Emrah SİMSEK, Alparslan YİLDİRİM, Abdullah INCİ, Onder DUZLU, Erdal YİLMAZ, Zuhal ONDER, Arif CİLOĞLU, Gokmen Zafer PEKMEZCİ

Türkiye’de Tatlı Su Kefali’nin (Squalius cephalus) Solungaçlarını Enfekte Eden Posthodiplostomum cuticola (von Nordmann, 1832) Dubois, 1936 (Trematoda: Diplostomidae) Metaserkerlerinin İlk Moleküler Karakterizasyonu

Posthodiplostomum Dubois, 1936 türleri dünya genelinde oldukça yaygınlık göstermektedirler. Avrupa’da yaygın olan P. cuticola türünün metaserkeri tatlı su balıklarında siyah nokta “black spot” hastalığına sebep olmaktadır. GenBank veri tabanında P. cuticola türüne ilişkin mitokondriyal ve ribozomal sekans kaydı sınırlıdır. Diğer yandan, Türkiye’de Posthodiplostomum cinsinde yer alan trematodların sekans karakterizasyonuna dair herhangi bir veri bulunmamaktadır. Bu çalışmada, tatlı su kefallerinde (Squalius cephalus) Posthodiplostomum metaserkerlerinin araştırılması ve tespit edilen izolatların mt-COI ve ITS-2 gen bölgelerinin moleküler karakterizasyonlarının yapılması amaçlanmıştır. İncelenen 123 balıktan ikisinin (%1,6) solungaçlarında Posthodiplostomum metaserkerleri saptanmıştır. Çalışmada tespit ettiğimiz izolatların (POSTC1-C2) (MN701652-3) ITS-2 gen bölgesine ait nükleotid dizilimleri, Çek Cumhuriyeti ve Macaristan’da farklı balık türlerinin deri, kas ve yüzgeçlerinden izole edilen P. cuticola izolatları ile %99,8-100 arasında identiklik göstermiştir. Yine POSTC1-C2 izolatlarının (MN700658-9) mt-COI gen bölgesine ait nükleotid dizilimleri, Litvanya’da Planorbis planorbis’te bulunan P. cuticola izolatı (serker) (KX931424) ile %99,1-98,9 oranında identik bulunmuştur. Bu çalışma ile Türkiye’de ilk kez tatlı su kefalinde (Squalius cephalus) bulunan P. cuticola izolatlarının moleküler ve filogenetik karakterizasyonları ortaya konulmuştur.

Anahtar Kelimeler:

ITS-2 ve mt-COI gen bölgeleri, Moleküler karakterizasyon, Posthodiplostomum cuticola, Squalius cephalus, Türkiye

First Molecular Characterization of Posthodiplostomum cuticola (von Nordmann, 1832) Dubois, 1936 (Trematoda: Diplostomidae) Metacercariae Infecting the Gills of Chubs (Squalius cephalus) in Turkey

Posthodiplostomum Dubois, 1936 species have worldwide distribution. Metacercariae of P. cuticola that is common in Europe causes “black spot” disease in freshwater fishes. A limited number of data on mitochondrial and ribosomal sequences of P. cuticola are available in the GenBank. In addition, there is no sequencing data on trematodes belonging to genus of Posthodiplostomum in Turkey. The aim of this study was to investigate the presence of Posthodiplostomum metacercariae in freshwater chubs (Squalius cephalus) and to reveal the molecular characterization of mt-COI and ITS-2 gene regions of the identified isolates. Posthodiplostomum metacercariae were detected in the gills of two (1.6%) out of 123 fish. The ITS-2 gene sequences of the isolates (POSTC1-C2) (MN701652-3) found in this study showed 99.8-100% identity with P. cuticola sequences isolated from skin, muscle and fins of different fish species in Czech Republic and Hungary. In addition, the mt-COI gene sequences of the POSTC1-C2 isolates (MN700658-9) showed 99.1-98.9% identity with P. cuticola isolate (cercariae) (KX931424) from Planorbis planorbis in Lithuanian. This study provides first molecular and phylogenetic characterization of P. cuticola isolates found in chubs (Squalius cephalus) in Turkey.

Keywords:

ITS-2 and mt-COI gene regions, Molecular characterization, Posthodiplostomum cuticola, Squalius cephalus, Turkey,

PDF

___

Boone EC, Laursen JR, Colombo RE, Meiners SJ, Romani MF, Keeney DB. Infection patterns and molecular data reveal host and tissue specificity of Posthodiplostomum species in centrarchid hosts. Parasitology. 2018; 145(11):1458-1468.
Dönges J. Der Lebenszyklus von Posthodiplostomum cuticola (v. Nordmann, 1832) Dubois, 1936 (Trematoda, Diplostomatidae). Z Parasıtenk. 1964; 24(2):169-248.
Güven A, Öztürk T. Metazoan parasite faunas of three Gobiid species (Actinopterygii: Gobiidae) inhabiting the lower Kızılırmak Delta in Samsun: A comparative study. Türkiye Parazitol Derg. 2018; 42:33-38.
Hoogendoorn C, Smit NJ, Kudlai O. Molecular and morphological characterisation of four diplostomid metacercariae infecting Tilapia sparrmanii (Perciformes: Cichlidae) in the North West Province, South Africa. Parasitol Res. 2019; 118:1403-1416.
Huelsenbeck JP, Ronquist F. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics. 2001; 17:754-755.
Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, Buxton S, Cooper A, Markowitz S, Duran C, Thierer T, Ashton B, Meintjes P, Drummond A. Geneious Basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012; 28:1647-9.
Kimura M. A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980; 16:111-20.
Kumar S, Stecher G, Tamura K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol. 2016; 33(7):1870-4.
Kvach Y, Jurajda P, Bryjová A, Trichkova T, Ribeiro F, Přikrylová I, Ondračková M. European distribution for metacercariae of the North American digenean Posthodiplostomum cf. minimum centrarchi (Strigeiformes: Diplostomidae). Parasitol Int. 2017; 66:635-642.
Lane RL, Morris, JE. Biology, prevention and effects of common grubs (digenetic trematodes) in freshwater fish. Tech. Bull. 2000; 115:1-6.
Locke SA, McLaughlin JD, Marcogliese DJ. DNA barcodes show cryptic diversity and a potential physiological basis for host specificity among Diplostomoidea (Platyhelminthes: Digenea) parasitizing freshwater fishes in the St. Lawrence River, Canada. Mol Ecol. 2010; 19:2813-2827.
López-Hernández D, Locke SA, de Melo AL, Rabelo ÉML, Pinto HA. Molecular, morphological and experimental assessment of the life cycle of Posthodiplostomum nanum Dubois, 1937 (Trematoda: Diplostomidae) from Brazil, with phylogenetic evidence of the paraphyly of the genus Posthodiplostomum Dubois, 1936. Infect Genet Evol 2018; 63:95-103.
Moszczynska A, Locke S, McLaughlin J, Marcogliese D, Crease T. Development of primers for the mitochondrial cytochrome c oxidase I gene in digenetic trematodes (Platyhelminthes) illustrates the challenge of barcoding parasitic helminths. Mol Ecol Resour. 2009; 9:75-82.
Nguyen TC, Li YC, Makouloutou P, Jimenez LA, Sato H. Posthodiplostomum sp. metacercariae in the trunk muscle of northern snakeheads (Channa argus) from the Fushinogawa River, Yamaguchi, Japan. J Vet Med Sci. 2012; 74;1367-1372.
Niewiadomska K. Superfamily Diplostomoidea Poirier, 1886, In: Keys to the Trematoda, Ed; Gibson DI, Jones A, Bray RA, 1th Ed., CAB International and The Natural History Museum, UK. 2002; pp. 159-242.
Ondračková M, Dávidová M, Gelnar M, Jurajda P. Susceptibility of Prussian carp infected by metacercariae of Posthodiplostomum cuticola (v. Nordmann, 1832) to fish predation. Ecol Res. 2006; 21:526-529.
Öztürk MO. Eber Gölü (Afyon)’ndeki Sazan (Cyprinus carpio L.)’ların metazoon parazitleri üzerine bir araştırma. Türkiye Parazitol Derg. 2005; 29(3):204-210.
Poulin R. Uneven distribution of cryptic diversity among higher taxa of parasitic worms. Biol Lett 2011; 7:241-244.
Skov J, Kania PW, Dalsgaard A, Jørgensen TR, Buchmann K. Life cycle stages of heterophyid trematodes in Vietnamese freshwater fishes traced by molecular and morphometric methods. Vet Parasitol. 2009; 160:66-75.
Stoyanov B, Georgieva S, Pankov P, Kudlai O, Kostadinova A, Georgiev BB. Morphology and molecules reveal the alien Posthodiplostomum centrarchi Hoffman, 1958 as the third species of Posthodiplostomum Dubois, 1936 (Digenea: Diplostomidae) in Europe. Syst Parasitol. 2017; 94:1-20.
Uzunay E, Soylu E. Sapanca Gölü’nde yaşayan Sazan (Cyprinus carpio Linnaeus, 1758) ve Karabalık (Vimba vimba Linnaeus, 1758)’ın metazoan parazitleri. Türkiye Parazitol Derg. 2006; 30(2):141-150.
Van Steenkiste N, Locke SA, Castelin M, Marcogliese DJ, Abbott CL. New primers for DNA barcoding of digeneans and cestodes (Platyhelminthes). Mol Ecol Resour. 2015; 15:945-952.