Characterization of Eimeria Species in Commercial Broilers by PCR Based on ITS1 Regions of rDNA
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Abstract
Background: Coccidiosis is an intestinal disease of chickens caused by various species of protozoan parasites within the genus Eimeria. Diagnosis and genetic characterization of different species of Eimeria are central to the prevention, surveillance, and control of coccidiosis. The aim of this study was to detect different chicken Eimeria species from several areas in Khuzestan, southwest Iran.
Methods: From February to September 2008, PCR assay as well as parasitological examinations was applied for the identification of field isolates of Eimeria parasites around Ahvaz, center of Khuzestan, southwest Iran. Data were analyzed by the Kappa statistic test.
Results: Eimeria maxima, E. necatrix, E. tenella, E. acervulina and E. mitis were detected in this study. The prevalence of Eimeria spp. was 31.5% (126 of 400) and E. tenella was the most prevalent species in Khuzestan. Based on the Kappa statistical test, a good correlation between the results of PCR and traditional biometrical methods was only observed for E. maxima.
Conclusion: The present study is the first on the prevalence of Eimeria species in Khuzestan, based on the molecular findings. We believe that traditional methods are not sufficiently reliable for specific diagnosis of Eimeria species in chickens and PCR based amplification of DNA sequence of parasite, could resolve this problem.
McDougald LR, Reid WM. Coccidiosis. In: Calnek BW, Barnes HJ, Beard, CW, McDougald LR., Saif MY, editors. Diseases of poultry. Iowa State University Press, Ames, IA; 1997. p: 865–883.
Shirley MW, Smith AL, Tomley FM. The biology of avian Eimeria with an emphasis on their control by vaccination. Adv Parasitol. 2005; 60: 285-330.
McDougald LR. Protozoal Infections. In: Saif YM, Barnes HJ, Glisson JR, Fadly AM, McDougald LR, Swayne DE, editors. Diseases of poultry. Iowa State University Press; 2003. p: 973-1023.
Long PL, Reid, WM. A guide for the diagnosis of coccidiosis in chickens. The University of Georgia, College of Agriculture Experiment Stations, Research report 1982; 404, p: 1–17.
Long PL, Joyner LP. Problems in the identification of species of Eimeria. J Protozool. 1984; 31(4): 535-41.
Meireles MV, Roberto LO, Riera RF. Identification of Eimeria mitis and Eimeria praecox in Broiler Feces Using Polymerase Chain Reaction. Brazilian J Poult Sci. 2004; 6(4): 249–252.
Morgan JA, Morris GM, Wlodek BM, Byrnes R, Jenner M, Constantinoiu CC, Anderson GR, Lew-Tabor AE, Molloy JB, Gasser RB, Jorgensen WK. Real-time] polymerase chain reaction (PCR) assays for the specific detection and quantification of seven Eimeria species that cause coccidiosis in chickens. Mol Cell Probes. 2009; 23(2): 83-89.
Schnitzler BE, Thebo P, Mattson JG, Tomley F, Shirley MW. Development of a diagnostic PCR assay for the detection and discrimination of four pathogenic Eimeria species of the chicken. Avian Pathol. 1998; 27(5): 490–497.
Conwa DP, McKenzie ME. Poultry coccidiosis, Diagnostic and testing procedures. 3th ed. Blackwell publishing; 2007; p: 7-11.
Haug A, Thebo P, Mattsson JG. A simplified protocol for molecular identification of Eimeria species in field samples. Vet Parasitol. 2007; 146(1-2): 35–45.
Dohoo I, Martin W, Stryhn H. Veterinary Epidemiologic Research. 1st ed. AVC Inc, Charlottetown, Canada; 2003; p: 85–120.
Stucki U, Braun R, Roditi I. Eimeria tenella: characterization of a 5S ribosomal RNA repeat unit and its use as a species specific probe. Exp Parasitol. 1993; 76(1): 68–75.
Tsuji N, Kawazu S, Ohta M, Kamio T, Isobe T, Shimura K, Fujisaki K. Discrimination of eight chicken Eimeria species using the two-step polymerase chain reaction. J Parasitol. 1997; 83(5): 966–970.
Molloy JB, Eaves FW, Jeston PJ, Minchin CM, Stewart NP, Lew AE, Jorgensen WK. Detection of Eimeria acervulina using the polymerase chain reaction. Avian Dis. 1998; 42(1): 119–123.
Lew AE, Anderson GR., Minchin CM, Jeston PJ, Jorgensen WK. Inter and intrastrain variation and PCR detection of the internal transcribed spacer 1 (ITS-1) sequences of Australian isolates of Eimeria species from chickens. Vet Parasitol. 2003; 112(1-2): 33–50.
Schnitzler BE, Thebo P, Tomley F, Uggla A, Shirley MW. PCR identification of chicken Eimeria: a simplified readout. Avian Pathol. 1999; 28(1): 89–93.
Su YC, Fei AC, Tsai FM. Differential diagnosis of five avian Eimeria species by polymerase chain reaction using primers derived from the internal transcribed spacer 1 (ITS-1) sequence. Vet Parasitol. 2003; 117(3): 221–227.
Gasser RB, Woods WG, Wood JM, Ashdown L, Richards G, Whithear KG. Automated, fluorescence-based approach for the specific diagnosis of chicken coccidiosis. Electrophoresis. 2001; 22(16), 3546–3550.
Lien YY, Sheu SC, Liu HJ, Chen SC, Tsai MY, Luo SC, Wu KC, Liu SS, Su HY. Cloning and nucleotide sequencing of the second internal transcribed spacer of ribosomal DNA for three species of Eimeria from chickens in Taiwan. Vet J. 2007; 173(1): 186–191.
Woods WG, Whithear KG, Richards DG, Anderson GR, Jorgensen WK, Gasser RB. Single-strand restriction fragment length polymorphism analysis of the second internal transcribed spacer (ribosomal DNA) for six species of Eimeria from chickens in Australia. Int J Parasitol. 2000; 30(9): 1019–1023.
Holmdahl OJM, Mattsson JG. Rapid and sensitive identification of Neospora caninum by in vitro amplification of the internal transcribed spacer 1. Parasitology. 1996; 112(2): 177-182.
Nowzari N, Yakhchali B, Rahbari S, Mouazeni Jula GH. Identification of Eimeria spp. isolated from poultry breeder farms in Iran by PCR. J fac Vet Med, Univ Tehran. 2004; 59(2): 125-130.
Razmi GR, Kalideri A. Prevalence of subclinical coccidiosis in broiler-chicken farms in the municipality of Mashhad, Khorasan, Iran. Prevent Vet Med. 2000; 44(3-4): 247-253.
Nematollahi A, Moghaddam GH, Farshbaf Pourabad R. Prevalence of Eimeria species among broiler chicks in Tabriz (Northwest of Iran). Mun Ent Zool. 2009; 4(1): 53-58.
Haug A, Gjevre AG, Thebo P, Mattsson JG, Kaldhusdal M. Coccidial infections in commercial broilers: epidemiological aspects and comparison of Eimeria species identification by morphometric and polymerase chain reaction techniques. Avian Pathol. 2008; 37(2): 161-70.
Nowzari N, Dinparast Djadid N, Rahbari S,Yakchali B, Kazemi B, Moazeni Jula G. Inter and intra-specific genetic variation of avian Eimeria isolated from Iran by random amplified polymorphic DNA- PCR. Vet Parasitol. 2005; 128(1-2): 59–64.
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| Issue | Vol 5 No 4 (2010) | |
| Section | Articles | |
| Keywords | ||
| Eimeria Poultry coccidiosis PCR ITS1 Microscopic examinations Iran | ||
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