Investigating the presence of infectious haematopoietic necrosis and infectious pancreatic necrosisvirus infections in rainbow trout (Oncorhynchus mykiss Walbaum, 1792)

Abstract

In this study, presence of infectious pancreatic necrosis and infectious haematopoietic necrosis (IHN) viruses were investigated in fresh water in commercial rainbow trout in middle and east Black Sea region (Samsun, Sinop, Ordu, Giresun, Trabzon, Rize, Amasya, Tokat, Gümüşhane). For this purpose, total of 229 isolation material from 168 portion rainbow trout of various sizes (200 to 500 g) and 61 immature fish (from 305 immature fish, weighing 3-10 g) were obtained from 32 farms in the region in December and March in 2006-2007 which period is water temperature under the 12 °C. All isolation materials were passaged in BF-2 (Bluegill fry-2) cell cultures for two times. While cell culture supernatants showed cytopathic effect (CPE) were tested by antigen-capture enzyme linked immunosorbent assay (Ag-ELISA), all isolation materials were tested by reverse transcriptase polymerase cahin reaction (RT-PCR). As a result, while IPN virus was detected in immature fish and portion rainbow trout in 10 farms in Ordu, Samsun, Tokat and Trabzon provinces, IHN virus was not detected. That cell culture isolation and RTPCR test are harmonious were revealed

Keywords

• CPE
• infectious haematopoietic necrosis
• infectious pancreatic necrosis
• RT-PCR

Gökkuşağı alabalıklarında (Oncorhynchus mykiss Walbaum, 1792) infeksiyöz pankreatik nekrozis ve infeksiyöz hematopoietik nekrozis virus enfeksiyonlarının varlığının araştırılması

Abstract

Bu çalışmada, Orta ve Doğu Karadeniz Bölgesinde (Samsun, Sinop, Ordu, Giresun, Trabzon, Rize, Amasya, Tokat, Gümüşhane) tatlı sulardaki ticari gökkuşağı alabalık çiftliklerinde IPN ve IHN viruslarının varlığı araştırıldı. Bu amaçla, 2006-2007 yılları Aralık-Mart aylarında (su sıcaklığının 12 °C’nin altına düştüğü dönem) bölgedeki 32 çiftlikten, 168’i (200-500 g) porsiyon balıklardan, 61’i (305 adet yavru alabalıktan, 3-10 g’lık yavru) yavru balıklardan olmak üzere toplam 229 izolasyon materyali temin edildi. İzolasyon materyallerinin BF-2 hücresinde iki pasajı yapıldı. Cytopathic effect (CPE) gösteren hücre kültür süpernatantları antijen enzyme linked immunosorbet assay (Ag-ELISA), izolasyon materyalleri ise reverse transcriptase polymerase chain reaction (RT-PCR) ile test edildi. Sonuç olarak, Ordu, Samsun, Tokat ve Trabzon illerinde bulunan 10 çiftlikte yavru ve porsiyon balıklardan IPN virusu tespit edilirken, IHN virusu tespit edilemedi. Hücre kültürü izolasyonu ve RT-PCR testlerinin birbiriyle uyumlu olduğu ortaya konuldu

Keywords

• CPE
• infeksiyöz hematopoetik nekrozis
• infeksiyöz pankreatik nekrozis
• RT-PCR

References

1. 1. Anonim (2005): İl ve Türlere Göre Avlanan Tatlı Su Ürünleri. Devlet İstatistik Enstitüsü Başkanlığı 2003 yıllık raporu.
2. 2. Anonim (2005): Kültür Balıkları Üretimi. Devlet İstatistik Enstitüsü Başkanlığı 2003 yıllık raporu.
3. 3. Anonim (2006): Infectious Pancreatic Necrosis. Manual of Diagnostic Tests for Aquatic Animals. Erişim: http://www.oie.int/eng/normes/fmanual/A_00025.htm. Erişim tarihi: 01.02.2009.
4. 4. Anonim (2006): Infectious Haematopoietic Necrosis. Manual of Diagnostic Tests for Aquatic Animals. Erişim: http://www.oie.int/eng/normes/fmanual/A_00019.htm. Erişim tarihi: 01.02.2009.
5. 5. Anonim (2007): 2004-2007 Epidemiyoloji Ünitesi Hastalık Çıkış Raporları. Samsun Veteriner Kontrol ve Araştırma Enstitüsü Hastalık Çıkış Raporları Arşivi.
6. 6. Anonim (2009): Birnaviridae. International Committee on Taxonomy of Viruses. Erişim: http://www.ncbi.nlm.nih.gov/ICTVdb/Ictv/fs_birna.htm. Erişim Tarihi: 06.02.2009.
7. 7. Anonim (2009): Rhabdoviridae. International Committee on Taxonomy of Viruses. Erişim: http://www.ncbi.nlm.nih.gov/ICTVdb/Ictv/fs_rhabd.htm. Erişim Tarihi: 06.02.2009.
8. 8. Arda M, Seçer S, Sarıeyyüpoğlu M (2002): Balık Hastalıkları. Medisan yayınevi, Ankara.

9. 9. Besse P, de Kinkelin P (1965): Sur l’existence en France de la nécrose pancréatique de la truite arc en ciel (Salmo gairdneri). B Acad Vet France, 38, 185-190.
10. 10. Blake SL, Schill WB, McAllister PE, Lee MK, Singer JT, Nicholson BL (1995): Detection and identification of aquatic birnaviruses by PCR assay. J Clin Microbiol, 33, 835-839.
11. 11. Bootland LM, Leong JC (1999): Infectious haematopoietic necrosis virus. 57-121. In: PTK Woo, DW Bruno (Eds), Fish Diseases and Disorders, Viral, Bacterial and Fungal Infections. CABI Publishing, Oxon.
12. 12. Bowers RM, Lapatra SE, Dhar AK (2008): Detection and quantitation of infectious pancreatic necrosis virus by real-time reverse transcriptase-polymerase chain reactionusing lethal anda non-lethal tissue sampling. J Virol Methods, 147, 226-234.
13. 13. Braňas MV, Morales JC, Estepa A (1994): A sandwich ELISA to detect VHSV and IPNV in turbot. Aquacult Int, 2, 206-217.
14. 14. Bruchhof B, Marguart O, Enzmann PJ (1995): Differential diagnosis of fish pathogenic rhabdoviruses by reverse transcriptase-dependent polymerase chain reaction. J Virol Methods, 55, 111-119.
15. 15. Candan A (2002): 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, 22, 45.
16. 16. Chomcyznski P, Sacchi N (1987): Single–Step Method of RNA Isolation by acid guanidinium thiocyanate-phenolchloroform extraction. Anal Biochem, 162, 156-159.
17. 17. Chikova V, Ilieva D (2004): Detection of infectious panreatic necrosis (IPN) vırus in rainbow trouts in Bulgaria. Bulg J Vet Med, 7, 129-135.
18. 18. Değirmenci U, Nemli E, Çağırgan H (2008): Türkiye’nin değişik bölgelerinden infeksiyöz pankreatik necrozis virusu izolasyonu. I. Ulusal Alabalık Sempozyumu, Isparta, s.35.
19. 19. Hill BJ (1992) Impact of Viral Diseases of Salmonid Fish in the European Community.48-59. In: T Kimura (Ed), Proceedings of the OJI International Symposium on Salmonid Diseases. Hokkaido University Press, Sapparo.
20. 20. Knüsel R, Bergmann SM, Einer-Jensen K, Casey J, Segner H, Wahli T (2007): Virus isolation vs RT-PCR: which method is more successful in detecting VHSV and IHNV in fish tissue sampled under field conditions? J Fish Dis, 30, 559-568.
21. 21. Liu Z, Teng Y, Liu H, Jiang Y, Xie X, Li H, Lv J, Gao L, He J, Shi X, Tian F, Yang J, Wie C (2008): Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay. J Virol Methods, 149, 103-109.
22. 22. Lorenzen E, Carstensen B, Olesen NJ (1999): Interlaboratory comparison of cell lines for susceptibility to three viruses: VHSV,IHNV and IPNV. Dis Aquat Org, 37, 81-88.
23. 23. Miller TA, Rapp J, Wastlhuber U, Hoffmann RW, Enzmann PJ (1998): Rapid and sensitive reverse transcriptase-polymerase chain reaction based detection and differential diagnosis of fish pathogenic rhabdoviruses in organ samples and cultured cells. Dis Aquat Org, 34, 13-20 .
24. 24. Nishizawa T, Savaş H, Işıdan H, Üstündağ C, Iwamoto H, Yoshimizu M (2006): Genotyping and pathogenicity of viral hemorrhagic septicemia virus from free-living turbot Psetta maxima in a Turkish coastal area of the black sea. Appl Environ Microbiol, 72, 2373-2378.
25. 25. Reno PW (1999): Infectious Pancreatic Necrosis and Associated Aquatic Birnaviruses. 1-55. In: PTK Woo, DW Bruno (Eds), Fish Diseases and Disorders, Viral, Bacterial and Fungal Infections. CABI Publishing, Oxon.
26. 26. Sweeney A (1996): Multiplex PCR Assay for the Simultaneus Detection of IPNV and IHNV. M.S. thesis, University of Maine, Orono, Maine.
27. 27. Toplu N, Albayrak H, Aydoğan A, Epikmen ET, Metin N (2008): Gökkuşağı alabalıklarının (Oncorhynchus mykiss Walbaum,1972) enfeksiyöz pankreatik nekrozunda (IPN) patolojik bulgular, apoptozis, immunohistokimyasal ve PCR tanı. IV. Ulusal Veteriner Patoloji Kongresi, Antalya, s. 149-150.
28. 28. Williams K, Blake S, Sweeney A, Singer JT, Nicholson BL (1999): Multiplex reverse transcriptase PCR assay for simultaneous detection of three fish viruses. J Clin Microbiol, 37, 4139-4141.
29. 29. Wolf K (1988) Fish Viruses and Fish Viral Diseases. Cornel University Press, Ithaca, New York.