D. and Galun R.), pp. 119–168. Pergamon Press, Oxford.CrossRefGoogle Scholar
  9. Laemli U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.CrossRefGoogle Scholar
  10. Maizel J. V. (1971) Polyacrylamide gel electrophoresis of viral proteins. Meth. Virol. 5, 179–246.CrossRefGoogle Scholar
  11. Merril C. R., Goldman D., Sedman S. A. and Elbert M. H. (1981) Ultrasensitive stain for proteins in Polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 21, 1437–1441.CrossRefGoogle Scholar
  12. Mongi A. O., Shapiro S. Z., Doyle J. J. and Cunningham M. P. (1986a) Immunization of rabbits with Rhipicephalus appendiculatus antigen-antibody complexes. Insect Sci. Applic. 7, 471–477.Google Scholar
  13. Mongi A. O., Shapiro S. Z., Doyle J. J. and Cunningham M. P. (1986b) Characterization of antigens from extracts of fed ticks using sera from rabbits immunized with extracted tick antigen and by successive tick infestation. Insect Sci. Applic. 1, 479–487.Google Scholar
  14. Ouchterlony O. (1958) Diffusion-in-gel methods for immunological analysis. Progress in Allergy 5, 1–78.PubMedPubMedCentralGoogle Scholar
  15. Porath J. (1974) General methods and coupling procedures. In Methods in Enzymology XXXIV (Edited by Jakoby W. B. and Wilcheck M.), pp. 13–29. Academic Press, New York.CrossRefGoogle Scholar
  16. Purnell R. E. and Joyner L. P. (1968) The development of Theileria parva in the salivary glands of the tick Rhipicephalus appendiculatus. Parasitology 58, 725–730.CrossRefGoogle Scholar
  17. Towbin H., Stachlin T. and Gordon J. (1979) Electrophoretic transfer of proteins from Polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Science (Washington) 76, 4350–4354.CrossRefGoogle Scholar
  18. Wharton R. H. (1976) Tick-borne livestock diseases and their vectors. V. Acaricide resistance and alternative methods of tick control. World Anim. Rev. 20, 8–15.Google Scholar
  19. Wikel S. K. and Allen J. R. (1976) Acquired resistance to ticks. I. Passive transfer of resistance. Immunology 30, 311–316.PubMedPubMedCentralGoogle Scholar
  20. Wikel S. K. and Allen J. R. (1982) Immunological basis of host resistance to ticks. In Physiology of Ticks (Edited by Obenchain F. D. and Galun R.), pp. 169–196. Pergamon Press, Oxford.CrossRefGoogle Scholar
  21. Willadsen P. and Riding G. A. (1979) Characterization of a proteolytic enzyme inhibitor with allergenic activity. Biochem. J. 17,41-47.CrossRefGoogle Scholar

Copyright information

© ICIPE 1990

Authors and Affiliations

  • Abdul O. Mongi
    • 1
  • C. A. Aganyo
    • 1
  1. 1.The International Centre of Insect Physiology and Ecology (ICIPE)NairobiKenya

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