Molecular Characterization of Fasciola spp. from a Donkey (Equus asinus) Using Partial Sequencing of cox1 and nad1
,
Abstract
Background: Fasciola hepatica as an important parasite affects health of humans and animals in some tropical and subtropical areas of the world, including Iran. Little is known about the molecular diversity of Fasciola in Equidae. Therefore, this study aimed to characterize the genetic polymorphisms among parasites.
Methods: Eight adult Fasciola spp. isolates were collected from a working donkey after necropsy in Shiraz, southwestern Iran, in 2018. Primarily, various parameters were measured morphologically. Subsequently, DNA was extracted from each fluke and molecular markers of cytochrome C oxidase (cox1) and NADH dehydrogenase 1(nad1) from individual Fasciola isolates were amplified using PCR assay and sequence data were employed for molecular and phylogenetic analysis. Genetic diversity between isolates was evaluated by comparing the sequences of these two mitochondrial regions.
Results: Based on the morphological and analyzed mitochondrial sequences, all of eight donkey isolates (100%) were identified as F. hepatica. Moreover, nine and five nucleotide polymorphisms were identified in the cox1and nad1 region sequences, respectively.
Conclusion: Accordingly, phylogenetic data revealed five and four haplotypes among donkey isolates based on the cox1and nad1 markers. Similarly, some of these haplotypes have been previously reported from different host species in Iran as well as all around the world.
1. Mas-Coma S, Bargues M, Valero M. Diagnosis of human fascioliasis by stool and blood techniques: update for the present global scenario. Parasitology. 2014;141(14):1918-46.
2. Mas-Coma S, Bargues M, Valero M. Human fascioliasis infection sources, their diversity, incidence factors, analytical methods and prevention measures. Parasitology. 2018;145(13):1665-99.
3. Takele B, Nibret E. Prevalence of gastrointestinal helminthes of donkeys and mules in and around Bahir Dar, Ethiopia. Ethiop Vet J. 2013;17(1):13.
4. Alves RM, Van Rensburg L, Van Wyk JA. Fasciola in horses in the Republic of South Africa: a single natural case of Fasciola hepatica and the failure to infest ten horses either with F. hepatica or Fasciola gigantica. Onderstepoort J Vet Res. 1988; 55(3):157-63.
5. Soykan E, Öge H. The prevalence of liver trematodes in equines in different cities of Turkey. Turkiye Parazitol Derg. 2012;36(3):152-5.
6. Getachew M, Innocent G, Trawford A, et al. Epidemiological features of fasciolosis in working donkeys in Ethiopia. Vet Parasitol. 2010;169(3-4):335-9.
7. Acici M, Bolukbas CS, Gurler AT, et al. Seroprevalence of fasciolosis in equines of the Black Sea Region, Turkey. J Equine Vet Sci. 2013;33(1):62-6.
8. Bargues MD, Gayo V, Sanchis J, et al. DNA multigene characterization of Fasciola hepatica and Lymnaea neotropica and its fascioliasis transmission capacity in Uruguay, with historical correlation, human report review and infection risk analysis. PLoS Negl Trop Dis. 2017;11(2):e0005352.
9. Mas‐Coma S, Valero MA, Bargues MD. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv Parasitol. 2009;69:41-146.
10. Quigley A, Sekiya M, Egan S, et al. Prevalence of liver fluke infection in Irish horses and assessment of a serological test for diagnosis of equine fasciolosis. Equine Vet J. 2017;49(2):183-8.
11. Shafiei R, Sarkari B, Sadjjadi SM, et al. Molecular and morphological characterization of Fasciola spp. isolated from different host species in a newly emerging focus of human fascioliasis in Iran. Vet Med Int. 2014;2014:405740.
12. Diyana J, Mahiza M I, Latiffah H, et al. Occurrence, Morphometric, and Molecular Investigation of Cat-tle and Buffalo Liver Adult Fluke in Peninsular Ma-laysia Main Abattoirs. J Parasitol Res. 2020;2020:5436846.
13. Gordy MA, Locke SA, Rawlings TA, et al. Molecu-lar and morphological evidence for nine species in North American Australapatemon (Sudarikov, 1959): a phylogeny expansion with description of the zygocercous Australapatemon mclaughlini n. sp. Parasitol Res. 2017;116(8): 2181-98.
14. Moazeni M, Sharifiyazdi H, Izadpanah A. Charac-terization of Fasciola hepatica genotypes from cattle and sheep in Iran using cytochrome C oxidase gene (CO1). Parasitol Res. 2012;110(6):2379-84.
15. Itagaki T, Kikawa M, Sakaguchi K, et al. Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. Parasitology. 2005;131(Pt 5):679-85.
16. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33(7):1870-4.
17. Haridy F, Morsy G, Abdou N, et al. Zoonotic fascioliasis in donkeys: ELISA (Fges) and postmortum examination in the Zoo, Giza, Egypt. J Egypt Soc Parasitol. 2007;37(3):1101-10.
18. Mas-Coma S, Rodriguez A, Bargues M, et al. Secondary reservoir role of domestic animals other than sheep and cattle in fascioliasis transmission in the Northern Bolivian Altiplano. Res Rev Parasitol. 1997;57(1):39-46.
19. WHO Report, Informal Meeting on use of triclabendazole in fascioliasis control. World Health Organization, Headquarters, Geneva, Switzerland. 2006;17:18.
20. Lin R, Dong S, 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. 2007;101(3):813-7.
21. Tadayon S, Sharifiyazdi H, Moazeni M, et al. Molecular differentiation of Fasciola species and characterization of genetic diversity of F. gigantica using NADH dehydrogenase I (ND1) gene in the endemic areas of Iran. Iran J Parasitol . 2015;10(1):9-18.
22. Amor N, Farjallah S, Said K, et al. First report of Fasciola hepatica in Equus caballus host species from Tunisia based on the ribosomal internal transcribed spacer regions. Turk J Vet Anim Sci. 2011;35(5):319-24.
23. Semyenova SK, Morozova EV, Chrisanfova GG, et al. Genetic differentiation in eastern European and western Asian populations of the liver fluke, Fasciola hepatica, as revealed by mitochondrial nad1 and cox1 genes. J Parasitol. 2006;92(3):525-30.
24. Valero M, Panova M, Mas-Coma S. Phenotypic analysis of adults and eggs of Fasciola hepatica by computer image analysis system. J Helminthol. 2005;79(3):217-25.
25. Yoshida Y, Nakanishi Y, Miyamoto M, et al. A case report of human infection with Fasciola sp. and its pharmacotherapy. Jpn J Parasitol. 1960;9:711-6.
26. Periago MV, Valero MA, El Sayed M, et al. First phenotypic description of Fasciola hepatica/Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infect Genet Evol. 2008;8(1):51-58.
27. Aryaeipour M, Bozorgomid A, Kazemi B, et al. Molecular and Morphometrical characterization of Fasciola species isolated from domestic ruminants in Ardabil Province, Northwestern Iran. Iran J Public Health. 2017; 46(3):318-325.
28. Yakhchali M, Malekzadeh-Viayeh R, Imani-Baran A, et al. Morphological and molecular discrimination of Fasciola species isolated from domestic ruminants of Urmia city, Iran. Iran J Parasitol. 2015; 10(1):46-55.
29. Sharifiyazdi H, Moazeni M, Rabbani F. Molecular characterization of human Fasciola samples in Gilan province, Northern Iran on the basis of DNA sequences of ribosomal and mitochondrial DNA genes. Comp Clin Pathol. 2012;21(5):889-94.
30. Yakhchali M, Bahramnejad K. Morphologic and morphometric variations of the adult and the eggs of frequent Fasciola species from domestic rumi-nants of North West of Iran. Iran J Vet Sci Technol. 2016; 7(2):75-83.
31. Valero MA, Darce NA, Panova M, et al. Relation-ships between host species and morphometric pat-terns in Fasciola hepatica adults and eggs from the Northern Bolivian Altiplano hyperendemic region. Vet Parasitol. 2001; 102(1-2):85-100.
32. Bozorgomid A, Rouhani S, Harandi MF, et al. Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies.Vet Parasitol Reg Stud Reports. 2020;19:100359.
33. Reaghi S, Haghighi A, Harandi MF, et al. 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. Vet World. 2016;9(9):1034-38.
34. Rouhani S, Raeghi S, Mirahmadi H, et al. Identification of Fasciola spp. in the east of Iran, based on the spermatogenesis and nuclear ribosomal DNA (ITS1) and mitochondrial (ND1) genes. Arch Clin Infect Dis. 2017;12(2):572-83.
35. Sarkari B, Parhoode M, Khabisi SA, et al. Genetic diversity of Fasciola spp. isolates from northern part of Iran: comparison with southwestern isolates. J Parasit Dis. 2017;41(3):768-72.
36. Constantine CC. Importance and pitfalls of molecular analysis to parasite epidemiology. Trends Parasitol. 2003;19(8):346-8.
2. Mas-Coma S, Bargues M, Valero M. Human fascioliasis infection sources, their diversity, incidence factors, analytical methods and prevention measures. Parasitology. 2018;145(13):1665-99.
3. Takele B, Nibret E. Prevalence of gastrointestinal helminthes of donkeys and mules in and around Bahir Dar, Ethiopia. Ethiop Vet J. 2013;17(1):13.
4. Alves RM, Van Rensburg L, Van Wyk JA. Fasciola in horses in the Republic of South Africa: a single natural case of Fasciola hepatica and the failure to infest ten horses either with F. hepatica or Fasciola gigantica. Onderstepoort J Vet Res. 1988; 55(3):157-63.
5. Soykan E, Öge H. The prevalence of liver trematodes in equines in different cities of Turkey. Turkiye Parazitol Derg. 2012;36(3):152-5.
6. Getachew M, Innocent G, Trawford A, et al. Epidemiological features of fasciolosis in working donkeys in Ethiopia. Vet Parasitol. 2010;169(3-4):335-9.
7. Acici M, Bolukbas CS, Gurler AT, et al. Seroprevalence of fasciolosis in equines of the Black Sea Region, Turkey. J Equine Vet Sci. 2013;33(1):62-6.
8. Bargues MD, Gayo V, Sanchis J, et al. DNA multigene characterization of Fasciola hepatica and Lymnaea neotropica and its fascioliasis transmission capacity in Uruguay, with historical correlation, human report review and infection risk analysis. PLoS Negl Trop Dis. 2017;11(2):e0005352.
9. Mas‐Coma S, Valero MA, Bargues MD. Fasciola, lymnaeids and human fascioliasis, with a global overview on disease transmission, epidemiology, evolutionary genetics, molecular epidemiology and control. Adv Parasitol. 2009;69:41-146.
10. Quigley A, Sekiya M, Egan S, et al. Prevalence of liver fluke infection in Irish horses and assessment of a serological test for diagnosis of equine fasciolosis. Equine Vet J. 2017;49(2):183-8.
11. Shafiei R, Sarkari B, Sadjjadi SM, et al. Molecular and morphological characterization of Fasciola spp. isolated from different host species in a newly emerging focus of human fascioliasis in Iran. Vet Med Int. 2014;2014:405740.
12. Diyana J, Mahiza M I, Latiffah H, et al. Occurrence, Morphometric, and Molecular Investigation of Cat-tle and Buffalo Liver Adult Fluke in Peninsular Ma-laysia Main Abattoirs. J Parasitol Res. 2020;2020:5436846.
13. Gordy MA, Locke SA, Rawlings TA, et al. Molecu-lar and morphological evidence for nine species in North American Australapatemon (Sudarikov, 1959): a phylogeny expansion with description of the zygocercous Australapatemon mclaughlini n. sp. Parasitol Res. 2017;116(8): 2181-98.
14. Moazeni M, Sharifiyazdi H, Izadpanah A. Charac-terization of Fasciola hepatica genotypes from cattle and sheep in Iran using cytochrome C oxidase gene (CO1). Parasitol Res. 2012;110(6):2379-84.
15. Itagaki T, Kikawa M, Sakaguchi K, et al. Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. Parasitology. 2005;131(Pt 5):679-85.
16. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016;33(7):1870-4.
17. Haridy F, Morsy G, Abdou N, et al. Zoonotic fascioliasis in donkeys: ELISA (Fges) and postmortum examination in the Zoo, Giza, Egypt. J Egypt Soc Parasitol. 2007;37(3):1101-10.
18. Mas-Coma S, Rodriguez A, Bargues M, et al. Secondary reservoir role of domestic animals other than sheep and cattle in fascioliasis transmission in the Northern Bolivian Altiplano. Res Rev Parasitol. 1997;57(1):39-46.
19. WHO Report, Informal Meeting on use of triclabendazole in fascioliasis control. World Health Organization, Headquarters, Geneva, Switzerland. 2006;17:18.
20. Lin R, Dong S, 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. 2007;101(3):813-7.
21. Tadayon S, Sharifiyazdi H, Moazeni M, et al. Molecular differentiation of Fasciola species and characterization of genetic diversity of F. gigantica using NADH dehydrogenase I (ND1) gene in the endemic areas of Iran. Iran J Parasitol . 2015;10(1):9-18.
22. Amor N, Farjallah S, Said K, et al. First report of Fasciola hepatica in Equus caballus host species from Tunisia based on the ribosomal internal transcribed spacer regions. Turk J Vet Anim Sci. 2011;35(5):319-24.
23. Semyenova SK, Morozova EV, Chrisanfova GG, et al. Genetic differentiation in eastern European and western Asian populations of the liver fluke, Fasciola hepatica, as revealed by mitochondrial nad1 and cox1 genes. J Parasitol. 2006;92(3):525-30.
24. Valero M, Panova M, Mas-Coma S. Phenotypic analysis of adults and eggs of Fasciola hepatica by computer image analysis system. J Helminthol. 2005;79(3):217-25.
25. Yoshida Y, Nakanishi Y, Miyamoto M, et al. A case report of human infection with Fasciola sp. and its pharmacotherapy. Jpn J Parasitol. 1960;9:711-6.
26. Periago MV, Valero MA, El Sayed M, et al. First phenotypic description of Fasciola hepatica/Fasciola gigantica intermediate forms from the human endemic area of the Nile Delta, Egypt. Infect Genet Evol. 2008;8(1):51-58.
27. Aryaeipour M, Bozorgomid A, Kazemi B, et al. Molecular and Morphometrical characterization of Fasciola species isolated from domestic ruminants in Ardabil Province, Northwestern Iran. Iran J Public Health. 2017; 46(3):318-325.
28. Yakhchali M, Malekzadeh-Viayeh R, Imani-Baran A, et al. Morphological and molecular discrimination of Fasciola species isolated from domestic ruminants of Urmia city, Iran. Iran J Parasitol. 2015; 10(1):46-55.
29. Sharifiyazdi H, Moazeni M, Rabbani F. Molecular characterization of human Fasciola samples in Gilan province, Northern Iran on the basis of DNA sequences of ribosomal and mitochondrial DNA genes. Comp Clin Pathol. 2012;21(5):889-94.
30. Yakhchali M, Bahramnejad K. Morphologic and morphometric variations of the adult and the eggs of frequent Fasciola species from domestic rumi-nants of North West of Iran. Iran J Vet Sci Technol. 2016; 7(2):75-83.
31. Valero MA, Darce NA, Panova M, et al. Relation-ships between host species and morphometric pat-terns in Fasciola hepatica adults and eggs from the Northern Bolivian Altiplano hyperendemic region. Vet Parasitol. 2001; 102(1-2):85-100.
32. Bozorgomid A, Rouhani S, Harandi MF, et al. Genetic diversity and distribution of Fasciola hepatica haplotypes in Iran: Molecular and phylogenetic studies.Vet Parasitol Reg Stud Reports. 2020;19:100359.
33. Reaghi S, Haghighi A, Harandi MF, et al. 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. Vet World. 2016;9(9):1034-38.
34. Rouhani S, Raeghi S, Mirahmadi H, et al. Identification of Fasciola spp. in the east of Iran, based on the spermatogenesis and nuclear ribosomal DNA (ITS1) and mitochondrial (ND1) genes. Arch Clin Infect Dis. 2017;12(2):572-83.
35. Sarkari B, Parhoode M, Khabisi SA, et al. Genetic diversity of Fasciola spp. isolates from northern part of Iran: comparison with southwestern isolates. J Parasit Dis. 2017;41(3):768-72.
36. Constantine CC. Importance and pitfalls of molecular analysis to parasite epidemiology. Trends Parasitol. 2003;19(8):346-8.
| Files | ||
| Issue | Vol 15 No 4 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v15i4.4860 | |
| Keywords | ||
| Fasciola Haplotype Donkey Iran | ||
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How to Cite
1.
ASADPOUR M, SHARIFIYAZDI H, MOAZENI M, MALEKPOUR SH. Molecular Characterization of Fasciola spp. from a Donkey (Equus asinus) Using Partial Sequencing of cox1 and nad1. Iran J Parasitol. 2020;15(4):549-558.
