ARMAĞAN ERDEM ÜTÜK, Fatma Çiğdem PİŞKİN, İBRAHİM BALKAYA

Molecular detection of anisakis pegreffii in horse mackerels (trachurus trachurus) sold for human consumption in Erzurum province of Turkey

Bu çalışma, Erzurum ilinde insan tüketimi için satılan istavrit balıklarında (Trachurus trachurus) tespit edilen altı parazitin tür tanısını yapmak amacıyla planlanmıştır. Nematod olarak belirlenen altı parazitten DNA ekstraksiyonu yapılmıştır. Tür tanısı, rDNA’nın ITS bölgelerinin (ITS1, 5.8 alt ünite rRNA ve ITS2) PCR ve PCR-RFLP analizleri ve mitokondrial sitokrom c oksidaz alt ünite 1 (mt-CO1) ve mt- CO2 genlerinin kısmi olarak sekanslanması ile yapılmıştır. PCR ve PCR-RFLP sonuçlarına göre tüm parazitlerin Anisakis pegreffii olduğu belirlenmiştir. Rastgele seçilen bir parazitin mt-CO1 geninin kısmi sekans sonucu A. simplex, iki parazitin mt-CO2 geninin kısmi sekans sonuçları ise A. pegreffii ile uyuşmuştur. Bu çalışma ile A. pegreffii Türkiye’de ilk kez moleküler olarak tespit edilmiştir.

Anahtar Kelimeler:

Anisakis, teşhis, anisakiyaz, Türkiye, Trachurus trachurus

Türkiye’nin Erzurum ilinde insan tüketimi için satılan istavrit balıklarında (trachurus trachurus) anisakis pegreffii’nin moleküler tanısı

This study was planned to identify six parasites detected in horse mackerels (Trachurus trachurus) sold for human consumption in Erzurum province. DNA extraction was performed on six parasites diagnosed as nematode. Species identification was done by PCR and PCR-RFLP analysis of ITS (ITS-1, 5.8S subunit rRNA gene and ITS-2) region of ribosomal DNA (rDNA) and partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) and mt-CO2 genes. According to the PCR and PCR-RFLP results, all parasites were identified as Anisakis pegreffii. Partial sequence of mt-CO1 gene of one randomly selected parasite was corresponding with A. simplex and mt-CO2 genes of two parasites were corresponding with A. pegreffii. With this study, A. pegreffii was molecularly detected for the first time in Turkey.

Keywords:

Anisakis, diagnosis, anisakiasis, Turkey, Trachurus trachurus,

PDF

___

1. Lymbery AJ, Cheah FY: Anisakid nematodes and anisakiasis. In, Murrell KD, Fried B (Eds): Food-Borne Parasitic Zoonoses, Fish and Plant Borne Parasites. 1st ed., pp. 185-209, Springer Science Business Media, LLC, New York, USA, 2007.
2. Sterling CR: Food-borne nematod infection. In, Ortega YR (Ed): Food- borne Parasites. 1st ed., pp. 135-153, Springer Science Business Media, LLC, New York, USA, 2006.
3. Anderson RC: Nematode parasites of vertebrates: Their development and transmission. 2nd ed., pp. 245-347, Cambridge, CAB International, 1992.
4. Mattiucci S, Nascetti G: Molecular systematics, phylogeny and ecology of anisakid nematodes of the genus Anisakis Dujardin, 1845: An update. Parasité, 13, 99-113, 2006.
5. Umerha A, Kawakami Y, Araki J, Uchida A: Multiplex PCR for the identification of Anisakis simplex sensu stricto, Anisakis pegreffii and the other anisakid nematodes. Parasitol Int, 57, 49-53, 2008.
6. Fang W, Xu S, Zhang S, Wang Y, Chen X, Luo D: Multiple primer PCR for the identification of Anisakid nematodes from Taiwan Strait. Exp Parasitol, 124, 197-201, 2010.
7. Umerha A, Kawakami Y, Matsui T, Araki J, Uchida A: Molecular identification of Anisakis simplex sensu stricto and Anisakis pegreffii (Nematoda: Anisakidae) from fish and cetacean in Japanase waters. Parasitol Int, 55, 267-271, 2006.
8. Umerha A, Kawakami Y, Ooi HK, Uchida A, Ohmae H, Sugiyama H: Molecular identification of Anisakis type l larvae isolated from hairtail fish off the coasts of Taiwan and Japan, Int J Food Microbiol, 143, 161-165, 2010.
9. S´anchez JM, Artacho-Reinoso ME, D´iaz-Gavil´an M, Valero-L´opez A: Structure of Anisakis simplex s.l. populations in a region sympatric for A. pegreffii and A. simplex s.s. Absence of reproductive isolation between both species. Mol Biochem Parasitol, 141, 155-162, 2005.
10. Mattiucci S, Paoletti M, Borrini F, Palumbo M, Palmieri RM, Gomes V, Casati A, Nascetti G: First molecular identification of the zoonotic parasite Anisakis pegreffii (Nematoda: Ascaridida) in paraffin-embedded granuloma taken from a case of human intestinal anisakiasis in Italy. BMC Infect Dis, 11, 1-6, 2011.
11. Kim KH, Eom KS, Park JK: The complete mitochondrial genome of Anisakis simplex (Ascaridida: Nematoda) and phylogenetic implications. Int J Parasitol, 36, 319-328, 2006.
12. Sambrook J, Fritsch EF, Maniatis T: Molecular cloning: A laboratory manual, 2nd ed., Cold Spring Harbor, NY, 1989.
13. Utuk AE, Simsek S, Koroglu E, McManus DP: Molecular genetic characterization of different isolates of Echinococcus granulosus in East and Southeast Regions of Turkey. Acta Tropica, 107, 192-194, 2008.
14. Umerha A, Kawakami Y, Araki J, Uchida A: Molecular identification of the etiological agent of the human anisakiasis in Japan. Parasitol Int, 56, 211-215, 2007.
15. Pontes T, Amelio SD, Costa G, Paggi L: Molecular characterization of larval anisakid nematodes from marine fishes of Madeira by a PCR-based approach, with evidence for a new species. J Parasitol, 9, 1430-1434, 2005.
16. Lee MH, Cheon DS, Choi C: Molecular genotyping of Anisakis species from Korean sea fish by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Food Cont, 20, 623-626, 2009.
17. Bowles J, Blair D, McManus DP: Genetic variants within the genus Echinococcus identified by mitochondrial DNA sequencing. Mol Biochem Parasitol, 54, 165-174, 1992.
18. Nadler SA, Hudspeth DSS: Phylogeny of the Ascaridoidea (Nematoda: Ascaridida) based on three genes and morphology: Hypotheses of structural and sequence evolution. J Parasitol, 86, 380-393, 2000.
19. Knudsen B, Knudsen T, Flensborg M, Sandmann H, Heltzen M, Andersen A, Dickenson M, Bardram J, Steffensen PJ, Mønsted S, Lauritzen T, Forsberg R, Thanbichler A, Bendtsen JD, Görlitz L, Rasmussen J, Tordrup D, Værum M, Ravn MN, Hachenberg C, Fisker E, Dekker P, Schultz J, Hein AMK, Sinding JB: CLC Main Workbench, Version 5.5, CLC bio, Finlandsgade 10-12, Katrinebjerg, 8200 Aarhus N, Denmark, 2007.
20. Santora M, Mattiucci S, Paoletti M, Liotta A, Uberti BD, Galiero G, Nascetti G: Molecular identification and pathology of Anisakis pegreffii (Nematode: Anisakidae) infection in the Mediterranean loggerhead sea turtle (Caretta caretta). Vet Parasitol, 174, 65-71, 2010.
21. Valentini A, Mattiucci S, Bondanelli P, Webb SC, Mignucci- Giannone AA, Colom-Llavina MM, Nascetti G: Genetic relationships among Anisakis species (Nematoda: Anisakidae) Inferred from mitochondrial cox2 sequences, and comparison with allozyme data. J Parasitol, 92, 156-166, 2006.
22. Quiazon KMA, Yoshinaga T, Ogawa K: Distribution of Anisakis species larvae from fishes of the Japanese waters. Parasitol Int, 60, 223- 226, 2011.
23. Abe N, Tominaga K, Kimata I: Usefulness of PCR-restriction fragment length polymorphism analysis of the internal transcribed spacer region of rDNA for identification of Anisakis simplex complex. Jpn J Infect Dis, 59, 60-62, 2006.
24. Suzuki J, Murata R, Hosaka M, Araki J: Risk factors for human Anisakis infection and association between the geographic origins of Scomber japonicus and anisakid nematodes. Int J Food Microbiol, 137, 88- 93, 2010.
25. Quiazon KMA, Yoshinaga T, Ogawa K: Morphological differences between larvae and in vitro-cultured adults of Anisakis simplex (sensu stricto) and Anisakis pegreffii (Nematoda: Anisakidae). Parasitol Int, 57, 483-489, 2008.
26. Noguera P, Collins C, Bruno D, Pert C, Turnbull A, McIntosh A, Lester K, Bricknell I, Wallace S, Cook P: Red vent syndrome in wild Atlantic salmon Salmo salar in Scotland is associated with Anisakis simplex sensu stricto (Nematoda: Anisakidae). Dis Aquat Org, 87, 199-215, 2009.
27. Cross MA, Collins C, Campbell N, Watts PC, Chubb JC, Cunningham CO, Hatfield EMC, MacKenzie K: Levels of intra-host and temporal sequence variation in a large COI sub-units from Anisakis simplex sensu stricto (Rudolphi 1809) (Nematoda: Anisakisdae): Implications for fisheries management. Mar Biol, 151, 695-702, 2007.
28. Akmirza A: Gökçeada civarında avlanan Tekir (Mullus surmuletus L) balığının metazoon parazitleri. İstanbul Üniv Vet Fak Derg, 26, 129-140, 2000.
29. Tuncel VA, Akmirza A: Karadeniz ve Marmara’da avlanan hamsi (Engraulis encrasicolus (Linnaeus, 1758), balığının endoparazitlerinin karşılaştırılması. İst Üniv Su Ürünleri Derg, 20, 17-26, 2006.
30. Oğuz MC, Güre H, Özdemir H, Öztürk MO, Savaş Y: Çanakkale ili kıyılarında yakalanan ekonomik öneme sahip bazı teleost balıklarda Anisakis simplex, Rudolphi 1809 araştırılması. Türkiye Parazitol Derg, 24, 431-434, 2000.