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Eimeria spp. of the domestic fowl: Analysis of genetic variability between species and strains using DNA polymorphisms amplified by arbitrary primers and denaturing gradient-gel electrophoresis


The genetic relatedness of 5Eimeria spp. of the domestic fowl, including 11 strains ofE. acervulina, 2 strains ofE. tenella and 1 precocious line ofE. acervulina, was assayed by means of random amplified polymorphic DNA (RAPD) and denaturing gradient-gel electrophoresis (DGGE). Seven different oligonucleotides were used to generate similarity coefficients for the species and strains ofEimeria infecting chickens. Between 1 and 13 DNA segments, depending on the species/strain-primer combination, were amplified with the various primers. Amplification products ranged in size from 0.16 to 3.8 kb.E. acervulina strains demonstrated two to four major common bands unique to the species. These strains also exhibited major and minor differences in their DNA patterns. Band-match analyses from both polyacrylamide and denaturing gradient gels were used to calculate similarity coefficients for theEimeria spp. and strains tested. Species differences, readily detected upon examination of DNA banding patterns, gave similarity coefficients of 4%–38% and 3%–18% when analyzed by polyacrylamide-and denaturing gradient-gel electrophoresis, respectively. A similar analysis ofE. acervulina strains yielded similarity coefficients of 55%–95% and 51%–85%, respectively. The differences observed between both species and strains were greater when the RAPD-assay products were analyzed via DGGE, indicating that a combination of these two techniques may provide a more stringent analysis of the genetic relatedness of these coccidia.

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  1. Barral V, This P, Imbert-Establer D, Combes C, Delseny M (1993) Genetic variability and evolution of theSchistosoma genome analysed by using random amplified polymorphic DNA markers. Mol Biochem Parasitol 59:211–222

  2. Chapman HD (1982) The use of enzyme electrophoresis for the identification of the species ofEimeria present in field isolates of coccidia. Parasitology 85:437–442

  3. Fernando MA, Pasternak JJ (1991)Eimeria spp. of the domestic fowl: resolution of chromosomes by field inversion gel electrophoresis. Exp Parasitol 72:306–310

  4. Fischer SG, Lerman LS (1983) DNA fragments differing by single base-pair substitutions are separated in denaturing gradient-gels: correspondence with melting theory. Proc Natl Acad Sci USA 80:1579–1583

  5. Goodwin PH, Annis SL (1991) Rapid identification of genetic variation and pathotype ofLeptosphaeria maculans by random amplified polymorphic DNA assay. Appl Environ Microbiol 57:2482–2486

  6. Gray M, Charpentier A, Walsh K, Wu P, Bender W (1991) Mapping point mutations in the drosophilarosy locus using denaturing gradient-gel blots. Genetics 127:139–149

  7. Jeffers TK (1975) AttenuatedEimeria tenella through selection for precociousness. J Parasitol 61:1083–1090

  8. Karim MJ, Hoare JA, Trees AJ (1991) Limited heterogeneity between strains ofEimeria tenella isolated from Britain and Bangladesh. Res Vet Sci 51:29–33

  9. Kučera J (1991) Enzyme variants ofEimeria parasitizing the domestic fowl and possibilities of species diagnostics. Folia Parasitol (Praha) 38:193–199

  10. Lee E-H, Fernando MA (1977) Characterization of coccidial DNA an aid in species identification. Z Parasitenkd 53:129–131

  11. Lerman LS, Fischer SG, Hurley I, Silverstein K, Lumelsky N (1984) Sequence-determined DNA separations. Annu Rev Biophys Bioeng 13:339–423

  12. Long PL, Joyner LR (1984) Problems in the identification of species ofEimeria. J Protozool 31:535–541

  13. Long PL, Reid WM (1982) A guide to the diagnosis of coccidiosis in chickens. University of Georgia Research Report 404. University of Georgia Press, Athens, Georgia

  14. Long PL, Millard BJ, Joyner LP, Norton CC (1976) A guide to laboratory techniques used in the study and diagnosis of avian coccidiosis. Folia Vet Lat 6:201–217

  15. McDougald LR, Jeffers TK (1976)Eimeria tenella (Sporozoa: Coccidia): gametogony following a single asexual generation. Science 192:258–259

  16. Myers RM, Maniatis T, Lerman LS (1987) Detection and localization of single base changes by denaturing gradient-gel electrophoresis. Methods Enzymol 155:501–527

  17. Nei M, Li WH (1979) Mathematical model for studying genetic variation in terms of restriction endonucleases. Proc Natl Acad Sci USA 76:5269–5273

  18. Procunier JD, Fernando MA, Barta JR (1993) Species and strain differentiation ofEimeria spp. of the domestic fowl using DNA polymorphisms amplified by arbitrary primers. Parasitol Res 79:98–102

  19. Remmler O, McGregor JK (1964) A method to facilitate isolation of single coccidial oocysts. J Parasitol 50:294

  20. Remmler O, McGregor JK (1964) A method to facilitate isolation of single coccidial oocysts. J Parasitol 50:294

  21. Shirley MW (1975) Enzyme variation inEimeria species of the chicken. Parasitology 71:369–376

  22. Shirley MW (1978) Electrophoretic variation of enzymes: a further marker for genetic studies of theEimeria. Z Parasitenkd 57:83–87

  23. Shirley MW, McDonald V, Chapman HD, Millard BJ (1984)Eimeria praecox: selection and characteristics of precocious lines. Avian Pathol 13:669–682

  24. Smith NC, Hunt M, Ellenrieder C, Shirley MW (1994) Detection of metabolic enzymes ofEimeria by ampholinepolyacrylamide gel isoelectric focusing. Parasitol Res 80:165–169

  25. Wang CC, Stotish RL (1975) Changes of nucleic acids and proteins in the oocysts ofEimeria tenella during sporulation. J Protozool 22:438–443

  26. Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res 18:7213–7218

  27. Welsh J, Petersen C, McClelland M (1991) Polymorphisms generated by arbitrarily primed PCR in the mouse: application to strain identification and genetic mapping. Nucleic Acids Res 19:303–306

  28. Williams JG, Kubelik AR, Livak KJ, Rafalski TA, Tingey SV (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535

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Correspondence to M. A. Fernando.

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Johnston, D.A., Fernando, M.A. Eimeria spp. of the domestic fowl: Analysis of genetic variability between species and strains using DNA polymorphisms amplified by arbitrary primers and denaturing gradient-gel electrophoresis. Parasitol Res 81, 91–97 (1995).

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  • Banding Pattern
  • Similarity Coefficient
  • Species Difference
  • Genetic Relatedness
  • Arbitrary Primer