Parasitology Research

, Volume 107, Issue 4, pp 1013–1017 | Cite as

Molecular prevalence and preponderance of Eimeria spp. among chickens in Tamil Nadu, India

  • S. Aarthi
  • Gopal Dhinakar RajEmail author
  • M. Raman
  • S. Gomathinayagam
  • K. Kumanan
Short Communication


Coccidosis is one of the most commonly prevalent and economically important parasitic diseases of poultry worldwide. Chicken coccidia are protozoan parasites of the genus Eimeria. This study aimed at analysing the molecular prevalence of seven species of Eimeria infecting chickens in Tamil Nadu, India. Tissue samples (caecum, rectum and upper and mid intestines) collected from chickens exhibiting symptoms of coccidiosis were used for DNA extraction, followed by amplification of the internal transcribed spacer (ITS) region of Eimeria genome with genus-specific primers and speciation in nested polymerase chain reaction (PCR) with species-specific primers. Of 43 tissue samples examined, 25 were positive in ITS PCR and all the seven species could be identified. However, the prevalence of each species varied. In broilers, Eimeria necatrix was present in all infected chickens with Eimeria brunetti, Eimeria tenella, Eimeria maxima and Eimeria acervulina present in more than 50% of infected chickens, while Eimeria praecox and Eimeria mitis were only present in 11% to 16%. Although only 7 samples were positive among layers, the prevalence was largely similar, but with a higher prevalence of E. praecox and E. mitis and a lower prevalence of E. tenella. Multiple infections were most common, with 2–6 Eimeria species infecting the same chickens. In order to estimate the preponderance of each infecting species of Eimeria, a random cloning technique was adopted. The genus-specific ITS PCR product was cloned in a TA vector and ten clones were randomly picked and used as template for amplification of all the seven genera of Eimeria. If the specific species of Eimeria is preponderant, then the frequency of the clones showing that species-specific PCR amplification would be higher. Using this method, the most preponderant species present in the rectum, mid and upper intestines of layers was assessed to be E. acervulina, E. brunetti and E. necatrix. E. acervulina was present in 60–90%, E. necatrix in 10–30% and E. brunetti in 10–20% of the clones screened, indicating that these species could be the most preponderant Eimeria species. Intervention strategies should aim at these species. This new method of estimating preponderance of infecting Eimeria species could be used to assess the relative importance of each species at the farm or region level instead of relying only on prevalence estimates.


Internal Transcribe Spacer Coccidiosis Polymerase Chain Reaction Inhibitor Eimeria Species Oocyst Wall 
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.



Internal transcribed spacer


Polymerase chain reaction



The authors thank Dr. Damer Blake, Chandra Subramaniam and Fiona Tomley of the Institute for Animal Health, Compton, UK for their help and suggestions. Part of the work was funded by the BBSRC, UK under the IndiaPA. The authors also thank the Tamil Nadu Veterinary and Animal Sciences University for all the facilities provided and the Avian Disease Laboratory, Namakkal for the samples.


  1. Allen PC, Fetterer RH (2002) Recent advances in biology and immunobiology of Eimeria species and in diagnosis and control of infection with these coccidian parasites of poultry. Clin Microbiol Rev 15:58–65CrossRefPubMedGoogle Scholar
  2. Barta JR, Martin SD, Liberator PA, Dashkevicz M, Anderson JW, Feighner SD, Elbrecht A, Perkins-Barrow A, Jenkins MC, Danforth HD, Ruff MD, Profous-juchelka H (1997) Phylogenetic relationships among eight Eimeria species infecting domestic fowl inferred using complete small subunit ribosomal DNA sequences. J Parasitol 83:262–271CrossRefPubMedGoogle Scholar
  3. Blake DP, Hesketh P, Archer A, Shirley MW, Smith AL (2006) Eimeria maxima: the influence of host genotype on parasite reproduction as revealed by quantitative real-time PCR. Int J Parasitol 36:97–105CrossRefPubMedGoogle Scholar
  4. Blake DP, Qin Z, Cai J, Smith AL (2008) Development and validation of real-time polymerase chain reaction assays specific to four species of Eimeria. Avian Pathol 37:89–94CrossRefPubMedGoogle Scholar
  5. Fernandez S, Costa C, Katsuyama M, Madeira AMBN, Gruber A (2003) A survey of the inter- and intra specific RAPD markers of Eimeria spp. of the domestic fowl and the development of reliable diagnostic tools. Parasitol Res 89:437–445PubMedGoogle Scholar
  6. Fernandez S, Katsuyama AM, Kashiwabara A, Madeira AMBN, Durham AM, Gruber A (2004) Characterization of SCAR markers of Eimeria spp. of domestic fowl and construction of a public relational database (The Eimeria SCARdb). FEMS Microbiol Lett 238:183–188PubMedGoogle Scholar
  7. Gasser RB, Woods WG, Wood JM, Ashdown L, Richards G, Whithear KG (2001) Automated, fluorescence-based approach for the specific diagnosis of chicken coccidiosis. Electrophoresis 22:3546–3550CrossRefPubMedGoogle Scholar
  8. Goldsmith M, Kiss C, Bradbury ARM, Tawfik DS (2007) Avoiding and controlling double transformation artifacts. Protein Eng Des Sel 20:315–318CrossRefPubMedGoogle Scholar
  9. Haug A, Thebo P, Mattson JG (2007) A simplified protocol for molecular identification of Eimeria species in field samples. Vet Parasitol 146:35–45CrossRefPubMedGoogle Scholar
  10. Karim MJ, Begum N (1995) Morphological and biological characterization of chicken Eimeria with special reference to species identification. Vet Rev 9–10Google Scholar
  11. Kawahara F, Taira K, Nagai S, Onaga H, Onuma M, Nunoya T (2008) Detection of five avian Eimeria species by species-specific real-time polymerase chain reaction assay. Avian Dis 52:652–656CrossRefPubMedGoogle Scholar
  12. Lew AE, Anderson GR, Minchin CM, Jeston PJ, Jorgensen WK (2003) Inter- and intra-strain variation and PCR detection of the internal transcribed spacer (ITS1) sequences of Australian isolates of Eimeria species from chickens. Vet Parasitol 112:33–50CrossRefPubMedGoogle Scholar
  13. Long PL, Joyner LP (1984) Problems in the identification of species of Eimeria. J Protozool 31:535–541PubMedGoogle Scholar
  14. Panda DN, Mishra A, Mishra SC, Panda BK (1997) Incidence of coccidiosis in broiler birds in and around Bhubaneswar, Orissa. Indian Vet J 74:430–431Google Scholar
  15. Rana N, Tikaram SM (1999) Comparative evaluation of some epidemiological parameters in coccidiosis in broiler chickens. Indian Vet J 76:382–384Google Scholar
  16. Saira Banu S, Raman M, Gomathinayagam S, Dhinakar Raj G (2009) Preponderance of Eimeria Species in commercial poultry in Tamil Nadu. Indian J Anim Sci 79:620–622Google Scholar
  17. Schnitzler BE, Thebo PL, Mattsson JG, Tomley FM, Shirley MW (1998) Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic Eimeria species of the chicken. Avian Pathol 27:490–497CrossRefPubMedGoogle Scholar
  18. Tsuji N, Kawazu S, Ohta M, Kamio T, Isobe T, Shimura K, Fujisaki K (1997) Discrimination of eight chicken Eimeria species using the two-step polymerase chain reaction. J Parasitol 83:966–970CrossRefPubMedGoogle Scholar
  19. Su Y-C, Fei AC-Y, Tsai FM (2003) Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1 (ITS-1) sequence. J Vet Parasitol 117:221–227CrossRefGoogle Scholar
  20. Zhao X, Duszynski DW, Loker ES (2001) A simplified method of DNA extraction for Eimeria species. J Microbiol Meth 44:131–137CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • S. Aarthi
    • 1
  • Gopal Dhinakar Raj
    • 1
    Email author
  • M. Raman
    • 2
  • S. Gomathinayagam
    • 2
  • K. Kumanan
    • 1
  1. 1.Department of Animal BiotechnologyMadras Veterinary College, Tamil Nadu Veterinary and Animal Sciences UniversityChennaiIndia
  2. 2.Department of Veterinary ParasitologyMadras Veterinary College, Tamil Nadu Veterinary and Animal Sciences UniversityChennaiIndia

Personalised recommendations