Genetic Analysis of the Mitochondrial rrnS Gene of Zoonotic Anisakis pegreffii (Nematoda: Anisakidae) Isolated from Micromesistius Poutassou (R.) in the Aegean Sea

Yıl 2021, , 266 - 269, 30.06.2021

Gökmen Zafer Pekmezci

https://doi.org/10.35229/jaes.913695

Cited By: 3

Öz

Ascaridoid nematodes were found in viscera of the blue whiting Micromesistius poutassou (Risso, 1826) from the Aegean Sea in the parasitological examination. Collected nematodes were morphologically identified as the third (L3) stage of Anisakis type I larvae and then subject to Restriction Fragment Length Polymorphism (RFLP) analysis of the internal transcribed spacer (ITS) region to identify the species. Randomly selected larvae were molecularly confirmed as Anisakis pegreffii by RFLP analysis. Subsequently, L3 of A. pegreffii were amplified and bi-directionally sequenced at the small subunit ribosomal RNA (rrnS) gene. The rrnS sequence of A. pegreffii (MT484284) had 100% identity with the rrnS gene of A. pegreffii (JX500050, LC222461, MF140359, MT312511, MT312512) which were found in fish and cetaceans hosts, Scomber japonicus, Conger myriaster, Neophocaena asiaeorientalis, and Stenella coeruleoalba, from the Mediterranean Sea, Japanese, Chinese, and Korean waters. This study provided the genetic analysis of zoonotic A. pegreffii from the Turkish marine waters based on the rrnS gene for the first time. This sequence (MT484284) can be used as the novel rrnS sequence of A. pegreffii in the genetic analysis for ascaridoid nematodes in the Mediterranean Sea.

Anahtar Kelimeler

Anisakis pegreffii, molecular characterization, mtDNA, rrnS gene, Turkish coast

Ege Denizi'ndeki Micromesistius poutassou'dan (R.) İzole Edilen Zoonotik Anisakis pegreffii'nin (Nematoda: Anisakidae) Mitokondriyal rrnS Geninin Genetik Analizi

Öz

Parazitolojik incelemede Ege Denizi'nden mavi mezgit Micromesistius poutassou (Risso, 1826) balığının iç organlarında askaridoid nematodlar bulundu. Toplanan nematodlar morfolojik olarak üçüncü (L3) dönem Anisakis tip I larva olarak tanımlandı ve daha sonra türleri tanımlamak için internal transcribed spacer (ITS) bölgesi Restriksiyon Parça Uzunluk Polimorfizm (RFLP) analizlerine tabi tutuldu. Rastgele seçilen larvalar RFLP analizleri ile Anisakis pegreffii olarak doğrulandı. Daha sonra A. pegreffii'nin L3'ünün küçük alt birim ribozomal RNA (rrnS) geni çoğaltıldı ve iki yönlü olarak sekanslandı. Anisakis pegreffii'nin rrnS sekansı (MT484284) Akdeniz, Japon, Çin ve Kore sularından Scomber japonicus, Congenaer asyager, Neophocaena asiaeorientalis ve Stenella coeruleoalba gibi balık ve deniz memelilerinde bulunan A. pegreffii'nin rrnS geni (JX500050, LC222461, MF140359, MT312511, MT312512) ile %100 benzerliğe sahipti. Bu çalışma ilk kez Türk deniz sularından zoonotik A. pegreffii'nin rrnS genine dayalı genetik analizini sağladı. Bu dizi (MT484284), Akdeniz'deki askaridoid nematodların genetik analizinde A. pegreffii'nin yeni rrnS dizisi olarak kullanılabilir.

Anahtar Kelimeler

Anisakis pegreffii, moleküler karakterizasyon, mtDNA, rrnS geni, Türk kıyıları

Kaynakça

• Altschul, S.F., Gish, W., Miller, W., Myers, E.W. & Lipman, D.J. (1990). Basic local alignment search tool. Journal of Molecular Biology, 215, 403-410. doi: 10.1016/S0022-2836(05)80360-2.
• Berland, B. (1961). Nematodes from some Norwegian marine fishes. Sarsia, 2, 1-50.
• D'Amelio, S., Barros, N.B., Ingrosso, S., Fauquier, D.A., Russo, R. & Paggi L. (2007). Genetic characterization of members of the genus Contracaecum (Nematoda: Anisakidae) from fish-eating birds from west-central Florida, USA, with evidence of new species. Parasitology, 134, 1041-1051. doi: 10.1017/S003118200700251X.
• D'Amelio, S., Mathiopoulos, K. D., Santos, C. P., Pugachev, O. N., Webb, S. C., Picanço, M. & Paggi, L. (2000). Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda: Ascaridoidea) defined by polymerase-chain-reaction-based restriction fragment length polymorphism. International Journal for Parasitology, 30(2), 223-226. doi: 10.1016/s0020-7519(99)00178-2.
• 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. (2012). Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28(12), 1647-1649.
• Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. (2018). MEGA X: Molecular evolutionary genetics analysis across computing platforms. Molecular Biology Evoluation, 35, 1547-1549. doi: 10.1093/molbev/msy096.
• Mattiucci, S., Cipriani, P., Levsen, A., Paoletti, M. & Nascetti, G. (2018). Molecular epidemiology of Anisakis and Anisakiasis: an ecological and evolutionary road map. Advances in Parasitology, 9, 93-263. doi: 10.1016/bs.apar.2017.12.001.
• Mattiucci, S. & Nascetti, G. (2008). Advances and trends in the molecular systematics of anisakid nematodes, with implications for their evolutionary ecology and host-parasite co-evolutionary processes. Advances in Parasitology, 66, 47-148. doi: 10.1016/S0065-308X(08)00202-9.
• Mattiucci, S., Cipriani, P., Webb, S. C., Paoletti, M., Marcer, F., Bellisario, B., Gibson, D. & Nascetti, G. (2014). Genetic and morphological approaches distinguish the three sibling species of the Anisakis simplex species complex, with a species designation as Anisakis berlandi n. sp. for A. simplex sp. C (Nematoda: Anisakidae). Journal of Parasitology, 100, 199-214. doi: 10.1645/12-120.1.
• Pekmezci, G.Z. (2019). Molecular characterization of Hysterothylacium aduncum (Nematoda: Raphidascarididae) larvae infecting Merlangius merlangus euxinus (Linnaeus, 1758) from the Turkish Black Sea coast based on mitochondrial small subunit ribosomal RNA gene analysis. Etlik Veteriner Mikrobiyoloji Dergisi, 30(1), 64-69. doi:10.35864/evmd.562427
• Pekmezci, G.Z., Onuk, E.E., Bolukbas, C.S., Yardimci, B., Gurler, A.T., Acici, M. & Umur, S. (2014). Molecular identification of Anisakis species (Nematoda: Anisakidae) from marine fishes collected in Turkish waters. Veterinary Parasitology, 201, 82-94. doi: 10.1016/j.vetpar.2014.01.005.
• Pekmezci, G.Z. & Onuk, E.E. (2020). Molecular characterization of Anisakis typica (Nematoda: Anisakidae) from the Mediterranean Sea coasts of Turkey: First mitochondrial rrnS sequence data. Journal of Anatolian Environmental and Animal Sciences, 5(2), 241-244. doi: 10.35229/jaes.738822
• Pekmezci, G. Z. & Yardimci, B. (2019). On the occurrence and molecular identification of Contracaecum larvae (Nematoda: Anisakidae) in Mugil cephalus from Turkish waters. Parasitology Research, 118(5), 1393-1402. doi: 10.1007/s00436-019-06278-x.
• Peter, A.J. & Maillard, C. (1988). Larves d’ascarides parasites de poisons en Mediterranee occidentale. Bulletin du Museum National d'histoire Naturelle, 10 (sect. A), 347-369.
• Simsek, E., Sürsal, N., Karademir, G.K. & Pekmezci, G.Z. (2021). First molecular characterization of Hysterothylacium fabri larvae (Nematoda: Raphidascarididae) in the Mediterranean Sea based on the small subunit ribosomal RNA gene sequence. Kocatepe Veterinary Journal, 14(1), 71-76. doi: 10.30607/kvj.832003
• Zhu, X., Gasser, R.B., Podolska, M. & Chilton, N.B. (1998). Characterization of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. International Journal for Parasitology, 28, 1911-1921. doi:10.1016/s0020-7519(98)00150-7