Molecular Characterization of Echinococcus granulosus Isolates Found in Cattle, Buffaloes, Sheep and Goats in Afyonkarahisar, Turkey

Year 2020, Volume: 13 Issue: 2, 152 - 160, 30.06.2020

Kürşat Kartal , Mustafa Köse , Metin Erdoğan

https://doi.org/10.30607/kvj.711873

Abstract

This study has been carried out to determine the genotypes of Echinococcus granulosus cysts in cattle, buffaloes, sheep and goats raised in Afyonkarahisar region. Cysts were collected from the internal organs of 258 animals, including 65 goats, 71 sheep, 119 cattle and 3 buffaloes infected with hydatid cysts. DNA was isolated from a total of 78 cysts from germinal membranes and protoscoleces extracted from cysts to identify the genotypes of E. granulosus in infected animals. PCR-RFLP was carried out using the Hin6I and StuI restriction enzymes in the ND1 gene and no polymorphism could be determined in all isolates. In the COX1 gene analysis, G1 strain known as domestic sheep strains and 18 different haplotypes were found in all isolates from cattle, buffaloes, sheep and goats. As a result, it was concluded that all isolates of the cattle, buffaloes, sheep and goats grown in Afyonkarahisar region determined in the analyses carried on the COX1 gene were G1 strain.

Keywords

Ruminants, Echinococcus granulosus, ND1, COX1, PCR-RFLP, DNA Sequencing

Supporting Institution

Afyon Kocatepe University Scientific Research Projects Coordination

Project Number

12. SAĞ. BİL. 01

Thanks

This study was supported by the Scientific Research Projects Coordination Unit of Afyon Kocatepe University (Project number: 12. SAG. BIL. 01).

Afyonkarahisar’da Sığır, Manda, Koyun ve Keçilerde Bulunan Echinococcus granulosus İzolatlarının Moleküler Karakterizasyonu

Abstract

Bu çalışma, Afyonkarahisar yöresinde yetiştirilen sığır, manda, koyun ve keçilerde bulunan Echinococcus granulosus kistlerinin genotiplerinin belirlenmesi amacıyla yapılmıştır. Araştırmada, hidatik kistle enfekte 65 keçi, 71 koyun, 119 sığır ve 3 manda olmak üzere toplam 258 hayvanın iç organlarından kistler toplanmıştır. Enfekte hayvanlardaki E. granulosus genotiplerini belirlemek amacıyla kistlerden çıkarılan germinal membran ve protoskolekslerden toplam 78 kistten DNA izole edilmiştir. ND1 gen bölgesi Hin6I ve StuI restriksiyon enzimleri kullanılarak PCR-RFLP yapılmış ve tüm izolatlarda polimorfizm belirlenememiştir. COX1 gen bölgesi analizinde sığır, manda, koyun ve keçilerden elde edilen izolatların tümünde evcil koyun suşu olarak bilinen G1 suşu ve 18 farklı haplotip bulunmuştur. Sonuç olarak, Afyonkarahisar yöresinde yetiştirilen sığır, manda, koyun ve keçilerde bulunan izolatların COX1 geninde yapılan genetik analizler sonucunda tüm izolatların G1 suşu olduğu kanısına varılmıştır.

Keywords

Ruminantlar, Echinococcus granulosus, ND1, COX1, PCR-RFLP, DNA Dizileme

Project Number

12. SAĞ. BİL. 01

References

• Aboelhadid SM, El-Dakhly KM, Yanai T, Fukushi H, Hasanin KM. Molecular Characterization of Echinococcus granulosus in Egyptian. Vet Parasitol. 2013; 193: 292-296.
• Adwan G, Adwan K, Bdir S, Abuseir S. Molecular Characterization of Echinococcus granulosus Isolated from Sheep in Palestine. Exp Parasitol. 2013; 134: 195-199.
• Ayaz E, Tınar R. Cestoda. In: Tınar R (ed) Helmintoloji, 1st edn. Nobel yayınevi, Ankara. 2006; pp 103-213.
• Barış İ, Şahin A, Bilir N, Kalyoncu F, Emri S, Barış B, Çapur S, Selçuk T. Hidatik Kist Hastalığı ve Türkiye’deki Konumu. Türkiye Akciğer Hastalıkları Vakfı Yayını No: 1 Ankara. 1989
• Bart JM, Abdukader M, Zhang YL, Lin RY, Wang YH, Nakao M, Ito A, Craig PS, Piarroux R, Vuitton DA, Wen H. Genotyping of Human Cystic Echinococcosis in Xinjiang, PR China. Parasitology. 2006a; 133(5): 571-579.
• Bart JM, Morariu S, Knapp J, Ilie MS, Pitulescu M, Anghel A, Cosoroaba I, Piarroux R. Genetic Typing of Echinococcus granulosus in Romania. Parasitol Res. 2006b; 98: 130-137.
• Beyhan YE, Umur Ş. Molecular Characterization and Prevalence of Cystic Echinococcosis in Slaughtered Water Buffaloes in Turkey. Vet Parasitol. 2011; 181: 174-179.
• Boom R, Sol CJA, Salimans MMM, Jansen CL, Wertheim-Van Dillen PME, Van Der Noorda J. Rapid and Simple Method for Purification of Nucleic Acids. J Clin Microbiol. 1990; 28(3): 495-503.
• Bowles J, McManus DP. Rapid discrimination of Echinoccus specises and strains using a polymerase chain reaction-based RFLP method. Mol Biochem Parasitol. 1993; 57: 231–240.
• Breyer I, Georgieva D, Kurdova R, Gottstein B. Echinococcusgranulosus Strain Typing in Bulgaria: The G1 Genotype is Predominant in Intermediate and Definitive Wild Hosts. Parasitol Res. 2004; 93: 127-130
• Cadona GA, Carmena D. A Review Of The Global Prevalence, Molecular Epidemiology and Economics Of Cystic Echinococcus In Production Animals. Vet Parasitol. 2013; 192: 10-32.
• Capuano F, Rinaldi L, Maurelli MP, Perugini AG, Veneziano V, Garippa G, Genchi C, Musella V, Cringoli G. Cystic echinococcocosis in water buffaloes: epidemiological survey and molecular evidence of ovine (G1) and buffalo (G3) strains. Vet Parasitol. 2006; 137: 262-268.
• Casulli A, Zeyhle E, Brunetti E, Pozio E, Meroni V, Genco F, Filice C. Molecular Evidence of the Camel Strain (G6 Genotype) of Echinococcusgranulosus in humans from Turkana, Kenya. Trans Roy Soc Trop Med Hyg. 2010; 104: 29-32.
• Dalimi A, Motamedi GH, Hosseni M, Mohammadian B, Malaki H, Ghamari Z, Far GF. Echinococcosis/ hydatidosis in western Iran. Vet Parasitol. 2002; 105: 161–171.
• Eckert J, Thompson RCA. IntraspesificVarition of Echinococcus granulosus and Related Species with Emphises on their Infectivity to Humans. Acta Trop. 1997; 64: 19-34.
• Gıcık Y, Arslan MÖ, Kara M, Köse M. Kars ilinde Kesilen Sığır ve Koyunlarda Kistik Ekinokokkozisin Yaygınlığı. Türkiye Parazitol Derg. 2004; 28(3): 136-139.
• Gonzalez LM, Daniel-Mwambete K, Montero E, Rosenzvit MC, McManus DP, Garate T, Cuesta-Bandera C. Further Molecular Discrimination of Spanish Strains of Echinococcus granulosus. Exp Parasitol. 2002; 102: 46-56
• Guo ZH, Kubo M, Kudo M, Nibe K, Horii Y, Nonaka N. Growth and Genotypes of Echinococcus granulosus Found in Cattle Imported from Australia and Fattened in Japan. Parasitol Int. 2011; 60: 498-502.
• Haag KL, Zaha A, Araujo AM, Gottstein B. Reduced Genetic Variability within Coding and Non-coding Regions of the Echinococcus multilocularis Genome. Parasitology. 1997; 115: 521-529.
• Hall TA. BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95/98/NT. Nucleic Acids Symposium Series. 1999; 41: 95-98.
• Hoss M, Paabo S. DNA extraction from Pleistocene bones by a silica-based purification method. Nucleic Acids Res. 1993; 21(16): 3913-3914.
• Jabbar A, Narankhajid M, Nolan MJ, Jex AR, Campbell BE, Gasser RB. A First İnsight into the Genotypes of Echinococcus granulosus from Humans in Mongolia. Mol Cell Probe. 2011; 25: 49-54.
• Kalendar R, Lee D, Schulman AH. Fast PCR Software for PCR Primer and Probe Design and Repeat Search. G3. 2009; 3(1): 1-14. Köse M, Sevimli FK. Prevalence of Cystic Echinococcosis in Slaughtered Cattle in Afyonkarahisar. Türkiye Parazitol Derg. 2008; 32(1): 27-30.
• Latif AA, Tanveer A, Maqbool A, Siddiqi N, Kyaw-Tanner M, Traub RJ. Morphological and Molecular Characterisation of Echinococcus granulosus in Livestock and Humans in Punjab, Pakistan. Vet Parasitol. 2010; 170: 44-49.
• Lavikainen A, Lehtinen MJ, Meri T, Hirvela-Koski V, Meri S. Molecular genetic characterization of the Fennoscandiancervidstrain, a new genotypic group (G10) of Echinococcusgranulosus. Parasitology. 2003; 127: 207-215.
• Markel EK, Voge M, John DT. Medical Parasitology. 8th Edn, WB Saunders Company, USA. 1999; pp 234-268.
• McManus DP, Wenbao Z, Li J, Bartley PB. Echinococcosis. Lancet. 2003; 326: 1295-1304.
• Merdivenci A. İstanbul Sokak Köpeklerinde Echinococcus granulosus (Batsch, 1786) Rudolphi, 1805. Mikrobiol Derg. 1963; 16(1): 23-28.
• Nakao M, McManus DP, Schantz PM, Craig PS, Ito A. A Molecular Phylogeny of the Genus Echinococcus Inferred from Complete Mitochondrial Genomes. Parasitology. 2007; 134: 713-722.
• Nakao M, Yangida T, Okomoto M, Knapp J, Nkouawa A, Sako Y, Ito A. State-of-the-Art Echinococcus and Taenia: Phylogenetic Taxanomy of Human Pathogenic Tapeworms an Its Application to Molecular Diagnosis. Infect Genet Evol. 2010; 10: 444-452.
• Pour AA, Hosseini SS, Shayan P. Comparative Genotyping of Echinococcusgranulosus Infecting Buffalo in Iran Using Cox1 Gene. Parasitol Res. 2011; 108: 1229-1234.
• Romig T, Dinkel A, Mackenstedt U. The Present Situation of Echinococcus in Europe. Parasitol Int. 2006; 55: 187-191.
• Saarma U, Jogisalu I, Moks E, Varcasia A, Lavikainen A, Oksanen A. A Novel pylogenyfort he Genus Echinococcus, Based on Nuclear Data, Challenges Relationships Based on Mitochondrial Evidence. Parasitology. 2009; 136: 317-328.
• Sanchez E, Caceres O, Naquira C, Miranda E, Samudio F, Fernandes O. Echinococcus granulosus Genotypes Circulating in Alpacas (Lama pacos) and Pigs (Susscrofa) from an Endemic Region in Peru. Mem Inst Oswaldo Cruz, Rio de Janeiro. 2012; 107: 275-278.
• Scott JC, Stefaniak J, Pawlowski ZS, McManus DP. Molecular Genetic Analysis of Human Cystic Hydatid Cases from Poland: Identification of a New Genotipic Group (G9) of Echinococcus granulosus. Parasitology. 1997; 114: 37-43.
• Şimşek S, Balkaya I, Köroğlu E. Epidemiological Survey and Molecular Characterization of Echinococcusgranulosus in Cattle in an Endemic Area Eastern Turkey. Vet Parasitol . 2010; 172: 347-349.
• Singh BB, Sharma JK, Ghatak S, Sharma R., Bal MS, Tuli A, Gill JPS. Molecular Epidemiology of Echinococcosis from Food Producing Animals in North India. Vet Parasitol. 2012; 186: 503-506.
• Snabel V, Altıntaş N, D’amelio S, Nakao M, Romig T, Yolasığmaz A, Güneş K, Turk M, Busi M, Huttner M, Sevcova D, Ito A, Altıntaş N, Dubinsky P. Cystic Echinococcosis in Turkey: Genetic Variability and First Record of the Pig Strain (G7) in the Country. Parasitol Res. 2009; 105(1): 145-154.
• Soriano SV, Pierangelii NB, Pianciola L, Mazzeo M, Lazzarini LE, Saiz MS, Kossman AV, Bergagna HFJ, Chartier K, Basualdo JA. Molecular characterization of Echinococcus canadensis G6 Genotype in Neuquen, Patagonia Argentina. Parasitol Int. 2010; 59: 626-628.
• Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) Software Version 4.0. Molecular Biology and Evolution 10.1093/molbev/msm092. 2007.
• Tajima F. Statistical Methods to Test for Nucleotide Mutation Hypothesis by DNA Polymorphism. Genetics . 1989; 123: 585-595.
• Thompson RCA. The Taxanomy, Phylogeny and Transmission of Echinococcus. Exp parasitol. 2008; 119: 439-446.
• Thompson RCA, McManus DP. Towards a Taxonomic Revision of the Genus Echinococcus. Trends Parasitol. 2002; 18: 452-457.
• Toparlak M, Tüzer E. Veteriner Helmintoloji. İ.Ü. Veteriner Fakültesi Parazitoloji Anabilim Dalı. 2000; pp 50-56.
• Unat EK, Yücel A, Atlaş K, Samastı M. Unat’ ın Tıp Parazitolojisi (5. Baskı). İ.Ü Cerrahpaşa Tıp Fak. Vakfı Yayın no: 15, İstanbul. 1995.
• Ütük AE, Şimşek S, Köroğlu E, McManus DP. Molecular Genetic Characterization of Different İsolates of Echinococcus granulosus in east and southeast regions of Turkey. Acta Trop. 2008; 107: 192-194.
• Varcasia A, Canu S, Kogkos A, Pipia AP, Scala A, Garippa G, Seimenis A. Molecular characterization of Echinococcus granulosus in sheep and goats of Peloponnesus, Greece. Parasitol Res. 2007; 101: 1135-1139.
• Vural G, Unsal BA, Gauci CG, Bagci O, Gıcık Y, Lightowlers MW. Variability in the Echinococcusgranulosus Cytochrome c oxidase 1 Mitochondrial Gene Sequencefrom Livestock in Turkey and a Re-appraisal of the G1-3 Genotype Cluster. Vet Parasitol. 2008; 154(3-4): 347-350.