Original Article

Detection and Molecular Characterization of Babesia canis vogeli and Theileria annulata in Free-Ranging Dogs and Ticks from Shahriar County, Tehran Province, Iran


Background: We aimed to detect and characterize vector-borne parasites of Babesia and Theileria in dog and ticks by PCR assay. Canine babesiosis is a significant tick-borne disease caused by different Babesia species.  As the infection has not been reported in Shahriar region Tehran, Iran, molecular techniques allowed us to identify tick-borne parasites in asymptomatic dogs.

Methods: The number of 40 dog peripheral blood samples and 27 skin attached ticks were analyzed by molecular PCR assay. The specific primers were used for detecting Babesia canis, B. gibsoni and T. annulata.

Results: B. c. vogeli was detected in 10 dog blood samples (25%). Additionally, T. annulata infection was identified in 13 dog blood samples (32.5%) and 18 isolated tick DNAs (66.7%). The results of PCR were confirmed by 18S rRNA and Tams1 gene sequence analyzing and have been registered in GenBank under following accession numbers for B. c. vogeli (MH793502) and T. annulata (MK105284).

Conclusion: The verification of T. annulata infection in free-ranging dogs and ticks shows dogs might be considered as important natural carriers/reservoirs for T. annulata in enzootic region for bovine theileriosis. The obtained data may be useful for veterinary practitioners and dog owners to aware of Babesia and Theileria infection in dog and tick to establish the effective preventive measures.

1. Uilenberg G, Franssen FF, Perie NM, et al. Three groups of Babesia canis distinguished and a proposal for nomenclature. Vet Q. 1989;11(1):33-40.
2. Zahler M, Schein E, Rinder H, et al. Characteristic genotypes discriminate between Babesia canis isolates of differing vector specificity and pathogenicity to dogs. Parasitol Res.1998; 84(7):544-548.
3. Criado-Fornelio A, Gonzalez-del-Rio MA, Buling-Sarana A, et al. Molecular characterization of a Babe-sia gibsoni isolate from a Spanish dog. Vet Parasitol. 2003; 117(1-2):123-129.
4. Schnittger L, Rodriguez AE, Florin-Christensen M, et al. Babesia: a world emerging. Infect Genet Evol. 2012; 12(8):1788–1809.
5. Lack JB, Reichard MV, Van Den Bussche RA. Phylogeny and evolution of the Piroplasmida as in-ferred from 18S rRNA sequences. Int J Parasitol. 2012; 42(4):353–363.
6. Uilenberg G. Babesia – a historical overview. Vet Parasitol. 2006; 138(1–2):3–10.
7. Inokuma H, Yoshizaki Y, Matsumoto K, et al. Mo-lecular survey of Babesia infection in dogs in Okina-wa, Japan. Vet Parasitol. 2004; 121(3–4): 341–346.
8. Irwin PJ, Hutchinson GW. Clinical and pathological findings of Babesia infection in dogs. Aust Vet J. 1991; 68(6):204–209.
9. Penzhorn BL. Why is Southern African canine babesiosis so virulent? An evolutionary perspective. Parasit Vectors. 2011; 4:51.
10. Criado A, Martinez J, Buling A, et al. New data on epizootiology and genetics of piroplasms based on sequences of small ribosomal subunit and cyto-chrome b genes. Vet Parasitol. 2006; 142(3-4):238-47.
11. Bigdeli M, Mashhady Rafie S, Namavari MM, et al.Report of Theileria annulata and Babesia canis infec-tions in dogs. Comparative Clin Pathol. 2012; 21(3):375–377.
12. Dixit P, Dixit AK, Varshney JP. Evidence of new pathogenic Theileria species in dogs. J Parasit Dis. 2010; 34(1):29-32.
13. Falkenö U, Tasker S, Osterman-Lind E, et al. Theil-eria annae in a young Swedish dog. Acta Vet Scand. 2013; 55(1):50.
14. Rosa CT, Pazzi P, Nagel S, et al. Theileriosis in six dogs in South Africa and its potential clinical signifi-cance. J S Afr Vet Assoc. 2014; 85(1):1114.
15. Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual, Volume 3, Cold Spring Harbor Laboratory, Cold Spring Harbor; NY, 1989. B.16.
16. Birkenheuer AJ, Levy MG, Breitschwerdt EB. Development and evaluation of a seminested PCR for detection and differentiation of Babesia gibsoni (Asian genotype) and B. canis DNA in canine blood samples. J Clin Microbiol. 2003; 41(9):4172-7.
17. Saitou N, and Nei M. The neighbor-joining meth-od: A new method for reconstructing phylogenetic trees. Mol Biol Evol.1987; 4:406-425.
18. Tamura K, Peterson D, Peterson N, et al. MEGA5: Molecular Evolutionary Genetics Analy-sis using Maximum Likelihood, Evolutionary Dis-tance, and Maximum Parsimony Methods. Mol Bi-ol Evol. 2011; 28(10): 2731-2739.
19. Solano-Gallego L, Baneth G. Babesiosis in dogs and cats - expanding parasitological and clinical spectra. Vet Parasitol. 2011; 181(1):48–60.
20. Jongejan F, and Uilenberg G. Ticks and control methods. Rev Sci Tech. 1994; 13(4):1201-1226.
21. Akhtardanesh B, Saberi M, Nurollahifard SR, et al. Molecular Detection of Babesia spp. in Tick-infested Dogs in Southeastern Iran. J Dis Glob Health. 2016; 8(2):72-77.
22. Noaman V, Abdigoudarzi M, Nabinejad AR. Abundance, diversity, and seasonal dynamics of hard ticks infesting cattle in Isfahan Province, Iran. Arch Razi Inst. 2017; 72(1):15-21.
23. Tavassoli M, Tabatabaei M, Mohammadi M, et al. PCR-based Detection of Babesia spp. Infection in Collected Ticks from Cattle in West and North-West of Iran. J Arthropod Borne Dis. 2013; 7(2):132-8.
24. Fatemian Z, Salehzadeh A, Sedaghat MM, et al. Hard tick (Acari: Ixodidae) species of livestock and their seasonal activity in Boyer-Ahmad and Dena cities of Kohgiluyeh and Boyer-Ahmad Province, Southwest of Iran. Vet World. 2018; 11(9):1357–1363.
25. Davari B, Nazari Alam F, Nasirian H, et al. Seasonal distribution and faunistic of ticks in the Alashtar county (Lorestan Province), Iran. Pan Afr Med J. 2017; 27:284.
26. Telmadarraiy Z, Oshaghi MA, Hosseini Vasouko-laei N, et al. First molecular detection of Theileria ovis in Rhipicephalus sanguineus tick in Iran. Asian Pac J Trop Med. 2012; 5(1):29-32.
27. Niak A, Anwar M, Khatibi S. Canine babesiosis in Iran. Trop Anim Health Prod. 1973; 5(3):200–201.
28. Raki A, Doosti A, Shahrani M. Detection of Babesia canis in the blood samples of dogs in Iran by PCR method. Int J Biosci. 2014; 4(1):118-124.
29. Razi Jalali MH, Mosallanejad B, Avizeh R, et al. Babesia infection in urban and rural dogs in Ahvaz district, Southwest of Iran. Arch Razi Inst. 2013; 68(1):37-42.
30. Solano-Gallego L, Trotta M, Carli E, et al. Babesia canis canis and Babesia canis vogeli clinicopathological findings and DNA detection by means of PCR-RFLP in blood from Italian dogs suspected of tick-borne disease. Vet Parasitol. 2008; 157(3-4):211–21.
31. Solano-Gallego L, Capri A, Pennisi MG, et al. Acute febrile illness is associated with Rickettsia spp infection in dogs. Parasites & Vectors. 2015; 8:216.
32. Kraje AC. Canine haemobartonellosis and babesio-sis. Compend Contin Educ Pract Vet. 2001; 23:310–8.
33. Irwin PJ. Canine babesiosis: from molecular taxon-omy to control. Parasit Vectors. 2009; Suppl 1:S4.
34. Solano-Gallego L, Sainz Á, Roura X, et al. A review of canine babesiosis: the European perspective. Parasit Vectors. 2016; 9:336.
35. Fukumoto S, Tamaki Y, Igarashi I, et al. Immuno-genicity and growth inhibitory efficacy of the prime-boost immunization regime with DNA fol-lowed by recombinant vaccinia virus carrying the P29 gene of Babesia gibsoni in dogs. Exp Parasitol. 2009; 123(4):296–301.
IssueVol 15 No 3 (2020) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v15i3.4196
Babesia canis vogeli Theileria annulata Polymerase chain reac-tion Dog Tick

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
HABIBI G, IMANI A, AFSHARI A, BOZORGI S. Detection and Molecular Characterization of Babesia canis vogeli and Theileria annulata in Free-Ranging Dogs and Ticks from Shahriar County, Tehran Province, Iran. Iran J Parasitol. 2020;15(3):321-331.