Identification and Phylogenetic Classification of Fasciola species Isolated from Sheep and Cattle by PCR-RFLP in Zabol, in Sistan and Baluchistan Province, Southeast Iran
-
Sedighe MIR
,
Mansour DABIRZADEH
,
Mohammad Bagher ROKNI
,
Mojgan ARYAEIPOUR
,
Mahdi KHOSHSIMA SHAHRAKI
,
Hakim AZIZI
Abstract
Background: The detection of Fasciola species in various geographical regions is essential for health policymaking. Here, we aimed to identify livestock (cattle and sheep) related Fasciola genotypes by restriction fragment length polymorphism PCR.
Methods: Seventy adult Fasciola flukes were collected from 70 infected livers of 35 cattle and 35 sheep slaughtered in Zabol abattoir, outh-east Iran (Jan-Jul 2017). Fasciola species were determined based on molecular features. For molecular detection, Fasciola ITS1 region was amplified and sequenced. A 700 bp fragment was amplified. These were digested with RasΙ enzyme. F. hepatica specific fragments were 47, 59, 68, 104, and 370, while those related to F. gigantica had 45, 55, 170, 370.
Results: The two main species of F. hepatica and F. gigantica are responsible for fasciolosis in sheep and cattle in our region. From 35 Fasciola isolated from cattle, 3 and 32 were F. hepatica and F. giagantica respectively. From 35 Fasciola isolated from sheep, 4 were F. hepatica and 31 were F. gigantica.
Conclusion: All Seventy Fasciola samples from two different hosts (cattle and sheep) were identified as either F. hepatica or F. gigantica by PCR-RFLP. Genotypic variability of Fasciola species was high in our region. It is recommended to assess molecular variation of Fasciola isolates in other host livestock.
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13. Ashrafi K, Valero MA, Panova M et al (2006). Phenotypic analysis of adults of Fasciola hepatica, F. gigantica and intermedi-ate forms from the endemic region of Gilan Iran. Parasitol Int, 55:249-260.
14. Maley LE, Marshall CR (1998). The coming of age of molecular systematics. Science, 279:505-6.
15. Hwang UW, Kim W (1999). General prop-erties and phylogenetic utilities of nuclear ribosomal DNA and mitochondrial DNA commonly used in molecular sys-tematics. Korean J Parasitol, 37:215-28.
16. Aryaeipour M, Rouhani S, Bandehpour M et al (2014). Genotyping and Phylogenetic Analysis of Fasciola spp. Isolated from sheep and Cattle Using PCR-RFLP in Ardabil Province, North western Iran. Iran J Public Health, 43(10):1364-1371.
17. Shoriki T, Ichikawa-Seki M, Suganuma K et al (2016). Novel methods for the molecu-lar discrimination of Fasciola spp. on the basis of nuclear protein –coding genes. Parasitol Int, 65(3):180-183.
18. Semyenova SK, Morozova EV, Chrisanfova GG et al (2006). Genetic differentiation in Eastern European and Western Asian populations of the liver fluke, Fasciola he-patica, as revealed by mitochondrial nad1 and cox1 genes. J Parasitol, 92:525–30.
19. Le TH, Blair D, McManus DP (2001). Complete DNA sequence and gene or-ganization of the mitochondrial genome of the liver fluke, Fasciola hepatica L. (Platy-helminthes; Trematoda). J Parasitol, 123:609–21.
20. Marcilla A, Bargues MD, Mas-Coma S (2002). A PCR-RFLP assay for the dis-tinction between Fasciola hepatica and Fasci-ola gigantica. Mol Cell Probes, 16:327–33.
21. Alasaad S, Soriguer RC, Abu-Madi M et al (2011). A fluorescence-based polymerase chain reaction-linked singlestrand con-formation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Parasitol Res, 108:1513–17.
22. Ai L, Dong SJ, Zhang WY et al (2010). Spe-cific PCR-based assays for the identifica-tion of Fasciola species: their develop-ment, evaluation and potential usefulness in prevalence surveys. Ann Trop Med Para-sitol, 104:65–72.
23. Gandomkar A, Khademolhosseini A (2009). Study on seasonal precipitation changes in the Zabol. J Amayesh Mohit; 2(6): 65-76.
24. Karimi A (2008). Genetic diagnosis of Fascio-la species based on 18S ribosomal DNA sequences. J Biol Sci, 8:1166–1173.
25. Saki J, Khademvatan S, Yousefi E (2011). Molecular identification of animal Fasciola isolates in Southwest of Iran. Aust J Basic Appl Sci, 5:1878-1883.
26. 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.
27. Mirahmadi H, Bigleri P, Sekandarpour S et al (2018). Molecular and phylogenetic char-acterisation of Fasciola spp. isolated from cattle and sheep in southeastern Iran. Bul-garian J Vet Med, 21(1): 86-93.
28. Shahbakhsh M, Nabavi R, Ganjali M (2016). Molecular Characterization of Fasciola Samples Using Sequences of Second In-ternal Transcribed Spacer-rDNA in dif-ferent Geographical Localities of Sistan and Balouchestan Province, Iran. Int J En-teric Pathog, 4(1):e33362.
29. Ghavami M.B, Rahimi P, Haniloo A, Mo-savinasab SN (2009). Genotypic and Phenotypic Analysis of Fasciola Isolates. Iran J Parasitol, 4: 61-670.
30. Rokni M, Mirhendi H, Behnia M, Jalalizand N (2010). Molecular characterization of Fasciola hepatica isolates by RAPD-PCR and ribosomal ITS1 sequencing. Iran Red Cresc Med J,12:27-32.
2. MAS CS, Bargues MD (1997). Human liver flukes: a review. Res Rev Parasitol, 57(3-4):145-218
3. Amor N, Halajian A, Farjallah S et al (2011). Molecular characterization of Fasciola spp. from the endemic area of north Iran based on nuclear ribosomal DNA se-quences. Exp Parasitol, 128(3):196 -204.
4. WHO (2012). Accelerating Work to Over-come the Global Impact of Neglected Tropical Diseases; A Roadmap for Im-plementation- Executive Summary.
5. Keiser J, Utzinger J (2009). Food-borne trematodiases. Clin Microbiol Rev, 22(3):466-86.
6. Rokni MB (2008). The present status of human helminthic diseases in Iran. Ann Trop Med Parasitol, 102(4): 283-295.
7. Massoud J (1989). Fascioliasis outbreak in man and drug test (triclabendazole) in Caspian littoral, northern part of Iran. Bull Soc Fran Parasitol, 8:438.
8. Asmar M, Milani A, Yadgari D, Amirkhani A (1991). Seroepidemiological investiga-tion of fascioliasis in north Iran. Med J IR Iran, 5:23-27.
9. Massoud J (1993). Present status of fasciolosis in Iran. Document SCH/SG/93/WP. Ge-neva: Word Health Organization.
10. World Health Organization (1995). Control of Food-borne Trematode Infections. No.849 Technical Report Series.
11. Rokni M, Massoud J, Neill S et al (2002). Diagnosis of Human fasciolosis in the Gilan Province of northern Iran: applica-tion of cathepsin L-ELISA. Diagn Microbi-ol Infect Dis, 44:175-179.
12. 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. Exp Parasitol, 124:209-13.
13. Ashrafi K, Valero MA, Panova M et al (2006). Phenotypic analysis of adults of Fasciola hepatica, F. gigantica and intermedi-ate forms from the endemic region of Gilan Iran. Parasitol Int, 55:249-260.
14. Maley LE, Marshall CR (1998). The coming of age of molecular systematics. Science, 279:505-6.
15. Hwang UW, Kim W (1999). General prop-erties and phylogenetic utilities of nuclear ribosomal DNA and mitochondrial DNA commonly used in molecular sys-tematics. Korean J Parasitol, 37:215-28.
16. Aryaeipour M, Rouhani S, Bandehpour M et al (2014). Genotyping and Phylogenetic Analysis of Fasciola spp. Isolated from sheep and Cattle Using PCR-RFLP in Ardabil Province, North western Iran. Iran J Public Health, 43(10):1364-1371.
17. Shoriki T, Ichikawa-Seki M, Suganuma K et al (2016). Novel methods for the molecu-lar discrimination of Fasciola spp. on the basis of nuclear protein –coding genes. Parasitol Int, 65(3):180-183.
18. Semyenova SK, Morozova EV, Chrisanfova GG et al (2006). Genetic differentiation in Eastern European and Western Asian populations of the liver fluke, Fasciola he-patica, as revealed by mitochondrial nad1 and cox1 genes. J Parasitol, 92:525–30.
19. Le TH, Blair D, McManus DP (2001). Complete DNA sequence and gene or-ganization of the mitochondrial genome of the liver fluke, Fasciola hepatica L. (Platy-helminthes; Trematoda). J Parasitol, 123:609–21.
20. Marcilla A, Bargues MD, Mas-Coma S (2002). A PCR-RFLP assay for the dis-tinction between Fasciola hepatica and Fasci-ola gigantica. Mol Cell Probes, 16:327–33.
21. Alasaad S, Soriguer RC, Abu-Madi M et al (2011). A fluorescence-based polymerase chain reaction-linked singlestrand con-formation polymorphism (F-PCR-SSCP) assay for the identification of Fasciola spp. Parasitol Res, 108:1513–17.
22. Ai L, Dong SJ, Zhang WY et al (2010). Spe-cific PCR-based assays for the identifica-tion of Fasciola species: their develop-ment, evaluation and potential usefulness in prevalence surveys. Ann Trop Med Para-sitol, 104:65–72.
23. Gandomkar A, Khademolhosseini A (2009). Study on seasonal precipitation changes in the Zabol. J Amayesh Mohit; 2(6): 65-76.
24. Karimi A (2008). Genetic diagnosis of Fascio-la species based on 18S ribosomal DNA sequences. J Biol Sci, 8:1166–1173.
25. Saki J, Khademvatan S, Yousefi E (2011). Molecular identification of animal Fasciola isolates in Southwest of Iran. Aust J Basic Appl Sci, 5:1878-1883.
26. 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.
27. Mirahmadi H, Bigleri P, Sekandarpour S et al (2018). Molecular and phylogenetic char-acterisation of Fasciola spp. isolated from cattle and sheep in southeastern Iran. Bul-garian J Vet Med, 21(1): 86-93.
28. Shahbakhsh M, Nabavi R, Ganjali M (2016). Molecular Characterization of Fasciola Samples Using Sequences of Second In-ternal Transcribed Spacer-rDNA in dif-ferent Geographical Localities of Sistan and Balouchestan Province, Iran. Int J En-teric Pathog, 4(1):e33362.
29. Ghavami M.B, Rahimi P, Haniloo A, Mo-savinasab SN (2009). Genotypic and Phenotypic Analysis of Fasciola Isolates. Iran J Parasitol, 4: 61-670.
30. Rokni M, Mirhendi H, Behnia M, Jalalizand N (2010). Molecular characterization of Fasciola hepatica isolates by RAPD-PCR and ribosomal ITS1 sequencing. Iran Red Cresc Med J,12:27-32.
| Files | ||
| Issue | Vol 48 No 5 (2019) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijph.v48i5.1811 | |
| Keywords | ||
| ITS1 PCR-RFLP Genotyping Fasciola hepatica Fasciola gigantica Iran | ||
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How to Cite
1.
MIR S, DABIRZADEH M, ROKNI MB, ARYAEIPOUR M, KHOSHSIMA SHAHRAKI M, AZIZI H. 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. 2019;48(5):934-942.
