Parasitology Research

, Volume 109, Issue 4, pp 1021–1028 | Cite as

A comparative study of karyotypes and chromosomal location of rDNA genes in important liver flukes Fasciola hepatica and Fascioloides magna (Trematoda: Fasciolidae)

  • Marianna Reblánová
  • Marta Špakulová
  • Martina Orosová
  • Ivica Králová-Hromadová
  • Eva Bazsalovicsová
  • Dušan Rajský
Original Paper


Chromosomal characteristics, i.e., number, size, morphology, and location of ribosomal DNA (rDNA) clusters were examined in two medically important liver flukes, Fasciola hepatica and Fascioloides magna (Fasciolidae), using conventional Giemsa staining and fluorescent in situ hybridization (FISH) with ribosomal 18S rDNA probe. A comparison of F. magna and F. hepatica karyotypes confirmed significant differences in all chromosomal features. Whilst the karyotype of F. hepatica comprised ten pairs of chromosomes (one metacentric and nine medium-sized subtelocentrics and submetacentrics; 2n = 20, n = 1 m + 5 sm + 4 st; TCL = 49.9 μm), the complement of F. magna was composed of 11 pairs of medium-sized subtelocentrics and submeta-metacentrics (2n = 22, n = 9 st + 1 sm + 1 sm-m; TCL = 35.2 μm). Noticeable differences were found mainly in length and morphology of first chromosome pair. It was metacentric and 9.0 μm long in F. hepatica while subtelocentric and 4.7 μm long in F. magna. Although FISH with rDNA probe revealed a single cluster of ribosomal genes in both species, conspicuous interspecific differences were displayed by chromosomal location of ribosomal loci (i.e., NORs). The signals were found on short arms of fifth homologous pair in F. hepatica; however, they were detected in pericentromeric regions of the long arms of tenth pair in F. magna. The observed cytogenetic differences were interpreted in terms of karyotype evolution of fasciolid flukes; F. hepatica may be regarded phylogenetically younger than F. magna. The present paper provides a pilot study on molecular cytogenetics within a group of hermaphroditic digenetic flukes.


Chromosome Pair Secondary Constriction Liver Fluke rDNA Signal Metacentric Pair 
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.



We are highly indebted to František Marec (Institute of Entomology, Biology Centre ASCR, České Budějovice, Czech Republic) and Tomáš Scholz (Institute of Parasitology, Biology Centre, České Budějovice, Czech Republic) for kind help and providing facilities during research stays of M. R. and M. O. We wish to thank Katarína Oberhauserová (University of Veterinary Medicine and Pharmacy, Department of Nutrition, Dietetics and Animal Breeding, Košice, Slovakia) for providing us with F. hepatica material. The work was supported by the Slovak Research and Development Agency under contracts LPP-0126-07 and APVV-51-062205, and by the Slovak Grant Agency VEGA (2/0148/09 and 2/0014/10). The publication has been created within realization of the project Centre of Excellence for Parasitology (Code ITMS: 26220120022), based on the support of the Operational Programme “Research and Development” funded from the European Fund of Regional Development (rate 0.3).


  1. Ali H, Ai L, Song HQ, Ali S, Lin RQ, Seyni B, Issa G, Zhu XQ (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 102:1021–1024PubMedCrossRefGoogle Scholar
  2. Bassi R (1875) Sulla cachessia ittero-vermicosa, o marciaia dei Cervi, causata dal Distomum magnum. Med Vet 4:497–515Google Scholar
  3. Bazsalovicsová E, Králová-Hromadová I, Špakulová M, Reblánová M, Oberhauserová K (2010) 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 47:76–82CrossRefGoogle Scholar
  4. Bombarová M, Marec F, Nguyen P, Špakulová M (2007) Divergent location of ribosomal genes in chromosomes of fish thorny-headed worms, Pomphorhynchus laevis and Pomphorhynchus tereticollis (Acanthocephala). Genetica 131:141–149PubMedCrossRefGoogle Scholar
  5. Bombarová M, Vítková M, Špakulová M, Koubková B (2009) Telomere analysis of platyhelminths and acanthocephalans by FISH and Southern hybridization. Genome 52:897–903PubMedCrossRefGoogle Scholar
  6. Fletcher HL, Hoey EM, Orr EM, Trudgett A, Fairweather I, Robinson MV (2004) The occurrence and significance of triploidy in the liver fluke, Fasciola hepatica. Parasitology 128:69–72PubMedCrossRefGoogle Scholar
  7. Foreyt WJ (1996) Susceptibility of bighorn sheep (Ovis canadensis) to experimentally induced Fascioloides magna infections. J Wild Dis 32:556–559Google Scholar
  8. Frydrychová R, Marec F (2002) Repeated losses of TTAGG telomere repeats in evolution of beetles (Coleoptera). Genetica 115:179–187PubMedCrossRefGoogle Scholar
  9. Fuková I, Nguyen P, Marec F (2005) Codling moth cytogenetics: karyotype, chromosomal location of rDNA, and molecular differentiation of sex chromosomes. Genome 48:1083–1092PubMedCrossRefGoogle Scholar
  10. Hirai H, LoVerde PT (1996) Identification of the telomeres on Schistosoma mansoni chromosomes by FISH. J Parasitol 82:511–512PubMedCrossRefGoogle Scholar
  11. Hirai H, Spotila LD, LoVerde PT (1989) Chromosomal localization of DNA repeat elements by in situ hybridization using biotinylated DNA probes. Exp Parasitol 69:175–188PubMedCrossRefGoogle Scholar
  12. Hirai H, Tanaka M, LoVerde PT (1993) Schistosoma mansoni: Chromosomal localization of female-specific genes and female-specific DNA element. Exp Parasitol 76:175–181PubMedCrossRefGoogle Scholar
  13. Hirai H, Taguchi T, Saitoh Y, Kawanaka M, Sugiyama H, Habe S, Okamoto M, Hirata M, Shimada M, Tiu WU, Lai K, Upatham ES, Agatsuma T (2000) Chromosomal differentiation of the Schistosoma japonicum complex. Int J Parasitol 30:441–452PubMedCrossRefGoogle Scholar
  14. Ichikawa M, Itagaki T (2010) Discrimination of the ITS1 types of Fasciola spp. based on a PCR-RFLP method. Parasitol Res 106:757–761PubMedCrossRefGoogle Scholar
  15. Jaros S, Jaros D, Wesolowska A, Zygner W, Wedrychowics H (2010) Blocking Fasciola hepatica´s energy metabolism—pilot study of vaccine potential of a novel gene—phosphoglycerate kinase. Vet Parasitol 172:229–237PubMedCrossRefGoogle Scholar
  16. Jones A (2005) Family Fasciolidae Railliet, 1895. In: Jones A, Bray RA, Gibson DI (eds) Keys to the Trematoda, vol 2, CABI Publishing and The Natural History Museum. London, UK, pp 79–86Google Scholar
  17. 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 94:58–67PubMedCrossRefGoogle Scholar
  18. Králová-Hromadová I, Štefka J, Špakulová M, Orosová M, Bombarová M, Hanzelová V, Bazsalovicsová E, Scholz T (2010) Intra-individual internal transcribed spacer 1 (ITS1) and ITS2 ribosomal sequence variation linked with multiple rDNA loci: a case of triploid Atractolytocestus huronensis, the monozoic cestode of common carp. Int J Parasitol 40:175–181PubMedCrossRefGoogle Scholar
  19. Levan A, Fredga K, Sandberg A (1964) Nomenclature for centromere position on chromosomes. Hereditas 52:201–220CrossRefGoogle Scholar
  20. Li J, He L (1988) Analysis of meiosis and karyotype of Fasciola hepatica. J First Millit Med Univ 1:11. doi: cnki: ISSN: 1000-2588.0.1988-01-020Google Scholar
  21. Li GQ, Jin JS, Wang PY (1988) A study on the chromosomes of Fasciola hepatica. Chin J Veter Sci Technol 6:11–16Google Scholar
  22. Lotfy WM, Brant SV, DeJong RJ, Le TH, Demiaszkiewicz A, Rajapakse RPVJ, Perera VBVP, Laursen JR, Loker ES (2008) Evolutionary origins, diversification and biogeography of liver flukes (Digenea, Fasciolidae). Am J Trop Med Hyg 79:248–255PubMedGoogle Scholar
  23. Mas-Coma S, Bargues MD, Valero MA (2005) Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol 35:1255–1278PubMedCrossRefGoogle Scholar
  24. Mas-Coma S, Valero MA, Bargues MD (2009) Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv Parasitol 69:41–146PubMedCrossRefGoogle Scholar
  25. McConville M, Hanna REB, Brennan GP, McCoy M, Edgar HWJ, McConnell S, Castillo R, Hernández-Campos A, Fairweather I (2010) Fasciola hepatica: disruption of spermatogenesis by the fasciolicide compound alpha. Parasitol Res 106:311–323PubMedCrossRefGoogle Scholar
  26. Nguyen P, Sahara K, Yoshido A, Marec F (2010) Evolutionary dynamics of rDNA clusters on chromosomes of moths and butterflies (Lepidoptera). Genetica 138:343–354PubMedCrossRefGoogle Scholar
  27. Orosová M, Králová-Hromadová I, Bazsalovicsová E, Špakulová M (2010a) Karyotype, chromosomal characteristics of multiple rDNA clusters and intragenomic variability of ribosomal ITS2 in Caryophyllaeides fennica (Cestoda). Parasitol Int 59:351–357PubMedCrossRefGoogle Scholar
  28. Orosová M, Marec F, Oros M, Xi BV, Scholz T (2010b) A chromosome study and localization of 18S rDNA in Khawia saurogobii (Cestoda: Caryophyllidea). Parasitol Res 106:587–593PubMedCrossRefGoogle Scholar
  29. Peng M, Ichinomiya M, Ohtori M, Ichikawa M, Shibahara T, Itagaki T (2009) Molecular characterization of Fasciola hepatica, Fasciola gigantica, and aspermic Fasciola sp. in China based on nuclear and mitochondrial DNA. Parasitol Res 105:809–815PubMedCrossRefGoogle Scholar
  30. Pybus MJ (2001) Liver flukes. In: Samuel WM, Pybus MJ, Kocan AA (eds) Parasitic diseases of wild mammals. Iowa State Press, Ames, pp 121–149CrossRefGoogle Scholar
  31. Rajský D, Čorba J, Várady M, Špakulová M, Cabadaj R (2002) Control of fascioloidosis (Fascioloides magna Bassi, 1875) in red deer and roe deer. Helminthologia 39:67–70Google Scholar
  32. Reblánová M, Špakulová M, Orosová M, Bazsalovicsová E, Rajský D (2010) A description of karyotype of giant liver fluke Fascioloides magna (Trematoda, Platyhelminthes) and a review of Fasciolidae cytogenetics. Helminthologia 47:69–75CrossRefGoogle Scholar
  33. Romanenko LN, Pleshanova NM (1975) Chromosome sets of Fasciola hepatica and Fasciola gigantica. Tr VIGIS (Teoreticheskie problemy veterinarnoy geľmintologii) 22:137–142Google Scholar
  34. Sanderson AR (1953) Maturation and probable gynogenesis in the liver fluke, Fasciola hepatica L. Nature 172(4368):110–112PubMedCrossRefGoogle Scholar
  35. Semyenova SK, Morozova EV, Chrisanfova GG, Gorokhov VV, Arkhipov IA, Moskvin AS, Movsessyan SO, Ryskov AP (2006) Genetic differentiation in eastern European and western Asian populations of the liver fluke, Fasciola hepatica, as revealed by mitochondrial Nad1 and Cox1 genes. J Parasitol 92:525–530PubMedCrossRefGoogle Scholar
  36. Špakulová M, Králová I (1991) Chromosomes of Fasciola hepatica (Digenea: Fasciolidae) from western Bohemia (CSFR). Helminthologia 28:197–200Google Scholar
  37. Špakulová M, Rajský D, Sokol J, Vodňanský M (2003) Giant liver fluke (Fascioloides magna), an important liver parasite of ruminants. Parpress, BratislavaGoogle Scholar
  38. Sripa B, Kaewkes S, Intapan PM, Maleewong W, Brindley PJ (2010) Food-borne trematodiases in Southeast Asia: epidemiology, pathology, clinical manifestation and control. Adv Parasitol 72:305–350PubMedCrossRefGoogle Scholar
  39. Tanaka M, Hirai H, LoVerde PT, Nagafuchi S, Franco GR, Simpson AJG, Pena SDJ (1995) Yeast artificial chromosome (YAC)-based genome mapping of Schistosoma mansoni. Mol Biochem Parasitol 69:41–51PubMedCrossRefGoogle Scholar
  40. Teofanova D, Kantzoura V, Walker S, Radoslavov G, Hristov P, Theodoropoulos G, Bankov I, Trudgett A (2011) Genetic diversity of liver flukes (Fasciola hepatica) from Eastern Europe. Infect Genet Evol 11:109–115PubMedCrossRefGoogle Scholar
  41. Walker SM, Prodöhl PA, Fletcher HL, Hanna REB, Kantzoura V, Hoey EM, Trudgett A (2007) Evidence for multiple mitochondrial lineages of Fasciola hepatica (liver fluke) within infrapopulations from cattle and sheep. Parasitol Res 101:117–125PubMedCrossRefGoogle Scholar
  42. White MJD (1973) Animal Cytology and Evolution. Cambridge University Press, New YorkGoogle Scholar
  43. Yin HZ, Ye BY (1990) Studies on the karyotypes of Fasciola spp. Chin J Parasitol Parasitic Dis 8:124–126Google Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Marianna Reblánová
    • 1
  • Marta Špakulová
    • 1
  • Martina Orosová
    • 1
    • 2
  • Ivica Králová-Hromadová
    • 1
  • Eva Bazsalovicsová
    • 1
  • Dušan Rajský
    • 3
  1. 1.Parasitological InstituteSlovak Academy of SciencesKošiceSlovakia
  2. 2.Institute of Parasitology, Biology Centre AS CRČeské BudějoviceCzech Republic
  3. 3.Faculty of ForestryTechnical University in ZvolenZvolenSlovakia

Personalised recommendations