Immunization of Cattle with Tick Salivary Gland Extracts

  • Ali Nikpay Department of Pathobiology, Amol University of Special Modern Technologies, Amol, Iran
  • Sedigheh Nabian Department of Parasitology, Faculty of Veterinary Medicine, Tehran, Iran
Keywords: Rhipicephalus (Boophilus) annulatus, Immunization, Salivary gland, Humoral immune responses, Tick


Background: Rhipicephalus (Boophilus) annulatus tick is one of the most important ectoparasite of cattle. Re­cently, several laboratories in the world have been concentrated on immunizing cattle against tick using various types of tissue extracts of ticks. The aim of this study was to evaluate the effect of immunization of cattle with tick salivary gland extract on biological parameters of ticks and humoral immune responses of cattle.Methods: Fourteen more dominant protein bands identified as immunogenic by Western-blot analysis were eluted from polyacrylamide gel. Test and control groups were injected three times with eluted proteins and sterile PBS (pH= 7.2) respectively with equivalent amount of adjuvant. After four weeks a tick challenge was performed. Fi­nally, biological parameters of collected engorged female ticks were recorded and humoral immune responses to immunization measured by ELISA.Results: The results indicated immunization of cattle resulted in reduction in mean tick counts, attachment, en­gorgement weights, feeding index, egg mass weight, hatchability and fertility index (respectively 63.1%, 62.6%, 30.2%, 36.4%, 40%, 78.7% and 13.3%) and increased duration of feeding, pre-oviposition and incubation period of eggs (respectively 8.6%, 45 and 31.34%). All changes were statistically significant (P< 0.05). Results showed an increase in antibody production of test group from the first week after immunization. The antibody level was boosted following tick infestation.Conclusion: This investigation indicates that immunization of cattle with these antigens could induce a protective immune response against Rh. (B.) annulatus tick that would be expected to provide a safe non-chemical means of tick control.


Brossard M, Fivaz V (1982) Ixodes ricinus L.: mast cells, basophils and eosinophils in the sequence of cellular events in the skin of the infested or reinfested rabbits. Parasitol. 85: 583–592.

Brown SJ, Shapiro SZ, Askenase PW (1984) Characterization of tick antigens in- ducing host immune resistance. I. Immunization of guinea pigs with Amblyomma americanum-derived sali- vary gland extracts and identification of an important salivary gland pro- tein antigen with guinea pig anti-tick antibodies. J Immunol. 133: 3319–3325.

Center for Cocoa biotechnology research and development (2007) Protein detection: Gel lectrophoresis, Isolation and renaturation of proteins from SDS- PAGE gels, The Laboratory of Jim Kadonaga at the University of Cali- fornia, San Diego. Available at: h/Protein%20Detection7.html#title.

Cobon GS, Moore JT, Johnston LAY, Willadsen P, Kemp DH, Sriskantha A, Riding GA, Rand KN (1996) DNA encoding a cell membrane gly- coprotein of a tick gut. US Patent and Trademark Office Appl. No. 325 071 435/240.2.

Dipeolu OO, Mongi AO, Essuman S, Amoo AO, Ndungu JN (1992) Studies on naturally acquired immunity to African ticks. II. Observations on cattle ex- posed to Rhipicephalus appendiculatus under varying periods of repeated infestations. Vet Parasitol. 41: 293–320.

Estrada-peña A, Bouattour A, Camicas JL, Walker AR (2004) Rhipicephalus (Boophilus) annulatus, In: Ticks of domestic animals in the Mediterranean region: a guide to identification of species. 1 Ed. Univ Zaragoza, Spain.

FAO (2004) Acaricide resistance: diagnosis, management and prevention, in: Guidelines Resistance Management and Integrated Parasite Control in Ruminants, Animal Production and Health Division, Agriculture Depart- ment, Food and Agriculture Organ- ization of the United Nations, Rome, Italy, pp. 25–77.

Hager DA, Burgess RR (1980) Elution of proteins from sodium dodecyl sulfate-polyacrylamide gels, removal of sodium dodecyl sulfate, and renaturation of enzymatic activity: results with sigma subunit of Esche- richia coli RNA polymerase, wheat germ DNA topoisomerase, and other enzymes. Anal Biochem. 109: 76–86.

Harlow E, Lane D (1988) Antibodies: A laboratory Manual. Cold Spring Har- bor Laboratory, New York.

Hudson L, Hay FC (1984) Practical Immu- nology. 3rd Ed. Blackwell Scientific Publication, Oxford, UK.

Hunkapiller MW, Lujan E, Ostrander F,Hood LE (1983) Isolation of mi- crogram quantities of proteins from polyacrylamide gels for amino acid sequence analysis. Methods Enzymol.91: 227–236.

Jittapalapong S, Jansawan W, Barriga OO, Stich RW (2004a) Reduced inci- dence of Babesia bigemina infection in cattle immunized against the cattle tick, Boophilus microplus. Ann N Y Acad Sci. 1026: 312–318.

Jittapalapong S, Jansawan W, Gingkaew A, Barriga OO, Stich RW (2004b) Pro- tection of dairy cows immunized with tick tissues against natural Boophilus microplus infestation in Thailand. Ann N Y Acad Sci. 1026: 289–294.

Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature.227: 680–685.

Manzano-Román R, García-Varas S, Encinas- Grandes A, Pérez-Sánchez R (2007) Purification and characterization of a 45-kDa concealed antigen from the midgut membranes of Ornithodoros erraticus that induces lethal anti-tick immune responses in pigs. Vet Parasitol. 145: 314–325.

Mattewson MD, Wilson RG, Hamment CA (1975) The development of resistance to certain organophosphorus and carbamate Ixodicides by the blue tick, Boophilus decoloratus, in Rho- desia. Bull Ent Res. 66: 553–560.

Murrell A, Barker SC (2003) Synonymy of Boophilus Curtice, 1891 with Rhipicephalus Koch, 1844 (Acari: Ixodidae). Systematic Parasitol. 56:169–172.

Nabian S, Rahbari S, Haddadzadeh H, Alidadi N (2005) A study on the effects of different temperatures on protein changes in the salivary glands of Hyalomma anatolicum anatolicum and Rhipicephalus (Boophilus) annulatus. Iranian J Vet Res Univ Shiraz. 6: 35–39.

Nari A, Hansen JW (1999) Resistance of ecto- and endo-parasites: current and future solutions. 67th General Ses- sion. International Committee, 17–21 May, OIE. Paris, pp. 17–21.

Nikpay A, Nabian S, Rahbari S, Shayan P, Alidadi N (2008) Detection of humoral immune response of calves to Boophilus Annulatus by ELISA. Irann J Parasitol. 3: 26–30.

Nikpay A, Nabian S, Taheri M (2012) Anal- ysis of immunogenic relevant pro- teins in Rhipicephalus (Boophilus) annulatus tick. J Arthropod-Borne Dis. 6: 36–44.

Norouzi F, Hashemitabar GR, Razmi GR (2007) Detection of midgut antigens of Hyalomma anatolicum anatolicum tick using SDS-PAGE and Western blot. Iran J Vet Res. 8: 166–169.

Rajput ZI, Hu SH, Chen WJ, Arijo AG, Xiao CW (2006) Importance of ticks and their chemical and immunological control in livestock. J Zhejiang Univ Science B. 7: 912–921.

Razmi GR, Tabar GRH, Naghibi A (2005) Immunization of rabbits against Hyalomma anatolicum anatolicum tick using antigen derived from whole tick and gut extract. Iran J Vet Res. 6: 23–26.

Tietz NW, Andresen BD (1986) Textbook of Clinical Chemistry. WB, Saunders, Philadelphia.Walker A (1994) The arthropods of humans and domestic animals: a guide to preliminary identification. Chapman and Hall, London, New York.

Wang H, Nuttall PA (1994) Comparison of the proteins in salivary glands, saliva and haemolymph of Rhipicephalus appendiculatus female ticks during feeding. Parasitol. 109(4): 517–523.

Wikel SK (1996) Host immunity to ticks.Annu Rev Entomol. 41: 1–22.

Wikel SK (1999a) Modulation of the host immune system by ectoparasitic arthropods: blood-feeding and tissue- dwelling arthropods manipulate host defenses to their advantages. Bio Science. 49: 311–320.

Wikel SK (1999b) Tick modulation of host immunity: an important factor in path- ogen transmission. Int J Parasitol. 29:851–859.

Willadsen P (1980) Immunity to ticks. Ad Parasitol. 18: 293–311.

Willadsen P (2006) Tick control: thoughts on a research agenda. Vet Parasitol.138: 161–168.

Willadsen P, Jongejan F (1999) Immunology of the tick-host interaction and the control of ticks and tick-borne diseases. Parasitol. 15: 258–262.

How to Cite
Nikpay A, Nabian S. Immunization of Cattle with Tick Salivary Gland Extracts. J Arthropod Borne Dis. 10(3):282-291.
Original Article