Department of Biology, College of Education for Girls, University of Mosul, Mosul, Iraq
Department of Physiology, Biochemistry, and Pharmacology, College of Veterinary Medicine, University of Mosul, Mosul, Iraq
Document Type : Research Paper
This study was conducted to investigate the 18S rRNA gene of Fasciola gigantica obtained from the liver of cattle live in Aqrah city, Iraq. Fifty-nine Fasciola flukes were collected through routine investigation from livers of naturally infected local cattle (42 cows), from May to August 2017, at the central slaughterhouse of Aqrah city, Kurdistan region of Iraq, the flukes were washed by PBS and then fixed in ethanol. Genomic DNA was extracted, and a 560 bp fragment was amplified by PCR, subsequent by sequencing of PCR products. A remarkable result of this project was the deposition of our gene isolate in GenBank (Accession No. MG786553). However, it was confirmed by the sequence results that isolate species was F. gigantica, and interestingly our samples sequences have alignment match of 100% with many international isolates, without genetic mutations or variations. It is concluded that molecular study could be utilized for both diagnosis and differential diagnosis of parasites with huge precise. Also, an 18S rRNA gene is a perfect fragment for molecular study and phylogenetic analysis of F. gigantica, also our samples have 100% alignment match with universal isolates.
- Yakhchali M, Malekzadeh R, Imani A, Mardani K. Morphological and molecular discrimination of Fasciola species isolated from domestic ruminants of Urmia city, Iran. Iran J Parasitol. 2015;10(1):46-55.
- Sah R, Khadka S, Lakhey PJ, Pradhan S, Shah NP, Singh YP, Mas S. Human case of F. gigantica-like infection, review of human fascioliasis reports in Nepal, and epidemiological analysis within the South-Central Asia. Acta Parasitol. 2018;63(3):435-443. DOI: 10.1515/ap-2018-0053.
- Al-Mahmood SS, Al-Sabaawy HB. Fasciolosis: grading the histopathological lesions in naturally infected bovine liver in Mosul city. Iraqi J Vet Med. 2019; 33(2): 379-387. DOI: 10.33899/ijvs.2019.125546.1066.
- El-Tahawy AS, Bazh EK, Khalafalla RE. Epidemiology of bovine fascioliasis in the Nile Delta region of Egypt: Its prevalence, evaluation of risk factors, and its economic significance. Vet World. 2017;10(10):1241-1249. DOI: 10.14202/vetworld.2017.1241-1249.
- Vara MP, Villa H, Martinez M, Rojo FA. Genetic heterogeneity of Fasciola hepatica isolates in the northwest of Spain. Parasitol Res. 2007;101(4):1003-1006. DOI: 10.1007/s00436-007-0574-z.
- Beesley NJ, Caminade C, Charlier J, Flynn RJ, Hodgkinson JE, Martinez A, Martinez M, Perez J, Rinaldi L, Williams DJL. Fasciola and fasciolosis in ruminants in Europe: Identifying research needs. Transbound Emerg Dis. 2017;65:199-216. DOI: 10.1111/tbed.12682.
- Aryaeipour M, Rouhani S, Bandehpour M, Mirahmadi H, Kazemi B, Rokni MB. Genotyping and phylogenetic analysis of Fasciola spp. isolated from sheep and cattle using PCR-RFLP in Ardabil province, Northwestern Iran. Iranian J Publ Health. 2014;43(10):1364-1371.
- Cwiklinski K, Dalton JP, Dufresne PJ, La Course J, Williams DJL, Hodgkinson J, Paterson S. The Fasciola hepatica genome: Gene duplication and polymorphism reveals adaptation to the host environment and the capacity for rapid evolution. Genome Biol. 2015;16:71-84. DOI: 10.1186/s13059-015-0632-2.
- Simsek S, Utuk AE, Balkaya I. Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and F. gigantica. Helminthologia. 2011;48:3-7. doi.org/10.2478/s11687-011-0001-y.
- McNulty SN, Tort JF, Rinaldi G, Fischer K, Rosa BA, Smircich P, Fontenla S, Choi YJ, Tyagi R, Hallsworth K, Mann VH, Kammili L, Latham PS, Dell’Oca N, Dominguez F, Carmona C, Fischer PU, Brindley PJ, Mitreva M. Genomes of Fasciola hepatica from the Americas reveal colonization with Neorickettsia endobacteria related to the agents of Potomac horse and human sennetsu fevers. PLoS Genet. 2017;13(1):1006537. DOI: 10.1371/journal.pgen.1006537.
- Walker SM, Prodohl P, Hoey EM, Fairweather I, Hanna RE, Brennan G, Trudgett A. Substantial genetic divergence between morphologically indistinguishable populations of Fasciola suggests the possibility of cryptic speciation. Int J Parasitol. 2012;42:1193-1199. DOI: 10.1016/j.ijpara.2012.10.007.
- Periago MV, Valero MA, El Sayed M, Ashrafi K, El Wakeel A, Mohamed MY, Desquesnes M, Curtale F, Mas S. First phenotypic description of Fasciola hepatica/ F. gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infect Genet Evol. 2008;8:51-8. DOI: 10.1016/j.meegid.2007.10.001.
- Choe SE, Nguyen TT, Kang TG, Kweon CH, Kang SW. Genetic analysis of Fasciola isolates from cattle in Korea based on second internal transcribed spacer (ITS-2) sequence of nuclear ribosomal DNA. Parasitol Res. 2011;109:833-839. DOI: 10.1007/s00436-011-2323-6.
- Ichikawa M, Tokashiki M, Opara MN, Iroh G, Hayashi K, Kumar UM, Itagaki T. Molecular characterization and phylogenetic analysis of F. gigantica from Nigeria. Parasitol Int. 2017;66:893-897. DOI: 10.1016/j.parint.2016.10.010.
- Martinez JM, Robles D, Rojo FA, Martinez M. Comparison of three different techniques to diagnose Fasciola hepatica infection in experimentally and naturally infected sheep. Vet Parasitol. 2012;190:80-6. DOI: 10.1016/j.vetpar.2012.06.002.
- Arifin MI, Hoglund J, Novobilsky A. Comparison of molecular and conventional methods for the diagnosis of Fasciola hepatica infection in the field. Vet Parasitol. 2016;232:8-11. DOI: 10.1016/j.vetpar.2016.11.003.
- Hamoo RN, Mustafa NG, Abdulraheem SA. Nad1 gene analysis of Echinococcus granulosus from sheep in Aqrah city, Iraq. Iraqi J Vet Med. 2019; 33(2): 341-345. DOI: 10.33899/ijvs.2019.162965.
- Farage HF. Human Fascioliasis in some countries of the Mediterranean Region. East Mediterran Health J. 1998;4:156-160.
- Al-Mayah SH. Studies on serology, biology, and control of F. gigantica Cobbold 1856 infections in Basrah [MSc dissertation]. Basrah: College of Science, University of Basrah, Basrah, Iraq; 2004. 188 p.
- Gzi MH, Rahma JH. Distribution of the parasite F. gigantica in the intermediate host in Al-Najaf Al-Ashraf governorate. J Thi-Qar Sci. 2012;3(2):39-47.
- Shahatha SS. Prevalence of F. gigantica among sheep in Al-Anbar governorate and study of effect for aqueous extract of Urtica urens L. on the viability of parasite. Anbar J Sci. 2013;7(3):33-38.
- Raina OK, Jacob SS, Sankar M, Bhattacharya D, Bandyopadyay S, Varghese A, Chamuah JK, Lalrinkima H. Genetic characterization of F. gigantica from different geographical regions of India by ribosomal DNA markers. J Parasit Dis. 2015;39(1):27-32. DOI: 10.1007/s12639-013-0276-7
- Karimi A. Genetic diagnosis of Fasciola species based on 18S ribosomal DNA gene. J Biol Sci. 2008;8(7):1166-1173. DOI: 10.3923/jbs.2008.1166.1173.
- Elkhtam AO, Khalafalla RE. Surveillance of helminthes and molecular phylogeny of F. gigantica infecting goats in Sadat district, Egypt. Int J Sci Res Sci Technol. 2016;2(4):188-192.
- Peng M, Ichinomiya M, Ohtori M, Ichikawa M, Shibahara T, Itagaki T. Molecular characterization of Fasciola hepatica, F. gigantica, and aspermic Fasciola spp. in China based on nuclear and mitochondrial DNA. Parasitol Res. 2009;105:809-815. DOI: 10.1007/s00436-009-1459-0.