Skip to main content
SpringerLink
Log in

Determination of ribosomal internal transcribed spacer 2 (ITS2) interspecific markers in Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi (Trematoda), parasites of wild and domestic ruminants

  • Published:
Helminthologia

Summary

The species-specific ribosomal internal transcribed spacer 2 (ITS2) markers were designed for PCR-based molecular differentiation of Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi, liver and stomach flukes of domestic and free living ruminants. Complete ITS2 sequences were obtained for D. dendriticum and P. cervi, for the later species, ITS2 structure was determined for the first time. Intraspecific variation within geographically distant populations was found to be either very low (F. hepatica; D. dendriticum) or even absent (F. magna; P. cervi). ITS2 regions with the absence of intraspecific polymorphisms but with interspecific sequence heterogeneity were applied for design of speciesspecific primers. The specificity of developed primers was tested on genomic DNA isolated from adult individuals of studied fluke species. Application of the primers is of particular value for molecular differentiation of morphologically hardly distinguishable F. hepatica, F. magna and P. cervi eggs after coprological examinations.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Alasaad, S., Huang, C. Q., Li, Q. Y., Granados, J. E., Garcia-Romero, C., Perez, J. M. Zhu, X. Q. (2007): Characterization of Fasciola samples from different host species and geographical localities in Spain by sequences of internal transcribed spacers of rDNA. Parasitol. Res., 101(5): 1245–1250. DOI: 10.1007/s00436-007-0628-2

    Article  CAS  PubMed  Google Scholar 

  • Ali, H., Ai, L., Song, H. Q., Ali, S., Lin, R. Q., Seyni, B., Issa, G., Zhu, X. Q. (2008): Genetic characterisation of Fasciola samples from different host species and geographical localities revealed the existence of F. hepatica and F. gigantica in Niger. Parasitol. Res., 102(5): 1021–1024. DOI: 10.1007/s00436-007-0870-7

    Article  CAS  PubMed  Google Scholar 

  • Anuracpreeda, P., Wanichanon, C., Sobhon, P. (2008): Paramphistomum cervi: Antigenic profile of adults as recognized by infected cattle sera. Exp. Parasitol., 118(2): 203–207. DOI:10.1016/j.exppara.2007.08.005

    Article  CAS  PubMed  Google Scholar 

  • Bildfell, R. J., Whipps, C. M., Gillin, C. M., Kent, M. L. (2007): DNA-based identification of hepatic trematode in an elk calf. J. Wildlife Dis., 43(4): 762–769

    CAS  Google Scholar 

  • Cunningham, C. O. (1997): Species variation within the internal transcribed spacer (ITS) region of Gyrodactylus (Monogenea: Gyrodactylidae) ribosomal RNA genes. J. Parasitol., 83(2): 215–219

    Article  CAS  PubMed  Google Scholar 

  • Dorchies, P. (2006): Flukes: Old parasites but new emergence. In Manuscript of 24th World Buiatrics Congress, Nice, France, 15–19 October, 2006. Ecole Nationale Vétérinaire, Toulouse, France, 2006

    Google Scholar 

  • Erensoy, A., Kuk, S. Ozden, M. (2009): Genetic identification of Fasciola hepatica by ITS-2 sequence of nuclear ribosomal DNA in Turkey. Parasitol. Res., 105(2): 407–412. DOI: 10. 1007/s00436-009-1399-8

    Article  PubMed  Google Scholar 

  • Erhardová-Kotrlá, B. (1971): The occurence of Fascioloides magna (Bassi, 1875) in Czechoslovakia. Praha, Czech Republic, Academia, 155 pp.

    Google Scholar 

  • Farjallah, S., Sanna, D., Amor, N., Ben Mehel, B., Piras, M. C., Merella, P., Casu, M., Curini-Galletti, M., Said, K., Garippa, G. (2009): Genetic characterization of Fasciola hepatica from Tunisia and Algeria based on mitochondrial and nuclear DNA sequences. Parasitol. Res., 105(6): 1617–1621. DOI 10.1007/s00436-009- 1601-z

    Article  PubMed  Google Scholar 

  • Hillis, D. M., Dixon, M. T. (1991): Ribosomal RNA: molecular evolution and phylogenetic inference. Q. Rev. Biol., 66(4): 411–453

    Article  CAS  PubMed  Google Scholar 

  • Huang, W. Y., He, B., Wang, C. R., Zhu, X. Q. (2004): Characterisation of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence. Vet. Parasitol., 120(1–2): 75–83. DOI: 10.1016/j.vetpar.2003.12.006

    Article  CAS  PubMed  Google Scholar 

  • Itagaki, T., Tsutsumi, K. (1998): Triploid form of Fasciola in Japan: Genetic relationships between Fasciola hepatica and Fasciola gigantica determined by ITS-2 sequence of nuclear rDNA. Int. J. Parasitol., 28(5): 777–781. DOI: 10.1016/S0020-7519(98)00037-X

    Article  CAS  PubMed  Google Scholar 

  • Itagaki, T., Kikawa, M., Sakaguchi, K., Shimo, J., Terasaki, K., Shibahara, T., Fukuda, K. (2005): Genetic characterization of parthenogenetic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. Parasitology, 131(5): 679–685. DOI: 10.1017/S0031182005008292

    Article  CAS  PubMed  Google Scholar 

  • Králová-Hromadová, I., Scholz, T., Shinn, A. P., Cunningham, C. O., Wootten, R., Hanzelová, V., Sommerville, C. (2003): A molecular study of Eubothrium rugosum (Batsch, 1786) (Cestoda: Pseudophyllidea) using ITS rDNA sequences, with notes on the distribution and intraspecific sequence variation of Eubothrium crassum (Bloch, 1779). Parasitol. Res., 89(6): 473–479. DOI: 10.1007/s00436-002-0779-0

    PubMed  Google Scholar 

  • Králová-Hromadová, I., Špakulová, M., Horáčková, E., Turčeková, L., Novobilský, A., Beck, R., Koudela, B., Marinculić, A., Rajský, D., Pybus, M. (2008): Sequence analysis of ribosomal and mitochondrial genes of the giant liver fluke Fascioloides magna (Trematoda: Fasciolidae): intraspecific variation and differentiation from Fasciola hepatica. J. Parasitol., 94(1): 58–67. DOI: 10.1645/GE-1324.1

    Article  PubMed  Google Scholar 

  • Lang, B.Z. (1977): Snail and mammalian hosts for Fasciola hepatica in eastern Washington. J. Parasitol., 63(5): 938–939

    Article  CAS  PubMed  Google Scholar 

  • Le, T. H., De, N. V., Agatsuma, T., Thi Nguyen, T. G., Nguyen, Q. D., Mcmanus, D. P., Blair, D. (2008): Human fascioliasis and the presence of hybrid/introgressed forms of Fasciola hepatica and Fasciola gigantica in Vietnam. Int. J. Parasitol., 38(6): 725–730. DOI: 10.1016/j.ijpara.2007.10.003

    Article  PubMed  Google Scholar 

  • Lotfy, W. M., Brant, S. V., Dejong, R. J., Le, T. H., Demiaszkiewicz, A., Rajapakse, R. P., Perera, V. B., Laursen, J. R., Loker, E. S. (2008): Evolutionary origins, diversification and biogeography of liver flukes (Digenea, Fasciolidae). Am. J. Trop. Med. Hyg., 79(2): 248–255

    CAS  PubMed  Google Scholar 

  • Mas-Coma, S., Funatsu, I. R., Bargues, M. D. (2001): Fasciola hepatica and lymnaeid snails occurring at very high altitude in South America. Parasitology, 123: 115–127

    Article  Google Scholar 

  • Mas-Coma, S., Bargues, M. D., Valero, M. A. (2005): Fascioliasis and other plant-borne trematode zoonoses. Int. J. Parasitol., 35(11–12): 1255–1278. DOI:10.1016/j.ijpara.2005.07.010

    Article  CAS  PubMed  Google Scholar 

  • Maurelli, M. P., Rinaldi, L., Capuano, F., Perugini, A. G., Veneziano, V., Cringoli, G. (2007): Characterization of the 28S and the second internal transcribed spacer of ribosomal DNA of Dicrocoelium dendriticum and Dicrocoelium hospes. Parasitol. Res., 101(5): 1251–1255. DOI: 10.1007/s00436-007-0629-1

    Article  CAS  PubMed  Google Scholar 

  • Minchella, D. J., Sorensen, R. E., Curtis, J., Bieberich, A. (1997): Molecular biology of trematodes: Approaches and Applications. In: Fried, B., Graczyk, T. K. (Eds), Advances in Trematode Biology, CRC Press, pp. 405–446

  • Nolan, J. M., Cribb, T. H. (2005): The use and implications of ribosomal DNA sequencing for the discrimination of digenean species. Adv. Parasitol., 60: 101–163. DOI: 10.1016/S0065-308X(05)60002-4

    Article  PubMed  Google Scholar 

  • Oberhauserová, K., Bazsalovicsová, E., Králová-Hromadová, I., Major, P., Reblánová, M. (2010): Molecular discrimination of eggs of cervid trematodes using the Teflon (PTFE) technique for eggshell disruption. Helminthologia, in press

  • Otranto, D., Traversa, D. (2003): Dicrocieliosis of ruminants: a little known fluke disease. Trends in Parasitol., 19(1): 12–15. DOI: 10.1016/S1471-4922(02)00009-0

    Article  Google Scholar 

  • Pritchard, G. C., Forbes, A. B., Williams, D. J., Salimi-Bejestani, M. R., Daniel, R. G. (2005): Emergence of fasciolosis in cattle in East Anglia. Vet. Rec., 157(19): 578–582

    CAS  PubMed  Google Scholar 

  • Pybus, M. J. (2001): Liver flukes. In: Samuel, W. M., Pybus, M. J., Kocan, A. A. (Eds) Parasitic diseases in wild mammals. Iowa State Press, Iowa City, pp. 121–149

    Chapter  Google Scholar 

  • Rolfe, P. F., Boray, J. C., Nichols, P., Collins, G. H. (1991): Epidemiology of paramphistomosis in cattle. Aust. Vet. J., 21(7): 813–819. DOI: 10.1016/0020-7519(91)90150-6

    CAS  Google Scholar 

  • Sanabria, R. E. F., Romero, J. R. (2008): Review and update of paramphistomosis. Helminthologia, 45(2): 64–68. DOI: 10.2478/s11687-008-0012-5

    Article  Google Scholar 

  • Sanabria, R. E. F., Martorelli, S. R., Romero, J. R. (2009): First report of Paramphistomum leydeni Näsmark, 1937 (Trematoda: Paramphistomidae) in Argentina, and reexamination of Cotylophoron cotylophorum sensu Racioppi et al. (1994). Helminthologia, 46(4): 225–229. DOI: 10.2478/s11687-009-0042-7

    Article  Google Scholar 

  • Špakulová, M., Rajský, D., Sokol, J., Vodňanský, M. (2003): Giant liver fluke (Fascioloides magna), an important parasite of ruminants. Bratislava, SR: PaRPRESS, 77 pp.

    Google Scholar 

  • Thompson, J. D., Higgins, D. G., Gibson, T. J. (1994): CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment throughout sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Res., 22(22): 4673–4680

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Parasitological Institute, Slovak Academy of Sciences, KoŠice, Slovakia

    E. Bazsalovicsová, I. Králová-Hromadová, M. Špakulová & M. Reblánová

  2. Department of Nutrition, Dietetics and Animal Breeding, University of Veterinary Medicine and Pharmacy, KoŠice, Slovakia

    K. Oberhauserová

Authors
  1. E. Bazsalovicsová
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I. Králová-Hromadová
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Špakulová
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M. Reblánová
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. K. Oberhauserová
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to E. Bazsalovicsová.

Additional information

Note: Nucleotide sequence data reported in this paper are available in the GenBank™, EMBL and DDBJ databases under the accession numbers HM026461 and HM026462.

About this article

Cite this article

Bazsalovicsová, E., Králová-Hromadová, I., Špakulová, M. et al. Determination of ribosomal internal transcribed spacer 2 (ITS2) interspecific markers in Fasciola hepatica, Fascioloides magna, Dicrocoelium dendriticum and Paramphistomum cervi (Trematoda), parasites of wild and domestic ruminants. Helminthologia 47, 76–82 (2010). https://doi.org/10.2478/s11687-010-0011-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11687-010-0011-1

Keywords

Search

Navigation