Abstract
Molecular characterization is important for discriminating Fasciola specimens having the deoxyribonucleic acid (DNA) sequences of Fasciola hepatica, Fasciola gigantica, and both Fasciola species, since three Fasciola forms coexist in Asian countries. We have developed a restriction fragment length polymorphism of amplified DNA (PCR–RFLP) of the nuclear ribosomal internal transcribed spacer 1 (ITS1) region in Fasciola species. The band patterns of the fragments digested with a restriction enzyme, Rsa I, were accurately distinguished among the three forms of Fasciola. Amplicons with the sequences of F. hepatica and F. gigantica were divided into fragments of about 360, 100, and 60 bp, and 360, 170, and 60 bp, respectively, and amplicons with the sequences of both Fasciola species yielded fragments of 360, 170, 100, and 60 bp. The results of PCR–RFLP completely coincided with those of sequence analysis, and thus PCR–RFLP is a useful technique for determining the ITS1 type in Fasciola species.
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References
Adlard RD, Barker SC, Blair D, Cribb TH (1993) Comparison of the second internal transcribed spacer (ribosomal DNA) from populations and species of Fasciolidae (Digenea). Int J Parasitol 23:423–425
Agatsuma T, Arakawa Y, Iwagami M, Honzako Y, Cahyaningsih U, Kang S-Y, Hong S-J (2000) Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. Parasitol Int 49:231–238
Ashrafi K, Valero MA, Panova M, Periago MV, Massoud J, Mas-Coma S (2006) Phenotypic analysis of adults of Fasciola hepatica, Fasciola gigantica and intermediate forms from the endemic region of Gilan, Iran. Parasitol Int 55:249–260
Chu JK, Kim YK (1967) Taxonomical study on the Fasciolidae in Korea. Kor J Parasitol 5:139–146
Itagaki H, Akane S (1959) Morphological study on the Japanese liver fluke, compared with the African specimens. Bull Azabu Vet Coll 6:115–123
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 28:777–781
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
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
Itagaki T, Sakaguchi K, Terasaki K, Sasaki O, Yoshihara S, Van Dung T (2009) Occurrence of spermic diploid and aspermic triploid forms of Fasciola in Vietnam and their molecular characterization based on nuclear and mitochondrial DNA. Parasitol Int 58:81–85
Kimura S, Shimizu A, Kawano J (1984) Morphological observation on liver fluke detected from naturally infected carabaos in the Philippines. Sci Rept Fac Agr Kobe Univ 16:353–357
Marcilla A, Bargues MD, Mas-Coma S (2002) A PCR–RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Mol Cell Probes 16:327–333
McGarry JW, Ortiz PL, Hodgkinson JE, Goreish I, Williams DJL (2007) PCR-based differentiation of Fasciola species (Trematoda: Fasciolidae), using primers based on RAPD-derived sequences. Ann Trop Med Parasitol 101:415–521
Moriyama N, Tsuji M, Seto T (1979) Three karyotypes and their phenotypes of Japanese liver flukes (Fasciola sp.). Jpn. J Parasitol 28:23–33 (in Japanese with English summary)
Oshima T, Akahane H, Shimazu T (1968) Patterns of the variation of the common liver fluke (Fasciola sp.) in Japan. I. Variations in the sizes and shapes of the worms and eggs. Jpn. J Parasitol 17:97–105 (in Japanese with English summary)
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 105:809–815
Reddy PV, Subramanyam S (1973) Chromosome studies in the liver fluke, Fasciola gigantica Cobbold, 1856, from Andra Pradesh. Curr Sci 42:288–291
Sakaguchi Y (1980) Karyotype and gametogenesis of the common liver fluke, Fasciola sp., in Japan. Jpn J Parasitol 29:507–513
Sanderson AR (1953) Maturation and probable gynogenesis in the liver fluke, Fasciola hepatica L. Nature 172:110–112
Terasaki K, Moriyama-Gonda N, Noda Y (1998) Abnormal spermatogenesis in the common liver fluke (Fasciola sp.) from Japan and Korea. J Vet Med Sci 60:1305–1309
Torgerson P, Claxton J (1999) Epidemiology and control. In: Dalton JP (ed) Fasciolosis. CABI Publishing, UK, pp 113–149
Varma AK (1953) On Fasciola indica n. sp. with some observations on F. hepatica and F. gigantica. J Helminthol 27:185–198
Watanabe S, Iwata S (1954) Fasciola species in Japan. J Jpn Vet Med Assoc 7:124–126 (in Japanese)
Acknowledgements
This study was supported in part by a grant-in-aid for Scientific Research (B) (no. 18405035) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Ichikawa, M., Itagaki, T. Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method. Parasitol Res 106, 757–761 (2010). https://doi.org/10.1007/s00436-010-1724-2
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DOI: https://doi.org/10.1007/s00436-010-1724-2