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Home / Equine Research Bank / J Arthropod Borne Dis / Genetic Variation of the β-tubulin Gene of Babesia cab...
Journal of arthropod-borne diseases2017; 11(3); 344-353;

Genetic Variation of the β-tubulin Gene of Babesia caballi Strains.

Abstract: Equine piroplasmosis is caused by two haemoprotozoan parasites: and . Negative economic impact on international trade has been associated to endemic sites. This is the reason why carrier detection requires reliable diagnostic methods. Various diagnostic modalities can be used alone or in combination including PCR. However, genetic variation of commonly used genes is still of debate. The aim of this research was to sequence the β-tubulin gene of a strain from Spain and to compare it with known β-tubulin sequences. Methods: DNA was isolated from a cryopreserved strain from Spain and acute and chronic carrier horses. Firstly, degenerated primer pairs were designed based on GenBank sequences of different and species for sequencing. The primers were redesigned to amplify both parasites, simultaneously. Finally, a species-specific primer pair for was designed and a Restriction Fragment Length Polymorphism-PCR (PCR-RFLP) assay performed to know the difference of known strains. Results: We provided new insights of the β-tubulin gene and a good molecular coverage of this gene, contributing with a number of useful primers to amplify and . Moreover, PCR-RFLP assays based on the exon II of this gene confirmed the causative strain in Spanish horses. Conclusions: We reported useful primer pairs for diagnostic and a new sequence of the β-tubulin gene of , which will facilitate the development of future assays and the detection of infected horses, preventing thus the spread of this disease worldwide.
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Publication Date: 2017-09-08 PubMed ID: 29322051PubMed Central: PMC5758630
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study revolves around the genetic variance of the β-tubulin gene in Babesia caballi strains, a parasite causing equine piroplasmosis. The researchers aim to sequence this gene from a Spanish strain and compare it with existing sequences, leading to the development of more reliable diagnostic methods.

Research Methodology

  • The researchers started by isolating DNA from a Babesia caballi strain cryopreserved in Spain and from horses that were acute and chronic carriers of the parasite.
  • Initial primer pairs were devised based on GenBank sequences from different Babesia and Theileria species for sequencing purposes.
  • These primers were then reformed to amplify both parasites simultaneously.
  • Finally, a species-specific primer pair for Babesia caballi was crafted and a PCR-RFLP (Polymerase Chain Reaction – Restriction Fragment Length Polymorphism) assay was executed to distinguish known Babesia caballi strains.

Key Findings

  • Researchers provided fresh insights into the β-tubulin gene and contributed a number of useful primers to amplify Babesia and Theileria.
  • Through PCR-RFLP assays based on the second exon of the β-tubulin gene, the study confirmed the causative Babesia caballi strain in Spanish horses.

Conclusion and Future Implications

  • The study contributed useful primer pairs for diagnostic purposes and a new sequence of the β-tubulin gene of Babesia caballi.
  • Such findings will aid in the development of future tests and the detection of infected horses, thereby preventing the global spread of equine piroplasmosis.

Cite This Article

APA
Montes-Cortés MG, Fernández-García JL, Martínez-Estéllez MÁH. (2017). Genetic Variation of the β-tubulin Gene of Babesia caballi Strains. J Arthropod Borne Dis, 11(3), 344-353.

Publication

Journal of arthropod-borne diseases
ISSN: 2322-1984
NlmUniqueID: 101593448
Country: Iran
Language: English
Volume: 11
Issue: 3
Pages: 344-353

Researcher Affiliations

Montes-Cortés, María Guadalupe
  • Parasitology and Parasitological Diseases, Veterinary Faculty, Universidad de Extremadura, Cáceres, Spain.
Fernández-García, José Luis
  • Genetics and Animal Breeding, Veterinary Faculty, Universidad de Extremadura, Cáceres, Spain.
Martínez-Estéllez, Miguel Ángel Habela
  • Parasitology and Parasitological Diseases, Veterinary Faculty, Universidad de Extremadura, Cáceres, Spain.

References

This article includes 37 references
  1. Rodríguez Bautista JL, Ikadai H, You M, Battsetseg B, Igarashi I, Nagasawa H, Fujisaki K. Molecular evidence of Babesia caballi (Nuttall and Strickland, 1910) parasite transmission from experimentally-infected SCID mice to the ixodid tick, Haemaphysalis longicornis (Neuman, 1901).. Vet Parasitol 2001 Dec 13;102(3):185-91.
    pubmed: 11777598doi: 10.1016/s0304-4017(01)00531-3google scholar: lookup
  2. Mehlhorn H, Schein E. Redescription of Babesia equi Laveran, 1901 as Theileria equi Mehlhorn, Schein 1998.. Parasitol Res 1998 Jun;84(6):467-75.
    pubmed: 9660136doi: 10.1007/s004360050431google scholar: lookup
  3. Uilenberg G. Babesia--a historical overview.. Vet Parasitol 2006 May 31;138(1-2):3-10.
    pubmed: 16513280doi: 10.1016/j.vetpar.2006.01.035google scholar: lookup
  4. De Waal DT, Van Heerden J. Equine Piroplasmosis. 1994.
  5. 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
  6. Bashiruddin JB, Cammà C, Rebêlo E. Molecular detection of Babesia equi and Babesia caballi in horse blood by PCR amplification of part of the 16S rRNA gene.. Vet Parasitol 1999 Jul;84(1-2):75-83.
    pubmed: 10435792doi: 10.1016/s0304-4017(99)00049-7google scholar: lookup
  7. Wise LN, Kappmeyer LS, Mealey RH, Knowles DP. Review of equine piroplasmosis.. J Vet Intern Med 2013 Nov-Dec;27(6):1334-46.
    pubmed: 24033559doi: 10.1111/jvim.12168google scholar: lookup
  8. Böse R, Jorgensen WK, Dalgliesh RJ, Friedhoff KT, de Vos AJ. Current state and future trends in the diagnosis of babesiosis.. Vet Parasitol 1995 Mar;57(1-3):61-74.
    pubmed: 7597794doi: 10.1016/0304-4017(94)03111-9google scholar: lookup
  9. Mans BJ, Pienaar R, Latif AA. A review of Theileria diagnostics and epidemiology.. Int J Parasitol Parasites Wildl 2015 Apr;4(1):104-18.
    pmc: PMC4356873pubmed: 25830110doi: 10.1016/j.ijppaw.2014.12.006google scholar: lookup
  10. 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
  11. Sumbria D, Das Singla L, Sharma A. Theileria equi and Babesia caballi infection of equids in Punjab, India: a serological and molecular survey.. Trop Anim Health Prod 2016 Jan;48(1):45-52.
    pubmed: 26387094doi: 10.1007/s11250-015-0917-1google scholar: lookup
  12. Nicolaiewsky TB, Richter MF, Lunge VR, Cunha CW, Delagostin O, Ikuta N, Fonseca AS, da Silva SS, Ozaki LS. Detection of Babesia equi (Laveran, 1901) by nested polymerase chain reaction.. Vet Parasitol 2001 Oct 31;101(1):9-21.
    pubmed: 11587829doi: 10.1016/s0304-4017(01)00471-xgoogle scholar: lookup
  13. 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
  14. Sumbria D, Singla LD, Kumar S, Sharma A, Dahiya RK, Setia R. Spatial distribution, risk factors and haemato-biochemical alterations associated with Theileria equi infected equids of Punjab (India) diagnosed by indirect ELISA and nested PCR.. Acta Trop 2016 Mar;155:104-12.
    pubmed: 26747007doi: 10.1016/j.actatropica.2015.12.020google scholar: lookup
  15. Kim CM, Blanco LB, Alhassan A, Iseki H, Yokoyama N, Xuan X, Igarashi I. Diagnostic real-time PCR assay for the quantitative detection of Theileria equi from equine blood samples.. Vet Parasitol 2008 Feb 14;151(2-4):158-63.
    pubmed: 18077095doi: 10.1016/j.vetpar.2007.10.023google scholar: lookup
  16. Ueti MW, Palmer GH, Kappmeyer LS, Scoles GA, Knowles DP. 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 2003 Dec;41(12):5803-9.
    pmc: PMC308990pubmed: 14662988doi: 10.1128/jcm.41.12.5803-5809.2003google scholar: lookup
  17. Bhoora R, Quan M, Zweygarth E, Guthrie AJ, Prinsloo SA, Collins NE. Sequence heterogeneity in the gene encoding the rhoptry-associated protein-1 (RAP-1) of Babesia caballi isolates from South Africa.. Vet Parasitol 2010 May 11;169(3-4):279-88.
    pubmed: 20138703doi: 10.1016/j.vetpar.2010.01.009google scholar: lookup
  18. Zamoto A, Tsuji M, Wei Q, Cho SH, Shin EH, Kim TS, Leonova GN, Hagiwara K, Asakawa M, Kariwa H, Takashima I, Ishihara C. Epizootiologic survey for Babesia microti among small wild mammals in northeastern Eurasia and a geographic diversity in the beta-tubulin gene sequences.. J Vet Med Sci 2004 Jul;66(7):785-92.
    pubmed: 15297749doi: 10.1292/jvms.66.785google scholar: lookup
  19. García-Bocanegra I, Arenas-Montes A, Hernández E, Adaszek L, Carbonero A, Almería S, Jaén-Téllez JA, Gutiérrez-Palomino P, Arenas A. Seroprevalence and risk factors associated with Babesia caballi and Theileria equi infection in equids.. Vet J 2013 Feb;195(2):172-8.
    pubmed: 22784418doi: 10.1016/j.tvjl.2012.06.012google scholar: lookup
  20. Habela MA, Gragera-Slikker A, Moreno A, Montes G, Sevilla R. Piroplasmosis equina: conocimiento y grado de concienciación de los productores de caballos Pura Raza Española. Revista Equinus 2005 11: 17– 34.
  21. Zweygarth E, Lopez-Rebollar LM, Nurton J, Guthrie AJ. Culture, isolation and propagation of Babesia caballi from naturally infected horses.. Parasitol Res 2002 May;88(5):460-2.
    pubmed: 12049465doi: 10.1007/s00436-002-0609-4google scholar: lookup
  22. Camacho AT, Guitian FJ, Pallas E, Gestal JJ, Olmeda AS, Habela MA, Telford SR 3rd, Spielman A. Theileria (Babesia) equi and Babesia caballi infections in horses in Galicia, Spain.. Trop Anim Health Prod 2005 May;37(4):293-302.
    pubmed: 15934637doi: 10.1007/s11250-005-5691-zgoogle scholar: lookup
  23. Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells.. Nucleic Acids Res 1988 Feb 11;16(3):1215.
    pmc: PMC334765pubmed: 3344216doi: 10.1093/nar/16.3.1215google scholar: lookup
  24. Fernández-García JL. The endangered Dama dama mesopotamica: genetic variability, allelic loss and hybridization signals. Contrib. Zool. 2012 81: 223– 233.
  25. Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0.. Mol Biol Evol 2007 Aug;24(8):1596-9.
    pubmed: 17488738doi: 10.1093/molbev/msm092google scholar: lookup
  26. Barbosa AM, Fernández-García JL, Carranza J. A new Marker for rapid sex identification of red deer (Cervus elaphus). Hystrix It J Mamm. 2009 20: 169– 172.
  27. 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
  28. Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.. Nucleic Acids Res 1997 Sep 1;25(17):3389-402.
    pmc: PMC146917pubmed: 9254694doi: 10.1093/nar/25.17.3389google scholar: lookup
  29. Sivakumar T, Tattiyapong M, Okubo K, Suganuma K, Hayashida K, Igarashi I, Zakimi S, Matsumoto K, Inokuma H, Yokoyama N. PCR detection of Babesia ovata from questing ticks in Japan.. Ticks Tick Borne Dis 2014 Apr;5(3):305-10.
    pubmed: 24572609doi: 10.1016/j.ttbdis.2013.12.006google scholar: lookup
  30. Criado-Fornelio A, Gónzalez-del-Río MA, Buling-Saraña A, Barba-Carretero JC. The "expanding universe" of piroplasms.. Vet Parasitol 2004 Feb 6;119(4):337-45.
    pubmed: 15154598doi: 10.1016/j.vetpar.2003.11.015google scholar: lookup
  31. Nagore D, García-Sanmartín J, García-Pérez AL, Juste RA, Hurtado A. Detection and identification of equine Theileria and Babesia species by reverse line blotting: epidemiological survey and phylogenetic analysis.. Vet Parasitol 2004 Aug 13;123(1-2):41-54.
    pubmed: 15265570doi: 10.1016/j.vetpar.2004.04.010google scholar: lookup
  32. Bhoora R, Franssen L, Oosthuizen MC, Guthrie AJ, Zweygarth E, Penzhorn BL, Jongejan F, Collins NE. Sequence heterogeneity in the 18S rRNA gene within Theileria equi and Babesia caballi from horses in South Africa.. Vet Parasitol 2009 Feb 5;159(2):112-20.
    pubmed: 19019541doi: 10.1016/j.vetpar.2008.10.004google scholar: lookup
  33. Munkhjargal T, Sivakumar T, Battsetseg B, Nyamjargal T, Aboulaila M, Purevtseren B, Bayarsaikhan D, Byambaa B, Terkawi MA, Yokoyama N, Igarashi I. Prevalence and genetic diversity of equine piroplasms in Tov province, Mongolia.. Infect Genet Evol 2013 Jun;16:178-85.
    pubmed: 23416256doi: 10.1016/j.meegid.2013.02.005google scholar: lookup
  34. Alanazi AD, Said AE, Morin-Adeline V, Alyousif MS, Slapeta J. Quantitative PCR detection of Theileria equi using laboratory workflows to detect asymptomatic persistently infected horses.. Vet Parasitol 2014 Dec 15;206(3-4):138-45.
    pubmed: 25450724doi: 10.1016/j.vetpar.2014.09.019google scholar: lookup
  35. Baneth G, Florin-Christensen M, Cardoso L, Schnittger L. Reclassification of Theileria annae as Babesia vulpes sp. nov.. Parasit Vectors 2015 Apr 8;8:207.
    pmc: PMC4393874pubmed: 25890372doi: 10.1186/s13071-015-0830-5google scholar: lookup
  36. El-Sayed SAES, Rizk MA, Terkawi MA, Mousa A, El Said ESES, Elsayed G, Fouda M, Yokoyama N, Igarashi I. Cocktail of Theileria equi antigens for detecting infection in equines. Asian Pac J Trop Biomed 2015 5: 977– 981.
  37. Mahmoud MS, El-Ezz NT, Abdel-Shafy S, Nassar SA, El Namaky AH, Khalil WK, Knowles D, Kappmeyer L, Silva MG, Suarez CE. Assessment of Theileria equi and Babesia caballi infections in equine populations in Egypt by molecular, serological and hematological approaches.. Parasit Vectors 2016 May 4;9:260.
    pmc: PMC4857240pubmed: 27146413doi: 10.1186/s13071-016-1539-9google scholar: lookup

Citations

This article has been cited 1 times.
  1. 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
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