Kayseri Yöresinde Sığırlarda Blastocystis’in Moleküler Prevalansı ve Filogenetik Karakterizasyonu

Yıl 2022, , 1 - 6, 31.03.2022

Ahmet Tavur Zuhal Önder

https://doi.org/10.30607/kvj.996557

Cited By: 1

Öz

Blastocystis insanlarda ve hayvanlarda en yaygın bulunan zoonotik parazitlerden biridir. Bu çalışmada, sığırlarda (Bos taurus) Blastocystis’in moleküler prevalansı ve alt tiplerinin belirlenmesi amaçlanmıştır. Kayseri yöresinde bulunan çeşitli kesimhanelerde kesilen toplam 150 sığırdan taze dışkı örnekleri toplanmıştır. Dışkı örneklerinden genomik DNA ekstraksiyonu yapılmış ve DNA örnekleri Blastocystis’in small subunit ribosomal RNA (SSU rRNA) geninin amplifikasyonunda kullanılmıştır. Tüm PCR pozitif ürünler alt tiplerin belirlenmesi için sekanslanmıştır. Elde edilen sekanslar uygun genetik yazılımlarla işlenerek genotipik yapıları ve sonrasında da filogenetik ilişkileri ortaya çıkarılmıştır. İncelemesi yapılan örneklerin SSU rRNA geninin PCR analizleri ile Blastocystis’in ortalama prevalansı %58,7 olarak belirlenmiştir. Blastocystis SSU rRNA geninin PCR ürünlerinin sekans analizleri sonucunda izolatların ruminant spesifik alt tip, ST10, içerisinde olduğu tespit edilmiştir. Filogenetik analizler çalışmada karakterize edilen ST10 izolatlarının daha önce sığırlarda identifiye edilen izolatlarla birlikte kümelendiğini ortaya koymuştur. Bu çalışma ile mezbahada kesilen sığırlarda Blastocystis sp.’nin moleküler karakterizasyonu ve alt tipi üzerine veriler elde edilmiştir.

Anahtar Kelimeler

Alt tip, Blastocystis, Filogenetik Karakterizasyon, Moleküler Prevalans, Sığır

Destekleyen Kurum

Erciyes Üniversitesi

Proje Numarası

TYL-2018-8643

Molecular Prevalence and Phylogenetic Characterization of Blastocystis in Cattle in Kayseri Province, Turkey

Öz

Blastocystis is one of the most common emerging zoonotic parasites in humans and animals. This study aimed to determine the molecular prevalence and subtype of Blastocystis in cattle (Bos taurus). A total of 150 fresh fecal samples were collected from slaughtered cattle from various slaughterhouses in Kayseri Province, Central Anatolia. Genomic DNA was extracted from all samples and used in PCR analyses of the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis. Blastocystis positive samples were sequenced for identify subtypes. Obtained sequences were assembled with suitable genetic software, then phylogenetic relationships were revealed. According to PCR analyses, overall prevalence of Blastocystis was determined as 58.7%. The sequence analyses of the PCR product gene revealed the presence of one known livestock-specific subtype, ST10. Phylogenetic analysis revealed that ST10 isolates the characterized in the study were clustered with isolates identified previously from cattle. Molecular characterization and subtype of Blastocystis sp. in slaughtered cattle in slaughterhouses were obtained data with this study.

Anahtar Kelimeler

Blastocystis, Cattle, Phylogenetic Characterization, Subtype, Molecular Prevalence

Proje Numarası

TYL-2018-8643

Kaynakça

• Abe N, Nagoshi M, Takami K, Sawano Y, Yoshikawa H. A survey of Blastocystis sp. in livestock, pets, and zoo animals in Japan. Vet Parasitol. 2002; 106: 203-212.
• Alfellani MA, Taner-Mulla D, Jacob AS, Imeede CA, Yoshikawa H, Stensvold CR, Clark CG. Genetic diversity of Blastocystis in livestock and zoo animals. Protist. 2013; 164: 497-509.
• Aynur ZE, Guclu O, Yıldız I., Aynur H, Ertabaklar H, Bozdogan B, Ertug S. Molecular characterization of Blastocystis in cattle in Turkey. Parasitol Res. 2019; 118: 1055-1059.
• Badparva E, Sadraee J, Kheirandish F. Genetic diversity of Blastocystis isolated from cattle in Khorramabad, Iran. Jundishapur J Microbiol. 2015; 8.
• Beyhan Y, Yilmaz H, Cengiz Z, Ekici A. Clinical significance and prevalence of Blastocystis hominis in Van, Turkey. SMJ. 2015; 36: 1118-1121.
• Cian A, El Safadi D, Osman M Moriniere R, Gantois N, Benamrouz-Vanneste S, Delgado-Viscogliosi P, Guyot K, Li L.L., Monchy S, No¨el C, Poirier P, Nourrisson C, Wawrzyniak I, Delbac F, Bosc S, Chab´e M, Petit T, Certad G, Viscogliosi E. Molecular epidemiology of Blastocystis sp. in various animal groups from two French zoos and evaluation of potential zoonotic risk. PLoS ONE 2017; 12: e0169659.
• Clark CG, van der Giezen M, Alfellani MA, Stensvold CR. Chapter One-Recent Developments in Blastocystis Research. In Advances in Parasitology (D. Rollinson, ed). Academic Press, 2013; pp. 1-32.
• Dogan N, Aydin M, Tuzemen NU, Dinleyici EC, Oguz I, Dogruman-Al F. Subtype distribution of Blastocystis spp. isolated from children in Eskisehir, Turkey. Parasitol Int. 2017; 66: 948-951.
• 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.
• Greige S, El-Safadi D, Khaled S, Gantois N, Baydouna B, Chemalyc M, Benamrouz-Vannestea S, Chabea M, Osman M, Certada G, Hamzeb M, Viscogliosia E. First report on the prevalence and subtype distribution of Blastocystis spp. in dairy cattle in Lebanon and assessment of zoonotic transmission. Acta Trop. 2019; 194: 23-29
• Koltas IS, Eroglu F. Subtype analysis of Blastocystis isolates using SSU rRNA-DNA sequencing in rural and urban population in southern Turkey. Exp Parasitol. 2016; 170:247-251.
• Koloren Z, Gulabi BB, Karanis P. Molecular identification of Blastocystis sp. subtypes in water samples collected from Black sea, Turkey. Acta Tropic. 2018;180: 58-68.
• Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016; 33: 1870-1874.
• Lee H, Lee SH, Seo MG, Kim HY, Kim JW, Lee YR, Kim JH, Kwon OD, Kwak D. Occurrence and genetic diversity of Blastocystis in Korean cattle. Vet Parasitol. 2018; 258: 70-73.
• Lee L, Chye T, Karmacharya B, Govind S. Blastocystis sp.: waterborne zoonotic organism, a possibility? Parasit Vectors 2012; 5: 130.
• Lee SH, Kim HY, Choi EW, Kim D. Causative agents and epidemiology of diarrhea in Korean native calves. J Vet Sci. 2019; 20.
• Li W, Zhou B, Li Juan, Zhu S, Li Jie. Single parabolic band behavior of thermoelectric p-type Cu 4 Mn 2 Te 4. J Alloy Compd. 2018; 753: 93-99.
• Librado P, Rozas J. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 2009; 25: 1451-1452.
• Malatyalı E, Ertabaklar H, Ertug S. Subtype distribution of Blastocystis spp. with DNA barcoding and evaluation of diagnostic methods. Mikrobiyol Bul. 2019; 53(3): 308-318.
• Malatyalı E, Başaran G, Gökçimen A, Ertabaklar H, Ertug S. First Molecular characterisation of Blastocystis from experimental rats in Turkey and comparison of the frequencies between obese and non-obese groups subtype distribution of Blastocystis spp. with DNA barcoding and evaluation of diagnostic methods. Turkiye Parazitol Derg. 2021; 45 (3), 165-170.
• Maloney JG, Lombard JE, Urie NJ, Shivley CB, Santin M. Zoonotic and genetically diverse subtypes of Blastocystis in US pre-weaned dairy heifer calves. Parasitol Res. 2019; 118: 575-582.
• Noradilah SA, Anuar TS, Moktar N, Lee IL, Salleh FM, Manap SNA, Mohtar NSH, Azrul SM, Abdullah WO, Nordin A, Abdullah SR. Molecular epidemiology of Blastocystis spp. in animals reared by the aborigines during wet and dry seasons in rural communities, Pahang, Malaysia Southeast. Asian J Trop Med Public Health. 2017; 48(6): 1151-1160.
• Onder Z, Yildirim A, Pekmezci D, Duzlu O, Pekmezci Z, Ciloglu A, Simsek E, Delibasi Kokcu N, Yetismis G, Ercan N, Inci A. Molecular identification and subtype distribution of Blastocystis sp. in farm and pet animals in Turkey. Acta Tropic. 2021; 220: 105939.
• Parkar U, Traub RJ, Vitali S, Elliot A, Levecke B, Robertson I, Geurden T, Steele J, Drake B, Thompson RCA. Molecular characterization of Blastocystis isolates from zoo animals and their animal-keepers. Vet Parasitol. 2010; 169: 8-17.
• Posada D. jModelTest: Phylogenetic Model Averaging. Mol Biol Evol. 2008; 25: 1253-1256.
• Ramírez JD, Sánchez A, Hernández C, Flórez C, Bernal MC, Giraldo JC, Reyes P, López MC, García L, Cooper PJ, Vicuña Y, Mongi F, Casero RD. Geographic distribution of human Blastocystis subtypes in South America. Infect Genet Evol. 2016; 41: 32-35. Ramírez JD, Sánchez LV, Bautista DC, Corredor AF, Flórez AC, Stensvold CR. Blastocystis subtypes detected in humans and animals from Colombia. Infect Genet Evol. 2014; 22: 223-228.
• Ren M, Song J, Yang F, Zou M, Wang P, Wang D, Zhang H, Zhao G, Lin Q. First genotyping of Blastocystis in yaks from Qinghai Province, northwestern China. Parasites Vectors 2019;12: 171.
• Salim HR, Kumar GS, Vellayan S, Mak JW, Anuar AK, Init I, Vennila GD, Saminathan R, Ramakrishnan K. Blastocystis in animal handlers. Parasitol Res. 1999; 85: 1032-1033.
• Santín M, Gómez-Muñoz MT, Solano-Aguilar G, Fayer R. Development of a new PCR protocol to detect and subtype Blastocystis spp. from humans and animals. Parasitol Res. 2011; 109: 205-212.
• Stensvold CR, Alfellani MA, Nørskov-Lauritsen S, Prip K, Victory EL, Maddox C, Nielsen HV, Clark CG. Subtype distribution of Blastocystis isolates from synanthropic and zoo animals and identification of a new subtype. Int J Parasitol. 2009; 39: 473-479.
• Stensvold CR, Clark CG. Current status of Blastocystis: A personal view. Parasitol Int. 2016; 65: 763-771. Stensvold CR, Clark CG. Pre-empting Pandora’s box: Blastocystis subtypes revisited. Trends Parasitol. 2020; 36: 229-232.
• Suwanti LT, Susana Y, Hastutiek P, Suprihati E, Lastuti NDR. Blastocystis spp. subtype 10 infected beef cattle in Kamal and Socah, Bangkalan, Madura, Indonesia. Vet World. 2020; 13: 231-237.
• Valença-Barbosa C, do Bomfim TCB, Teixeira BR, Gentile R, Neto SF da C, Magalhães BSN, Balthazar D de A., Silva FA, Biot R, Levy CM d’Avila, Santos HLC. Molecular epidemiology of Blastocystis isolated from animals in the state of Rio de Janeiro, Brazil. PLoS One 2019; 14.
• Wang J, Gong B, Yang F, Zhang W, Zheng Y, Liu A. Subtype distribution and genetic characterizations of Blastocystis in pigs, cattle, sheep and goats in northeastern China’s Heilongjiang Province. Infect Genet Evol. 2018; 57: 171-176.
• Yoshikawa H, Abe N, Wu Z. Genomic polymorphism among Blastocystis isolates and development of PCR-based identification of zoonotic isolates. J Eukaryotic Microbiology 2003; 50: 710-711.
• Yoshikawa H, Abe N, Wu Z. PCR-based identification of zoonotic isolates of Blastocystis from mammals and birds. Microbiology 2004; 150: 1147-1151.
• Yoshikawa H, Koyama Y, Tsuchiya E, Takami K. Blastocystis phylogeny among various isolates from humans to insects. Parasitol Int. 2016; 65: 750-759.
• Zhao GH, Hu XF, Liu TL, Hu RS, Yu ZQ, Yang WB, Wu YL, Yu SK, Song JK. Molecular characterization of Blastocystis sp. in captive wild animals in Qinling Mountains. Parasitol Res. 2017; 116: 2327-2333.
• Zhu W, Tao W, Gong B, Yang H, Li Y, Song M, Lu Y, Li W. First report of Blastocystis infections in cattle in China. Vet Parasitol. 2017; 246: 38-42.