Determination of the Infectivity of Cryopreserved Theileria annulata Sporozoites in Tick Derived Stabilates Iran Ak-93 Strain, by In Vivo and In Vitro Methods

  • Hossein MODIRROUSTA Department of Wild Life Research, Razi Vaccine and Serum Research Institute, Agriculture Research, Education and Extension Organization, Karaj, Iran
  • Gholamreza HABIBI Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Agriculture Re-search, Education and Extension Organization, Karaj, Iran
  • Parviz SHAYAN Department of Parasitology, School of Veterinary, Tehran University, Tehran, Iran
  • Asghar AFSHARI Department of Parasite Vaccine Research and Production, Razi Vaccine and Serum Research Institute, Agriculture Re-search, Education and Extension Organization, Karaj, Iran
  • Ali MIRJALILI Department of Biotechnology, Razi Vaccine and Serum Research Institute, Agriculture Research, Education and Exten-sion Organization, Karaj, Iran
  • Mohamad ABDIGOUDARZI Department of Parasitology, Razi Vaccine and Serum Research Institute, Agriculture Research, Education and Extension Organization, Karaj, Iran
Keywords:
Theileria annulata, Hyalomma a. anatolicum, Tick stabilate, RT-PCR

Abstract

Background: The protozoan parasite Theileria annulata is the causative agent of tropical theileriosis in cattle. Vaccination is recommended by administration of attenuated schizont-infected cell lines. The expected protective immunity post-vaccination can be demonstrated by challenge test through inoculation of highly virulent infective sporozoites. The aim of this study was to produce Hyalomma anatolicum anatolicum tick infected with T. annulata (local strain) for preparation of tick-derived sporozoite stabilates for molecular characterization and infectivity test assay.Methods: A local T. annulata strain was used for experimental infection of calves. A field isolate of H. a. anatolicum was isolated, laboratory-reared and infected by blood-feeding on Theileria infected above-mentioned calves. The infectivity of calf, tick and prepared stabilate were confirmed by clinical signs of theileriosis, microscopic inspection, RT-PCR and in vitro cell culture. Results: The tick stabilate was prepared and cryopreserved in liquid nitrogen. The infectivity of the tick stabilate was verified by in vivo bioassay, in vitro cell culture infection, microscopic inspection in salivary glands and RT-PCR assay. The in vitro produced cell line in this study was characterized by T. annulata Cytochrome b gene analyzing.Conclusion: The infectivity of a new prepared tick-derived sporozoite stabilate was confirmed in susceptible calves; by microscopically, post mortem, tick microscopic and molecular assays. Moreover, naïve PBMCs were transformed and proliferated by T. annulata infected tick stabilate to immortal T. annulata schizont infected cell line. The potent infective sporozoite tick derived stabilate could be used for vaccine efficacy and challenge test as well as in vaccine development.

References

1. OIE Terrestrial Manual, 2018. Chapter 3.4.14. Theileriosis. pp: 1185-1209 https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.04.14_THEILIERIOSIS.pdf
2. Biglari P, Bakhshi H, Chinikar S et al. Hyalomma anatolicum as the Main Infesting Tick in an Im-portant Livestock Rearing Region, Central Ar-ea of Iran. Iran J Public Health. 2018;47(5):742-749.
3. Arjmand Yamchi J, Tavassoli M. Survey on infection rate, vectors and molecular identifica-tion of Theileria annulata in cattle from North West, Iran. J Parasit Dis. 2016;40(3):1071-6.
4. Haque M, Jyoti, Singh NK, Rath SS. Preva-lence of Theileria annulata infection in Hyalomma anatolicum anatolicum in Punjab state, India. J Parasit Dis. 2010;34(1):48-51.
5. Flach EJ, Ouhelli H, Waddington D, el Hasnaoui M. Prevalence of Theileria in the tick Hyalomma detritum detritum in the Doukkala re-gion, Morocco. Med Vet Entomol. 1993;7(4):343-50.
6. Aktas M, Dumanli N, Angin M. Cattle infesta-tion by Hyalomma ticks and prevalence of Theil-eria in Hyalomma species in the east of Turkey. Vet Parasitol. 2004;119(1):1-8.
7. Shaw MK. Cell invasion by Theileria sporozo-ites. Trends Parasitol. 2003;19(1):2-6.
8. Minjauw B, McLeod A. Tick-borne diseases and poverty. The impact of ticks and tick-borne diseases on the livelihood of small-scale and marginal livestock owners in India and eastern and southern Africa. Edinburgh: Cen-tre for Tropic al Veterinary, Medicine, Univer-sity of Edinburgh, UK; 2003.
9. Babiuk LA. Vaccination: a management tool in veterinary medicine. Vet J. 2002;164(3):188-201.
10. Gill BS, Bhattacharyulu Y, Kaur D, Singh A. Immunization of cattle against tropical theiler-iosis (Theileria annulata infections) by infection-treatment method. Ann Rech Vet. 1977; 8: 285-292.
11. Cunningham MP, Brown CG, Burridge MJ, Purnell RE. Cryopreservation of infective par-ticles of Theileria parva. Int J Parasitol. 1973;3(5):583-7.
12. Hoogstraal H, Wassef HY, Büttiker W. Ticks (Acarina) of Saudi Arabia Fam. Argasidae, Ix-odidae. Fauna of Saudi Arabia. 1981; 3, 25-110 .
13. Hoogstraal, H. Ticks (Acarina) of Saudi Arabia Fam. Argasidae, Ixodidae. Fauna of Saudi Ara-bia. 1981; 6, 6170-174.
14. Edwards KT, Goddard J, Varela-Stokes AS. Examination of the Internal Morphology of the Ixodid Tick, Amblyomma maculatum Koch, (Acari: Ixodidae); a “How-to” Pictorial Dissec-tion Guide. Mississippi Entomological Associa-tion. 2009; 2: 28-39.
15. Irvin AD, Cunningham AP, Young AS. Ad-vances in the Control of Theileriosis: Interna-tional Conference Proceedings (Current Topics in Veterinary Medicine). Mertinus Nijhoff Pub-lishers, The Hague.1981; 73-175.
16. Spooner RL, Innes EA, Glass EJ et al. Bovine mononuclear cell lines transformed by Theileria parva or Theileria annulata express different sub-population markers. Parasite Immunol. 1988;10(6):619-29.
17. Kurtti TJ, Munderloh UG, Irvin AD, Büscher G. Theileria parva: early events in the develop-ment of bovine lymphoblastoid cell lines per-sistently infected with macroschizonts. Exp Parasitol. 1981;52(2):280-90.
18. Habibi G. In silico analysis of Ta9 gene poly-morphism in an Iranian Theileria annulata schi-zont-infected cell line S15 vaccine strain and native isolates. Archives of Razi Institute. 2016; 71(2): 71-80.
19. Singh DK, Jagdish S, Gautam OP, Dhar S. Infectivity of ground-up tick supernates pre-pared from Theilerai annulata infected Hyalomma anatolicum anatolicum. Trop Anim Health Prod. 1979;11(2):87-90.
20. Ouhelli H, Innes EA, Brown CG et al. The effect of dose and line on immunisation of cat-tle with lymphoblastoid cell infected lines with Theileria annulata. Vet Parasitol. 1989;31(3-4):217-28.
21. Samish M, Pipano E. Relative infectivity of Theileria annulata (Dschunkowsky and Luhs, 1904) stabilate derived from female and male Hyalomma excavatum (Koch, 1844) ticks. Para-sitology. 1978;77(3):375-9.
22. Samish M, Pipano E. Transmission of Theileria annulata (Dschunkowsky & Luhs, 1904) by Hy-alomma excavatum (Koch, 1844). Parasitology. 1983;86 (Pt 2):269-74.
23. Marcotty T, Speybroeck N, Berkvens D et al. In vitro titration of Theileria parva tick derived stabilates. Parasitology. 2004;128(Pt 2):131-7.
Published
2019-12-29
How to Cite
1.
MODIRROUSTA H, HABIBI G, SHAYAN P, AFSHARI A, MIRJALILI A, ABDIGOUDARZI M. Determination of the Infectivity of Cryopreserved Theileria annulata Sporozoites in Tick Derived Stabilates Iran Ak-93 Strain, by In Vivo and In Vitro Methods. Iran J Parasitol. 14(4):572-583.
Section
Original Article(s)