MOLECULAR IDENTIFICATION AND PHYLOGENETIC ANALYSIS OF FASCIOLA SPECIES ISOLATED FROM SHEEP AND CATTLE USING THE ITS2 rDNA MARKER IN DUHOK PROVINCE, KURDISTAN REGION, IRAQ
Keywords:
ITS2, PCR, Fasciola species, Phylogenetic analysis, Ruminants, Iraq
Abstract
The aims of the current study were molecular identification and phylogenetic analysis of Fasciola species isolated from infected cattle and sheep using the ITS2 region of rDNA as a molecular marker in Duhok province, Kurdistan region, Iraq. DNA was extracted from 40 adult flukes isolated from the livers of infected sheep (23) and cattle (17) slaughtered at the abattoirs of Duhok Province. The amplification of the rDNA ITS2 region of these 40 specimens was performed by PCR using designed primers. ITS2 fragments of the rDNA with a length of 314 bp were successfully amplified. From these isolates, 5 amplicons of ITS2 from both species (sheep and cattle) were selected for sequencing. The blast alignment of NCBI-GenBank revealed four sequences of five amplicons belonging to Fasciola hepatica and one belonging to Fasciola gigantica, both with 100% similarity. The genetic distance between F. hepatica and F. gigantica was 0.0127. Phylogenetic analysis clustered the sequences of this study into two main clades (F. hepatica and F. gigantica) in comparison with the corresponding sequences of Fasciola species worldwide. The study concluded that ITS2 rDNA sequences were appropriate to identify both Fasciola species isolated from the infected animals, which were identical to those of neighboring countries (Iran and Turkey). The primers for the ITS2 rDNA region that were designed for the current study can be useful for the diagnosis of animal and human fasciolosis
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References
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Alasaad, S., Li, Q.Y., Lin, R. Q, Martín-Atance P., Granados, J. E., Diez- Banos, P., Perez J. M., Zhu, X. Q. (2008). Genetic variability among samples from different host species and geographical localities in Spain revealed by the novel SRAP marker. Parasitology Research, 03(1):181–186. DOI: 10.1007/s00436-008-0952-1.
Ai, L., Chen, M-X., Alasaad, S. et al (2011). Genetic characterization, species differentiation and detection of Fasciola spp. by molecular approaches. Parasite Vectors, 4:101.
Alves, D. P, Carneiro, M. B., Martins, I. V .F., Bernardo, C. C., Donatele, D. M., Pereira Junior, O. S., Almeida, B. R., Avelar, B. R., Leao A. G. C. (2011). Distribution and factors associated with Fasciola hepatica infection in cattle in the south of Espírito Santo State, Brazil. The Journal of Venomous Animals and Toxins including Tropical Diseases, 7(3) :271-276.
Amer, S., ElKhatam A., Zidan, S., Feng, Y., and Xiao L. (2016). Identity of Fasciola spp. in sheep in Egypt. Parasites and vectors, 9 (1): 623-631. DOI:10.1186/s13071-016-1898-2.
Amor, N., Halajian, A., Farjallah, S., Merella, P., Said, K., Ben Slimane, B. (2011). Molecular characterization of Fasciola spp. from the endemic area of north Iran based on nuclear ribosomal DNA sequences. Experimental Parasitology, 128(3):196 -204.
Ashrafi, K., Valero, M. A., Panova, M., 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. Parasitology International, 55(4):249–260. DOI: 10.1016/j.parint.2006.06.003.
Bargues, M. D., Halajian, A., Artigas, P., Luus-Powell, W.J., Valero, M.A. and Mas-Coma, S. (2022) Paleobiogeographical origins of Fasciola hepatica and F. gigantica in light of new DNA sequence characteristics of F. nyanzae from hippopotamus. Front. Veterinary Science. 9:990872. doi: 10.3389/fvets.2022.990872
Dinnik, J., and Dinnik, N. (1961). On the Morphology and Life History of Fasciola nyanzae Leiper, 1910 from the Hippopotamus. Journal of Helminthology, 35(S1), 53-62. Published online by Cambridge University Press: 05 June 2009. DOI: https://doi.org/10.1017/S0022149X00017570
El-Rahimy, H. H., Mahgoub, A. M., El-Gebaly, N. S, Mousa, W.M., Antably, A. S. (2012). Molecular, biochemical, and morphometric characterization of Fasciola species potentially causing zoonotic disease in Egypt. Parasitology Research, 111:1103-1111.
Felsenstein J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39:783-791.
Galavani, H., Gholizadeh, S., Hazrati Tappeh, K. (2016). Genetic characterization of Fasciola isolates from west Azerbaijan province Iran based on ITS1 and ITS2 sequence of ribosomal DNA. Iranian journal of parasitology, 11(1):52-64. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835470
Ghavami, M. B., Rahimi, P., Haniloo, A., Mosavinasab, S. N. (2009). Genotypic and phenotypic analysis of Fasciola isolates. Iranian journal of parasitology, 4(3):61-70.
Hasanpour, H., Falak, R., Naddaf, S. R., Mas-Coma, S., Rokni, M.B., Badirzadeh, A., Mokhtarian, K., Mohebali, M., Jafarpour Azami, S., Fadavi, A., Gharagozlou, M. J., Mohammad, K., Mowlavi, G. (2020) Molecular Characterization of Fasciola spp. from Some Parts of Iran. Iran Journal Public Health, 49(1):157-166.
Haridwal, S., Malatji, M. P., Mukaratirwa, S. (2021). Morphological and molecular characterization of Fasciola hepatica and Fasciola gigantica phenotypes from co-endemic localities in Mpumalanga and KwaZulu-Natal provinces of South Africa. Food Waterborne Parasitology, 3(22):e00114. https://doi.org/10.1016/j.fawpar.2021.e00114
Huang, W. Y., He, B., Wang, C. R., Zhu, X. Q. (2004). Characterization of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence. Veterinary parasitology, 120 (1-2): 75-83. DOI: 10.1016/j.vetpar.2003.12.006
Karimi, A. (2008). Genetic diagnosis of Fasciola species based on 18S ribosomal DNA sequences. Journal of Biological Sciences, 8:1166–1173.
Khan, M.K., Sajid, M.S., Riaz, H. et al. (2013). The global burden of fasciolosis in domestic animals with an outlook on the contribution of new approaches for diagnosis and control. Parasitology Research, 112: 2421–2430. https://doi.org/10.1007/s00436-013-3464-6
Khanjari, A., Bahonar, A., Fallah, S., Bagheri, M., Alizadeh, A., Fallah, M., Khanjari, Z. (2014). Prevalence of fasciolosis and dicrocoelio-sis in slaughtered sheep and goats in Amol Abattoir, Mazandaran, northern Iran. Asian Pacific Journal of Tropical Disease, 4(2): 120-124.
Lotfy, W. M., Brant, S. V., DeJong, R. J., Le, T. H., Demiaszkiewicz, A., Rajapakse, R. P., Perera, V. B., Laursen, J. R., Loker, E. S. (2008). Evolutionary origins, diversification, and biogeography of liver flukes (Digenea, Fasciolidae). American journal tropical medicine hygiene 79(2):248–255. DOI:10.4269/AJTMH.2008.79.248
Lotfy, W. M., El-Morshedy, H. N., Abou El-Hoda, M., El-Tawila, M. M., Omar, E. A., Farag, H. F. (2002) Identification of the Egyptian species of Fasciola. Veterinary Parasitology, 103(4): 323–332. https://doi.org/10.1016/S0304-4017(01)00613-6.
Lotfy, W. M., Hillyer, G.V. (2003). Fasciola species in Egypt. Experimental pathology and parasitology. 6, 9–22.
Mahami-Oskouei, M., Dalimi, A., Forouzandeh-Moghadam, M., Rokni, M. (2011). Molecular identification and differentiation of Fasciola isolates using PCR- RFLP method based on internal transcribed spacer (ITS1, 5.8S rDNA, ITS2). Iranian journal of parasitology. Aug;6(3):35-42. PMID: 22347295; PMCID: PMC3279892.
Marcilla, A., Bargues, M. D., Mas-Coma, S. A. (2002). PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Molecular and Cellular Probes 16(5): 327–333. DOI: 10.1006/mcpr.2002.0429
Mas-Coma, S., Bargues, M. D. (1997). Human liver flukes: a review. Research and Reviews in Parasitology, 57(3-4): 145-218.
Mas-Coma, S., Bargues, M. D., Esteban, J. G. (1999). Human fascioliasis. In: Dalton JP. Fascioliasis. Wallingford: CAB International, p. 411-434.
Mas-Coma, S., Valero, M. A., Bargues, M. D. (2009). Chapter 2 Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Advances in Parasitology 69:141-146.
Mas-Coma, S., Bargues, M. D., Valero, M. A. (2005). Fascioliasis and other plant-borne trematode zoonoses. International journal for parasitology, 35(11-12):1255-1278.
Mir S., Dabirzadeh M., Rokni M. B., Aryaeipour, M., Shahraki, M. K., Azizi, H. (2019). Identification and phylogenetic classification of Fasciola species isolated from sheep and cattle by PCR-RFLP in Zabol, in Sistan and Baluchistan province, Southeast Iran. Iranian journal of public health, 48 (5):934-942. https://pubmed.ncbi.nlm.nih.gov/31523651
Mohammed, A. B., Mero, W. M. S., Nerway, C. A. (2022). Molecular identification of Fasciola spp. isolated from domestic animals based on DNA sequencing of the nuclear ribosomal ITS1 -ITS2 markers, Kurdistan region, Iraq. Pakistan Veterinary Journal, 42(2):246-250. http://dx.doi.org/10.29261/pakvetj/2021.066
Nerway, C. A., Mero, W. M. S., Mohammed, A. B. (2021). Prevalence of Fasciola spp. among slaughtered livestock in Zakho city, Duhok governorate - Iraq. Academic Journal of Nawroz University, 10(2):199–206. https://doi.org/10.25007/ajnu.v10n2a1034
Nguyen, T. B. N., Van De, N., Nguyen, T. K.L., Quang, H. H., Doan, H. T. T., Agatsuma, T., Le, T. H., (2018). Distribution status of hybrid types in large liver flukes, Fasciola species (Digenea: Fasciolidae), from ruminants and humans in Vietnam. Korean Journal of Parasitology, 56(5):453-461.
Periago, M. V., Valero, M. A., El Sayed, M., Ashrafi, K., El Wakeel, A., Mohamed, M. Y. and Mas-Coma, S. (2008). First phenotypic description of Fasciola hepatica/Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infection. Genetics and Evolution journal. 8(1):51-58.
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. Parasitology Research, 105(3): 809-815
Prasad, P. K., Tandon, V., Biswal, D. K., Goswami, L. M., Chatterjee, A. (2008). Molecular identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae) based on the ribosomal internal transcribed spacer regions. Parasitology Research. 103(6):1247–1255. DOI: 10.1007/s00436-008-1121-2
Qureshi, A. W., Tanveer, A., Qureshi, S. W., Maqbool, A., Gill, T. J., Ali, S. A. (2005). Epidemiology of human fasciolosis in rural areas of Lahore, Pakistan. Punjab University Journal of Zoology, 20:159-168.
Ramadhan, C. A., (2021). Prevalence of Fasciola species and their molecular characterization using ITS1 and ITS2 in Duhok province, Kurdistan region, Iraq. M.Sc. Thesis, Faculty of Science, Zakho university.
Raoof H. S., Marif, H. F., Rahman, H. S., Omar, M., Sheikh, B., San Ahmed, A. M. (2020). Molecular characterization and phylogenetic analysis of Fasciola species in sheep and goats in Sulaymaniyah Province, Northern Iraq. Journal of Zankoy Sulaimani, 22(1):297-305. https://www.researchgate.net/publication/342926652
Rokni, M., Mirhendi, H., Mizani, A., et al. (2010). Identification and differentiation of Fasciola hepatica and F. gigantica using a simple PCR-restriction enzyme method. Experimental Parasitology, 124:209-213.
Saki, J., Khademvatan, S., Yousefi, E. (2011). Mo-lecular identification of animal Fasciola isolates in Southwest of Iran. Australian Journal of Basic and Applied Sciences, 5:1878-1883.
Shafiei, R., Sarkari, B., Moshfe, A. (2013). A Consistent PCR-RFLP Assay Based on ITS-2 Ribosomal DNA for Differentiation of Fasciola Species. Iranian Journal of Basic Medical Sciences, 16(12):1266-1269.
Simsek, A., Utuk,A. E., and Balkaya, I. (2011). Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica. Helminthologia Journal, 48(1):3-7.
Tamura, K., Stecher, G., and Kumar, S. (2021). MEGA 11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology Evolution, 38(7):3022-3027. https://doi.org/10.1093/molbev/msab120.
Turhan, O., Korkmaz, M., Saba, R., Kabaaaliogu, A., Inan, D., Mamikoglu, L. (2006). Seroepidemiology of fascioliasis in the Antalya region and uselessness of eosinophil count as a surrogate marker and portable ultrasonography for epidemiological surveillance. Infezioni in Medicina, 14:208-212.
World Health Organization (WHO). Trematode infections/fascioliasis/fascioliasis diagnosis/en/. 2013. http://www.who.int/foodborne
Abdulwahed, T. Kh., Al-Amery A. M. (2019). Morphological and molecular study of Fasciola spp. in sheep in Alkut city. International Journal of Bioscience, 14(1):121-130. http://dx.doi.org/10.12692/ijb/14.1.121-130
Alasaad, S., Li, Q.Y., Lin, R. Q, Martín-Atance P., Granados, J. E., Diez- Banos, P., Perez J. M., Zhu, X. Q. (2008). Genetic variability among samples from different host species and geographical localities in Spain revealed by the novel SRAP marker. Parasitology Research, 03(1):181–186. DOI: 10.1007/s00436-008-0952-1.
Ai, L., Chen, M-X., Alasaad, S. et al (2011). Genetic characterization, species differentiation and detection of Fasciola spp. by molecular approaches. Parasite Vectors, 4:101.
Alves, D. P, Carneiro, M. B., Martins, I. V .F., Bernardo, C. C., Donatele, D. M., Pereira Junior, O. S., Almeida, B. R., Avelar, B. R., Leao A. G. C. (2011). Distribution and factors associated with Fasciola hepatica infection in cattle in the south of Espírito Santo State, Brazil. The Journal of Venomous Animals and Toxins including Tropical Diseases, 7(3) :271-276.
Amer, S., ElKhatam A., Zidan, S., Feng, Y., and Xiao L. (2016). Identity of Fasciola spp. in sheep in Egypt. Parasites and vectors, 9 (1): 623-631. DOI:10.1186/s13071-016-1898-2.
Amor, N., Halajian, A., Farjallah, S., Merella, P., Said, K., Ben Slimane, B. (2011). Molecular characterization of Fasciola spp. from the endemic area of north Iran based on nuclear ribosomal DNA sequences. Experimental Parasitology, 128(3):196 -204.
Ashrafi, K., Valero, M. A., Panova, M., 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. Parasitology International, 55(4):249–260. DOI: 10.1016/j.parint.2006.06.003.
Bargues, M. D., Halajian, A., Artigas, P., Luus-Powell, W.J., Valero, M.A. and Mas-Coma, S. (2022) Paleobiogeographical origins of Fasciola hepatica and F. gigantica in light of new DNA sequence characteristics of F. nyanzae from hippopotamus. Front. Veterinary Science. 9:990872. doi: 10.3389/fvets.2022.990872
Dinnik, J., and Dinnik, N. (1961). On the Morphology and Life History of Fasciola nyanzae Leiper, 1910 from the Hippopotamus. Journal of Helminthology, 35(S1), 53-62. Published online by Cambridge University Press: 05 June 2009. DOI: https://doi.org/10.1017/S0022149X00017570
El-Rahimy, H. H., Mahgoub, A. M., El-Gebaly, N. S, Mousa, W.M., Antably, A. S. (2012). Molecular, biochemical, and morphometric characterization of Fasciola species potentially causing zoonotic disease in Egypt. Parasitology Research, 111:1103-1111.
Felsenstein J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution, 39:783-791.
Galavani, H., Gholizadeh, S., Hazrati Tappeh, K. (2016). Genetic characterization of Fasciola isolates from west Azerbaijan province Iran based on ITS1 and ITS2 sequence of ribosomal DNA. Iranian journal of parasitology, 11(1):52-64. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4835470
Ghavami, M. B., Rahimi, P., Haniloo, A., Mosavinasab, S. N. (2009). Genotypic and phenotypic analysis of Fasciola isolates. Iranian journal of parasitology, 4(3):61-70.
Hasanpour, H., Falak, R., Naddaf, S. R., Mas-Coma, S., Rokni, M.B., Badirzadeh, A., Mokhtarian, K., Mohebali, M., Jafarpour Azami, S., Fadavi, A., Gharagozlou, M. J., Mohammad, K., Mowlavi, G. (2020) Molecular Characterization of Fasciola spp. from Some Parts of Iran. Iran Journal Public Health, 49(1):157-166.
Haridwal, S., Malatji, M. P., Mukaratirwa, S. (2021). Morphological and molecular characterization of Fasciola hepatica and Fasciola gigantica phenotypes from co-endemic localities in Mpumalanga and KwaZulu-Natal provinces of South Africa. Food Waterborne Parasitology, 3(22):e00114. https://doi.org/10.1016/j.fawpar.2021.e00114
Huang, W. Y., He, B., Wang, C. R., Zhu, X. Q. (2004). Characterization of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence. Veterinary parasitology, 120 (1-2): 75-83. DOI: 10.1016/j.vetpar.2003.12.006
Karimi, A. (2008). Genetic diagnosis of Fasciola species based on 18S ribosomal DNA sequences. Journal of Biological Sciences, 8:1166–1173.
Khan, M.K., Sajid, M.S., Riaz, H. et al. (2013). The global burden of fasciolosis in domestic animals with an outlook on the contribution of new approaches for diagnosis and control. Parasitology Research, 112: 2421–2430. https://doi.org/10.1007/s00436-013-3464-6
Khanjari, A., Bahonar, A., Fallah, S., Bagheri, M., Alizadeh, A., Fallah, M., Khanjari, Z. (2014). Prevalence of fasciolosis and dicrocoelio-sis in slaughtered sheep and goats in Amol Abattoir, Mazandaran, northern Iran. Asian Pacific Journal of Tropical Disease, 4(2): 120-124.
Lotfy, W. M., Brant, S. V., DeJong, R. J., Le, T. H., Demiaszkiewicz, A., Rajapakse, R. P., Perera, V. B., Laursen, J. R., Loker, E. S. (2008). Evolutionary origins, diversification, and biogeography of liver flukes (Digenea, Fasciolidae). American journal tropical medicine hygiene 79(2):248–255. DOI:10.4269/AJTMH.2008.79.248
Lotfy, W. M., El-Morshedy, H. N., Abou El-Hoda, M., El-Tawila, M. M., Omar, E. A., Farag, H. F. (2002) Identification of the Egyptian species of Fasciola. Veterinary Parasitology, 103(4): 323–332. https://doi.org/10.1016/S0304-4017(01)00613-6.
Lotfy, W. M., Hillyer, G.V. (2003). Fasciola species in Egypt. Experimental pathology and parasitology. 6, 9–22.
Mahami-Oskouei, M., Dalimi, A., Forouzandeh-Moghadam, M., Rokni, M. (2011). Molecular identification and differentiation of Fasciola isolates using PCR- RFLP method based on internal transcribed spacer (ITS1, 5.8S rDNA, ITS2). Iranian journal of parasitology. Aug;6(3):35-42. PMID: 22347295; PMCID: PMC3279892.
Marcilla, A., Bargues, M. D., Mas-Coma, S. A. (2002). PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Molecular and Cellular Probes 16(5): 327–333. DOI: 10.1006/mcpr.2002.0429
Mas-Coma, S., Bargues, M. D. (1997). Human liver flukes: a review. Research and Reviews in Parasitology, 57(3-4): 145-218.
Mas-Coma, S., Bargues, M. D., Esteban, J. G. (1999). Human fascioliasis. In: Dalton JP. Fascioliasis. Wallingford: CAB International, p. 411-434.
Mas-Coma, S., Valero, M. A., Bargues, M. D. (2009). Chapter 2 Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Advances in Parasitology 69:141-146.
Mas-Coma, S., Bargues, M. D., Valero, M. A. (2005). Fascioliasis and other plant-borne trematode zoonoses. International journal for parasitology, 35(11-12):1255-1278.
Mir S., Dabirzadeh M., Rokni M. B., Aryaeipour, M., Shahraki, M. K., Azizi, H. (2019). Identification and phylogenetic classification of Fasciola species isolated from sheep and cattle by PCR-RFLP in Zabol, in Sistan and Baluchistan province, Southeast Iran. Iranian journal of public health, 48 (5):934-942. https://pubmed.ncbi.nlm.nih.gov/31523651
Mohammed, A. B., Mero, W. M. S., Nerway, C. A. (2022). Molecular identification of Fasciola spp. isolated from domestic animals based on DNA sequencing of the nuclear ribosomal ITS1 -ITS2 markers, Kurdistan region, Iraq. Pakistan Veterinary Journal, 42(2):246-250. http://dx.doi.org/10.29261/pakvetj/2021.066
Nerway, C. A., Mero, W. M. S., Mohammed, A. B. (2021). Prevalence of Fasciola spp. among slaughtered livestock in Zakho city, Duhok governorate - Iraq. Academic Journal of Nawroz University, 10(2):199–206. https://doi.org/10.25007/ajnu.v10n2a1034
Nguyen, T. B. N., Van De, N., Nguyen, T. K.L., Quang, H. H., Doan, H. T. T., Agatsuma, T., Le, T. H., (2018). Distribution status of hybrid types in large liver flukes, Fasciola species (Digenea: Fasciolidae), from ruminants and humans in Vietnam. Korean Journal of Parasitology, 56(5):453-461.
Periago, M. V., Valero, M. A., El Sayed, M., Ashrafi, K., El Wakeel, A., Mohamed, M. Y. and Mas-Coma, S. (2008). First phenotypic description of Fasciola hepatica/Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infection. Genetics and Evolution journal. 8(1):51-58.
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. Parasitology Research, 105(3): 809-815
Prasad, P. K., Tandon, V., Biswal, D. K., Goswami, L. M., Chatterjee, A. (2008). Molecular identification of the Indian liver fluke, Fasciola (Trematoda: Fasciolidae) based on the ribosomal internal transcribed spacer regions. Parasitology Research. 103(6):1247–1255. DOI: 10.1007/s00436-008-1121-2
Qureshi, A. W., Tanveer, A., Qureshi, S. W., Maqbool, A., Gill, T. J., Ali, S. A. (2005). Epidemiology of human fasciolosis in rural areas of Lahore, Pakistan. Punjab University Journal of Zoology, 20:159-168.
Ramadhan, C. A., (2021). Prevalence of Fasciola species and their molecular characterization using ITS1 and ITS2 in Duhok province, Kurdistan region, Iraq. M.Sc. Thesis, Faculty of Science, Zakho university.
Raoof H. S., Marif, H. F., Rahman, H. S., Omar, M., Sheikh, B., San Ahmed, A. M. (2020). Molecular characterization and phylogenetic analysis of Fasciola species in sheep and goats in Sulaymaniyah Province, Northern Iraq. Journal of Zankoy Sulaimani, 22(1):297-305. https://www.researchgate.net/publication/342926652
Rokni, M., Mirhendi, H., Mizani, A., et al. (2010). Identification and differentiation of Fasciola hepatica and F. gigantica using a simple PCR-restriction enzyme method. Experimental Parasitology, 124:209-213.
Saki, J., Khademvatan, S., Yousefi, E. (2011). Mo-lecular identification of animal Fasciola isolates in Southwest of Iran. Australian Journal of Basic and Applied Sciences, 5:1878-1883.
Shafiei, R., Sarkari, B., Moshfe, A. (2013). A Consistent PCR-RFLP Assay Based on ITS-2 Ribosomal DNA for Differentiation of Fasciola Species. Iranian Journal of Basic Medical Sciences, 16(12):1266-1269.
Simsek, A., Utuk,A. E., and Balkaya, I. (2011). Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica. Helminthologia Journal, 48(1):3-7.
Tamura, K., Stecher, G., and Kumar, S. (2021). MEGA 11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology Evolution, 38(7):3022-3027. https://doi.org/10.1093/molbev/msab120.
Turhan, O., Korkmaz, M., Saba, R., Kabaaaliogu, A., Inan, D., Mamikoglu, L. (2006). Seroepidemiology of fascioliasis in the Antalya region and uselessness of eosinophil count as a surrogate marker and portable ultrasonography for epidemiological surveillance. Infezioni in Medicina, 14:208-212.
World Health Organization (WHO). Trematode infections/fascioliasis/fascioliasis diagnosis/en/. 2013. http://www.who.int/foodborne
Published
2023-01-24
How to Cite
REKANI, A. M. A., & MERO, W. M. (2023). MOLECULAR IDENTIFICATION AND PHYLOGENETIC ANALYSIS OF FASCIOLA SPECIES ISOLATED FROM SHEEP AND CATTLE USING THE ITS2 rDNA MARKER IN DUHOK PROVINCE, KURDISTAN REGION, IRAQ. Journal of Duhok University, 25(2), 286-297. https://doi.org/10.26682/ajuod.2022.5.2.26
Section
Agriculture and Veterinary Science
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