Yıl: 2020 Cilt: 44 Sayı: 1 Sayfa Aralığı: 36 - 42 Metin Dili: Türkçe DOI: 10.4274/tpd.galenos.2020.6851 İndeks Tarihi: 20-10-2020

Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu

Öz:
Amaç: Bu çalışmada sağlıklı sığırlarda Enterocytozoon bieneusi’nin moleküler prevalansının ve genotiplerinin belirlenmesiamaçlanmıştır.Yöntemler: Çalışmada, Ekim 2017 ve Mart 2018 tarihleri arasında Sivas yöresindeki sağlıklı görünümlü 150 sığırdan dışkıörnekleri toplanmış ve genomik DNA (gDNA) izolasyonları yapılmıştır. Elde edilen gDNA izolatları, E. bieneusi’nin identifikasyonuamacıyla, ITS rRNA gen bölgesini amplifiye eden Nested PZR ile işlenmiştir. E. bieneusi pozitif izolatların ITS rRNA gen bölgesiamplikonları genotiplendirme ve filogenetik analizler için sekanslanmıştır. Elde edilen sekanslar uygun genetik yazılımlarlaişlenerek genotipik yapıları ve sonrasında da filogenetik ilişkileri ortaya çıkarılmıştır.Bulgular: Nested PZR ile incelemesi yapılan örneklerden 29’u (%19,3) E. bieneusi yönünden pozitif bulunmuştur. Sekansanalizleri sonucu beş ayrı genotip belirlenmiştir. En yaygın genotip olarak bulunan ERUSS1 genotipi ile ERUSS2-4 genotipleriyakın genotipler olarak karakterize edilmiş ve ilk kez rapor edilmiştir. İki izolatın ise Almanya’da bir sığırdan rapor edilmiş olan Ngenotipinde olduğu belirlenmiş ve diğer genotiplerden daha farklı olduğu görülmüştür. Filogenetik analizler çalışmada karakterizeedilen tüm genotiplerin genogrup 2’de yer aldığını ortaya koymuştur.Sonuç: Bu çalışma ile Türkiye’de sığırlarda E. bieneusi enfeksiyonları üzerine ilk moleküler epidemiyolojik veriler elde edilmiştir.
Anahtar Kelime:

Konular: Mühendislik, Biyotıp Parazitoloji Veterinerlik Biyofizik

Molecular Prevalence and Phylogenetic Characterization of Enterocytozoon bieneusi in Healthy Cattle

Öz:
Objective: In this study, it was aimed to determine the molecular prevalence and genotypes of Enterocytozoon in healthy cattle. Methods: Fecal samples were collected from 50 cattle in Sivas between October 2017 and March 2018 and genomic DNA (gDNA) isolations were performed. gDNA isolates were processed by Nested PCR specifically amplifying ITS rRNA gene region to identify E. bieneusi. ITS rRNA region of E. bieneusi positive isolates were sequenced for genotyping and phylogenetic analyzes. Obtained sequences were assembled with appropriative genetic software, then phylogenetic relationships were revealed. Results: According to Nested PCR analyses, 29 (19.3%) out of totally examined samples were found positive for E. bieneusi. As a result of the sequence analyses, five distinct genotypes were determined. The most frequent genotype ERUSS1 and the other ERUSS2-4 genotypes were characterized as close to each other, which was reported for the first time in the world. Two isolates were determined in N genotype that was reported from cattle in Germany and were more different from the other genotypes. Phylogenetic analysis revealed that all the genotypes characterized in the study belonged to the genogroup 2. Conclusion: First molecular epidemiological data on E. bieneusi in cattle from Turkey were obtained with this study
Anahtar Kelime:

Konular: Mühendislik, Biyotıp Parazitoloji Veterinerlik Biyofizik
Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık
  • 1. Mathis A, Weber R, Deplazes P. Zoonotic potential of the microsporidia. Clin Microbiol Rev. 2005;18:423-445.
  • 2. Vávra J, Lukeš J. Microsporidia and ‘the art of living together’. Adv Parasitol. 2013;82:253-319.
  • 3. Keeling P. Five questions about microsporidia. PLoS Pathogens. 2009;5:e1000489.
  • 4. Santin M, Fayer R. Microsporidiosis: Enterocytozoon bieneusi in domesticated and wild animals. Res Vet Sci. 2011;90:363-371.
  • 5. Matos O, Lobo ML, Xiao L. Epidemiology of Enterocytozoon bieneusi Infection in Humans. J Parasitol Res. 2012;2012:981424.
  • 6. Thellier M, Breton J. Enterocytozoon bieneusi in human and animals, focus on laboratory identification and molecular epidemiology. Parasite. 2008;15:349-358.
  • 7. Santin M, Fayer R. Enterocytozoon bieneusi genotype nomenclature based on the internal transcribed spacer sequence: a consensus. J Eukar Microbiol. 2009;56:34-8.
  • 8. Jiang Y, Tao W, Wan Q, Li Q, Yang Y, Lin Y, et al. Zoonotic and potentially host-adapted Enterocytozoon bieneusi genotypes in sheep and cattle in northeast China and an increasing concern about the zoonotic importance of previously considered ruminant-adapted genotypes. Appl Environ Microbiol. 2015;81:3326-3335.
  • 9. Zhao W, Yu S, Yang Z, Zhang Y, Zhang L, Wang R, et al. Genotyping of Enterocytozoon bieneusi (Microsporidia) isolated from various birds in China. Infec Gen Evol. 2016;40:151-154.
  • 10. Karim MR, Dong H, Li T, Yu F, Li D, Zhang L, et al. Predomination and new genotypes of Enterocytozoon bieneusi in captive nonhuman primates in zoos in China: high genetic diversity and zoonotic significance. PloS One. 2015;10:e0117991.
  • 11. Deng L, Li W, Zhong Z, Gong C, Liu X, Huang X, et al. Molecular characterization and multilocus genotypes of Enterocytozoon bieneusi among horses in southwestern China. Parasit Vectors. 2016;9:561.
  • 12. Li W, Deng L, Yu X, Zhong Z, Wang Q, Liu X, et al. Multilocus genotypes and broad host-range of Enterocytozoon bieneusi in captive wildlife at zoological gardens in China. Parasit Vectors. 2016;9:395.
  • 13. Karim MR, Wang R, Dong H, Zhang L, Li J, Zhang S, et al. Genetic polymorphism and zoonotic potential of Enterocytozoon bieneusi from nonhuman primates in China. Appl Environ Microbiol. 2014;80:1893-8.
  • 14. Li N, Xiao L, Wang L, Zhao S, Zhao X, Duan L, et al. Molecular surveillance of Cryptosporidium spp., Giardia duodenalis, and Enterocytozoon bieneusi by genotyping and subtyping parasites in wastewater. PLoS Neg Trop Dis. 2012;6:e1809.
  • 15. Jiang Y, Tao W, Wan Q, Li Q, Yang Y, Lin Y, et al. Zoonotic and potentially host-adapted Enterocytozoon bieneusi genotypes in sheep and cattle in Northeast China and an increasing concern about the zoonotic importance of previously considered ruminant-adapted genotypes. Appl Environ Microbiol. 2015; 81: 3326-3335.
  • 16. Del Coco VF, Cordoba MA, Bilbao G, de Almeida Castro P, Basualdo JA, Santín M. First report of Enterocytozoon bieneusi from dairy cattle in Argentina. Vet Parasitol. 2014;199:112-5.
  • 17. Zhao W, Zhang W, Yang F, Zhang L, Wang R, Cao J, et al. Enterocytozoon bieneusi in dairy cattle in the northeast of china: genetic diversity of ITS gene and evaluation of zoonotic transmission potential. J Eukar Microbiol. 2015;62:553-560.
  • 18. Tang C, Cai M, Wang L, Guo Y, Li N, Feng Y, et al. Genetic diversity within dominant Enterocytozoon bieneusi genotypes in pre-weaned calves. Parasit Vectors. 2018;11:170.
  • 19. Li J, Luo N, Wang C, Qi M, Cao J, Cui Z, et al. Occurrence, molecular characterization and predominant genotypes of Enterocytozoon bieneusi in dairy cattle in Henan and Ningxia, China. Parasit Vectors. 2016;9:142.
  • 20. Ma J, Li P, Zhao X, Xu H, Wu W, Wang Y, et al. Occurrence and molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi in dairy cattle, beef cattle and water buffaloes in China. Vet Parasitol. 2015;207:220-7.
  • 21. Qi M, Jing B, Jian F, Wang R, Zhang S, Wang H, et al. Dominance of Enterocytozoon bieneusi genotype J in dairy calves in Xinjiang, Northwest China. Parasitol Int. 2017;66:960-3.
  • 22. Wang XT, Wang RJ, Ren GJ, Yu ZQ, Zhang LX, Zhang SY, et al. Multilocus genotyping of Giardia duodenalis and Enterocytozoon bieneusi in dairy and native beef (Qinchuan) calves in Shaanxi province, northwestern China. Parasitol Res.2016;115:1355-1361.
  • 23. Fayer R, Santin M, Macarisin D. Detection of concurrent infection of dairy cattle with Blastocystis, Cryptosporidium, Giardia, and Enterocytozoon by molecular and microscopic methods. Parasitol Res. 2012;111:1349-1355.
  • 24. Juránková J, Kamler M, Kovarcík K, Koudela B. Enterocytozoon bieneusi in Bovine Viral Diarrhea Virus (BVDV) infected and noninfected cattle herds. Res Vet Sci. 2013;94:100-4.
  • 25. Santin M, Fayer R. A longitudinal study of Enterocytozoon bieneusi in dairy cattle. Parasitol Res. 2009;105:141-4.
  • 26. Sak B, Brady D, Pelikanova M, Květoňová D, Rost M, Kostka M, et al. Unapparent microsporidial infection among immunocompetent humans in the Czech Republic. J Clin Microbiol. 2011;49:1064-1070.
  • 27. Zhang X, Wang Z, Su Y, Liang X, Sun X, Peng S, et al. Identification and genotyping of Enterocytozoon bieneusi in China. J Clin Microbiol. 2011;49:2006-8.
  • 28. Çetinkaya Ü, Yazar S, Kuk S, Sivcan E, Kaynar L, Arslan D, et al. The high prevalence of Encephalitozoon intestinalis in patients receiving chemotherapy and children with growth retardation and the validity of real-time PZR in its diagnosis. Turk J Medical Sci. 2016;46:1050-8.
  • 29. Oğuz İ, Doğruman AF, Mumcuoğlu İ. İshalli olgularda microsporidia sıklığının Calcofluor Beyazı ve Uvitex 2B kemolüminesans boyama yöntemleriyle araştırılması ve tiplendirilmesi. 18. Ulusal Parazitoloji Kongresi, 2013 28 Eylül-4 Ekim; Denizli. Turkiye; 2013.
  • 30. Özkoç S, Bayram Delibaş S, Akısü Ç. Evaluation of pulmonary microsporidiosis in iatrogenically immunosuppressed patients. Tuberk Toraks. 2016;64:9-16.
  • 31. Duzlu O, Yildirim A, Onder Z, Çiloglu A, Yetismis G, Inci A. Prevalence and Genotyping of Microsporidian Parasites in Dogs in Turkey: Zoonotic Concerns. J Eukaryot Microbiol. 2019;66:771-777.
  • 32. Pekmezci D, Pekmezci GZ, Yildirim A, Duzlu O, Inci A. Molecular Detection of Zoonotic Microsporidia in Domestic Cats in Turkey: A Preliminary Study. Acta Parasitol. 2019;64:13-18.
  • 33. Ercan N. Tavuklarda Enterocytozoon Bieneusi ve Encephalitozoon spp. türlerinin moleküler prevalansı, genotiplendirilmesi ve insan sağlığı açısından risk potansiyelleri. Doktora Tezi. Erciyes Üniversitesi Sağlık Bilimleri Enstitüsü. Kayseri. 2019.
  • 34. Buckholt MA, Lee JH, Tzipori S. Prevalence of Enterocytozoon bieneusi in swine: an 18-month survey at a slaughterhouse in Massachusetts. Appl Environ Microbiol. 2002;68:2595-2599.
  • 35. Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, et al. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012;28:1647-9.
  • 36. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980;16:111-120.
  • 37. Nei M. Phylogenetic analysis in molecular evolutionary genetics. Ann Rev Gen.1996;30:371-403.
  • 38. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016;33:1870-4.
  • 39. Posada D. jModelTest: phylogenetic model averaging. Mol Biol Evol. 2008;25:1253-6.
  • 40. Guindon S, Gascuel O. A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol. 2003;52:696-704.
  • 41. da Silva Fiuza VR, Lopes CW, de Oliveira FC, Fayera R, Santin M. New findings of Enterocytozoon bieneusi in beef and dairy cattle in Brazil. Vet Parasitol. 2016;216:46-51.
  • 42. Fayer R, Santin M, Trout JM. First detection of microsporidia in dairy calves in North America. Parasitol Res. 2003;90:383-386.
  • 43. Fiuza VR, Lopes CW, Cosendey RI, de Oliveira FC, Fayerd R, Santin M. Zoonotic Enterocytozoon bieneusi genotypes found in brazilian sheep. Res Vet Sci. 2016;107:196-201.
  • 44. Santin M, Vecino JA, Fayer R. A zoonotic genotype of Enterocytozoon bieneusi in horses. J Parasitol. 2010;96:157-161.
  • 45. Sak B, Salat J, Horka H, Saková K, Ditrich O. Antibodies enhance the protective effect of CD4+ T lymphocytes in SCID mice perorally infected with Encephalitozoon cuniculi. Parasite Immunol. 2006;28:95-9.
  • 46. Sulaiman IM, Fayer R, Lal AA, Trout JM, Schaefer FW 3rd, Xiao L. Molecular characterization of microsporidia indicates that wild mammals harbor host-adapted Enterocytozoon spp. as well as human-pathogenic Enterocytozoon bieneusi. Appl Environ Microbiol. 2003;69:4495-4501.
  • 47. Santin M, Cortes Vecino JA, Fayer R. Enterocytozoon bieneusi genotypes in dogs in Bogota, Colombia. Am J Trop Med Hyg. 2008;79:215-217.
  • 48. Santin M, Dargatz D, Fayer R. Prevalence and genotypes of Enterocytozoon bieneusi in weaned beef calves on cow-calf operations in the USA. Parasitol Res. 2012;110:2033-2041.
  • 49. Karim MR, Dong H, Yu F, Jian F, Zhang L, Wang R, et al. Genetic diversity in Enterocytozoon bieneusi isolates from dogs and cats in China: host specificity and public health implications. J Clin Microbiol. 2014;52:3297- 3302.
  • 50. Du SZ, Zhao GH, Shao JF, Fang YQ, Tian GR, Zhang LX, et al. Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi in captive non-human primates in Qinling Mountains. Korean J Parasitol. 2015;53:395-402.
  • 51. Peng XQ, Tian GR, Ren GJ, Yu ZQ, Lok JB, Zhang LX, et al. Infection rate of Giardia duodenalis, Cryptosporidium spp. and Enterocytozoon bieneusi in cashmere, dairy and meat goats in China. Infec Gen Evol. 2016;41:26-31.
  • 52. Ye J, Xiao L, Wang Y, Guo Y, Roellig DM, Feng Y. Dominance of Giardia duodenalis assemblage A and Enterocytozoon bieneusi genotype BEB6 in sheep in Inner Mongolia, China. Vet Parasitol. 2015;210:235-239.
  • 53. Zhang XX, Jiang J, Cai YN, Wang CF, Xu P, Yang GL, et al. Molecular characterization of Enterocytozoon bieneusi in domestic rabbits (Oryctolagus cuniculus) in Northeastern China. Korean J Parasitol. 2016;54:81-85.
  • 54. Stensvold CR, Beser J, Ljungstrom B, Troell K, Lebbad M. Low hostspecific Enterocytozoon bieneusi genotype BEB6 is common in Swedish lambs. Vet Parasitol. 2014;205:371-374.
  • 55. Fayer R, Santin M, Trout JM. Enterocytozoon bieneusi in mature dairy cattle on farms in the eastern United States. Parasitol Res. 2007;102:15- 20.
  • 56. Wang L, Xiao L, Duan L, Ye J, Guo Y, Guo M. et al. Concurrent infections of Giardia duodenalis, Enterocytozoon bieneusi, and Clostridium difficile in children during a cryptosporidiosis outbreak in a pediatric hospital in China. PLoS Neg Trop Dis. 2013;7:e2437.
APA biigin t, uslug s, Karademir G, Atelge M, Yetismis G, YILDIRIM A (2020). Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. , 36 - 42. 10.4274/tpd.galenos.2020.6851
Chicago biigin tuğba,uslug sadullah,Karademir Gupse Kübra,Atelge Mubeccel,Yetismis Gamze,YILDIRIM Alparslan Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. (2020): 36 - 42. 10.4274/tpd.galenos.2020.6851
MLA biigin tuğba,uslug sadullah,Karademir Gupse Kübra,Atelge Mubeccel,Yetismis Gamze,YILDIRIM Alparslan Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. , 2020, ss.36 - 42. 10.4274/tpd.galenos.2020.6851
AMA biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. . 2020; 36 - 42. 10.4274/tpd.galenos.2020.6851
Vancouver biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. . 2020; 36 - 42. 10.4274/tpd.galenos.2020.6851
IEEE biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A "Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu." , ss.36 - 42, 2020. 10.4274/tpd.galenos.2020.6851
ISNAD biigin, tuğba vd. "Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu". (2020), 36-42. https://doi.org/10.4274/tpd.galenos.2020.6851
APA biigin t, uslug s, Karademir G, Atelge M, Yetismis G, YILDIRIM A (2020). Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. Türkiye Parazitoloji Dergisi, 44(1), 36 - 42. 10.4274/tpd.galenos.2020.6851
Chicago biigin tuğba,uslug sadullah,Karademir Gupse Kübra,Atelge Mubeccel,Yetismis Gamze,YILDIRIM Alparslan Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. Türkiye Parazitoloji Dergisi 44, no.1 (2020): 36 - 42. 10.4274/tpd.galenos.2020.6851
MLA biigin tuğba,uslug sadullah,Karademir Gupse Kübra,Atelge Mubeccel,Yetismis Gamze,YILDIRIM Alparslan Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. Türkiye Parazitoloji Dergisi, vol.44, no.1, 2020, ss.36 - 42. 10.4274/tpd.galenos.2020.6851
AMA biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. Türkiye Parazitoloji Dergisi. 2020; 44(1): 36 - 42. 10.4274/tpd.galenos.2020.6851
Vancouver biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu. Türkiye Parazitoloji Dergisi. 2020; 44(1): 36 - 42. 10.4274/tpd.galenos.2020.6851
IEEE biigin t,uslug s,Karademir G,Atelge M,Yetismis G,YILDIRIM A "Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu." Türkiye Parazitoloji Dergisi, 44, ss.36 - 42, 2020. 10.4274/tpd.galenos.2020.6851
ISNAD biigin, tuğba vd. "Sağlıklı Sığırlarda Enterocytozoon bieneusi’nin Moleküler Prevalansı ve Filogenetik Karakterizasyonu". Türkiye Parazitoloji Dergisi 44/1 (2020), 36-42. https://doi.org/10.4274/tpd.galenos.2020.6851