Metin GÜRÇAY, Ayşe PARMAKSIZ, Ahmet SAİT, Selehattin GÜRÇAY

Determination of nucleotide partial sequence of VP2 region of infectious pancreatic necrosis virus isolated from ovarian fluid of rainbow trout (Oncorhynchus mykiss, walbaum 1792) in Turkey

Bu çal ı ş mada, gökku ş a ğ ı alabal ı ğ ı (Oncorhynchus mykiss, Walbaum 1792) ovarial s ı vı ları ndanizole edilen, İnfeksiyöz Pankreatik Nekrozis Virüsünün (IPNV) viral proteininin VP2 bölgesinin kı sminükleotid sekans ı belirlendi. Nükleotid sekans ı n ı n daha önceden dünyada izole edilmi ş IPNVizolatları ile genetik yakı nl ı ğ ı araş tı rı ld ı . Bu amaç için, Elazı ğ bölgesindeki gökku ş a ğ ı alabal ı kları n ı novarial s ı vı ları ndan IPNV izole edildi, RT-PCR metoduyla identifiye edildi ve bu izolatı n viralproteininin VP2 bölgesinin kı smi nükleotid sekans analizi yap ı ld ı . Bu izolatı n nükleotid ve bundançı karı lan aminoasit sekans verisi GenBankda, KC525214 ELAZI Ğ eri ş im numaras ı ile yay ı nland ı .Bu izolat için filogenetik analiz yap ı ld ı . KC525214 ELAZI Ğ isolatı n ı n, filogenetik analiz sonucundaGenogroup 5 içinde bulundu ğu ve Fransa, Norveç, Danimarka, Tayvan, İspanya ve Amerikaizolatları ile serotip A2 grubunda bulunan izolatlarla yak ı ndan ili ş kili oldu ğ unu ortaya çı kard ı . Bubulgular, ovarian s ı v ı ları n transferleri, IPNVnin ta ş ı nma ve yay ı lmas ı na arac ı l ı k etti ğine i ş aretetmektedir. Bu çal ı ş ma ile, Türkiyede IPNV viral proteininin, VP2 bölgesinin kı smi nükleotiddizinanalizi gerçekle ş tirilmi ş oldu. Bu çal ı ş ma sonuçları gelecekte olu ş abilecek IPNV salg ı nları veepidemiyolojik çal ı ş malar için önem arz etmektedir.

Türkiye de Gökkuşağı alabalığı (Oncorhynchus mykiss walbaum, 1792) ovarial sıvılarından izole edilmiş infeksiyöz pankreatik nekrozis virüsünün, VP2 bölgesinin kısmi nükleotid dizin analizinin belirlenmesi

In this study, partial nucleotide sequence VP2 region of Infectious Pancreatic Necrosis Virus(IPNV) viral protein was detected from isolated ovarian fluid of rainbow trout (Oncorhynchus mykissWalbaum,1792) and its genetic relation was investigated to the worldwide IPNV isolates. For thispurpose, IPNV was isolated from ovarial fluid from fish cages belon to private enterprises in Elazi ğprovince and identified by RT-PCR method and subjected to partial nucleotide sequence for VP2region viral protein for this isolate. The nucleotide and deduced amino acid sequence data reportedin this isolate have been deposited in GenBank with the following accession number: KC525214ELAZI Ğ. Phylogenetic analysis was made for this isolate. Phylogenetic analysis revealed thatKC525214 ELAZI Ğ isolate was clustering within Genogroup 5 clade most closely related to theSerotype A2 together with French, Norwegian, Danish, Taiwanese, Spanish and American isolates. The findings indicated that the transfer of ovarian fluids is associated with the transmission andspread of IPNV. The determination of partial nucleotide sequence VP2 region viral protein for IPNVwas performed with this study in Turkey. These results are of importance for furtherepidemiological studies and IPNV outbreaks in Turkey.

PDF

___

1. Dobos P. The molecular biology of infectious pancreatic necrosis virüs (IPNV). Ann Rev Fish Dis 1995; 5: 25-54.
2. Sadasiv EC. Immunological and pathological responses of salmonids to infectious pancreatic necrosis virus (IPNV). Ann Rev Fish Dis 1995; 5: 209-223.
3. McAllister PE, Bebak J. Infectious pancreatic necrosis virus in the environment: Relationship to effluent from aquaculture facilities. J Fish Dis 1997; 20: 201-207.
4. Rivas CC, Cepeda CP, Dopazo B, Novoa MN, Barja JL. Marine environment as reservoir of birnaviruses from poikilothermic animals. Aquaculture 1993; 115: 83-194.
5. Espinoza E, Farı ´as G, Soler M, Kuznar J. Identity between infectious pancreatic necrosis virus VR-299 and a Chilean isolate. Intervirology 1985; 24: 58-60.
6. Anonymous. Infectious haematopoietic necrosis. Chapter 2.3.4, Manual of Diagnostic Tests for Aquatic Animals, 2012: 300-313.
7. Dopoza CP, Barja JL. Diagnosis and identification of IPNV in salmonids by molecular methods. In, Cunningham CO (Editor). Molecular Diagnosis of Salmonid Disease. Netherlands: Kluwer Academic Publishers, 2002: 23-48.
8. Williams K, Blake S, Sweeney A, Singer JT, Nicholson BL. Multiplex reverse transcriptase PCR assay for simultaneous detection of three fish viruses. J Clin Microbiol 1999; 37: 4139-4141.
9. Hall TA. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows. Nucleic Acids Symposium Series 1999; 41: 95-98.
10. Blake SL, Ma JY, Caporale DA, Jairath S, Nicholson BL. Phylogenetic relationships of aquatic birnaviruses based on deduced aminoacid sequences of genome segment A cDNA. Dis Aqua Org 200; 145: 89-102.
11. Tamura K, Dudley J, Nei M, Kumar S. MEGA4. Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007; 24: 1596-1599.
12. Sanju´an R, Nebot MR, Chirico N, Mansky LM, Belshaw R. Viral mutation rates . J Virol 2010; 84: 9733-9748.
13. Gürçay M, Turan T, Parmaks ı z A. A study on the presence of Infectious Pancreatic Necrosis Virus infections in farmed rainbow trout (Oncorhynchus mykiss Walbaum, 1792) in Turkey. Kafkas Univ Vet Fak Derg 2013; 19: 141-146.
14. Hill B J, Way K. Serological classification of infectious pancreatic necrosis (IPN) virus and other aquatic birnaviruses. Ann Rev Fish Dis 1995; 5: 55-77.
15. Heppell J, Berthiaume L, Tarrab E, Lecomte J, Arella M. Evidence of genomic variations between infectious pancreatic necrosis virus strains determined by restriction fragment profiles. J Gen Virol 1992; 73: 2863-2870.
16. Cutrin JM, Barja JL, Nicholson BL,et al. Restriction fragment length polymorphism and sequence analysis: Anapproach for genotyping infectious pancreatic necrosis virus reference strainsand other aquabirnaviruses isolated from northwestern Spain. Appl Envi-ron Microbiol 2004; 70: 1059-1067.
17. Zhang CX, Suzuki S. Comparison of the RNA polymerase genes of marine birnavirus strains and other birnaviruses. Arch Virol 2003;148: 745-758.
18. Nishizawa T, Kinoshita S, Yoshimizu M. An approach for genogrouping of Japanese isolates of aquabirnaviruses in a new genogroup, VII, based on the VP2/NS junction region. J Gen Virol 2005; 86: 1973-1978.
19. Havarstein LS, Kalland KH, Christie KE, Endresen C: Sequence of the large double-stranded RNA segment of the N1 strain of infectious pancreatic necrosis virus: A comparison with other Birnaviridae J Gen Virol, 1990; 71: 299-308.
20. Barrera-Mej´ ı a M, Sim´on-Mart´ı nez J, Ulloa-Arvizu R. Salgado-Miranda C, Soriano-Vargas E. Molecular characterization of the VP1 gene of a Mexican isolate of infectious pancreatic necrosis virus. Can J fish Aqua Sci 2010; 74: 218-222.
21. Zhang CX, Suzuki S: Aquabirnaviruses isolated from marine organisms form a distinct genogroup from other aquabirnaviruses. J Fish Dis 2004; 27: 633-643.
22. Candan A. First report on the diagnosis of infectious pancreatic necrosis (IPN) based on reverse transcription polymerase chain reaction (RT-PCR) in Turkey, Bull Eur Ass Fish Pathol 2002; 22: 45-48.
23. Albayrak H, Özan E. Gökku ş a ğı alabal ı kları nda (Oncorhynchus mykiss Walbaum, 1792) infeksiyöz pankreatik nekrozis ve infeksiyöz hematopoietik nekrozis virus enfeksiyonları n ı n varl ı ğ ı n ı n ara ş tı rı lmas ı . Ankara Üniv Vet Fak Derg 2010; 57: 125-129.
24. Ogut H, Altuntas C. Occurrence and prevalence of infectious pancreatic necrosis virus in rainbow trout (Oncorhynchus mykiss) cultured in cages in the Black Sea. Aqua Res 2012; 43: 1550-1556.
25. Silim A, Elazhary MASY, Lagace A. Susceptibility of trouts of different species and origins to various isolates of infectious pancreatic necrosis virus. Can J fish Aqua Sci 1982; 39: 1580-1584.
26. Reno PW. Infectious pancreatic and associated aquatic birnaviruses. In: Woo PTK, Bruno DW. (Editors): Fish Diseases and Disorders. UK: CAB International, Oxfordshire, 1999: 1-55.
27. Ogut H, Altuntas C, Parlak R. Viral surveillance of cultured rainbow trout in the eastern Black Sea, Turkey. J Aquat Anim Health 2013; 25: 27-35.