Genetic Identification of Echinococcus granulosus in Slaughtered Domestic Animals from Two Northeastern Iranian Cities Using HRM and Sequencing Techniques
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Abstract
Background: Hydatidosis is one of the common parasitic diseases common to both humans and animals with global spread. The disease is very important from both public health and economic perspectives. The parasite presents with highly diverse genetic features, including a different genotype which affect the life cycle of the parasite, its transmission routes, pathogenicity, immunogenicity, response to drugs, epidemiology and disease control. Therefore, we aimed to identify the genotype of hydatid cysts isolated from slaughtered animals in Sabzevar and Neyshabour slaughterhouses by sequencing and high-resolution melting curve analysis.
Methods: Out of 35 hydatid cyst samples, including 19 from sheep (15 liver and 4 lung samples) and 16 from cattle (9 liver and 7 lung samples) were included in this study. After the extraction of protoscoleces and DNA, a high-resolution melting curve analysis method was used to identify the genotype of the samples using pre-determined G1, G3, and G6 standard samples.
Results: Out of 35 hydatid cyst samples, 89.5% G1 and 10.5% G6 were detected in sheep samples, and 93.7% G1 and 6.3% G6 were detected in cattle samples. Results showed in the Sabzevar City, dominant genotype there was G1 and COX1 gene considered a perfect biomarker in a molecular identification and phylogenetic study of this parasite.
Conclusion: G1 was the most prevalent in samples of hydatid cysts (both sheep and cattle) in Sabzevar, while in Neyshabour samples, G1 and G6 were the most dominant types.
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22. Gholami S, Jahandar H, Abastabar M, et al. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran. Iran J Parasitol. 2016;11(2):186-194.
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24. Karamian M, Haghighi F, Hemmati M. Genotyping of Cystic Hydatidosis Agents in Birjand, Eastern Iran. Mod Care J. 2017; 14(3):e64531.
25. Sharbatkhori M, Mirhendi H, Jex AR et al. Genetic categorization of Echinococcus granulosus from humans and herbivorous hosts in Iran using an integrated mutation scanning‐phylogenetic approach. Ele-ctrophoresis. 2009; 30(15):2648-55.
26. Vahedi A, Mahdavi M, Ghazanchaei A et al. Genotypic characteristics of hydatid cysts isolated from humans in East Azerbaijan Province (2011-2013). J Anal Res Clin Med. 2014; 2(3): 152-157.
27. Gholami Sh, Sosari M, Fakhar M et al. Molecular characterization of Echinococcus granulosus from Hydatid Cysts isolated from human and animals in Golestan Province, North of Iran. Iran J Parasitol. 2012; 7(4):8-16.
28. Sadjjadi SM, Mikaeili F, Karamian M, et al. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans. Int J Parasitol. 2013;43(11):875-7.
29. Abolhasani M, Ebrahimzadeh E, Borji H. Morphometrical and Molecular Identification of Echinococcus granulosus Genotypes in peri-urban wild dogs from an endemic focus in Northwest of Iran. Arch Razi Inst. 2024; 79(4): 721-726.
30. Arbabi M, Pestechian N, Tavasol Khamseh H, et al. Molecular and genotyping identification of Echinococcus granulosus from camel and dog isolates in Isfahan, Iran (2015-2016). KAUMS Journal (FEYZ). 2017; 21(2):134-41.
31. Shamsi M DA, Khosravi A, Ghafarifar F. Identification and sequencing of genotypes of Echinococcus granulosus isolates in stray dogs using Cox gene in Ilam, western Iran. Journal of Ilam University of Medical Sciences. 2015;23(2)(10):111-9.
32. Jafarzadeh F RS, Abad M, Vafaei F. Genotyping and phylogenetic identification of Echinococcus granulosus from paraffin-embedded tissues of surgical hydatid cyst. Journal of North Khorasan University of Medical Sciences. 2018;9(4):15-19.
33. Abedi B, Maghsood AH, Khansarinejad B, et al. Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial COX1 gene, in the Markazi province, Iran. J Parasit Dis. 2019;43(4):592-6.
34. Rostami S, Talebi S, Babaei Z, et al. High resolution melting technique for molecular epidemiological studies of cystic echinococcosis: Differentiating G1, G3, and G6 genotypes of Echinococcus granulosus sensu lato. Parasitol Res. 2013; 112(10): 3441-7.
2. Jenkins DJ, Roming T, Thompson RCA. Emergence/re-emergence of Echinococcus SPP. Aglobal update. Int J Parasitol. 2005; 35(11-12):1205-19.
3. da Silva AM. Human echinococcosis: a neglected disease. Gastroenterol Res Pract. 2010; 2010:583297.
4. Casulli A. Recognising the substantial burden of neglected pandemics cystic and alveolar echinococcosis. Lancet Glob Health. 2020;8:e470–e471.
5. Thompson RC. Biology and Systematics of Echinococcus. Adv Parasitol. 2017; 95:65-109.
6. Eckert J, Deplazes P. Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev. 2004;17(1):107-35.
7. Eckert, J, Gemmell, M. A, Meslin, François-Xavier, Pawlowski, Z. S & World Health Organization. (2001). WHO/OIE manual on echinococcosis in humans and animals : a public health problem of global concern / edited by J. Eckert ... [et al.]. World Organisation for Animal Health.
8. Słomka M, Sobalska-Kwapis M, Wachulec M, Bartosz G, Strapagiel D. High Resolution Melting (HRM) for High-Throughput Genotyping-Limitations and Caveats in Practical Case Studies. Int J Mol Sci. 2017;18(11):2316.
9. Chou L, Huang SJ, Hsieh C, et al. A High Resolution Melting Analysis-Based Genotyping Toolkit for the Peach (Prunus persica) Chilling Requirement. Int J Mol Sci. 2020;21(4):1543.
10. Elyasi H, Farbodnia T, Javaheri E. Genetic Characteristics of Echinococcus granulosus from Fixed Paraffin-Embedded Tissue Samples in Human Isolates Based on the High-Resolution Melting Point Analysis in Sabzevar, Northeast Iran. Iran J Parasitol. 2024;19(3):305-313.
11. Orsten S, Boufana B, Ciftci T, et al. Human cystic echinococcosis in Turkey: a preliminary study on DNA polymorphisms of hydatid cysts removed from confirmed patients. Parasitol Res. 2018;117(4):1257-1263.
12. Berenji F, Shamsian SA, Nouri Daloee M, et al. Genotyping of Echinococcus granulosus Isolates from Human in Khorasan Province, North-Eastern Iran. Iran J Parasitol. 2019;14(1):52-58.
13. Nikmanesh B, Mirhendi H, Ghalavand Z, et al. Genotyping of Echinococcus granulosus Isolates from Human Clinical Samples Based on Sequencing of Mitochondrial Genes in Iran, Tehran. Iran J Parasitol. 2014;9(1):20-7
14. Abedi B, Maghsood AH, Khansarinejad B, et al. Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial COX1 gene, in the Markazi province, Iran. J Parasit Dis. 2019;43(4):592-596.
15. Dalimi A, Shamsi M, Khosravi A, Ghaffarifar F. Genotyping Echinococcus granulosus from Canine Isolates in Ilam Province, West of Iran. Iran J Parasitol. 2017;12(4):614-621.
16. Borji H, Bahar MM, Naghibi A, Masoom SHF. Genotyping of human cystic echinococcosis in northeastern Iran. Ann Parasitol. 2018;64(4):323-329.
17. Pezeshki A, Akhlaghi L, Sharbatkhori M, et al. Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol. 2013;87(4):387-91
18. Babaei Z, Taherkhani R, Barazesh A, et al. Genotyping and phylogenetic analysis of hydatid cysts isolated from livestock in Bushehr province, Iran. J Parasit Dis. 2021;45(1):197-203.
19. Zarei Z, Ghalehbin BM, Akhoundi B, et al. Genotyping of Echinococcus granulosus isolated from canine in Northwest Iran. J Parasit Dis. 2023; 47:757–761.
20. Shahbazi AE, Saidijam M, Maghsood AH, et al. Genotyping of Fresh and Parafinized Human Hydatid Cysts Using NAD1 and COX1 Genes in Hamadan Province, West of Iran. Iran J Parasitol. 2020;15(2):259-265.
21. Salehi M, Yaghfori S, Bahari P, et al. Molecular Characterization of Echinococcus granulosus Sensu Lato from Livestock in North Khorasan Province, Iran. Iran J Parasitol. 2018;13(4):577-586.
22. Gholami S, Jahandar H, Abastabar M, et al. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran. Iran J Parasitol. 2016;11(2):186-194.
23. Ghabdian S, Borji H, Naghibi A. Molecular identification of Echinococcus granulosus strain in stray dogs from Northeastern Iran. Vet Parasitol Reg Stud Reports. 2017; 9:6-8.
24. Karamian M, Haghighi F, Hemmati M. Genotyping of Cystic Hydatidosis Agents in Birjand, Eastern Iran. Mod Care J. 2017; 14(3):e64531.
25. Sharbatkhori M, Mirhendi H, Jex AR et al. Genetic categorization of Echinococcus granulosus from humans and herbivorous hosts in Iran using an integrated mutation scanning‐phylogenetic approach. Ele-ctrophoresis. 2009; 30(15):2648-55.
26. Vahedi A, Mahdavi M, Ghazanchaei A et al. Genotypic characteristics of hydatid cysts isolated from humans in East Azerbaijan Province (2011-2013). J Anal Res Clin Med. 2014; 2(3): 152-157.
27. Gholami Sh, Sosari M, Fakhar M et al. Molecular characterization of Echinococcus granulosus from Hydatid Cysts isolated from human and animals in Golestan Province, North of Iran. Iran J Parasitol. 2012; 7(4):8-16.
28. Sadjjadi SM, Mikaeili F, Karamian M, et al. Evidence that the Echinococcus granulosus G6 genotype has an affinity for the brain in humans. Int J Parasitol. 2013;43(11):875-7.
29. Abolhasani M, Ebrahimzadeh E, Borji H. Morphometrical and Molecular Identification of Echinococcus granulosus Genotypes in peri-urban wild dogs from an endemic focus in Northwest of Iran. Arch Razi Inst. 2024; 79(4): 721-726.
30. Arbabi M, Pestechian N, Tavasol Khamseh H, et al. Molecular and genotyping identification of Echinococcus granulosus from camel and dog isolates in Isfahan, Iran (2015-2016). KAUMS Journal (FEYZ). 2017; 21(2):134-41.
31. Shamsi M DA, Khosravi A, Ghafarifar F. Identification and sequencing of genotypes of Echinococcus granulosus isolates in stray dogs using Cox gene in Ilam, western Iran. Journal of Ilam University of Medical Sciences. 2015;23(2)(10):111-9.
32. Jafarzadeh F RS, Abad M, Vafaei F. Genotyping and phylogenetic identification of Echinococcus granulosus from paraffin-embedded tissues of surgical hydatid cyst. Journal of North Khorasan University of Medical Sciences. 2018;9(4):15-19.
33. Abedi B, Maghsood AH, Khansarinejad B, et al. Genotyping of Echinococcus granulosus isolates from livestock based on mitochondrial COX1 gene, in the Markazi province, Iran. J Parasit Dis. 2019;43(4):592-6.
34. Rostami S, Talebi S, Babaei Z, et al. High resolution melting technique for molecular epidemiological studies of cystic echinococcosis: Differentiating G1, G3, and G6 genotypes of Echinococcus granulosus sensu lato. Parasitol Res. 2013; 112(10): 3441-7.
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| Issue | Vol 20 No 3 (2025) | |
| Section | Original Article(s) | |
| Keywords | ||
| Hydatid cyst Genotyping Sequencing Iran | ||
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How to Cite
1.
Elyasi H, Hajari AH, Javaheri E. Genetic Identification of Echinococcus granulosus in Slaughtered Domestic Animals from Two Northeastern Iranian Cities Using HRM and Sequencing Techniques. Iran J Parasitol. 2025;20(3):346-355.
