Main Article Content

Muhammad Arif Khan
Muhammad Ali Abdullah Shah
Aayesha Riaz
Muhammad Shoaib
Abdul Asim Farooq
Muhammad Kamran
Zahid Manzoor


Eimeria, Poultry, Coccidiosis, Polymerase Chain Reaction, Molecular characterization, Phylogeny


The purpose of this study was to isolate the gene of Eimeria tenella species for molecular characterization and analysis. In this study, the evolutionary relationship of microneme gene2 of Eimeria tenella was confirmed by isolation, molecular characterization and phylogenetic analysis. The birds reared for this purpose were infected with oocyst of Eimeria tenella. Samples were collected from caecal lesions that appeared at the postmortem of birds. Oocysts were isolated using different egg floatation techniques. Analysis of EtMic gene 2 was performed by polymerase chain reaction (PCR). All the samples collected from the caecal lesion showed positive results for EtMic2 gene. The samples showing the best result for the EtMic2 gene were sent for sequencing at Macrogen ® Laboratory Korea. It was revealed from the phylogenetic analysis that its similarity index was 97.9% to all worldwide related sequences of the EtMic2 gene. It was revealed from the analysis that isolated gene of EtMic 2 will provide better opportunities for the development of DNA vaccine against different strains of Eimeria tenella in Pakistan.

Abstract 65 | Pdf Downloads 9


1. Abbas, G., Khan, S. H., Hassan, M., Mahmood, S., Naz, S., & Gilani, S. S. 2015. Incidence of poultry diseases in different seasons in Khushab district, Pakistan. Journal of Advanced Veterinary and Animal Research, 2(2): 141-145.
2. Alexander, D. J., Campbell, G., Manvell, R. J., Collins, M. S., Parsons, G., & McNulty, M. S. (1992). Characterisation of an antigenically unusual virus responsible for two outbreaks of Newcastle disease in the Republic of Ireland in 1990. The Veterinary record, 130(4), 65-68. doi:10.1136/vr.130.4.65
3. Ali, H., Naqvi, F., & Tariq, N. (2014). Prevalence of Coccidiosis and its Association with Risk Factors in Poultry of Quetta, Pakistan. Asian Journal of Applied Sciences (ISSN: 2321–0893), 2(04).
4. Miska, K. B. (2005). Cross protection studies with Eimeria maxima strains. Parasitology research, 97(3), 179-185.
5. Bachaya, H. A., Raza, M. A., Khan, M. N., Iqbal, Z., Abbas, R. Z., Murtaza, S., & Badar, N. (2012). Predominance and detection of different Eimeria species causing coccidiosis in layer chickens. J Anim Plant Sci, 22(3), 597-600.
6. Bansal A, Singh S, More KR, Hans D., & Nangalia, K. (2013) Characterization of Plasmodium falciparum calcium-dependent protein kinase 1 (PfCDPK1) and its role in microneme secretion during erythrocyte invasion. Journal of Biological Chemistry 288:1590–1602.
7. Blake, D. P. (2015). Eimeria genomics: where are we now and where are we going?. Veterinary parasitology, 212(1-2), 68-74.
8. Carmina Nogareda, Jose A. Moreno, Eduardo Angulo, Gerhard Sandmann, Manuel Portero, Teresa Capell. (2015). Carotenoid-enriched transgenic corn delivers bioavailable carotenoids to poultry and protects them against coccidiosis. Plant Biotechnology Journal, 14(1), 15-18.
9. Chapman, H, D., Cherry., T. E., Danforth, H. D., Richards, G., & Shirley, M. W.. (2002). Sustainable coccidiosis control in poultry production: the role of live vaccines. Int J Parasito, 32(5), 617–629.
10. Clark, J. D., Oakes, R. D., Redhead, K., Crouch, C. F., Francis, M. J., Tomley, F. M., & Blake, D. P. (2012). Eimeria species parasites as novel vaccine delivery vectors: anti-Campylobacter jejuni protective immunity induced by Eimeria tenella-delivered CjaA. Vac, 30(16), 2683-2688.
11. Conway, D. P., McKenzie, M. E., & Dayton, A. D. (1990). Relationship of coccidial lesion scores and weight gain in infections of Eimeria acervulina, E. maxima and E. tenella in broilers. Avian Pathology, 19(3), 489-49.
12. Carruthers, V. B., Tomley, F.M., (2008). Microneme proteins in apicomplexans. In: Molecular Mechanisms of Parasite Invasion. Springer, pp. 33–45.
13. Dexing, M., Ma, M. C., Pan, L., Li, G., Hong, H., Cai J., & Ren. X. (2010). Vaccination of chickens with DNA vaccine encoding Eimeria acervulina 3-1E and chicken IL-15 offers protection against homologous challenge. Exp Parasitol, 127, 208-214.
14. Farooq, M., Saliha, U., Munir, M., & Khan, Q. M. (2014). Biological and genotypic characterization of the Newcastle disease virus isolated from disease outbreaks in commercial poultry farms in northern Punjab, Pakistan. Virology Reports, 3-4, 30-39. doi:
15. Huang, J., Zhang, Z., Li, M., Song, X., Ruofeng, Y., Xu, L., & Li X. (2015). Eimeria maxima microneme protein 2 delivered as DNA vaccine and recombinant protein induces immunity against experimental homogenous challenge. Parasitol Int, 64, 408-416.
16. Jamil, A. M., Mansoor, M., Khan, M. K., & Mansoor, M. (2016). Prevalence of coccidiosis in broiler chickens in District Dera Ismail Khan , Pakistan, 3(3), 41–45.
17. Kumar, S., Garg, R., Moftah, A., Clark, E. L., Macdonald, S. E., Chaudhry, A. S., & Blake, D. P. (2014). An optimised protocol for molecular identification of Eimeria from chickens. Veterinary parasitology, 199(1-2), 24-31.
18. Lai, L., Bumstead, J., Liu, Y., Garnett, J., Campanero,Rhodes, M.A., Blake, D.P. & Matthews, S. (2011). The role of sialyl glycan recognition in host tissue tropism of the avian parasite Eimeria tenella. PLoS Pathogens, 7, e1002296.
19. Lillehoj, H. S., Park, D. W., & Lee, S. H. (2009). Protective effect of hyperimmune egg yolk IgY antibodies against Eimeria tenella and Eimeria maxima infections. Vet Parasitol, 163(1–2), 123-126.
20. Mehran, M. H., Ahad, O., Mohammad, N., Fariborz, A., & Omid, N. (2011). Prevalence of Eimeria species in scavenging native chickens of Shiraz, Iran. African Journal of Microbiology Research, 5(20), 3296–3299.
21. Munir, M., Cortey, M., Abbas, M., Qureshi, Z. U., Afzal, F., Shabbir, M. Z., Berg, M. (2012). Biological characterization and phylogenetic analysis of a novel genetic group of Newcastle disease virus isolated from outbreaks in commercial poultry and from backyard poultry flocks in Pakistan. Infect Genet Evol, 12(5), 1010-1019. doi:10.1016/j.meegid.2012.02.015.
22. Pedersen, J. C., Senne, D. A., Woolcock, P. R., Kinde, H., King, D. J., Wise, M. G., & Seal, B. S. (2004). Phylogenetic relationships among virulent Newcastle disease virus isolates from the 2002-2003 outbreak in California and other recent outbreaks in North America. Journal of Clinical Microbiology, 42(5), 2329-2334.
23. Saouros S, Dou Z, Henry M, Marchant J, & Carruthers VB. (2012) Microneme protein 5 regulates the activity of Toxoplasma subtilisin 1 by mimicking a subtilisin prodomain. Journal of Biological Chemistry, 287:36029–36040.
24. Shah Muhammad Ali A., Xu Lixin, Yan R., & Song, X. (2014). The DNA induced protective immunity with chicken interferon gamma against poultry coccidiosis. Parasitology Research, (107), 747–750.
25. Shah, M. A. A. (2013). DNA vaccines as sustainable coccidiosis control strategies in chickens. Science Letter, 1-4.
26. Song, X. K., Xu, L., Yan, R., Huang, R., Shah M. A. A., & Li. X. R. (2009). The optimal immunization procedure of DNA vaccine pcDNA-TA4-IL-2 of Eimeria tenella and its cross immunity to Eimeria necatrix and Eimeria acervulina. Veterinary Parasitology, 159, 30-36.
27. Song, X., Xiaofang, Z., Lixin X., Ruofeng, Y., & Xiangrui L. (2017). Immune protection duration and efficacy stability of DNA vaccine encoding Eimeria tenella TA4 and chicken IL-2 against coccidiosis. Veterinary Science, 111, 31–35.
28. Wang, M., Wu, S. Q., Liu, Q., Zhu, Y.J., Suo, X. & Jiang, J. S. (2004) Construction of DNA vaccines and their induced protective immunity against experimental Eimeria tenella infection. Parasitology Research, 94, 332-336.
29. Zaman, M. A., Iqbal, Z., Abbas, R. Z., & Khan, M. N. (2012). Anticoccidial activity of herbal complex in broiler chickens challenged with Eimeria tenella.Parasitology, 139(2), 237-43.
30. Zang, Z., Liu, L., Huang, J., , S., Lu, M., Song, X., Xu, L., Yan, R. & Li X. (2016). The The molecular characterization and immune protection of microneme 2 of Eimeria acervulina. Veterinary Parasitology, 215, 96-105.

Most read articles by the same author(s)

1 2 > >>