Parasitology Research

, Volume 109, Issue 1, pp 147–162

Ticks’ response to feeding on host immunized with glandular extracts of Rhipicephalus sanguineus females fed for 2, 4, and 6 days. I. Inactivity or early degeneration of salivary glands?

  • Karim Christina Scopinho Furquim
  • Maria Izabel Camargo Mathias
  • Letícia Maria Gráballos Ferraz Hebling
  • Gislaine Cristina Roma
  • Gervásio Henrique Bechara
Original Paper


The present study histologically analyzed the salivary glands of Rhipicephalus sanguineus females fed for 2, 4, and 6 days in hosts which had been previously immunized with glandular extracts obtained from females from this same species in different periods of feeding, having as main objective verify the action of these extracts in the secretor cycle of these glands. For this, glandular extract of females fed for 2 days (SGE2), glandular extract of females fed for 4 days (SGE4), and glandular extract of females fed for 6 days (SGE6) extracts were obtained from salivary glands of R. sanguineus females fed for 2, 4, and 6 days respectively. Then, New Zealand White naive rabbits were inoculated either with extracts (test group = TG), or with a mixture of phosphate buffer and Freund's complete adjuvant (control group 2 = CG2). Each inoculated rabbit (TG and CG2) and non-inoculated (control group 1 = CG1) was posteriorly infested with 15 couples of fasting R. sanguineus from which the salivary glands had been collected from females fed for 2, 4, and 6 days. The results revealed that the resistance the hosts had acquired by the immunization with the extracts affected differently the secretory activity of the glandular cells. It was verified that the resistance to SGE2 and SGE4 extracts acted in the cells of acini II and III, being c1 and c5 from II and d from III inactivated due to the action of SGE2 and c1 and c4 from II and f from III inactivated by the action of SGE4. As for the resistance to SGE6 the effect was only on cells of acini II (c1, c3 e c4), which were also inactivated. In addition, the hosts’ resistance to SGE2SGE6 extracts made the degenerative process earlier in comparison to CG1. On the other hand, the resistance to the extracts did not influence the characteristics of the degenerative process normally found in salivary glands. The assynchronism of the degenerative process was maintained—acini III were always the most affected and I the less affected. The structural cell alterations, such as cytoplasmic vacuolation, nuclear alterations and formation of apoptotic bodies which characterize the occurrence of atypical apoptosis were also maintained in the glands of individuals from TG making it clear that the immunization of the hosts with glandular extracts SGE2–SGE6 had influenced the glandular physiology of R. sanguineus, which is an important piece of information in the search for a way to control these ectoparasites.


  1. Almeida APG, Bechara GH, Varma RMG (1994) Cross-reactivity between hard tick antigens. Braz J Med Biol Res 27:697–707PubMedGoogle Scholar
  2. Balashov YuS (1983) An atlas of ixodid tick ultrastructure. In: Raikhel AS, Hoogstraal II (eds) Entomological Society of America. Special Publication, Lanham, pp 99–128Google Scholar
  3. Bechara GH, Szabó MPJ, Ferreira BR, Garcia MV (1995) Rhipicephalus sanguineus tick in Brazil: feeding and reproductive aspects under laboratorial conditions. Rev Bras Parasitol Vet 4(2):61–66Google Scholar
  4. Binnington KC (1978) Sequential changes in salivary gland structure during attachment and feeding of the cattle tick Boophilus microplus. Int J Parasitol 8:97–115PubMedCrossRefGoogle Scholar
  5. Caperucci D, Bechara GH, Camargo-Mathias MI (2009) Histopathology and ultrastructure features of the midgut of adult females of the tick Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) in various feeding stages and submitted to three infestations. Ultrastruct Pathol 33:249–259PubMedCrossRefGoogle Scholar
  6. Caperucci D, Bechara GH, Camargo-Mathias MI (2010) Ultrastructure features of the midgut of the female adult Amblyomma cajennense ticks Fabricius, 1787 (Acari: Ixodidae) in several feeding stages and subjected to three infestations. Micron. doi:10.1016/j.micron.2010.05.015 PubMedGoogle Scholar
  7. Coutinho MTZ, Bueno LL, Sterzik A, Fujiwara RT, Botelho JR, de Maria M, Genaro O, Linardi PM (2005) Participation of Rhipicephalus sanguineus (Acari: Ixodidae) in the epidemiology of canine leishmaniasis. Vet Parasitol 128:149–155PubMedCrossRefGoogle Scholar
  8. Dantas-Torres F (2008) Canine vector-borne deseases in Brazil. Parasit Vectors 1(25):1–17Google Scholar
  9. Demma LJ, Traeger MS, Nicholson WL, Paddock CD, Blau DM, Eremeeva ME, Dasch GA, Levin ML, Singleton J, Zaki SR, Cheek JE, Swerflow DL, McQuiston JH (2005) Rocky mountain spotted fever from an unexpected tick vector in Arizona. N Engl J Med 353(6):587–594PubMedCrossRefGoogle Scholar
  10. Fawcett DW, Binnington KC, Voight WR (1986) The cell biology of the ixodid tick salivary gland. In: Sauer JR, Hair JA (eds) Morphology, physiology and behavioral biology of ticks. Ellis Horwood, Chichester, pp 22–45Google Scholar
  11. Ferreira BR, Machado RZ, Bechara GH (1996) Western blot analysis of tick antigens from a Rhipicephalus sanguineus unfed larval extract and identification of antigenic sites in tick sections using immunohistochemistry. A comparative study between resistant and susceptible host species. Vet Parasitol 62:161–174PubMedCrossRefGoogle Scholar
  12. Furquim KCS, Bechara GH, Camargo-Mathias MI (2008) Death by apoptosis in salivary glands of females of the tick Rhipicephalus sanguineus (Latreille, 1806) (Acari, Ixodidae). Exp Parasitol 119:152–163CrossRefGoogle Scholar
  13. Geczy F, Nayghton MA, Clegg JB, Heweton RW (1971) Esterases and a carbohydrate-spliting enzyme in the saliva of the cattle tick, Boophilus microplus. J Parasitol 57:437–438PubMedCrossRefGoogle Scholar
  14. Gill HS, Walker AR (1987) The salivary glands of Hyalomma anatolicum anatolicum: nature of salivary gland components and their role in tick attachment and feeding. Int J Parasitol 18(1):83–93CrossRefGoogle Scholar
  15. Gill HS, Boid R, Ross CA (1986) Isolation and characterization of salivary antigens from Hyalomma anatolicum anatolicum. Parasite Immunol 8:11–25PubMedCrossRefGoogle Scholar
  16. Jittapalapong S, Stich RW, Gordon JC, Wittum TE, Barriga OO (2000a) Performance of female Rhipicephalus sanguineus (Acari: Ixodidae) fed dogs exposed to multiple infestations or immunization with tick salivary gland or midgut tissues. J Med Entomol 37(4):601–611PubMedCrossRefGoogle Scholar
  17. Jittapalapong S, Stich RW, Gordon JC, Bremer CA, Barriga OO (2000b) Humoral immune response of dogs immunized with salivary gland, midgut, or repeated infestations with Rhipicephalus sanguineus. Ann N Y Acad Sci 916:283–288PubMedCrossRefGoogle Scholar
  18. Jittapalapong S, Phichitrasilp T, Chanphao H, Rerkamnuychoke W, Stich RW (2008) Immunization with tick salivary gland extracts. Ann N Y Acad Sci 1149:200–204PubMedCrossRefGoogle Scholar
  19. Lomas LO (1993) The influence of a male-derived protein factor on salivary gland degeneration in the ixodid tick, Amblyomma hebraeum Koch. Ph.D Thesis, University of AlbertaGoogle Scholar
  20. Lomas LO, Gelman D, Kaufman WR (1998) Ecdysteroid regulation of salivary gland degeneration in the ixodid tick Amblyomma hebraeum: a reconciliation of in vivo and in vitro observations. Gen Comp Endocrinol 109:200–211PubMedCrossRefGoogle Scholar
  21. Mao H, Macblain WA, Kaufman WR (1995) Some properties of the ecdysteroid receptor in the salivary gland of the ixodid tick, Amblyomma hebraeum. Gen Comp Endocrinol 99:340–348PubMedCrossRefGoogle Scholar
  22. Marzouk AS, Darwish ZEA (1994) Changes in the salivary glands of female Hyalomma (Hyalomma) dromedarii during and after feeding. J Egypt Soc Parasitol 24(1):39–57PubMedGoogle Scholar
  23. Monteiro GER, Bechara GH (2008) Cutaneus basophilia in the resistance of goats to Amblyomma cajennense nymphs after repeated infestations. Ann N Y Acad Sci 1149:221–225PubMedCrossRefGoogle Scholar
  24. Monteiro GER, Bechara GH, Franzin AM, de Miranda Santos IKF (2010) Antigen-presenting in draining lymph nodes of goads repeatedly infested by the Cayenne tick Amblyomma cajennense nymphs. Exp Appl Acarol. doi:10.1007/s10493-010-9380-x PubMedGoogle Scholar
  25. Nunes PH, Bechara GH, Camargo-Mathias MI (2010) Secretory process of salivary glands of female Amblyomma cajennense Fabricius, 1787 (Acari: Ixodidae) ticks fed on resistant rabbits. Exp Appl Acarol. doi:10.1007/s10493-010-9388-2 PubMedGoogle Scholar
  26. Pegram RG, Clifford CM, Walker JB, Keirans JE (1987a) Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). I. R. sulcatus Neuman 1908 and R. turanicus Pomerantsev 1936. Syst Parasitol 10:3–26CrossRefGoogle Scholar
  27. Pegram RG, Keirans JE, Clifford CM, Walker JB (1987b) Clarification of the Rhipicephalus sanguineus group (Acari, Ixodoidea, Ixodidae). II. R. sanguineus (Latreille, 1806) and related species. Syst Parasitol 10:27–44CrossRefGoogle Scholar
  28. Ribeiro JMC, Makoul GT, Levine J, Robinson DK, Spilman A (1985) Antihaemostatic, antiinflammatory and immunosupressive properties of the saliva of a tick, Ixodes dammini. J Exp Med 161:332–344PubMedCrossRefGoogle Scholar
  29. Rosell-Davies R, Coons LB (1989) Relationship between feeding mating, vitellogenin production and vitellogenesis in the tick Dermacentor variabilis. Exp Appl Acarol 7:95–105CrossRefGoogle Scholar
  30. Sanders ML, Scott AL, Glass GE, Schwartz BS (1996) Salivary gland changes and host antibody responses associated with feeding of male lone star ticks (Acari: Ixodidae). J Med Entomol 33(4):628–634PubMedGoogle Scholar
  31. Sauer JR, Essenberg RC, Bowman AS (2000) Salivary glands in ixodid ticks: control and mechanism of secretion. J Insect Physiol 46:1069–1078PubMedCrossRefGoogle Scholar
  32. Sedmak JJ, Grossberg SE (1977) A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250. Anal Biochem 79:544–552Google Scholar
  33. Sonenshine DE (1991) Biology of ticks. Oxford University Press, New YorkGoogle Scholar
  34. Szabó MPJ, Bechara GH (1997) Immunisation of dogs and guinea pigs against Rhipicephalus sanguineus ticks using gut extract. Vet Parasitol 68:283–294PubMedCrossRefGoogle Scholar
  35. Till WM (1961) A contribution to the anatomy and histology of the brown ear tick Rhipicephalus appendiculatus Neumann, vol 6. Memoirs of the Entomological Society of Southern Africa, PretoriaGoogle Scholar
  36. Veronez VA, de Castro MB, Bechara GH, Szabó MPJ (2010) Histopathology of Rhipicephalus sanguineus (Acari: Ixodidae) ticks fed on resistant hosts. Exp Appl Acarol 50:151–161PubMedCrossRefGoogle Scholar
  37. Walker A, Fletcher JD, Gill HS (1985) Structural and histochemical changes in the salivary glands of Rhipicephalus appendiculatus during feeding. Int J Parasitol 15(1):81–100PubMedCrossRefGoogle Scholar
  38. Walker JB, Keirans JE, Horak IG (2000) The genus Rhipicephalus (Acari, Ixodidae): a guide to the brown ticks of the world. Cambridge University Press, CambridgeGoogle Scholar
  39. Weiss BL, Kaufman WR (2001) The relationship between ‘critical weight’ and 20-hydroxyecdysone in the female ixodid tick Amblyomma hebraeum. J Insect Physiol 47:1261–1267PubMedCrossRefGoogle Scholar
  40. Wikel SK (1981) The induction of host resistance to tick infestation with salivary gland antigen. Am J Trop Med Hyg 30(1):284–288PubMedGoogle Scholar
  41. Wikel SK (1999) Tick modulation of host immunity: an important factor in pathogen transmission. Int J Parasitol 29:851–859PubMedCrossRefGoogle Scholar
  42. Wikel SK, Graham JE, Allen JR (1978) Acquired resistance to ticks. IV. Skin reactivity and in vitro lymphocyte responsiveness to salivary gland antigen. Immunology 34:257–263PubMedGoogle Scholar
  43. Zhou J, Gong H, Zhou Y, Xuan X, Fujisaki K (2006) Identification of a glycine-rich protein from the tick Rhipicephalus haemaphysaloides and evaluation of its vaccine potential against tick feeding. Parasitol Res 100:77–84PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Karim Christina Scopinho Furquim
    • 1
  • Maria Izabel Camargo Mathias
    • 2
  • Letícia Maria Gráballos Ferraz Hebling
    • 2
  • Gislaine Cristina Roma
    • 2
  • Gervásio Henrique Bechara
    • 1
  1. 1.Departamento de Patologia VeterináriaFCAV, UNESPJaboticabalBrazil
  2. 2.Departamento de BiologiaInstituto de Biociências, UNESPRio ClaroBrazil

Personalised recommendations