Investigation of Mitochondrial Cytb Gene Region of Both Echinococcus granulosus Eggs from Dogs and Cystic Echinococcosis Isolates Obtained from Sheep and Cattle by Molecular Methods
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Abstract
Background: We aimed to determine the common Echinococcus granulosus genotypes in Ağrı, Türkiye and to obtain information on the transmission of this parasite.
Methods: Cystic echinococcosis samples from 100 slaughtered cattle and 100 slaughtered sheep and faecal samples from 200 stray dogs were included in 2021. Collected cyst fluid samples and faces were examined microscopically. DNA was isolated from the germinal membrane of the cysts and from the parasite eggs in the stool samples. The mitochondrial cytb gene region of the parasite was amplified by PCR. Genotypes were determined using the Basic Local Alignment Search Tool (BLAST) after sequence analysis of PCR amplicons.
Results: The highest percentage of cysts was found in the lungs of sheep and the liver of cattle. In addition, 75% of sheep cysts and 25.6% of cattle cysts were fertile. Taenia spp./Echinococcus spp. eggs were found in 6% of the faeces of 200 dogs examined microscopically. E. granulosus eggs were detected in 4 out of 50 stool samples analysed by PCR. All samples analysed by sequence analysis were identified as E. granulosus s.s. G1 genotype. Sequence comparison revealed revealed one or more-point mutations in different regions of the five samples.
Conclusion: E. granulosus s.s. G1 genotype, known as sheep strain, is common in the Ağrı, Türkiye. The controlled slaughter of livestock, especially sheep, and the avoidance of feeding hydatid cyst organs to dogs, together with public education, were necessary to prevent the spread of the disease.
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5. McManus DP, Zhang W, Li J, Bartley PB. Echinococcosis. Lancet. 2003; 362(9392):1295-1304.
6. Casulli A, Massolo A, Saarma U, et al. Species and genotypes belonging to Echinococcus granulosus sensu lato complex causing human cystic echinococcosis in Europe (2000–2021): a systematic review. Parasit Vectors. 2022;15(1):109.
7. Šnábel V, Kuzmina TA, Antipov AA, et al. Molecular study of Echinococcus granulosus cestodes in Ukraine and the first genetic identification of Echinococcus granulosus sensu stricto (G1 genotype) in the country. Acta Parasitol. 2022;67:244-254.
8. Willis, HH. A Simple Levitation Method for the Detection of Hookworm Ova. Med J Aust. 1921;2:75-376.
9. Zhong X, Wang N, Hu D, at al. Sequence analysis of cytb gene in Echinococcus granulosus from Western China. Korean J Parasitol. 2014;52(2):205-209.
10. Olive DM, Bean P. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol. 1999;37(6):1661-1669.
11. Akkaş Ö, Özgökçe M, Aydemir S, Dündar İ, Ekici A. Genotyping of Echinococcus granulosus Human Isolates Obtained from the Puncture, Aspiration, Injection, and Re-Aspiration (PAIR) Process in Türkiye. Iran J Parasitol. 2023;18(4):427-434.
12. Tahiri S, Naoui H, Iken M, Azelmat S, Khallayoune M, Bouchri M, et al. Genotyping of human Echinococcus granulosus cyst in Morocco. J Parasit Dis. 2019;43(4):560-565.
13. Khan A, Ahme H, Simsek S, Liu H, Yin J, Wang Y, et al., Molecular characterization of human Echinococcus isolates and the first report of E. canadensis (G6/G7) and E. multilocularis from the Punjab Province of Pakistan using sequence analysis. BMC Infect Dis. 2020;20(1):1-10.
14. Shahabi S, Sarkari B, Barazesh A. Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Türkiye and Iran, based on mitochondrial nad5 gene differentiation. Parasit Vectors. 2021;14(1):361.
15. Fadhil SA, A'aiz NN. Genotyping of cystic echinococcosis isolates from clinical samples of human and domestic animals. Iraqi J Vet Sci. 2016; 30(2):33-39.
16. Alvarez Rojas CA, Kronenberg PA, Aitbaev S, et al. Genetic diversity of Echinococcus multilocularis and Echinococcus granulosus sensu lato in Kyrgyzstan: The A2 haplotype of E. multilocularis is the predominant variant infecting humans. PLoS Negl Trop Dis. 2020;14(5): e0008242.
17. Abushhewa MH, Nolan MJ, Jex AR, Campbell BE, Jabbar A, Gasser RB. Genetic classification of Echinococcus granulosus cysts from humans, cattle and camels in Libya using mutation scanning-based analysis of mitochondrial loci. Mol Cell Probes. 2010;24(6):346-351.
18. Šoba B, Gašperšič Š, Keše D, Kotar T. Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia. Pathogens. 2020;9(7):562.
19. Joanny G, Mehmood N, Dessi G, Tampon C, Nonnis F, Hosri C, et al. Cystic echinococcosis in sheep and goats of Lebanon. Parasitology. 2021;148(7):871-878.
20. Varcasia A, Canu S, Lightowlers MW, Scala A, Garippa G. Molecular characterization of Echinococcus granulosus strains in Sardinia. Parasitol Res. 2006;98(3):273-277.
21. Issa HS, Abdel-Hafez SK, Hijjawi NS, Al-Qaoud KM. Molecular Characterization of Echinococcus granulosus sensu stricto Cysts of Domestic Ruminants in Jordan. Jordan J Biol Sci. 2018;11(3):301-306.
22. Rosenzvit M, Zhang LH, Kamenetzky L, Canova S, Guarnera E, McManus D. Genetic variation and epidemiology of Echinococcus granulosus in Argentina. Parasitology. 1999;118(Pt 5):523-530.
23. Abdybekova AM, Zhang Z, Sultanov AA, Abdibayeva AA, Zhaksylykova AA, Junisbayeva SM, et al. Genotypes of Echinococcus isolated from domestic livestock in Kazakhstan. J Helminthol. 2020;94:e69.
24. Faruk MO, Siddiki AMA, Masuduzzaman M, Chowdhury S, Hossain MA. Identification and molecular characterization of Echinococcus granulosus from domestic goat in Chittagong, Bangladesh. Trop Biomed. 2017;34(4):831-840.
25. Heidari Z, Sharbatkhori M, Mobedi I, Mirhendi SH, Nikmanesh B, Sharifdini M, et al. Echinococcus multilocularis and Echinococcus granulosus in canines in North-Khorasan Province, northeastern Iran, identified using morphology and genetic characterization of mitochondrial DNA. Parasit Vectors. 2019;12(1):606.
26. Arbabi M, Hooshyar H, Delavari M, Pestechian N. Genotypes Identification of Echinococcus granulosus isolated from iranian dogs and camels using three polymerase Chain reaction-based methods of cox1 gene. Int Arch Nurs Health Care. 2021;8(2):104-110.
27. Mulinge E, Magambo J, Odongo D, Njenga S, Zeyhle E, Mbae C, et al. Molecular characterization of Echinococcus species in dogs from four regions of Kenya. Vet Parasitol. 2018;255:49-57.
28. Mehmood S, Simsek S, Celik F, Kesik HK, Kilinc SG, Ahmed H. Molecular survey on cattle and sheep hydatidosis and first detection of Echinococcus canadensis (G6/G7) in sheep in Turkey. Parasitology. 2020;147(9):1055-1062.
29. Kesik HK, Celik F, Simsek S, Kilinc SG, Ahmed H, Gul A. Molecular characterization and haplotype analyses of lung hydatid cyst isolates of cattle and first report of Echinococcus canadensis (G6/G7) in cattle isolates in Turkey. Acta Parasitol. 2021;66:1538-1547.
30. Kesik HK, Celik F, Kilinc SG, Simsek S, Ahmed H, Shen Y, et al. Genetic Diversity and Haplotype Analysis of Cattle Hydatid Cyst Isolates Using Mitochondrial Markers in Türkiye. Pathogens. 2022;11(5):519.
31. Gökpınar S, Değirmenci R,Yıldız K. Genotyping of Echinococcus granulosus obtained from cattle slaughtered in Kırıkkale Province. Ankara Üniv Vet Fak Derg. 2017;64:51-54.
32. Oguz B, Ozdal N, Kilinc OO, Deger, MS. Preliminary studies on the prevalence and genotyping of Echinococcus granulosus infection in stray dogs in Van Province, Turkey. J Vet Res, 2018; 62(4):497-502.
33. Utuk, AE, Simsek, S, Koroglu, E and McManus, DP. Molecular genetic characterization of different isolates of Echinococcus granulosus in east and southeast regions of Turkey. Acta Trop, 2008; 107:192–194.
34. Avcioglu H, Guven E, Balkaya I, et al. The situation of echinococcosis in stray dogs in Turkey: the first finding of Echinococcus multilocularis and Echinococcus ortleppi. Parasitology. 2021;148(9):1092-1098.
2. Ertabaklar H, Yıldız İ, Malatyalı E, Tileklioğlu E, Çalışkan SÖ, Ertuğ S. Retrospective Analysis of Cystic Echinococcosis Results in Aydın Adnan Menderes University Training and Research Hospital Parasitology Laboratory Between 2005 and 2017. Turkiye Parazitol Derg. 2019;43:118-22.
3. Akyol, ÇV. Hidatidoz ve halk sağlığı yönünden önemi. J Fac Vet Med. 2001;20:137-142.
4. Demir P, Mor N. Seasonal Distribution and Economic Importance of Cystic Echinococcosis in Cattle Slaughtered at Kars Municipal Abattoir. Turkiye Parazitol Derg. 2011;35:185-188.
5. McManus DP, Zhang W, Li J, Bartley PB. Echinococcosis. Lancet. 2003; 362(9392):1295-1304.
6. Casulli A, Massolo A, Saarma U, et al. Species and genotypes belonging to Echinococcus granulosus sensu lato complex causing human cystic echinococcosis in Europe (2000–2021): a systematic review. Parasit Vectors. 2022;15(1):109.
7. Šnábel V, Kuzmina TA, Antipov AA, et al. Molecular study of Echinococcus granulosus cestodes in Ukraine and the first genetic identification of Echinococcus granulosus sensu stricto (G1 genotype) in the country. Acta Parasitol. 2022;67:244-254.
8. Willis, HH. A Simple Levitation Method for the Detection of Hookworm Ova. Med J Aust. 1921;2:75-376.
9. Zhong X, Wang N, Hu D, at al. Sequence analysis of cytb gene in Echinococcus granulosus from Western China. Korean J Parasitol. 2014;52(2):205-209.
10. Olive DM, Bean P. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol. 1999;37(6):1661-1669.
11. Akkaş Ö, Özgökçe M, Aydemir S, Dündar İ, Ekici A. Genotyping of Echinococcus granulosus Human Isolates Obtained from the Puncture, Aspiration, Injection, and Re-Aspiration (PAIR) Process in Türkiye. Iran J Parasitol. 2023;18(4):427-434.
12. Tahiri S, Naoui H, Iken M, Azelmat S, Khallayoune M, Bouchri M, et al. Genotyping of human Echinococcus granulosus cyst in Morocco. J Parasit Dis. 2019;43(4):560-565.
13. Khan A, Ahme H, Simsek S, Liu H, Yin J, Wang Y, et al., Molecular characterization of human Echinococcus isolates and the first report of E. canadensis (G6/G7) and E. multilocularis from the Punjab Province of Pakistan using sequence analysis. BMC Infect Dis. 2020;20(1):1-10.
14. Shahabi S, Sarkari B, Barazesh A. Echinococcus granulosus sensu stricto G1 is the predominant genotype in human and livestock isolates from Türkiye and Iran, based on mitochondrial nad5 gene differentiation. Parasit Vectors. 2021;14(1):361.
15. Fadhil SA, A'aiz NN. Genotyping of cystic echinococcosis isolates from clinical samples of human and domestic animals. Iraqi J Vet Sci. 2016; 30(2):33-39.
16. Alvarez Rojas CA, Kronenberg PA, Aitbaev S, et al. Genetic diversity of Echinococcus multilocularis and Echinococcus granulosus sensu lato in Kyrgyzstan: The A2 haplotype of E. multilocularis is the predominant variant infecting humans. PLoS Negl Trop Dis. 2020;14(5): e0008242.
17. Abushhewa MH, Nolan MJ, Jex AR, Campbell BE, Jabbar A, Gasser RB. Genetic classification of Echinococcus granulosus cysts from humans, cattle and camels in Libya using mutation scanning-based analysis of mitochondrial loci. Mol Cell Probes. 2010;24(6):346-351.
18. Šoba B, Gašperšič Š, Keše D, Kotar T. Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia. Pathogens. 2020;9(7):562.
19. Joanny G, Mehmood N, Dessi G, Tampon C, Nonnis F, Hosri C, et al. Cystic echinococcosis in sheep and goats of Lebanon. Parasitology. 2021;148(7):871-878.
20. Varcasia A, Canu S, Lightowlers MW, Scala A, Garippa G. Molecular characterization of Echinococcus granulosus strains in Sardinia. Parasitol Res. 2006;98(3):273-277.
21. Issa HS, Abdel-Hafez SK, Hijjawi NS, Al-Qaoud KM. Molecular Characterization of Echinococcus granulosus sensu stricto Cysts of Domestic Ruminants in Jordan. Jordan J Biol Sci. 2018;11(3):301-306.
22. Rosenzvit M, Zhang LH, Kamenetzky L, Canova S, Guarnera E, McManus D. Genetic variation and epidemiology of Echinococcus granulosus in Argentina. Parasitology. 1999;118(Pt 5):523-530.
23. Abdybekova AM, Zhang Z, Sultanov AA, Abdibayeva AA, Zhaksylykova AA, Junisbayeva SM, et al. Genotypes of Echinococcus isolated from domestic livestock in Kazakhstan. J Helminthol. 2020;94:e69.
24. Faruk MO, Siddiki AMA, Masuduzzaman M, Chowdhury S, Hossain MA. Identification and molecular characterization of Echinococcus granulosus from domestic goat in Chittagong, Bangladesh. Trop Biomed. 2017;34(4):831-840.
25. Heidari Z, Sharbatkhori M, Mobedi I, Mirhendi SH, Nikmanesh B, Sharifdini M, et al. Echinococcus multilocularis and Echinococcus granulosus in canines in North-Khorasan Province, northeastern Iran, identified using morphology and genetic characterization of mitochondrial DNA. Parasit Vectors. 2019;12(1):606.
26. Arbabi M, Hooshyar H, Delavari M, Pestechian N. Genotypes Identification of Echinococcus granulosus isolated from iranian dogs and camels using three polymerase Chain reaction-based methods of cox1 gene. Int Arch Nurs Health Care. 2021;8(2):104-110.
27. Mulinge E, Magambo J, Odongo D, Njenga S, Zeyhle E, Mbae C, et al. Molecular characterization of Echinococcus species in dogs from four regions of Kenya. Vet Parasitol. 2018;255:49-57.
28. Mehmood S, Simsek S, Celik F, Kesik HK, Kilinc SG, Ahmed H. Molecular survey on cattle and sheep hydatidosis and first detection of Echinococcus canadensis (G6/G7) in sheep in Turkey. Parasitology. 2020;147(9):1055-1062.
29. Kesik HK, Celik F, Simsek S, Kilinc SG, Ahmed H, Gul A. Molecular characterization and haplotype analyses of lung hydatid cyst isolates of cattle and first report of Echinococcus canadensis (G6/G7) in cattle isolates in Turkey. Acta Parasitol. 2021;66:1538-1547.
30. Kesik HK, Celik F, Kilinc SG, Simsek S, Ahmed H, Shen Y, et al. Genetic Diversity and Haplotype Analysis of Cattle Hydatid Cyst Isolates Using Mitochondrial Markers in Türkiye. Pathogens. 2022;11(5):519.
31. Gökpınar S, Değirmenci R,Yıldız K. Genotyping of Echinococcus granulosus obtained from cattle slaughtered in Kırıkkale Province. Ankara Üniv Vet Fak Derg. 2017;64:51-54.
32. Oguz B, Ozdal N, Kilinc OO, Deger, MS. Preliminary studies on the prevalence and genotyping of Echinococcus granulosus infection in stray dogs in Van Province, Turkey. J Vet Res, 2018; 62(4):497-502.
33. Utuk, AE, Simsek, S, Koroglu, E and McManus, DP. Molecular genetic characterization of different isolates of Echinococcus granulosus in east and southeast regions of Turkey. Acta Trop, 2008; 107:192–194.
34. Avcioglu H, Guven E, Balkaya I, et al. The situation of echinococcosis in stray dogs in Turkey: the first finding of Echinococcus multilocularis and Echinococcus ortleppi. Parasitology. 2021;148(9):1092-1098.
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| Issue | Vol 19 No 2 (2024) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v19i2.15860 | |
| Keywords | ||
| Cytb Echinococcus granulosus Genotyping Türkiye | ||
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How to Cite
1.
Yildiz R, Aydemir S, Halidi AG, Unlu AH, Yilmaz H. Investigation of Mitochondrial Cytb Gene Region of Both Echinococcus granulosus Eggs from Dogs and Cystic Echinococcosis Isolates Obtained from Sheep and Cattle by Molecular Methods. Iran J Parasitol. 2024;19(2):247-254.
