Molecular Characterization of Animal Fasciola Spp. Isolates from Lorestan Province, Western Iran
-
Peyman Heydarian
,
Vahid Jajarmi
,
Adel Spotin
,
Keyhan Ashrafi
,
Mehdi Mohebali
,
Mojgan Aryaeipour
,
Arezoo Bozorgomid
,
Elham Hajialilo
,
Mohammad Javad Abbaszadeh Afshar
,
Mandana Fadaei Tehrani
,
Mohammad Bagher Rokni
Abstract
Background: We aimed to detect the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Lorestan Province, western Iran using PCR-RFLP method. Besides, we evaluated the genetic diversity indices, sequencing and phylogenetic analysis using mitochondrial gene (ND1 and CO1).
Methods: PCR-RFLP analysis of ribosomal ITS1 fragment by RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (18 sheep, 21 cattle, and 17goats) was conducted. The samples were sequenced. Sequences were evaluated using BLAST software and the parasite species were identified with similarity percentage and overlap with the species registered in the gene bank. Then similarity and diversity of intra-species and intra-species diversity of Fasciola species were calculated.
Results: In Lorestan, based on RFLP pattern, 93% (52) of the Fasciola spp. isolates had a RFLP pattern related to F. hepatica and 7% (4) were F. gigantica. No hybrid forms were detected. The CO1 gene could clarify 19 haplotypes against ND1 gene that found 22 haplotypes among livestock. Sequencing results of the mtDNA showed intra-species identity 98. 5%-100% and Intra-species-diversity: 0-1.5% compared to the GenBank sequences.
Conclusion: Using PCR-RFLP method, two species of F. hepatica and F. gigantica, were present in Lorestan Province, but F. hepatica was more prevalent. Mitochondrial genes could better test variability indices in different hosts than ribosomal genes, consequently among mitochondrial genes, the ND1 gene could better examine differences and similarities than CO1.
1. Mas-Coma S, Valero MA, Bargues MD (2019). Fascioliasis. In: Digenetic Trematodes. Ed(s): Springer, pp. 71-103.
2. WHO (2013). Sustaining the drive to overcome the global impact of neglected tropical diseases: second WHO report on neglected diseases. ed. World Health Organization.
3. Torgerson PR, de Silva NR, Fèvre EM, et al (2014). The global burden of foodborne parasitic diseases: an update. Trends Parasitol, 30:20-26.
4. Mas-Coma S, Bargues MD, Valero M (2005). Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol, 35:1255-1278.
5. Ashrafi K, Valero M, Panova M, Periago 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. Parasitol Int, 55:249-260.
6. Aryaeipour M, Bozorgomid A, Kazemi B, Behnia M, Azizi H, Rokni MB (2017). Molecular and morphometrical characterization of Fasciola species isolated from domestic ruminants in Ardabil Province, Northwestern Iran. Iran J Public Health, 46:318-325.
7. Alasaad S, Soriguer RC, Abu-Madi M, et al (2011). A TaqMan real-time PCR-based assay for the identification of Fasciola spp. Vet Parasitol, 179:266-271.
8. Ali H, Ai L, Song H, Ali S, Lin R, Seyni B, Issa G, Zhu X (2008). Genetic characterisation of Fasciola samples from different host species and geographical localities revealed the existence of F. hepatica and F. gigantica in Niger. Parasitol Res, 102:1021-1024.
9. Hosseini-Safa A, Rokni MB, Mosawi SH, Heydarian P, Azizi H, Davari A, Aryaiepour M (2019). High-resolution melting analysis as an appropriate method to differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health, 48:501-507.
10. Itagaki T, Tsutsumi K-I, Ito K, Tsutsumi Y (1998). Taxonomic status of the Japanese triploid forms of Fasciola: comparison of mitochondrial ND1 and COI sequences with F. hepatica and F. gigantica. J Parasitol:445-448.
11. Aryaeipour M, Rouhani S, Bandehpour M, Mirahmadi H, Kazemi B, Rokni MB (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, 43:1364-1371.
12. Bozorgomid A, Nazari N, Rahimi H, et al (2016). Molecular characterization of animal Fasciola spp. isolates from Kermanshah, Western Iran. Iran J Public Health, 45:1315-21.
13. Hasanpour H, Falak R, Naddaf SR, et al (2020). Molecular characterization of Fasciola spp. From some parts of Iran. Iran J Public Health, 49:157-166.
14. 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, 5.8 S rDNA, ITS2). Iran J Parasitol, 6:35-42.
15. Rokni MB, Mirhendi H, Mizani A, et al (2010). Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method. Exp Parasitol, 124:209-213.
16. Ashrafi K (2015). The status of human and animal fascioliasis in Iran: a narrative review article. Iran J Parasitol, 10:306-328.
17. Hall TA (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic acids symposium series. [London]: Information Retrieval Ltd., c1979-c2000., pp. 95-98.
18. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 28:2731-39.
19. Librado P, Rozas J (2009). DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25:1451-52.
20. Shahnazi M, Ebadi M, Abbaspoor Z, Hajialilo E, Javadi A, Heydarian P, Saraei M, Alizadeh S (2020). Molecular characterization of Fasciola and Dicrocoelium species isolated from ruminant livestock in Qazvin, Iran. Infectious Disorders Drug Targets, 20(5):737-742.
21. Anh DN, Anh LT, Tuan LQ, et al (2018). Identification of Fasciola species isolates from Nghe An province, Vietnam, based on ITS1 Sequence of ribosomal DNA using a simple PCR-RFLP method. J Parasitol Res, 2018: 2958026.
22. Ichikawa M, Itagaki T (2010). Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method. Parasitol Res, 106:757-761.
23. Sedighe M, Dabirzadeh M, Rokni MB, Aryaeipour M, Shahraki MK, 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. Iran J Public Health, 48:934-942.
24. Siribat P, Dekumyoy P, Komalamisra C, Sumruayphol S, Thaenkham U (2018). Molecular identification of Fasiola spp. representative samples from Thailand based on PCR-RFLP. J Trop Med Parasitol, 41:1-7.
25. Farjallah S, Sanna D, Amor N, et al (2009). Genetic characterization of Fasciola hepatica from Tunisia and Algeria based on mitochondrial and nuclear DNA sequences. Parasitol Res, 105:1617.
26. Rahimi P, Ghavami M, Haniloo A, Nourian A, Biglari A (2009). Identification of Fasciola Species by PCR-RFLP Assay. ZUMS Journal, 16:41-48.
27. Shafiei R, Sarkari B, Sadjjadi SM, Mowlavi GR, Moshfe A (2014). 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.
28. Shahbazi A, Akbarimoghaddam M, Izadi S, Ghazanchaii A, Jalali N, Bazmani A (2011). Identification and genetic variation of Fasciola species from Tabriz, North-Western Iran. Iran J Parasitol, 6:52-59.
29. Sharifiyazdi H, Moazeni M, Rabbani F (2012). 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 Path, 21:889-894.
30. Agatsuma T, Arakawa Y, Iwagami M, et al (2000). Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. Parasitol Int, 49:231-238.
31. Amor N, Farjallah S, Salem M, Lamine DM, Merella P, Said K, Slimane BB (2011). Molecular characterization of Fasciola gigantica from Mauritania based on mitochondrial and nuclear ribosomal DNA sequences. Exp Parasitol, 129:127-136.
32. Chaudhry U, Van Paridon B, Shabbir M, Shafee M, Ashraf K, Yaqub T, Gilleard J (2016). Molecular evidence shows that the liver fluke Fasciola gigantica is the predominant Fasciola species in ruminants from Pakistan. J Helminthol, 90:206-213.
33. Amer S, ElKhatam A, Zidan S, Feng Y, Xiao L (2016). Identity of Fasciola spp. in sheep in Egypt. Parasites & Vectors, 9:623.
34. Ashrafi K, Massoud J, Holakouei K, et al (2004). Evidence suggesting that Fasciola gigantica might be the most prevalent causal agent of fascioliasis in northern Iran. Iran J Public Health, 33:31-37.
35. Itagaki T, Kikawa M, Terasaki K, Shibahara T, 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, 67:1115-1118.
36. 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.
37. Chaichanasak P, Ichikawa M, Sobhon P, 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, 61:545-549.
38. Dar Y, Amer S, Mercier A, Courtioux B, Dreyfuss G (2012). Molecular identification of Fasciola spp.(digenea: Fasciolidae) in Egypt. Parasite, 19:177-82.
39. Ichikawa M, Bawn S, Maw NN, et al (2011). Characterization of Fasciola spp. in Myanmar on the basis of spermatogenesis status and nuclear and mitochondrial DNA markers. Parasitol Int, 60:474-479.
40. Simsek S, Utuk A, Balkaya I (2011). Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica. Helminthologia, 48:3-7.
41. Constantine CC (2003). Importance and pitfalls of molecular analysis to parasite epidemiology. Trends Parasitol, 19:346-348.
42. Ichikawa-Seki M, Ortiz P, Cabrera M, Hobán C, Itagaki T (2016). Molecular characterization and phylogenetic analysis of Fasciola hepatica from Peru. Parasitol Int, 65:171-174.
43. Ichikawa-Seki M, Tokashiki M, Opara MN, et al (2017). Molecular characterization and phylogenetic analysis of Fasciola gigantica from Nigeria. Parasitol Int, 66:893-897.
44. Itagaki T, Kikawa M, Sakaguchi K, Shimo J, Terasaki K, Shibahara T, 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.
45. Reaghi 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 the North-East of Iran. Vet World, 9:1034-38.
46. Carnevale S, Malandrini JB, Pantano ML, et al (2017). First genetic characterization of Fasciola hepatica in Argentina by nuclear and mitochondrial gene markers. Vet Parasitol, 245:34-38.
47. Rouhani S, Raeghi S, Mirahmadi H, Harandi MF, Haghighi A, Spotin A (2017). 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, 12:e57283.
2. WHO (2013). Sustaining the drive to overcome the global impact of neglected tropical diseases: second WHO report on neglected diseases. ed. World Health Organization.
3. Torgerson PR, de Silva NR, Fèvre EM, et al (2014). The global burden of foodborne parasitic diseases: an update. Trends Parasitol, 30:20-26.
4. Mas-Coma S, Bargues MD, Valero M (2005). Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol, 35:1255-1278.
5. Ashrafi K, Valero M, Panova M, Periago 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. Parasitol Int, 55:249-260.
6. Aryaeipour M, Bozorgomid A, Kazemi B, Behnia M, Azizi H, Rokni MB (2017). Molecular and morphometrical characterization of Fasciola species isolated from domestic ruminants in Ardabil Province, Northwestern Iran. Iran J Public Health, 46:318-325.
7. Alasaad S, Soriguer RC, Abu-Madi M, et al (2011). A TaqMan real-time PCR-based assay for the identification of Fasciola spp. Vet Parasitol, 179:266-271.
8. Ali H, Ai L, Song H, Ali S, Lin R, Seyni B, Issa G, Zhu X (2008). Genetic characterisation of Fasciola samples from different host species and geographical localities revealed the existence of F. hepatica and F. gigantica in Niger. Parasitol Res, 102:1021-1024.
9. Hosseini-Safa A, Rokni MB, Mosawi SH, Heydarian P, Azizi H, Davari A, Aryaiepour M (2019). High-resolution melting analysis as an appropriate method to differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health, 48:501-507.
10. Itagaki T, Tsutsumi K-I, Ito K, Tsutsumi Y (1998). Taxonomic status of the Japanese triploid forms of Fasciola: comparison of mitochondrial ND1 and COI sequences with F. hepatica and F. gigantica. J Parasitol:445-448.
11. Aryaeipour M, Rouhani S, Bandehpour M, Mirahmadi H, Kazemi B, Rokni MB (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, 43:1364-1371.
12. Bozorgomid A, Nazari N, Rahimi H, et al (2016). Molecular characterization of animal Fasciola spp. isolates from Kermanshah, Western Iran. Iran J Public Health, 45:1315-21.
13. Hasanpour H, Falak R, Naddaf SR, et al (2020). Molecular characterization of Fasciola spp. From some parts of Iran. Iran J Public Health, 49:157-166.
14. 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, 5.8 S rDNA, ITS2). Iran J Parasitol, 6:35-42.
15. Rokni MB, Mirhendi H, Mizani A, et al (2010). Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR-restriction enzyme method. Exp Parasitol, 124:209-213.
16. Ashrafi K (2015). The status of human and animal fascioliasis in Iran: a narrative review article. Iran J Parasitol, 10:306-328.
17. Hall TA (1999). BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic acids symposium series. [London]: Information Retrieval Ltd., c1979-c2000., pp. 95-98.
18. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011). MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol, 28:2731-39.
19. Librado P, Rozas J (2009). DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25:1451-52.
20. Shahnazi M, Ebadi M, Abbaspoor Z, Hajialilo E, Javadi A, Heydarian P, Saraei M, Alizadeh S (2020). Molecular characterization of Fasciola and Dicrocoelium species isolated from ruminant livestock in Qazvin, Iran. Infectious Disorders Drug Targets, 20(5):737-742.
21. Anh DN, Anh LT, Tuan LQ, et al (2018). Identification of Fasciola species isolates from Nghe An province, Vietnam, based on ITS1 Sequence of ribosomal DNA using a simple PCR-RFLP method. J Parasitol Res, 2018: 2958026.
22. Ichikawa M, Itagaki T (2010). Discrimination of the ITS1 types of Fasciola spp. based on a PCR–RFLP method. Parasitol Res, 106:757-761.
23. Sedighe M, Dabirzadeh M, Rokni MB, Aryaeipour M, Shahraki MK, 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. Iran J Public Health, 48:934-942.
24. Siribat P, Dekumyoy P, Komalamisra C, Sumruayphol S, Thaenkham U (2018). Molecular identification of Fasiola spp. representative samples from Thailand based on PCR-RFLP. J Trop Med Parasitol, 41:1-7.
25. Farjallah S, Sanna D, Amor N, et al (2009). Genetic characterization of Fasciola hepatica from Tunisia and Algeria based on mitochondrial and nuclear DNA sequences. Parasitol Res, 105:1617.
26. Rahimi P, Ghavami M, Haniloo A, Nourian A, Biglari A (2009). Identification of Fasciola Species by PCR-RFLP Assay. ZUMS Journal, 16:41-48.
27. Shafiei R, Sarkari B, Sadjjadi SM, Mowlavi GR, Moshfe A (2014). 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.
28. Shahbazi A, Akbarimoghaddam M, Izadi S, Ghazanchaii A, Jalali N, Bazmani A (2011). Identification and genetic variation of Fasciola species from Tabriz, North-Western Iran. Iran J Parasitol, 6:52-59.
29. Sharifiyazdi H, Moazeni M, Rabbani F (2012). 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 Path, 21:889-894.
30. Agatsuma T, Arakawa Y, Iwagami M, et al (2000). Molecular evidence of natural hybridization between Fasciola hepatica and F. gigantica. Parasitol Int, 49:231-238.
31. Amor N, Farjallah S, Salem M, Lamine DM, Merella P, Said K, Slimane BB (2011). Molecular characterization of Fasciola gigantica from Mauritania based on mitochondrial and nuclear ribosomal DNA sequences. Exp Parasitol, 129:127-136.
32. Chaudhry U, Van Paridon B, Shabbir M, Shafee M, Ashraf K, Yaqub T, Gilleard J (2016). Molecular evidence shows that the liver fluke Fasciola gigantica is the predominant Fasciola species in ruminants from Pakistan. J Helminthol, 90:206-213.
33. Amer S, ElKhatam A, Zidan S, Feng Y, Xiao L (2016). Identity of Fasciola spp. in sheep in Egypt. Parasites & Vectors, 9:623.
34. Ashrafi K, Massoud J, Holakouei K, et al (2004). Evidence suggesting that Fasciola gigantica might be the most prevalent causal agent of fascioliasis in northern Iran. Iran J Public Health, 33:31-37.
35. Itagaki T, Kikawa M, Terasaki K, Shibahara T, 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, 67:1115-1118.
36. 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.
37. Chaichanasak P, Ichikawa M, Sobhon P, 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, 61:545-549.
38. Dar Y, Amer S, Mercier A, Courtioux B, Dreyfuss G (2012). Molecular identification of Fasciola spp.(digenea: Fasciolidae) in Egypt. Parasite, 19:177-82.
39. Ichikawa M, Bawn S, Maw NN, et al (2011). Characterization of Fasciola spp. in Myanmar on the basis of spermatogenesis status and nuclear and mitochondrial DNA markers. Parasitol Int, 60:474-479.
40. Simsek S, Utuk A, Balkaya I (2011). Molecular differentiation of Turkey cattle isolates of Fasciola hepatica and Fasciola gigantica. Helminthologia, 48:3-7.
41. Constantine CC (2003). Importance and pitfalls of molecular analysis to parasite epidemiology. Trends Parasitol, 19:346-348.
42. Ichikawa-Seki M, Ortiz P, Cabrera M, Hobán C, Itagaki T (2016). Molecular characterization and phylogenetic analysis of Fasciola hepatica from Peru. Parasitol Int, 65:171-174.
43. Ichikawa-Seki M, Tokashiki M, Opara MN, et al (2017). Molecular characterization and phylogenetic analysis of Fasciola gigantica from Nigeria. Parasitol Int, 66:893-897.
44. Itagaki T, Kikawa M, Sakaguchi K, Shimo J, Terasaki K, Shibahara T, 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.
45. Reaghi 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 the North-East of Iran. Vet World, 9:1034-38.
46. Carnevale S, Malandrini JB, Pantano ML, et al (2017). First genetic characterization of Fasciola hepatica in Argentina by nuclear and mitochondrial gene markers. Vet Parasitol, 245:34-38.
47. Rouhani S, Raeghi S, Mirahmadi H, Harandi MF, Haghighi A, Spotin A (2017). 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, 12:e57283.
| Files | ||
| Issue | Vol 51 No 8 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v51i8.10271 | |
| Keywords | ||
| Fascioliasis Fasciola Polymerase chain reaction Iran | ||
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How to Cite
1.
Heydarian P, Jajarmi V, Spotin A, Ashrafi K, Mohebali M, Aryaeipour M, Bozorgomid A, Hajialilo E, Abbaszadeh Afshar MJ, Fadaei Tehrani M, Rokni MB. Molecular Characterization of Animal Fasciola Spp. Isolates from Lorestan Province, Western Iran. Iran J Public Health. 2022;51(8):1847-1856.
