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Parasitology research2008; 103(5); 1145-1150; doi: 10.1007/s00436-008-1108-z

Diagnosis of Babesia caballi and Theileria equi infections in horses in Sudan using ELISA and PCR.

Abstract: The purpose of this study was to estimate the prevalence of equine piroplasmosis in Sudan. The presence of antibodies against Babesia caballi and Theileria equi was determined in serum samples obtained from 158 horses raised in different locations in Sudan by enzyme-linked immunosorbent assay (ELISA). The B. caballi 48-kDa and the T. equi EMA-2 purified recombinant proteins were used as antigens in the ELISA test. Results showed that seven (4.4%) were positive for B. caballi and 80 (63.5%) were positive for T. equi. Polymerase chain reaction (PCR) assays have been applied using primers targeting the B. caballi 48-kDa merozoite antigen, the T. equi SSUrRNA and the T. equi EMA-1 genes. PCR performed on 131 blood spots in filter paper revealed that 33 (25.2%) samples were positive for T. equi but no positives were found for B. caballi. It is concluded that equine piroplasmosis is endemic in the country. This is the first study on serological and molecular epidemiological diagnosis on equine piroplasmosis in Sudan.
Publication Date: 2008-07-10 PubMed ID: 18618143DOI: 10.1007/s00436-008-1108-zGoogle Scholar: Lookup
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  • Journal Article

Summary

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The research conducted a test on horses in Sudan to determine the prevalence of equine piroplasmosis, a disease caused by the parasites Babesia caballi and Theileria equi. They employed both an enzyme-linked immunosorbent assay (ELISA) and a polymerase chain reaction (PCR) technique to identify positive cases and found the disease to be endemic in the country.

Key Aims and Methods

  • The primary objective of this study was to figure out how prevalent equine piroplasmosis was in Sudan. Equine piroplasmosis is a parasitic disease affecting horses, caused by Babesia caballi and Theileria equi.
  • The researchers collected and analyzed serum samples from 158 different horses across various locations in Sudan.
  • In order to detect the presence of antibodies against these parasites, the team used an enzyme-linked immunosorbent assay (ELISA).
  • In order to identify the specific antigens of the parasites, a B. caballi 48-kDa merozoite antigen and the T. equi EMA-2 purified recombinant proteins were used.
  • Polymerase chain reaction (PCR) assays were utilized to analyze 131 blood spots, with the target being B. caballi 48-kDa merozoite antigen, the T. equi SSUrRNA, and the T. equi EMA-1 genes. PCR is a method used to amplify a specific DNA sequence to produce enough for analysis.

Major Findings

  • Using ELISA, about 4.4% (7 horses) were found to carry antibodies of B. caballi and 63.5% (80 horses) for T. equi.
  • When tested with PCR assays, 25.2% (33 samples) were found to be positive for T. equi, but none were found positive for B. caballi.
  • The results from this study strongly suggest that equine piroplasmosis is endemic in Sudan.

Conclusion and Relevance

  • This research provided crucial insight into the prevalence and spread of equine piroplasmosis in Sudan.
  • These findings are significant considering this is the first study on serological and molecular epidemiological diagnosis on equine piroplasmosis in Sudan. It reveals the urgent need to implement control measures to arrest the spread of this disease.

Cite This Article

APA
Salim BO, Hassan SM, Bakheit MA, Alhassan A, Igarashi I, Karanis P, Abdelrahman MB. (2008). Diagnosis of Babesia caballi and Theileria equi infections in horses in Sudan using ELISA and PCR. Parasitol Res, 103(5), 1145-1150. https://doi.org/10.1007/s00436-008-1108-z

Publication

ISSN: 0932-0113
NlmUniqueID: 8703571
Country: Germany
Language: English
Volume: 103
Issue: 5
Pages: 1145-1150

Researcher Affiliations

Salim, B O M
  • Faculty of Veterinary Medicine, University of Khartoum, Khartoum, Sudan.
Hassan, S M
    Bakheit, M A
      Alhassan, A
        Igarashi, I
          Karanis, P
            Abdelrahman, M B

              MeSH Terms

              • Animals
              • Antibodies, Protozoan / blood
              • Babesia
              • Babesiosis / blood
              • Babesiosis / diagnosis
              • Babesiosis / epidemiology
              • Babesiosis / veterinary
              • Enzyme-Linked Immunosorbent Assay / veterinary
              • Horse Diseases / blood
              • Horse Diseases / diagnosis
              • Horse Diseases / epidemiology
              • Horses
              • Polymerase Chain Reaction / veterinary
              • Prevalence
              • Sudan / epidemiology
              • Theileria
              • Theileriasis / blood
              • Theileriasis / diagnosis
              • Theileriasis / epidemiology

              References

              This article includes 29 references
              1. Ikadai H, Xuan X, Igarashi I, Tanaka S, Kanemaru T, Nagasawa H, Fujisaki K, Suzuki N, Mikami T. 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 1999 Nov;37(11):3475-80.
              2. Knowles DP Jr, Perryman LE, Goff WL, Miller CD, Harrington RD, Gorham JR. A monoclonal antibody defines a geographically conserved surface protein epitope of Babesia equi merozoites.. Infect Immun 1991 Jul;59(7):2412-7.
              3. Bakheit MA, Seitzer U, Mbati PA, Ahmed JS. Serological diagnostic tools for the major tick-borne protozoan diseases of livestock.. Parassitologia 2007 May;49 Suppl 1:53-62.
                pubmed: 17691608
              4. Holman PJ, Hietala SK, Kayashima LR, Olson D, Waghela SD, Wagner GG. Case report: field-acquired subclinical Babesia equi infection confirmed by in vitro culture.. J Clin Microbiol 1997 Feb;35(2):474-6.
                pubmed: 9003619doi: 10.1128/jcm.35.2.474-476.1997google scholar: lookup
              5. Alhassan A, Pumidonming W, Okamura M, Hirata H, Battsetseg B, Fujisaki K, Yokoyama N, Igarashi I. Development of a single-round and multiplex PCR method for the simultaneous detection of Babesia caballi and Babesia equi in horse blood.. Vet Parasitol 2005 Apr 20;129(1-2):43-9.
                pubmed: 15817201doi: 10.1016/j.vetpar.2004.12.018google scholar: lookup
              6. 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 Sep;41(9):4172-7.
              7. Rüegg SR, Torgerson P, Deplazes P, Mathis A. 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 2007 Jul;134(Pt 7):939-47.
                pubmed: 17306055doi: 10.1017/S0031182007002405google scholar: lookup
              8. Ikadai H, Nagai A, Xuan X, Igarashi I, Tsugihiko K, Tsuji N, Oyamada T, Suzuki N, Fujisaki K. Seroepidemiologic studies on Babesia caballi and Babesia equi infections in Japan.. J Vet Med Sci 2002 Apr;64(4):325-8.
                pubmed: 12014577doi: 10.1292/jvms.64.325google scholar: lookup
              9. Kappmeyer LS, Perryman LE, Knowles DP Jr. A Babesia equi gene encodes a surface protein with homology to Theileria species.. Mol Biochem Parasitol 1993 Nov;62(1):121-4.
                pubmed: 8114813doi: 10.1016/0166-6851(93)90185-zgoogle scholar: lookup
              10. Friedhoff KT, Soulé C. An account on equine babesioses.. Rev Sci Tech 1996 Sep;15(3):1191-201.
                pubmed: 9025154doi: 10.20506/rst.15.3.972google scholar: lookup
              11. Knowles DP, Kappmeyer LS, Perryman LE. Genetic and biochemical analysis of erythrocyte-stage surface antigens belonging to a family of highly conserved proteins of Babesia equi and Theileria species.. Mol Biochem Parasitol 1997 Dec 1;90(1):69-79.
                pubmed: 9497033doi: 10.1016/s0166-6851(97)00150-3google scholar: lookup
              12. Brüning A, Phipps P, Posnett E, Canning EU. Monoclonal antibodies against Babesia caballi and Babesia equi and their application in serodiagnosis.. Vet Parasitol 1997 Jan;68(1-2):11-26.
                pubmed: 9066047doi: 10.1016/s0304-4017(96)01074-6google scholar: lookup
              13. Papadopoulos B, Brossard M, Perié NM. Piroplasms of domestic animals in the Macedonia region of Greece. 3. Piroplasms of small ruminants.. Vet Parasitol 1996 May;63(1-2):67-74.
                pubmed: 8792581doi: 10.1016/0304-4017(95)00846-2google scholar: lookup
              14. Zaugg JL, Lane VM. Efficacy of buparvaquone as a therapeutic and clearing agent of Babesia equi of European origin in horses.. Am J Vet Res 1992 Aug;53(8):1396-9.
                pubmed: 1510317
              15. Friedhoff KT, Tenter AM, Müller I. Haemoparasites of equines: impact on international trade of horses.. Rev Sci Tech 1990 Dec;9(4):1187-94.
                pubmed: 2132711
              16. Alhassan A, Govind Y, Tam NT, Thekisoe OM, Yokoyama N, Inoue N, Igarashi I. Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis.. Parasitol Res 2007 Apr;100(5):1165-8.
                pubmed: 17216488doi: 10.1007/s00436-006-0430-6google scholar: lookup
              17. Xuan X, Nagai A, Battsetseg B, Fukumoto S, Makala LH, Inoue N, Igarashi I, Mikami T, Fujisaki K. Diagnosis of equine piroplasmosis in Brazil by serodiagnostic methods with recombinant antigens.. J Vet Med Sci 2001 Oct;63(10):1159-60.
                pubmed: 11714038doi: 10.1292/jvms.63.1159google scholar: lookup
              18. Rampersad J, Cesar E, Campbell MD, Samlal M, Ammons D. A field evaluation of PCR for the routine detection of Babesia equi in horses.. Vet Parasitol 2003 May 30;114(2):81-7.
                pubmed: 12781470doi: 10.1016/s0304-4017(03)00129-8google scholar: lookup
              19. Brüning A. Equine piroplasmosis an update on diagnosis, treatment and prevention.. Br Vet J 1996 Mar;152(2):139-51.
                pubmed: 8680838doi: 10.1016/s0007-1935(96)80070-4google scholar: lookup
              20. Criado-Fornelio A, Martinez-Marcos A, Buling-Saraña A, Barba-Carretero JC. Molecular studies on Babesia, Theileria and Hepatozoon in southern Europe. Part II. Phylogenetic analysis and evolutionary history.. Vet Parasitol 2003 Jun 11;114(3):173-94.
                pubmed: 12788253doi: 10.1016/s0304-4017(03)00141-9google scholar: lookup
              21. Edelhofer R, Kanout A, Schuh M, Kutzer E. Improved disease resistance after Babesia divergens vaccination.. Parasitol Res 1998;84(3):181-7.
                pubmed: 9521006doi: 10.1007/s004360050380google scholar: lookup
              22. Avarzed A, De Waal DT, Igarashi I, Saito A, Oyamada T, Toyoda Y, Suzuki N. Prevalence of equine piroplasmosis in Central Mongolia.. Onderstepoort J Vet Res 1997 Jun;64(2):141-5.
                pubmed: 9352563
              23. Knowles DP Jr, Kappmeyer LS, Stiller D, Hennager SG, Perryman LE. Antibody to a recombinant merozoite protein epitope identifies horses infected with Babesia equi.. J Clin Microbiol 1992 Dec;30(12):3122-6.
              24. Krause PJ. Babesiosis diagnosis and treatment.. Vector Borne Zoonotic Dis 2003 Spring;3(1):45-51.
                pubmed: 12804380doi: 10.1089/153036603765627451google scholar: lookup
              25. Persing DH, Conrad PA. Babesiosis: new insights from phylogenetic analysis.. Infect Agents Dis 1995 Dec;4(4):182-95.
                pubmed: 8665084
              26. Calder JA, Reddy GR, Chieves L, Courtney CH, Littell R, Livengood JR, Norval RA, Smith C, Dame JB. Monitoring Babesia bovis infections in cattle by using PCR-based tests.. J Clin Microbiol 1996 Nov;34(11):2748-55.
              27. Frerichs WM, Holbrook AA, Johnson AJ. Equine piroplasmosis: production of antigens for the complement-fixation test.. Am J Vet Res 1969 Aug;30(8):1337-41.
                pubmed: 5816365
              28. Cacciò S, Cammà C, Onuma M, Severini C. The beta-tubulin gene of Babesia and Theileria parasites is an informative marker for species discrimination.. Int J Parasitol 2000 Oct;30(11):1181-5.
                pubmed: 11027785doi: 10.1016/s0020-7519(00)00105-3google scholar: lookup
              29. de Waal DT. Equine piroplasmosis: a review.. Br Vet J 1992 Jan-Feb;148(1):6-14.
                pubmed: 1551016doi: 10.1016/0007-1935(92)90061-5google scholar: lookup

              Citations

              This article has been cited 18 times.
              1. Galon EM, Macalanda AM, Garcia MM, Ibasco CJ, Garvida A, Ji S, Zafar I, Hasegawa Y, Liu M, Ybañez RH, Umemiya-Shirafuji R, Ybañez A, Claveria F, Xuan X. Molecular Identification of Selected Tick-Borne Protozoan and Bacterial Pathogens in Thoroughbred Racehorses in Cavite, Philippines. Pathogens 2021 Oct 13;10(10).
                doi: 10.3390/pathogens10101318pubmed: 34684266google scholar: lookup
              2. Khogali RKI, Hassan SM. Drop-off rhythms and survival of Hyalomma anatolicum (Acari: Ixodidae) fed on crossbred (Friesians x Zebu) calves in Sudan. Heliyon 2021 Sep;7(9):e07930.
                doi: 10.1016/j.heliyon.2021.e07930pubmed: 34541348google scholar: lookup
              3. Kamani J, Bártová E, Kašpárková N, Mohammed SJ, Budíková M, Sedlák K. Seroprevalence of Theileria equi, Babesia caballi, and Trichinella spp. infections in horses and donkeys from Nigeria, West Africa. Trop Anim Health Prod 2021 May 24;53(3):338.
                doi: 10.1007/s11250-021-02737-9pubmed: 34031761google scholar: lookup
              4. Tirosh-Levy S, Gottlieb Y, Fry LM, Knowles DP, Steinman A. Twenty Years of Equine Piroplasmosis Research: Global Distribution, Molecular Diagnosis, and Phylogeny. Pathogens 2020 Nov 8;9(11).
                doi: 10.3390/pathogens9110926pubmed: 33171698google scholar: lookup
              5. Onyiche TE, Taioe MO, Molefe NI, Biu AA, Luka J, Omeh IJ, Yokoyama N, Thekisoe O. Equine piroplasmosis: an insight into global exposure of equids from 1990 to 2019 by systematic review and meta-analysis. Parasitology 2020 Nov;147(13):1411-1424.
                doi: 10.1017/S0031182020001407pubmed: 32741382google scholar: lookup
              6. Onyiche TE, Taioe MO, Ogo NI, Sivakumar T, Biu AA, Mbaya AW, Xuan X, Yokoyama N, Thekisoe O. Molecular evidence of Babesia caballi and Theileria equi in equines and ticks in Nigeria: prevalence and risk factors analysis. Parasitology 2020 Sep;147(11):1238-1248.
                doi: 10.1017/S0031182020000992pubmed: 32605687google scholar: lookup
              7. Elata A, Mossaad E, Satti R, Matar N, Ohari Y, Xuan X, Inoue N, Suganuma K. Serological and molecular detection of selected hemoprotozoan parasites in donkeys in West Omdurman, Khartoum State, Sudan. J Vet Med Sci 2020 Mar 5;82(3):286-293.
                doi: 10.1292/jvms.19-0534pubmed: 31969541google scholar: lookup
              8. Onyiche TE, Suganuma K, Igarashi I, Yokoyama N, Xuan X, Thekisoe O. A Review on Equine Piroplasmosis: Epidemiology, Vector Ecology, Risk Factors, Host Immunity, Diagnosis and Control. Int J Environ Res Public Health 2019 May 16;16(10).
                doi: 10.3390/ijerph16101736pubmed: 31100920google scholar: lookup
              9. Montes Cortés MG, Fernández-García JL, Habela Martínez-Estéllez MÁ. Seroprevalence of Theileria equi and Babesia caballi in horses in Spain. Parasite 2017;24:14.
                doi: 10.1051/parasite/2017015pubmed: 28497743google scholar: lookup
              10. Haghi MM, Etemadifar F, Fakhar M, Teshnizi SH, Soosaraei M, Shokri A, Hajihasani A, Mashhadi H. Status of babesiosis among domestic herbivores in Iran: a systematic review and meta-analysis. Parasitol Res 2017 Apr;116(4):1101-1109.
                doi: 10.1007/s00436-016-5368-8pubmed: 28054180google scholar: lookup
              11. Mans BJ, Pienaar R, Latif AA. A review of Theileria diagnostics and epidemiology. Int J Parasitol Parasites Wildl 2015 Apr;4(1):104-18.
                doi: 10.1016/j.ijppaw.2014.12.006pubmed: 25830110google scholar: lookup
              12. Malekifard F, Tavassoli M, Yakhchali M, Darvishzadeh R. Detection of Theileria equi and Babesia caballi using microscopic and molecular methods in horses in suburb of Urmia, Iran. Vet Res Forum 2014 Spring;5(2):129-33.
                pubmed: 25568706
              13. Onmaz AC, Beutel RG, Schneeberg K, Pavaloiu AN, Komarek A, van den Hoven R. Vectors and vector-borne diseases of horses. Vet Res Commun 2013 Mar;37(1):65-81.
                doi: 10.1007/s11259-012-9537-7pubmed: 23054414google scholar: lookup
              14. Salim B, Bakheit MA, Kamau J, Nakamura I, Sugimoto C. Nucleotide sequence heterogeneity in the small subunit ribosomal RNA gene within Theileria equi from horses in Sudan. Parasitol Res 2010 Jan;106(2):493-8.
                doi: 10.1007/s00436-009-1691-7pubmed: 19953269google scholar: lookup
              15. Khaing Y, Htun LL, Linn KS, Kyaw WO, Nwae TT, Chel HM, Win SY, Murata S, Nakao R, Nonaka N, Bawm S. Microscopic examination of haemoparasites and the first molecular detection of Theileria equi in horses in Myanmar. Parasitol Res 2025 Apr 21;124(4):42.
                doi: 10.1007/s00436-025-08488-ypubmed: 40257584google scholar: lookup
              16. Sadeddine R, Righi S, Saidani K, Benakhla A. First Molecular Characterization of Theileria equi from Northeastern Algeria. Acta Parasitol 2025 Mar 6;70(2):66.
                doi: 10.1007/s11686-025-01006-1pubmed: 40050535google scholar: lookup
              17. Ramadan RM, Taha NM, Auda HM, Elsamman EM, El-Bahy MM, Salem MA. Molecular and immunological studies on Theileria equi and its vector in Egypt. Exp Appl Acarol 2024 Aug;93(2):439-458.
                doi: 10.1007/s10493-024-00933-4pubmed: 38967736google scholar: lookup
              18. Mohammad-Naseri A, Shokrani H, Rahmani-Shahraki A. Equine Piroplasmosis in Asymptomatic Horses of Western Iran: Comparison of Microscopic Examination and Multiplex PCR. Acta Parasitol 2024 Mar;69(1):813-818.
                doi: 10.1007/s11686-024-00804-3pubmed: 38424400google scholar: lookup