First Molecular Characterization of Hysterothylacium Fabri Larvae (Nematoda: Raphidascarididae) in the Mediterranean Sea Based on the Small Subunit Ribosomal RNA Gene Sequence

Öz

Hysterothylacium fabri is commonly reported in various Mediterranean fish. However, there is no data on the mitochondrial ribosomal RNA (rrnS) gene sequence of H. fabri larvae in the Mediterranean Sea waters. Therefore, we aimed to reveal molecular characterization of H. fabri based on the rrnS gene sequence in the current study. Firstly, the fourth stage of H. fabri larvae were identified at parasitological examination from Mullus barbatus in the Mediterranean Sea. Morphologically identified H. fabri larvae herein were also supported by the sequencing of the nuclear ribosomal ITS gene regions. In the next step, the rrnS gene of these larvae was molecularly analysed. Our H. fabri isolate (MK886659) showed 96.42% identity with H. fabri reported from China (MF140349) based on the rrnS gene. The nucleotide difference based on the rrnS gene between Chinese isolate (MF140349) and our H. fabri isolate (MK886659) was determined as 3.2%. Consequently, for the first time, the mitochondrial rrnS gene sequencing of H. fabri from the Mediterranean waters were performed in the current study.

Anahtar Kelimeler

• Hysterothylacium fabri
• Mediterranean Sea
• Mitochondrial rrnS gene
• Molecular characterization

Akdeniz’deki Hysterothylacium Fabri (Nematoda: Raphidascarididae) Larvasının İlk Kez Küçük Alt Ünite Ribozomal RNA Gen Bölgesinin Sekansına Göre Moleküler Karakterizasyonu

Öz

Hysterothylacium fabri sıklıkla farklı türdeki Akdeniz balıklarında bildirilmektedir. Ancak, Akdeniz sularındaki H. fabri larvasının mitokondriyal ribozomal RNA (rrnS) gen bölgesinin sekansı üzerine herhangi bir veri bulunmamaktadır. Bu yüzden, bu çalışmada, H. fabri’nin rrnS gen bölgesinin sekansına göre moleküler karakterizasyonunu ortaya koymayı amaçladık. İlk olarak, Akdeniz'deki Mullus barbatus'tan dördüncü dönem H. fabri larvaları parazitolojik muayenede teşhis edildi. Burada morfolojik olarak teşhis edilen H. fabri, nükleer ribozomal ITS gen bölgesinin DNA dizi analizi ile de desteklendi. Bir sonraki aşamada ise bu larvaların rrnS geni moleküler olarak analiz edildi. RrnS gen bölgesine göre, H. fabri izolatımız (MK886659) Çin’den rapor edilen H. fabri (MF140349) ile %96,42 oranında benzerlik gösterdi. Çin (MF140349) izolatı ile H. fabri (MK886659) izolatımız arasındaki nükleotid farklılığı ise rrnS genine göre %3,2 olarak belirlendi. Sonuç olarak bu araştırma ile ilk kez Akdeniz sularından H. fabri'nin mitokondriyal rrnS sekanslaması gerçekleştirildi.

Anahtar Kelimeler

• Hysterothylacium fabri
• Akdeniz
• Mitokondriyal rrnS gen
• Moleküler karakterizasyon

Kaynakça

1. Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 1997; 25:3389-402.
2. Barcala E, Ramilo A, Ortega N, Pico G, Abollo E, Pascual S, Munoz P. Occurrence of Anisakis and Hysterothylacium larvae in commercial fish from Balearic Sea (Western Mediterranean Sea). Parasitol Res. 2018; 117:4003-4012.
3. Chen HX, Zhang LP, Gibson DI, Lu L, Xu Z, Li HT, Ju HD, Li L. Detection of ascaridoid nematode parasites in the important marine food-fish Conger myriaster (Brevoort) (Anguilliformes: Congridae) from the Zhoushan Fishery, China. Parasit Vectors. 2018; 11:274.
4. Costa A, Graci S, Cammilleri G, Buscemi MD, Collura R, Vella A, Ferrantelli V. Molecular Identification of Hysterothylacium spp. In Fishes From the Southern Mediterranean Sea (Southern Italy). J Parasitol. 2018; 104:398-406.
5. D'Amelio S, Barros N, Ingrosso S, Fauquier D, Russo R, Paggi L. 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. 2007; 134:1041.
6. da Fonseca MCG, Knoff M, Felizardo NN, Di Azevedo MIN, Torres EJL, Gomes DC, Iñiguez AM, de São Clemente SC. Integrative taxonomy of Anisakidae and Raphidascarididae (Nematoda) in Paralichthys patagonicus and Xystreurys rasile (Pisces: Teleostei) from Brazil. Int J Food Microbiol. 2016; 235:113-124.
7. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution. 1985; 39:783-791.
8. Gazzonis AL, Cavallero S, Zanzani SA, Olivieri E, Malandra R, Ranghieri V, D'Amelio S, Manfredi MT. Anisakis sp. and Hysterothylacium sp. larvae in anchovies (Engraulis encrasicolus) and chub mackerel (Scomber colias) in the Mediterranean Sea: Molecular identification and risk factors. Food Control. 2017; 80:366-373.

9. Guo YN, Xu Z, Zhang LP, Hu YH, Li L. Occurrence of Hysterothylacium and Anisakis nematodes (Ascaridida: Ascaridoidea) in the tanaka's snailfish Liparis tanakae (Gilbert & Burke) (Scorpaeniformes: Liparidae). Parasitol Res. 2014; 113:1289-300.
10. 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.
11. Keskin E, Koyuncu CE, Genc E. Molecular identification of Hysterothylacium aduncum specimens isolated from commercially important fish species of Eastern Mediterranean Sea using mtDNA cox1 and ITS rDNA gene sequences. Parasitol Int. 2015; 64:222-228.
12. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol Biol Evol. 2016; 33:1870-4.
13. Li L, Lü L, Nadler SA, Gibson DI, Zhang LP, Chen HX, Zhao WT, Guo YN. Molecular phylogeny and dating reveal a terrestrial origin in the Early Carboniferous for ascaridoid nematodes. Syst Biol. 2018; 67:888-900.
14. Martin-Sanchez J, Diaz M, Artacho ME, Valero A. Molecular arguments for considering Hysterothylacium fabri (Nematoda: Anisakidae) a complex of sibling species. Parasitol Res. 2003; 89:214-20.
15. Moravec F, Justine JL. Anisakid nematodes (Nematoda: Anisakidae) from the marine fishes Plectropomus laevis Lacepede (Serranidae) and Sphyraena qenie Klunzinger (Sphyraenidae) off New Caledonia, including two new species of Hysterothylacium Ward & Magath, 1917. Syst Parasitol. 2015; 92:181-95.
16. Pekmezci GZ. 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 Vet Mikrobiol Derg. 2019; 30:64-69.
17. Pekmezci GZ, Yardimci B, Onuk EE, Umur S. Molecular characterization of Hysterothylacium fabri (Nematoda: Anisakidae) from Zeus faber (Pisces: Zeidae) caught off the Mediterranean coasts of Turkey based on nuclear ribosomal and mitochondrial DNA sequences. Parasitol Int. 2014; 63:127-31.
18. Petter AJ, Maillard C. Larves d’ascarides parasites de poissons en Méditerranée occidentale. Bull Mus Natl Hist Nat. 1988; 10:347-369.
19. Roca-Gerones X, Montoliu I, Godinez-Gonzalez C, Fisa R, Shamsi S. Morphological and genetic characterization of Hysterothylacium Ward & Magath, 1917 (Nematoda: Raphidascarididae) larvae in horse mackerel, blue whiting and anchovy from Spanish Atlantic and Mediterranean waters. J Fish Dis. 2018; 41:1463-1475.
20. Simsek E, Ciloglu A, Yildirim A, Pekmezci GZ. Identification and molecular characterization of Hysterothylacium (Nematoda: Raphidascarididae) larvae in bogue (Boops boops L.) from the Aegean Sea, Turkey. Kafkas Üniv Vet Fak Derg. 2018; 24.
21. Szostakowska B, Myjak P, Kur J,Sywula T. Molecular evaluation of Hysterothylacium auctum (Nematoda, Ascaridida, Raphidascarididae) taxonomy. Acta Parasitol. 2001; 46:194-201.
22. Tedesco P, Gustinelli A, Caffara M, Patarnello P, Terlizzi A, Fioravanti ML. Hysterothylacium fabri (Nematoda: Raphidascarididae) in Mullus surmuletus (Perciformes: Mullidae) and Uranoscopus scaber (Perciformes: Uranoscopidae) from the Mediterranean. J Parasitol. 2018; 104:262-274.
23. Zhang K, Xu Z, Chen HX, Guo N, Li L. Anisakid and raphidascaridid nematodes (Ascaridoidea) infection in the important marine food-fish Lophius litulon (Jordan) (Lophiiformes: Lophiidae). Int J Food Microbiol. 2018; 284:105-111.
24. Zhu X, Gasser RB, Podolska M, Chilton NB. Characterisation of anisakid nematodes with zoonotic potential by nuclear ribosomal DNA sequences. Int J Parasitol. 1998; 28:1911-21.