Parasitology Research

, Volume 102, Issue 1, pp 63–68 | Cite as

Detection and molecular characterization of Babesia caballi and Theileria equi isolates from endemic areas of Brazil

  • Alexandra Heim
  • Lygia M. F. Passos
  • Múcio F. B. Ribeiro
  • Lívio M. Costa-Júnior
  • Camila V. Bastos
  • Dagmar D. Cabral
  • Jörg Hirzmann
  • Kurt Pfister
Original Paper


Blood samples were collected from 487 adult horses, including 83 pregnant mares, at a slaughterhouse located in Araguari, Minas Gerais State, Brazil. For each blood sample, the packed cell volume (PCV) was determined, and Giemsa-stained smears were microscopically examined for the presence of hemoparasites. The plasma was examined by the indirect fluorescent antibody test for detection of antibodies against Babesia caballi and Theileria equi. In addition, DNA was extracted and analyzed by a multiplex real-time polymerase chain reaction (PCR), specific for B. caballi and T. equi. Products of PCR were sequenced and compared with each other and with known sequences. The serological results showed a total prevalence of 91.0% for T. equi and 83.0% for B. caballi, while by PCR, prevalences of 59.7% for T. equi and 12.5% for B. caballi were observed. However, no correlations were seen between positivity (neither by serology nor by PCR) and PCV values. As expected, the microscopic examination of blood smears showed low sensitivity in detecting the infections when compared to the PCR. Only 35 out of 570 blood smears were positive, with parasitemias below 0.1%. No congenital transmission was detectable. As far as sequencing is concerned, no differences were seen among the isolates of each species nor among them and known sequences available. These results confirm, by molecular methods, the high prevalence rates of T. equi and B. caballi infections in carrier horses in Brazil. However, no diversity was observed among the isolates within the studied regions.


Blood Smear Packed Cell Volume Babesia Indirect Fluorescent Antibody Test Babesiosis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



The authors thank the personnel and the veterinarians at the slaughterhouse Pomar in Araguari for allowing the collection of samples, Mr. Ricardo Canesso Dalla Rosa and Ms. Ana Paula Ferreira for the technical assistance in the laboratory, Ms. Denise Guethlin for assistance in the statistical analysis, the Brazilian agency CAPES (Project 182/04) and the German agency DAAD for the financial support for exchanging scientists and students, and the Brazilian National Council for Scientific and Technological Development (CNPq) for awarding scholarships. The experiments carried out in the present study comply with the current laws of Brazil and Germany.


  1. Alhassan A, Pumidonming W, Okamura M, Hirata H, Battsetseg B, Fujisaki K, Yokoyama N, Igarashi I (2005) Development of a single-round and multiplex PCR method for the simultaneous detection of Babesia caballi and Babesia equi in horse blood. Vet Parasitol 129:43–49PubMedCrossRefGoogle Scholar
  2. Allsopp MTEP, Lewis BD, Penzhorn BL (2007) Molecular evidence for transplacental transmission of Theileria equi from carrier mares to their apparently healthy foals. Vet Parasitol 148:130–136PubMedCrossRefGoogle Scholar
  3. Bahiruddin JB, Camma C, Rebelo E (1999) Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene. Vet Parasitol 84:75–83CrossRefGoogle Scholar
  4. Battsetseg B, Lucero S, Xuan X, Claveria FG, Inoue N, Alhassan A, Kanno T, Igarashi I, Nagasawa H, Mikami T, Fujisaki K (2002) Detection of natural infection of Boophilus microplus with Babesia equi and Babesia caballi in Brazilian horses using nested polymerase chain reaction. Vet Parasitol 107:351–357PubMedCrossRefGoogle Scholar
  5. Böse R, Jorgensen WK, Dalgliesh RJ, Freidhoff KT, de Vos AJ (1995) Current state and future trends in the diagnosis of babesiosis. Vet Parasitol 57:61–74PubMedCrossRefGoogle Scholar
  6. Cunha CW (1993) Babesiose equina: Padronização da reação de imunofluorescência para sorodiagnóstico e levantamento epidemiológico em eqüinos puro sangue inglês (Thesis, Universidade Federal de Pelotas, RS, Brazil)Google Scholar
  7. de Waal DT (1992) Equine piroplasmosis: a review Br Vet J 148:6–13PubMedGoogle Scholar
  8. de Waal DT, van Heerden J (1994) In: Coetzer JAW, Thompson GR, Tustin RC (eds) Infectious diseases of livestock (with special reference to Southern Africa), vol 1. Oxford University Press, Cape Town, pp 295–304Google Scholar
  9. Donnelly J, Phipps LP (1982) Evidence of maternal antibodies to Babesia equi and Babesia caballi in foals of seropositive mares. Equine Vet J 14:126–128PubMedCrossRefGoogle Scholar
  10. Friedhoff KT (1988) Transmission of Babesia. In: Ristic M (ed) Babesiosis of domestic animals and man. CRC, Boca Raton, pp 23–52Google Scholar
  11. Ginther OS (1967) Reproductive biology of the mare—basic and applied aspects. Equiservices, Cross Plains, WI. 396Google Scholar
  12. Guimaraes AM, Lima JD, Ribeiro MFB, Camargos ERS, Bozzi IA (1998a) Ultrastructure of sporogony in Babesia equi in salivary glands of adult female Boophilus microplus ticks. Parasitol Res 84:69–74PubMedCrossRefGoogle Scholar
  13. Guimaraes AM, Lima JD, Ribeiro MFB (1998b) Sporogony and experimental infection of Babesia equi by Boophilus microplus ticks. Parasitol Res 84:323–327PubMedCrossRefGoogle Scholar
  14. Heuchert CMS, Giulli V Jr, Athaide DF, Böse R, Friedhoff KT (1999) Seroepidemiologic studies on Babesia equi and Babesia caballi infections in Brazil. Vet Parasitol 85:1–11PubMedCrossRefGoogle Scholar
  15. Ikadai H, Xuan X, Igarashi I, Tanaka S, Kanemaru T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T (1999) Cloning and expression of a 48-kilodalton Babesia caballi merozoite rhoptry protein and potential use of the recombinant antigen in an enzyme-linked immunosorbent assay. J Clin Microbiol 37:3475–3480PubMedGoogle Scholar
  16. Nicolaiewsky TB, Richter MF, Lunge VR, Cunha CW, Delagostin O, Ikuta N, Fonseca AS, Silva SS, Ozaki LS (2001) Detection of Babesia equi (Laveran, 1901) by nested polymerase chain reaction. Vet Parasitol 101:9–21PubMedCrossRefGoogle Scholar
  17. Passos LMF, Ribeiro MFB, Anderegg PI, Böse R (1999) Serological diagnosis of Babesia equi and B. caballi in pregnant mares. Arq Bras Med Vet Zootec 51:527–530CrossRefGoogle Scholar
  18. Pfeifer Barbosa I, Böse R, Peymann B, Friedhoff KT (1995) Epidemiological aspects of equine babesioses in a herd of horses in Brazil. Vet Parasitol 58:1–8PubMedCrossRefGoogle Scholar
  19. Phipps LP, Otter A (2004) Transplacental transmission of Theileria equi in two foals born and reared in the United Kingdom. Vet Rec 154:406–408PubMedGoogle Scholar
  20. Posnett ES, Ambrosio RE (1991) DNA Probes for the detection of Babesia caballi. Parasitol 102:357–365CrossRefGoogle Scholar
  21. Rampersad J, Cesar E, Campell MD, Samlal M, Ammons D (2003) A field evaluation of PCR for the routine detection of Babesia equi in horses. Vet Parasitol 114:81–87PubMedCrossRefGoogle Scholar
  22. Ribeiro MFB, Lima JD (1989) Diagnóstico sorológico de babesiose equine por Babesia equi em Minas Gerais. Seminário Brasileiro de Parasitologia Veterinária. IV, Bagé, p 111Google Scholar
  23. Ribeiro MFB, Saito JF, Pimentel PV (1995) Babesiose equina. I. Primo-infecção de potros em área endêmica. Arq Bras Med Vet Zootec 47:641–647Google Scholar
  24. Ribeiro MFB, Costa JO, Guimaraes AM (1999) Epidemiological aspects of Babesia equi in horses in Minas Gerais, Brazil. Vet Res Commun 23:385–390PubMedCrossRefGoogle Scholar
  25. Roby TO, Anthony DW (1963) Transmission of equine piroplasmosis by the tropical tick Dermacentor nitens (Neumann). J Am Vet Med Assoc 142:768–769PubMedGoogle Scholar
  26. Rüegg SR, Torgerson P, Deplazes P, Mathis A (2007) Age-dependent dynamics of Theileria equi and Babesia caballi infections in southwest Mongolia based on IFAT and/or PCR prevalence data from domestic horses and ticks. Parasitology 134:939–947PubMedCrossRefGoogle Scholar
  27. Tenter AM, Friedhoff KT (1986) Serodiagnosis of experimental and natural Babesia equi and B. caballi infections. Vet Parasitol 20:49–61PubMedCrossRefGoogle Scholar
  28. Ueti MW, Palmer GH, Kappmeyer LS, Scoles GA, Knowles DP (2003) Expression of equi merozoite antigen 2 during development of Babesia equi in the midgut and salivary gland of the vector tick Boophilus microplus. J Clin Microbiol 41:5803–5809PubMedCrossRefGoogle Scholar
  29. Uilenberg G (2006) Babesia-a historical overview. Vet Parasitol 138:3–10PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Alexandra Heim
    • 1
  • Lygia M. F. Passos
    • 1
    • 2
  • Múcio F. B. Ribeiro
    • 3
  • Lívio M. Costa-Júnior
    • 3
    • 4
  • Camila V. Bastos
    • 3
  • Dagmar D. Cabral
    • 5
  • Jörg Hirzmann
    • 1
  • Kurt Pfister
    • 1
  1. 1.Institute for Comparative Tropical Medicine and ParasitologyLudwig Maximilian University MunichMunichGermany
  2. 2.Departamento de Medicina Veterinária PreventivaEscola de Veterinária-UFMGBelo HorizonteBrazil
  3. 3.Departamento de ParasitologiaICB-UFMGBelo HorizonteBrazil
  4. 4.Centro de Ciências Agrárias e AmbientaisCCAA-UFMASão LuísBrazil
  5. 5.Universidade Federal de UberlândiaUberlândiaBrazil

Personalised recommendations