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Parasitology Research

, Volume 114, Issue 12, pp 4363–4370 | Cite as

Genetic characterization of livestock and human hydatid cyst isolates from northwest Iran, using the mitochondrial cox1 gene sequence

  • Mehdi Farhadi
  • Asghar Fazaeli
  • Ali HanilooEmail author
Original Paper

Abstract

Cystic echinococcosis (CE), caused by larval stages of the tapeworm Echinococcus granulosus, is one of the most important zoonoses distributed worldwide. Genotype analysis of the parasite isolates from various hosts is required to better understand the host specificity and transmission routes. The aim of this study was to identify the genotypes of E. granulosus isolated from humans and domestic animals from northwest of Iran (Zanjan Province) using the mitochondrial cox1 gene sequence. A total of 86 hydatid cysts including 49 sheep and 28 cattle isolates from the slaughterhouse and nine human isolates from surgical wards of local hospitals were collected. The isolates were subjected to DNA extraction, PCR amplification, and sequence. Eighty-two (95.35 %) isolates, including 47 sheep, 26 cattle, and all nine human isolates, were determined as G1 genotype, and the remaining four (4.65 %), including two sheep and two cattle isolates, were identified as G3 genotype. From the cox1 sequence data, 13 different haplotypes (10 G1s and three G3s) were detected and named as EGH1–EGH13 (GenBank accession numbers, KP859559–KP859571). EGH1 was the major variant among the haplotypes, and it was identified in 46 (53.49 %) isolates (31 sheep, 14 cattle, and one human). Alignment of the partial cox1 sequences showed 12 point mutations including seven (58.3 %) synonymous and five (41.7 %) non-synonymous substitutions. Based on the results, G1 was the major genotype of E. granulosus in northwest of Iran affecting sheep, cattle, and humans. In addition, a minor group of G3 genotype was found to be circulating in this region.

Keywords

Hydatid cyst Genotype Cox1 gene Northwest Iran 

Notes

Acknowledgments

This work was supported by a grant (no. A-10-95-1) from the Research Vice-Chancellery of Zanjan University of Medical Sciences (ZUMS). The authors would like to express their gratitude and appreciation to the staff working in the slaughterhouse and the operating rooms of the hospitals in Zanjan for their assistance in collecting the samples required for the study. Special thanks to Dr. Fasihi Harandi for his kind gift of the standard DNA strains as well as constructive comments. We would also like to extend our gratitude to Dr. Hassan Roknizadeh for his helpful comments in bioinformatics analysis. Last but not least, we would like to thank Dr. Mehrdad Pedram (Dept. of Medical Genetics and Molecular Medicine, ZUMS) for careful editing of the manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

436_2015_4673_MOESM1_ESM.docx (334 kb)
ESM 1 (DOCX 333 kb)

References

  1. Abushhewa MH, Abushhiwa MH, Nolan MJ, Jex AR, Campbell BE, Jabbar A, Gasser RB (2010) Genetic classification of Echinococcus granulosus cysts from humans, cattle and camels in Libya using mutation scanning-based analysis of mitochondrial loci. Mol Cell Probes 24:346–351CrossRefPubMedGoogle Scholar
  2. Alvarez Rojas CA, Romig T, Lightowlers MW (2014) Echinococcus granulosus sensu lato genotypes infecting humans—review of current knowledge. Int J Parasitol 44:9–18CrossRefPubMedGoogle Scholar
  3. Amin Pour A, Hosseini SH, Shayan P (2011) Comparative genotyping of Echinococcus granulosus infecting buffalo in Iran using cox1 gene. Parasitol Res 108:1229–1234CrossRefGoogle Scholar
  4. Bardonnet K, Piarroux R, Dia L, Schneegans F, Beurdeley A, Godot V, Vuitton D (2002) Combined eco-epidemiological and molecular biology approaches to assess Echinococcus granulosus transmission to humans in Mauritania: occurrence of the ‘camel’ strain and human cystic echinococcosis. Trans R Soc Trop Med Hyg 96:383–386CrossRefPubMedGoogle Scholar
  5. Bowles J, Blair D, McManus DP (1992) Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Mol Biochem Parasitol 54:165–173CrossRefPubMedGoogle Scholar
  6. Bowles J, Blair D, McManus DP (1994) Molecular genetic characterization of the cervid strain (‘northern form’) of Echinococcus granulosus. Parasitology 109:215–221CrossRefPubMedGoogle Scholar
  7. Busi M, Šnábel V, Varcasia A, Garippa G, Perrone V, De Liberato C, D’Amelio S (2007) Genetic variation within and between G1 and G3 genotypes of Echinococcus granulosus in Italy revealed by multilocus DNA sequencing. Vet Parasitol 150:75–83CrossRefPubMedGoogle Scholar
  8. Capuano F, Rinaldi L, Maurelli M, Perugini A, Veneziano V, Garippa G, Genchi C, Musella V, Cringoli G (2006) Cystic echinococcosis in water buffaloes: epidemiological survey and molecular evidence of ovine (G1) and buffalo (G3) strains. Vet Parasitol 137:262–268CrossRefPubMedGoogle Scholar
  9. Casulli A, Manfredi MT, La Rosa G, Di Cerbo AR, Genchi C, Pozio E (2008) Echinococcus ortleppi and E. granulosus G1, G2 and G3 genotypes in Italian bovines. Vet Parasitol 155:168–172CrossRefPubMedGoogle Scholar
  10. Casulli A, Interisano M, Sreter T, Chitimia L, Kirkova Z, La Rosa G, Pozio E (2012) Genetic variability of Echinococcus granulosus sensu stricto in Europe inferred by mitochondrial DNA sequences. Infect Genet Evol 12:377–383CrossRefPubMedGoogle Scholar
  11. Craig PS, McManus DP, Lightowlers MW, Chabalgoity JA, Garcia HH, Gavidia CM, Gilman RH, Gonzalez AE, Lorca M, Naquira C (2007) Prevention and control of cystic echinococcosis. Lancet Infect Dis 7:385–394CrossRefPubMedGoogle Scholar
  12. Eckert J, Deplazes P (2004) Biological, epidemiological, and clinical aspects of echinococcosis, a zoonosis of increasing concern. Clin Microbiol Rev 17:107–135PubMedCentralCrossRefPubMedGoogle Scholar
  13. Eftekhar M, Athari A (2007) Five years survey of human hydatidosis in Iran. National Congress of Hydatid Cyst, p 38, Yasouj University of Medical Sciences, Yasuj, Iran, 9–10 AprilGoogle Scholar
  14. Espinoza S, Salas AM, Vargas A, Freire V, Diaz E, Sánchez G, Venegas J (2014) Detection of the G3 genotype of Echinococcus granulosus from hydatid cysts of Chilean cattle using COX1and ND1mitochondrial markers. Parasitol Res 113:139–147CrossRefPubMedGoogle Scholar
  15. Fasihi Harandi M, Hobbs R, Adams P, Mobedi I, Morgan-Ryan U, Thompson R (2002) Molecular and morphological characterization of Echinococcus granulosus of human and animal origin in Iran. Parasitology 125:367–373CrossRefPubMedGoogle Scholar
  16. Haag K, Ayala F, Kamenetzky L, Gutierrez A, Rosenzvit M (2004) Livestock trade history, geography, and parasite strains: the mitochondrial genetic structure of Echinococcus granulosus in Argentina. J Parasitol 90:234–239CrossRefPubMedGoogle Scholar
  17. Hajialilo E, Harandi MF, Sharbatkhori M, Mirhendi H, Rostami S (2012) Genetic characterization of Echinococcus granulosus in camels, cattle and sheep from the south-east of Iran indicates the presence of the G3 genotype. J Helminthol 86:263–270CrossRefPubMedGoogle Scholar
  18. Haniloo A, Farhadi M, Fazaeli A, Nourian A (2013) Genotype characterization of hydatid cysts isolated from Zanjan using PCR-RFLP technique. J Zanjan Univ Med Sci 21:57–65Google Scholar
  19. Hüttner M, Nakao M, Wassermann T, Siefert L, Boomker JD, Dinkel A, Sako Y, Mackenstedt U, Romig T, Ito A (2008) Genetic characterization and phylogenetic position of Echinococcus felidis (Cestoda: Taeniidae) from the African lion. Int J Parasitol 38:861–868CrossRefPubMedGoogle Scholar
  20. Jamali R, Ghazanchaei A, Asgharzadeh M (2004) Identification and characterization of Echinococcus granulosus by PCR-RFLP technique in Tabriz district. J Parasit Dis 28:69–72Google Scholar
  21. Kamenetzky L, Gutierrez AM, Canova SG, Haag KL, Guarnera EA, Parra A, García GE, Rosenzvit MC (2002) Several strains of Echinococcus granulosus infect livestock and humans in Argentina. Infect Genet Evol 2:129–136CrossRefPubMedGoogle Scholar
  22. Kedra AH, Swiderski Z, Tkach VV, Dubinsky P, Pawlowski Z, Stefaniak J, Pawlowski J (1999) Genetic analysis of Echinococcus granulosus from humans and pigs in Poland, Slovakia and Ukraine. A multicenter study. Acta Parasitol 44:248–254Google Scholar
  23. Khademvatan S, Yousefi E, Rafiei A, Rahdar M, Saki J (2013) Molecular characterization of livestock and human isolates of Echinococcus granulosus from south-west Iran. J Helminthol 87:240–244CrossRefPubMedGoogle Scholar
  24. Khalifa NO, Khater HF, Fahmy HA, Radwan ME, Afify JS (2014) Genotyping and phylogenetic analysis of cystic echinococcosis isolated from camels and humans in Egypt. Am J Epidemiol Infect Dis 2:74–82CrossRefGoogle Scholar
  25. Kia EB, Rahimi H, Sharbatkhori M, Talebi A, Harandi MF, Mirhendi H (2010) Genotype identification of human cystic echinococcosis in Isfahan, central Iran. Parasitol Res 107:757–760CrossRefPubMedGoogle Scholar
  26. Lavikainen A, Lehtinen M, Meri T, Hirvelä-Koski V, Meri S (2003) Molecular genetic characterization of the Fennoscandian cervid strain, a new genotypic group (G10) of Echinococcus granulosus. Parasitology 127:207–215CrossRefPubMedGoogle Scholar
  27. Lotfi M (1999) Hydatid cyst diseases. Sahab Press, TehranGoogle Scholar
  28. M’rad S, Oudni-M’rad M, Filisetti D, Mekki M, Nouri A, Sayadi T, Candolfi E, Azaiez R, Mezhoud H, Babba H (2010) Molecular identification of Echinococcus granulosus in Tunisia: first record of the Buffalo strain (G3) in human and bovine in the country. Open Vet Sci J 4:27–30CrossRefGoogle Scholar
  29. Maillard S, Benchikh-Elfegoun M, Knapp J, Bart J, Koskei P, Gottstein B, Piarroux R (2007) Taxonomic position and geographical distribution of the common sheep G1 and camel G6 strains of Echinococcus granulosus in three african countries. Parasitol Res 100:495–503CrossRefPubMedGoogle Scholar
  30. Mario L, Takano K, Brochado JF, Costa CV, Soares AG, Yamano K, Yagi K, Katoh Y, Takahashi K (2011) Infection of humans and animals with Echinococcus granulosus (G1 and G3 strains) and E. ortleppi in Southern Brazil. Vet Parasitol 177:97–103CrossRefGoogle Scholar
  31. McManus D, Bowles J (1996) Molecular genetic approaches to parasite identification: their value in diagnostic parasitology and systematics. Int J Parasitol 26:687–704CrossRefPubMedGoogle Scholar
  32. McManus D, Thompson R (2003) Molecular epidemiology of cystic echinococcosis. Parasitology 127:S37–S51CrossRefPubMedGoogle Scholar
  33. Nakao M, McManus D, Schantz P, Craig P, Ito A (2007) A molecular phylogeny of the genus Echinococcus inferred from complete mitochondrial genomes. Parasitology 134:713–722CrossRefPubMedGoogle Scholar
  34. Nakao M, Li T, Han X, Ma X, Xiao N, Qiu J, Wang H, Yanagida T, Mamuti W, Wen H (2010) Genetic polymorphisms of Echinococcus tapeworms in China as determined by mitochondrial and nuclear DNA sequences. Int J Parasitol 40:379–385PubMedCentralCrossRefPubMedGoogle Scholar
  35. Nikmanesh B, Mirhendi H, Ghalavand Z, Alebouyeh M, Sharbatkhori M, EshratBeigom K, Mohebali M, Eghbali M, Rokni MB (2014) Genotyping of Echinococcus granulosus isolates from human clinical samples based on sequencing of mitochondrial genes in Iran, Tehran. Iran J Parasitol 9:20–27PubMedCentralPubMedGoogle Scholar
  36. Okamoto M, Bessho Y, Kamiya M, Kurosawa T, Horii T (1995) Phylogenetic relationships within Taenia taeniaeformis variants and other taeniid cestodes inferred from the nucleotide sequence of the cytochromec oxidase subunit I gene. Parasitol Res 81:451–458CrossRefPubMedGoogle Scholar
  37. Okamoto M, Nakao M, Blair D, Anantaphruti MT, Waikagul J, Ito A (2010) Evidence of hybridization between Taenia saginata and Taenia asiatica. Parasitol Int 59:70–74CrossRefPubMedGoogle Scholar
  38. Parsa F, Harandi MF, Rostami S, Sharbatkhori M (2012) Genotyping Echinococcus granulosus from dogs from Western Iran. Exp Parasitol 132:308–312CrossRefPubMedGoogle Scholar
  39. Pednekar RP, Gatne ML, Thompson RA, Traub RJ (2009) Molecular and morphological characterisation of Echinococcus from food producing animals in India. Vet Parasitol 165:58–65CrossRefPubMedGoogle Scholar
  40. Pezeshki A, Akhlaghi L, Sharbatkhori M, Razmjou E, Oormazdi H, Mohebali M, Meamar A (2013) Genotyping of Echinococcus granulosus from domestic animals and humans from Ardabil Province, northwest Iran. J Helminthol 87:387–391CrossRefPubMedGoogle Scholar
  41. Rokni MB (2009) Echinococcosis/hydatidosis in Iran. Iran J Parasitol 4:1–16Google Scholar
  42. Rostami Nejad M, Taghipour N, Nochi Z, Nazemalhosseini Mojarad E, Mohebbi S, Fasihi Harandi M, Zali M (2012) Molecular identification of animal isolates of Echinococcus granulosus from Iran using four mitochondrial genes. J Helminthol 86:485–492CrossRefPubMedGoogle Scholar
  43. Sánchez E, Cáceres O, Náquira C, Garcia D, Patiño G, Silvia H, Volotão AC, Fernandes O (2010) Molecular characterization of Echinococcus granulosus from Peru by sequencing of the mitochondrial cytochrome C oxidase subunit 1 gene. Mem Inst Oswaldo Cruz 105:806–810CrossRefPubMedGoogle Scholar
  44. Scott J, Stefaniak J, Pawlowski Z, McManus D (1997) Molecular genetic analysis of human cystic hydatid cases from Poland: identification of a new genotypic group (G9) of Echinococcus granulosus. Parasitology 114:37–43CrossRefPubMedGoogle Scholar
  45. Shahnazi M, Hejazi H, Salehi M, Andalib AR (2011) Molecular characterization of human and animal Echinococcus granulosus isolates in Isfahan, Iran. Acta Trop 117:47–50CrossRefPubMedGoogle Scholar
  46. Sharbatkhori M, Harandi MF, Mirhendi H, Hajialilo E, Kia EB (2011) Sequence analysis of cox1 and nad1 genes in Echinococcus granulosus G3 genotype in camels (Camelus dromedarius) from central Iran. Parasitol Res 108:521–527CrossRefPubMedGoogle Scholar
  47. Sharma M, Fomda BA, Mazta S, Sehgal R, Singh BB, Malla N (2013a) Genetic diversity and population genetic structure analysis of Echinococcus granulosus sensu stricto complex based on mitochondrial DNA signature. PLoS One 8:e82904PubMedCentralCrossRefPubMedGoogle Scholar
  48. Sharma M, Sehgal R, Fomda BA, Malhotra A, Malla N (2013b) Molecular characterization of Echinococcus granulosus cysts in north Indian patients: identification of G1, G3, G5 and G6 genotypes. PLoS Negl Trop Dis 7:e2262PubMedCentralCrossRefPubMedGoogle Scholar
  49. Simsek S, Kaplan M, Ozercan IH (2011) A comprehensive molecular survey of Echinococcus granulosus in formalin-fixed paraffin-embedded tissues in human isolates in Turkey. Parasitol Res 109:411–416CrossRefPubMedGoogle Scholar
  50. Thompson RA (2008) The taxonomy, phylogeny and transmission of Echinococcus. Exp Parasitol 119:439–446CrossRefPubMedGoogle Scholar
  51. Thompson RA, McManus DP (2002) Towards a taxonomic revision of the genus Echinococcus. Trends Parasitol 18:452–457CrossRefPubMedGoogle Scholar
  52. Utuk AE, Simsek S, Koroglu E, McManus DP (2008) Molecular genetic characterization of different isolates of Echinococcus granulosus in east and southeast regions of Turkey. Acta Trop 107:192–194CrossRefPubMedGoogle Scholar
  53. Vural G, Baca AU, Gauci CG, Bagci O, Gicik Y, Lightowlers MW (2008) Variability in the Echinococcus granulosus cytochrome C oxidase 1 mitochondrial gene sequence from livestock in Turkey and a re-appraisal of the G1–3 genotype cluster. Vet Parasitol 154:347–350CrossRefPubMedGoogle Scholar
  54. Yanagida T, Mohammadzadeh T, Kamhawi S, Nakao M, Sadjjadi SM, Hijjawi N, Abdel-Hafez SK, Sako Y, Okamoto M, Ito A (2012) Genetic polymorphisms of Echinococcus granulosus sensu stricto in the Middle East. Parasitol Int 61:599–603CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Department of Parasitology and Mycology, School of MedicineZanjan University of Medical SciencesShahrak KarmandanIran

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