Development of a qPCR to diagnose the genus Eimeria in bovines
DOI:
https://doi.org/10.5433/1679-0359.2020v41n6p2695Keywords:
qPCR, Eimeria, coccidiosis, 18S rRNA, cattleAbstract
Bovine coccidiosis is caused by protozoa of the genus Eimeria. These protozoa mainly affect young animals, causing a decrease in production and consequent economic losses. The routine diagnosis is made through morphological observation of the oocysts, which has several limitations. The objective of the present study was to develop a qPCR technique for the diagnose of Eimeria spp. in cattle. For this purpose, the 18S rRNA region of the DNA of these parasites was selected, since it is a region with low variability among the species. The qPCR was developed using the SYBR Green, resulting in a PCR with a high sensitivity, able to amplify samples containing only one oocyst of Eimeria spp. of bovines. The feasibility of using qPCR in the diagnosis of the Eimeria Genus is demonstrated in this study, once this technique shows to be less laborious and needs less skills for diagnostic training when compared to the technique conventionally used in theroutine (micromorphometry).
Downloads
References
Almeida, V. A., Magalhães, V. C. S., Muniz, E. S., Neta, & Munhoz, A. D. (2011). Frequency of species of the genus Eimeria in naturally infected cattle in Southern Bahia, Northeast Brazil. Revista Brasileira de Parasitologia Veterinária, 20(1), 78-81. doi: 10.1590/S1984-29612011000100017
Bangoura, B., Mundt, H. C., Schmäschke, R., Westphal, B., & Daugschies, A. (2011). Prevalence of Eimeria bovis and Eimeria zuernii in German cattle herds and factors influencing oocyst excretion. Parasitology Research, 109(2), 129-138. doi: 10.1007/s00436-011-2569-z
Bruhn, F. R. P., Lopes, M. A., Demeu, F. A., Perazza, C. A., Pedrosa, M. F., & Guimarães, A. M. (2011). Frequency of species of Eimeria in females of the Holstein Friesian breed at the post-weaning stage during autumn and winter. Revista Brasileira de Parasitologia Veterinária, 20(4), 303-307. doi: 10. 1590/S1984-29612011000400008
Bruhn, F. R. P., Silva, F. A., Jr., Carvalho, A. H. O., Orlando, D. R., Rocha, C. M. B. M., & Guimarães, A. M. (2012). Ocorrences of Eimeria spp. and gastrointestinal nematodes in dairy calves in southern Minas Gerais, Brazil. Revista Brasileira de Parasitologia Veterinária, 21(2), 171-175. doi: 10.1590/S1984-296 12012000200019
Daugschies, A., Imarom, S., Ganter, M., & Bollwahn, W. (2004). Prevalence of Eimeria spp. in sows at piglet producing farms in Germany. Journal of Veterinary Medicine, 51(3), 135-139. doi: 10.1111/j. 1439-0450.2004.00734.x
Florião, M. M., Lopes, B. B., Berto, B. P., & Lopes, C. W. G. (2016). New approaches for morphological diagnosis of bovine Eimeria species: a study on a subtropical organic dairy farm in Brazil. Tropical Animal Health and Production, 48(3), 577-584. doi: 10.1007/s11250-016-0998-5
Hillesheim, L. O., & Freitas, F. L. C. (2016). Ocorrência de eimeriose em bezerros criados em propriedades de agricultura familiar - nota científica. Ciência Animal Brasileira, 17(3), 472-481. doi: 10.1590/1089-6891v17i333327
Jonsson, N. N., Piper, E. K., Gray, C. P., Deniz, A., & Constantinoiu, C. C. (2011). Efficacy of toltrazuril 5% suspension against Eimeria bovis and Eimeria zuernii in calves and observations on the associated immunopathology. Parasitology Research, 109(1), 113-128. doi: 10.1007/s00436-011-2408-2
Kawahara, F., Zhang, G., Mingala, C. N., Tamura, Y., Koiwa, M., Onuma, M., & Nunoya, T. (2010). Genetic analysis and development of species-specific PCR assays based on ITS1 region of rRNA in bovine Eimeria parasites. Veterinary Parasitology, 174(1-2), 49-57. doi: 10.1016/j.vetpar.2010.08.001
Khademvatan, S., Neisi, N., Maraghi, S., & Saki, J. (2011). Diagnosis and identification of Leishmania spp. from Giemsa-stained slides, by real-time PCR and melting curve analysis in south-west of Iran. Annals of Tropics Medicine & Parasitology, 105(8), 559-565. doi: 10.1179/2047773211Y.0000000014
Kokozawa, T., Ichikawa-Seki, M., & Itagaki, T. (2013). Determination of phylogenetic relationships among Eimeria species, which parasitize cattle, on the basis of nuclear 18S rDNA sequence. The Journal of Veterinary Medicine Science, 75(11), 1427-1431. doi: 10.1292/jvms.13-0273
Koutny, H., Joachim, A., Tichy, A., & Baumgartner, W. (2012). Bovine Eimeria species in Austria. Parasitology Research, 110(5), 1893-1901. doi: 10.1007/s00436-011-2715-7
Lentze, T., Hofer, D., Gottstein, B., Gaillard, C., & Busato, A. (1999). Prevalence and importance of endoparasites in calves raised in Swiss cow-calf farms (Article in German.) Deutsche Tierärztliche Wochenschrift, 106(7), 275-281.
Marshall, R. N., Catchpole, J., Green, J. A., & Webster, K. A. (1998). Bovine coccidiosis in calves following turnout. Veterinary Record, 143(13), 366-367. doi: 10.1136/vr.143.13.366
Matjila, P. T., & Penzhorn, B. L. (2002). Occurrence and diversity of bovine coccidia at three localities in South Africa. Veterinary Parasitology, 104(2), 93-102. doi: 10.1016/s0304-4017(01)00605-7
Nicolas, L., Milon, G., & Prina, E. (2002). Rapid differentiation of Old World Leishmania species by LightCycle polymerase chain reaction and melting curve analysis. Journal of Microbiological Methods, 51(3), 295-299. doi: 10.1016/s0167-7012(02)00099-4
Ogedengbe, J. D., Hanner, R. H., & Barta, J. R. (2011). DNA barcoding identifies Eimeria species and contributes to the phylogenetics of coccidian parasites (Eimeriorina, Apicomplexa, Alveolata). International Journal for Parasitology, 41(8), 843-850. doi: 10.1016/j.ijpara.2011.03.007
Rebouças, M. M., Grasso, L. M. P. S., Spósito, E. Fa, Amaral, V., Santos, S. M., & Silva, D. M. (1994). Prevalência e distribuição de protozoários do gênero Eimeria (apicomplexa: Eimeriidae) em bovinos nos municípios de Altinópolis, Taquaritinga, São Carlos, Guaíra - Estado de São Paulo, Brasil. Revista Brasileira de Parasitologia Veterinária, 3(2), 125-130.
Rehman, T. U., Khan, M. N., Sajid, M. S., Abbas, R. Z., Arshad, M., Iqbal, Z., & Iqbal, A. (2011). Epidemiology of Eimeria and associated risk factors in cattle of district Toba Tek Singh, Pakistan. Parasitology Research, 108(5), 1171-1177. doi: 10.1007/s00436-010-2159-5
Sambrook, J., Fritshc, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual (2a ed.). New York: Cold Spring Harbor Laboratory Press.
Stewart, I. D., Smith, R. P., & Ellis-Iversen, J. (2008). Eimeria species in cattle on farms in England and Wales. Veterinary Record, 162(15), 482-483. doi: 10.1136/vr.162.15.482
Svensson, C. (2000). Excretion of Eimeria alabamensis oocysts in grazing calves and young stock. Journal of Veterinary Medicine, 47(2), 105-110. doi: 10.1046/j.1439-0450.2000.00324.x
Svensson, C., Hooshmand-Rad, P., Pehrson, B., Törnquist, M., & Uggla, A. (1993). Excretion of Eimeria oocysts in calves during their first three weeks after turn-out to pasture. Acta Veterinaria Scandinavica, 34(2), 175-182.
Svensson, C., Uggla, A., & Pehrson, B. (1994). Eimeria alabamensis infection as a cause of diarrhoea in calves at pasture. Veterinary Parasitology, 53(1-2), 33-43. doi: 10.1016/0304-4017(94)90014-0
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2020 Semina: Ciências Agrárias

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Semina: Ciências Agrárias adopts the CC-BY-NC license for its publications, the copyright being held by the author, in cases of republication we recommend that authors indicate first publication in this journal.
This license allows you to copy and redistribute the material in any medium or format, remix, transform and develop the material, as long as it is not for commercial purposes. And due credit must be given to the creator.
The opinions expressed by the authors of the articles are their sole responsibility.
The magazine reserves the right to make normative, orthographic and grammatical changes to the originals in order to maintain the cultured standard of the language and the credibility of the vehicle. However, it will respect the writing style of the authors. Changes, corrections or suggestions of a conceptual nature will be sent to the authors when necessary.