Veterinary World

     Open access and peer reviewed journal  

ISSN (Online): 2231-0916


Home l Editorial board l Instructions for authors l Reviewer guideline l Open access policy l Archives l FAQ

Open Access

Research (Published online: 29-09-2016)

20. Molecular characterization of Fasciola hepatica and phylogenetic analysis based on mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I and cytochrome oxidase subunit I) genes from the North-East of Iran - Saber Raeghi, Ali Haghighi, Majid Fasihi Harandi, Adel Spotin, Kourosh Arzamani and Soheila Rouhani

Veterinary World, 9(9): 1034-1038



   doi: 10.14202/vetworld.2016.1034-1038



Saber Raeghi: Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;

Ali Haghighi: Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;

Majid Fasihi Harandi: Research Center for Hydatid disease in Iran, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran;

Adel Spotin: Department of Parasitology, Medical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran;

Kourosh Arzamani: Vector-borne Diseases Research Center, North Khorasan University of Medical Sciences, Bojnurd, Iran;

Soheila Rouhani: Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran;


Received: 03-06-2016, Accepted: 17-08-2016, Published online: 29-09-2016


Corresponding author: Soheila Rouhani, e-mail:

Citation: Raeghi S, Haghighi A, Harandi MF, Spotin A, Arzamani K, Rouhani S (2016) Molecular characterization of Fasciola hepatica and phylogenetic analysis based on mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I and cytochrome oxidase subunit I) genes from North-East of Iran, Veterinary World, 9(9): 1034-1038.

Aim: Fascioliasis is one of the most zoonotic diseases with global extension. As the epidemiological distribution of Fasciola may lead to various genetic patterns of the parasite, the aim of this study is to identify Fasciola hepatica based on spermatogenesis, and phylogenetic analysis using mitochondrial (nicotiamide adenine dinucleotide dehydrogenase subunit I [ND1] and cytochrome oxidase subunit I) gene marker.

Materials and Methods: In this study, 90 F. hepatica collected from 30 cattle at slaughterhouse located in three different geographical locations in the North-East of Iran were evaluated based on spermatogenetic ability and internal transcribed spacer 1 gene restriction fragment length polymorphism pattern. Genetic diversity and phylogenetic relationship using mtDNA gene marker for the isolates from the North-East of Iran, and other countries were then analyzed.

Results: Partial sequences of mtDNA showed eight haplotypes in both genes. The phylogenic analysis using neighbor joining as well as maximum likelihood methods showed similar topologies of trees. Pairwise fixation index between different F. hepatica populations calculated from the nucleotide data set of ND1 gene are statistically significant and show the genetic difference.

Conclusion: F. hepatica found in this region of Iran has different genetic structures through the other Fasciola populations in the world.

Keywords: Cytochrome oxidase subunit I, Fasciola hepatica, Iran, molecular characterization, nicotiamide adenine dinucleotide dehydrogenase subunit I, phylogenic.

1. Mas-Coma, S., Bargues, M.D. and Valero, M.A. (2005) Fascioliasis and other plant-borne trematode zoonoses. Int. J. Parasitol., 35(11): 1255-1278.
2. Olaechea, F., Lovera, V., Larroza, M., Raffo, F. and Cabrera, R. (2011) Resistance of Fasciola hepatica against triclabendazole in cattle in Patagonia (Argentina). Vet. Parasitol., 178(3): 364-366.
3. Rokni, M.B., Lotfy, W.M., Ashrafi, K. and Murrell, K.D. (2014) Fasciolosis in the MENA region. In: Neglected Tropical Diseases-Middle East and North Africa. Springer, Vienna. p59-90.
4. Ashrafi, K., Valero, M.A., Panova, M., Periago, M.V., Massoud, J. and 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(4): 249-260.
5. Peng, M., Ichinomiya, M., Ohtori, M., Ichikawa, M., Shibahara, T. and 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(3): 809-815.
6. Terasaki, K., Itagaki, T., Shibahara, T., Noda, Y. and Moriyama-Gonda, N. (2001) Comparative study of the reproductive organs of Fasciola groups by optical microscope. J. Vet. Med. Sci., 63(7): 735-742.
7. Adlard, R.D., Barker, S.C., Blair, D. and Cribb, T.H. (1993) Comparison of the second internal transcribed spacer (ribosomal DNA) from populations and species of Fasciolidae (Digenea). Int. J. Parasitol., 23(3): 423-425.
8. Ichikawa, M. and Itagaki, T. (2010) Discrimination of the ITS1 types of Fasciola spp. based on a PCR-RFLP method. Parasitol. Res., 106(3): 757-761.
9. Itagaki, T., Kikawa, M., Sakaguchi, K., Shimo, J., Terasaki, K., Shibahara, T. and Fukuda, K. (2005) Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. Parasitology, 131: 679-685.
10. Mas‐Coma, S., Valero, M.A. and Bargues, M.D. (2009) Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv. Parasitol., 69: 41-146.
11. Ashrafi, K. (2015) The status of human and animal Fascioliasis in Iran: A narrative review article. Iran. J. Parasitol., 10(3): 306.
PMid:26622287 PMCid:PMC4662732
12. Hayashi, K., Ichikawa-Seki, M., Mohanta, U.K., Singh, T.S., Shoriki, T., Sugiyama, H. and Itagaki, T. (2015) Molecular phylogenetic analysis of Fasciola flukes from Eastern India. Parasitol. Int., 64(5): 334-338.
13. Mohanta, U.K., Ichikawa-Seki, M., Shoriki, T., Katakura, K. and Itagaki, T. (2014) Characteristics and molecular phylogeny of Fasciola flukes from Bangladesh, determined based on spermatogenesis and nuclear and mitochondrial DNA analyses. Parasitol. Res., 113(7): 2493-2501.
14. Itagaki, T., Kikawa, M., Terasaki, K., Shibahara, T. and Fukuda, K. (2005) 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(11): 1115-1118.
15. Tamura, K., Stecher, G., Peterson, D., Filipski, A. and Kumar, S. (2013) MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol., 30(12): 2725-2729.
PMid:24132122 PMCid:PMC3840312
16. Rozas, J., Sánchez-DelBarrio, J.C., Messeguer, X. and Rozas, R. (2003) DnaSP, DNA polymorphism analyses by the coalescent and other methods. Bioinformatics, 19(18): 2496-2497.
17. Excoffier, L. and Lischer, H.E. (2010) Arlequin suite ver 3.5: A new series of programs to perform population genetics analyses under linux and windows. Mol. Ecol. Resour., 10(3): 564-567.
18. Reynolds, J., Weir, B.S. and Cockerham, C.C. (1983) Estimation of the coancestry coefficient: Basis for a short-term genetic distance. Genetics, 105(3): 767-779.
PMid:17246175 PMCid:PMC1202185
19. Periago, M.V., Valero, M.A., Panova, M. and Mas-Coma, S. (2006) Phenotypic comparison of allopatric populations of Fasciola hepatica and Fasciola gigantica from European and African bovines using a computer image analysis system (CIAS). Parasitol. Res., 99(4): 368-378.
20. Chaichanasak, P., Ichikawa, M., Sobhon, P. and Itagaki, T. (2012) Identification of Fasciola flukes in Thailand based on their spermatogenesis and nuclear ribosomal DNA, and their intraspecific relationships based on mitochondrial DNA. Parasitol. Int., 61(4): 545-549.
21. Hashimoto, K., Watanobe, T., Liu, C.X., Init, I., Blair, D., Ohnishi, S. and Agatsuma, T. (1997) Mitochondrial DNA and nuclear DNA indicate that the Japanese Fasciola species is F. gigantica. Parasitol. Res., 83(3): 220-225.
22. Rokni, M.B., Mirhendi, H., Mizani, A., Mohebali, M., Sharbatkhori, M., Kia, E.B., Abdoli, H. and Izadi, S. (2010) Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method. Exp. Parasitol., 124(2): 209-213.
23. Shafiei, R., Sarkari, B. and Moshfe, A. (2013) A consistent PCR-RFLP assay based on ITS-2 ribosomal DNA for differentiation of Fasciola species. Iran. J. Basic Med. Sci., 16(12): 1266-1269.
PMid:24570833 PMCid:PMC3933804
24. Aryaeipour, M., Rouhani, S., Bandehpour, M., Mirahmadi, H., Kazemi, B. and Rokni, M.B. (2014) Genotyping and phylogenetic analysis of Fasciola spp. Isolated from sheep and cattle using PCR-RFLP in Ardabil province, Northwestern Iran. Iran. J. Public Health, 43(10): 1364-1371.
PMid:26060698 PMCid:PMC4441889
25. Galavani, H., Gholizadeh, S. and Tappeh, K.H. (2016) Genetic characterization of Fasciola isolates from West Azerbaijan province Iran based on ITS1 and ITS2 sequence of ribosomal DNA. Iran. J. Parasitol., 11(1): 52-64.
PMid:27095969 PMCid:PMC4835470
26. Yakhchali, M., Malekzadeh-Viayeh, R., Imani-Baran, A. and Mardani, K. (2015) Morphological and molecular discrimination of Fasciola species isolated from domestic ruminants of Urmia City, Iran. Iran. J. Parasitol., 10(1): 46-55.
PMid:25904945 PMCid:PMC4403539