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Sequence analysis of the first internal transcribed spacer of rDNA supports the existence of the intermediate Fasciola between F. hepatica and F. gigantica in mainland China

Parasitology Research volume 101, pages 813–817 (2007)Cite this article

Abstract

In the present study, a polymerase chain reaction-linked single-strand conformation polymorphism (PCR-SSCP) approach combined with DNA sequencing was used to characterise samples of Fasciola spp. from different host species and geographical locations in mainland China. The first internal transcribed spacer (ITS-1) of ribosomal DNA (rDNA) was amplified by PCR from individual Fasciola and analysed by SSCP. SSCP analyses displayed three different banding profiles that allowed the identification of all Fasciola samples examined into three groups: Fasciola hepatica, F. gigantica and the “intermediate” Fasciola. Then, the ITS-1 rDNA was sequenced from representative Fasciola samples, and analysis of the complete ITS-1 sequences supported the identification of all Fasciola samples by SSCP approach. The length of the ITS-1 sequences was 422 bp for all Fasciola samples sequenced. Although there was no variation in length or composition of the ITS-1 sequences among multiple specimens within each of the taxa, F. hepatica and F. gigantica differed by 1.2% in their ITS-1 sequences, whereas the “intermediate” Fasciola was unique, in which two different ITS-1 sequences exist in the rDNA array within a single Fasciola worm. One of the sequences is identical to that of F. hepatica, and the other is identical to that of F. gigantica. This study demonstrated that PCR-SSCP analysis of the ITS-1 rDNA followed by selective sequencing provides a reliable approach for the accurate identification of Fasciola spp., and also supports the existence of the “intermediate” Fasciola between F. hepatica and F. gigantica in mainland China.

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References

  • Agatsuma T, Arakawa Y, Iwagami M, Honzako Y, Cahyaningsih U, Kang SY, Hong SJ (2000) Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. Parasitol Int 49:231–238

    PubMed  Article  CAS  Google Scholar 

  • Chilton NB, Gasser RB, Beveridge I (1995) Differences in a ribosomal DNA sequence of morphologically indistinguishable species within the Hypodontus macropi complex (Nematoda: Strongyloidea). Int J Parasitol 25:647–651

    PubMed  Article  CAS  Google Scholar 

  • Gasser RB, Hu M, Chilton NB, Campbell BE, Jex AJ, Otranto D, Cafarchia C, Beveridge I, Zhu XQ (2006) Single-strand conformation polymorphism (SSCP) for the genetic analysis and identification of pathogens. Nat Protoc 1:3121–3128

    PubMed  Article  CAS  Google Scholar 

  • Haseeb AN, el-Shazly AM, Arafa MA, Morsy AT (2002) A review on fascioliasis in Egypt. J Egypt Soc Parasitol 32:317–354

    PubMed  Google Scholar 

  • Huang WY, He B, Wang CR, Zhu XQ (2004) Characterisation of Fasciola species from mainland China by ITS-2 ribosomal DNA sequence. Vet Parasitol 120:75–83

    PubMed  Article  CAS  Google Scholar 

  • Ishii Y, Nakamura-Uchiyama F, Nawa Y (2002) A praziquantel-ineffective fascioliasis case successfully treated with triclabendazole. Parasitol Int 51:205–209

    PubMed  Article  Google Scholar 

  • Itagaki T, Tsutsumi KI (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 28:777–781

    PubMed  Article  CAS  Google Scholar 

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

    PubMed  Article  CAS  Google Scholar 

  • Itagaki T, Kikawa M, Terasaki K, Shibahara T, Fukuda K (2005b) Molecular characterization of parthenogenic Fasciola sp. in Korea on the basis of DNA sequences of ribosomal ITS1 and mitochondrial NDI gene. J Vet Med Sci 67:1115–1118

    PubMed  Article  CAS  Google Scholar 

  • Jousson O, Bartoli P, Zaninetti L, Pawlowski J (1998) Use of the ITS rDNA for elucidation of some life-cycles of Mesometridae (Trematoda, Digenea). Int J Parasitol 28:1403–1411

    PubMed  Article  CAS  Google Scholar 

  • Jousson O, Bartoli P, Pawlowski J (1999) Molecular identification of developmental stages in Opecoelidae (Digenea). Int J Parasitol 29:1853–1858

    PubMed  Article  CAS  Google Scholar 

  • Kendall SB (1965) Relationships between the species of Fasciola and their molluscan hosts. Adv Parasitol 3:59–98

    PubMed  Article  CAS  Google Scholar 

  • Li AX, D’Amello S, Paggl L, He F, Gasser RB, Lun ZR, Abollo E, Turchetto M, Zhu XQ (2005) Genetic evidence for the existence of sibling species within Contracaecum rudolphii Hartwich, 1964 and the validity of Contracaecum septentrionale Kreis, 1955 (Nematoda: Anisakidae). Parasitol Res 96:361–366

    PubMed  Article  Google Scholar 

  • Luton K, Walker D, Blair D (1992) Comparisons of ribosomal internal transcribed spacers from two congeneric species of flukes (Platyhelminthes: Trematoda: Digenea). Mol Biochem Parasitol 56:323–327

    PubMed  Article  CAS  Google Scholar 

  • Mas-Coma S, Esteban JG, Bargues MD (1999) Epidemiology of human fascioliasis: a review and proposed new classification. Bull World Health Organ 77:340–346

    PubMed  CAS  Google Scholar 

  • Mas-Coma S, Bargues MD, Valero MA (2005) Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol 35:1255–1278

    PubMed  Article  CAS  Google Scholar 

  • Nolan MJ, Cribb TH (2005) The use and implications of ribosomal DNA sequencing for the discrimination of digenean species. Adv Parasitol 60:101–163

    PubMed  Article  Google Scholar 

  • Spithill TW, Dalton JP (1998) Progress in development of liver fluke vaccines. Parasitol Today 14:224–228

    PubMed  Article  CAS  Google Scholar 

  • Yamaguti S (1958) Systema helminthum, vol I. The digenetic trematodes of vertebrates. Interscience, New York

    Google Scholar 

  • Zhu XQ, D’Amelio S, Paggi L, Gasser RB (2000) Assessing sequence variation in the internal transcribed spacers of ribosomal DNA within and among members of the Contracaecum osculatum complex (Nematoda: Ascaridoidea: Anisakidae). Parasitol Res 86:677–683

    PubMed  Article  CAS  Google Scholar 

  • Zhu XQ, D’Amelio S, Palm HW, Paggi L, George-Nascimento M, Gasser RB (2002) SSCP-based identification of members within the Pseudoterranova decipiens complex (Nematoda: Ascaridoidea: Anisakidae) using genetic markers in the internal transcribed spacers of ribosomal DNA. Parasitology 124:615–623

    PubMed  Article  CAS  Google Scholar 

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Acknowledgements

Project support was provided in part by a grant from the South China Agricultural University (grant no. 2004K024) to RQL, and a grant from the National Natural Science Foundation of China (grant no. 30560111) to WYH. The experiments comply with the current laws of the countries in which the experiments were performed.

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Affiliations

  1. College of Veterinary Medicine, South China Agricultural University, 483 Wushan Street, Tianhe District, Guangzhou, Guangdong Province, 510642, People’s Republic of China

    R. Q. Lin, H. Q. Song & X. Q. Zhu

  2. Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai, 201106, People’s Republic of China

    S. J. Dong

  3. College of Animal Science and Technology, Southwest University, Chongqing, 400715, People’s Republic of China

    K. Nie

  4. College of Animal Science and Technology, Heilongjiang August-First Land Reclamation University, Daqing, Heilongjiang Province, 163319, People’s Republic of China

    C. R. Wang

  5. School of Life Sciences, Zhongshan (Sun Yat-Sen) University, 135 West Xingang Street, Haizhu District, Guangzhou, Guangdong Province, 510275, People’s Republic of China

    A. X. Li

  6. College of Animal Science and Technology, Guangxi University, Nanning, Guangxi Zhuang Nationality Autonomous Region, 530004, People’s Republic of China

    W. Y. Huang

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  1. R. Q. Lin
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  2. S. J. Dong
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  3. K. Nie
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  4. C. R. Wang
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  5. H. Q. Song
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  6. A. X. Li
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  7. W. Y. Huang
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  8. X. Q. Zhu
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Correspondence to X. Q. Zhu.

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Lin, R.Q., Dong, S.J., Nie, K. et al. Sequence analysis of the first internal transcribed spacer of rDNA supports the existence of the intermediate Fasciola between F. hepatica and F. gigantica in mainland China. Parasitol Res 101, 813–817 (2007). https://doi.org/10.1007/s00436-007-0512-0

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  • DOI: https://doi.org/10.1007/s00436-007-0512-0

Keywords

  • Liver Fluke
  • SSCP Analysis
  • Selective Sequencing
  • rDNA Array
  • Digenean Trematode