immunosorbent assay) and RT-PCR (reverse-transcription polymerase chain reaction. No virus was isolated in the isolation studies. The supernatants from passage and isolation samples were checked nucleic acid and viral antigen of VHSV by RT-PCR and Ag-ELISA. Neither nucleic acid nor viral antigen was detected from cell culture supernatants and tissue extracts. KEY WORDS: Rainbow trout. Viral haemorrhagic septicemia. RT-PCR. Ag-ELISA. CPE �������������������� 1. ALTARA, J., (1963). Resolutions on Item A, viral hemorrhagic septicemia. Permanent Commision for the Study of Diseases of Fish, ������������������������������������������������� Epizot, 59, 298-99 ������������������������������, Estepa, A., (1994). A sandwich ELISA to detect VHSV and IPNV in turbot. Aquaculture International , 2, 206-17 3. BRUCHHOF, B., Marguart, O., Enzmann, P.J., (1995). Differential diagnosis of fish pathogenic rhabdoviruses by reverse transcriptase-dependent polymerase chain reaction. J Virol Methods , 55, 111-9 4. BRUNSON, R., True, K., Yancey, J., (1989). VHS virus isolated at Makah national Fish Hatchery. Fish Health Sect. Am Fish Soc Newslett , 17, 3-4 5. CASTRIC, J., de Kinkelin, P., (1980). Occurence of viral haemorrhagic septicaemia in rainbow trout (Salmo gairdneri) Richardson reared in sea-water. J Fish Dis , 3, 21-7 6. CHICO, V., Gomez, N., Estepa, A., Perez, L., (2006). Rapid detection and quantitation of viral hemorrhagic septicemia virus in experimentally challanged rainbow trout by real-time RT- PCR. J Virol Methods , 132, 154-9 ��� ������������� ���� �������� ���� �������� ������–Step Method of RNA Isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, Anal Biochem , 162, 156-9 8. DE KINKELIN, P., Scherrer, R., (1970). Le virus d’Egtved I.Stabilite developpement et structure du virus de la souche danoise F1. Ann Rech Vet , 1, 17-30 9. DE KINKELIN, P., (1983). Viral haemorrhagic septicaemia. In: Anderson DP, Dorson M, Dubourget P (eds) Antigens of fish pathogens. Fondation Marcel Merieux, Lyon, France, Pp: 51-62 10.DIXON, P.F., Feist, S., Kehoe, E., Parry, L., Stone, D.M., Way, K., (1997). Isolation of viral haemrrhagic septicaemia virus from Atlantic herring Clupea harengus from the English Channel. Dis Aquat Dis , 30, 81-9 11.EINER-JENSEN, K., Olesen, N.J., Lorenzen, N., Jørgensen, P.E.V., (1995). Use of the polymerase chain reaction (PCR) to differentiate serologically similar viral haemorrhagic septicaemia (VHS) virus isolates from Europe and America. Vet Res , 26, 464-9 12.ELSAYED, E., Faisal, M., Thomas, M., Whelan, G., Batts, W., Winton, J., (2006). Isolation of viral haemorrhagic septicaemia virus from muskellunge, Esox masquinongy (Mitchill), in Lake St. Clair, Michigan, USA reveals a new sublineage of the North American genotype. J Fish Dis , 29, 611-9 13.FERMENTAS., (2004). RevertAid TM First Strand cDNA Synthesis Kit #K1622. http://www.fermentas.com/profiles/kits/pdf/revertaid1622.pdf]. Revision date: 15.05.2004 14.FIJAN, N., Sulimanovic, D., Bearzotte, M., Muzinic, D., Zwillenberg, L.O., Chilmonczyk, S., Vautherot, J.F., de Kinkelin, P., (1983). Some properties of the epithelioma papulosum cyprinid (EPC) cell line from carp Cyprinus Carpio. Ann Virol (Inst Pasteur) , 134E, 207-20 15.FRYER, J.L., Yuhsa, A., Pilcher, K.S., (1965). The invitro cultivation of tissue and cells of Pacific salmon and steelhead trout. Ann NY Acad Sci , 126, 566-86 16.GRAVELL, M., Malsberger, R.G., (1965). A permanent cell line from the fathead minnow (Pimephales promelas). Ann NY Acad Sci , 126, 555-65 17.GUILLOU, J.P., Merle, G., Henault, S., Hattenberger, A.M., (1999). Detection du virus de la septicemie hemorragique virale (SHV) de la truitearc-en-ciel Oncorhynchus mykiss par une transcription inverse suivie d’une amplification en chaine par polymerase. Validations diagnostiques. Vet Res , 30, 49-60 340
18.HOPPER, K., (1989). The isolation of VHSV from chinook salmon at Glenwood Springs, Orcas Islands, Washington. Am Fish Soc Fish Health Sect Newsl ,17, 1-2 19.HØRLYCK, V., Mellergård, S., Dalsgaard, I., Jørgensen, P.E.V., (1984). Occurence of VHS in danish maricultured rainbow trout. Bull Eur Assoc Fish Pathol , 4(1), 11- 3 20.IIDA, H., Mori, K., Nishizawa, T., Arimoto, M., Muroga, K., (2003). Fate of viral hemorrhagic septicemia virus in Japanese flounder Paralichthys olivaceus challenged by immersion. Fish Pathol , 38, 87-91 21.JENSEN, M.H., (1963). Preperation of fish tissue cultures for virus research. Bull Off int Epizot , 59, 131-134 22.JENSEN, N.J., Bloch, B., Larsen, J.L., (1979). The Ulcus syndrome in cod Gadus morhua III. A preliminary virological report. Nord Vet Med , 31, 436-42 23.JENSEN, K.E., Ahrens, P., Forsberg, R., Lorenzen, N., (2004). Evolution of the fish rhabdovirus viral haemorrhagic septicaemia virus. J Gen Virol , 85, 1167-79 24.JØRGENSEN, P.E.V., Olesen, N.J., (1987). Cod ulcus syndrome rhabdovirus is indistinguishable from the egtved (VHS) virus. Bull Eur Ass Fish Pathol , 7(3), 73-4 25.KNUSEL, R., Bergmann, S.M., 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(9), 559-68 26.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(1), 103-9 27.LORENZEN, E., Carstensen, B., Olesen, N.J., (1999). Inter-laboratory comparison of cell lines for susceptibility to three viruses: VHSV,IHNV and IPNV. Dis Aquat Org, 37, 81-8 28.MCALLISTER, P.E., (1979). Fish viruses and viral infections. In: Fraenkel Conrat H, Wagner RR (eds) Comprehensive virology. Plenum publishing Corp. Newyork, 14, 401-70 29.MCALLISTER, P.E., (1990). Fish Disease Leaflet 83. Viral Hemorrhagic Septicemia of Fishes. U.S. Fish and Wildlife Service, National Fisheries Research Center-Leetown, National Fish Health Research Laboratory, West Virginia. http://www.lsc.usgs.gov/fhb/leaflets/83.asp.Revision date: 06/02/2004 30.MEYERS, T.R., Winton, J.R., (1995). Viral hemorrhagic septicemia virus in North America. Annu Rev Fish Dis , 5, 3-24 31.MEYERS, T.R., Short, S., Lipson, K., (1999). Isolation of the North American strain of viral hemorrhagic septicemia virus (VHSV) associated with epizootic mortality in two new host species of Alaska marine fish. Dis Aquat Org, 38, 81-6 32.MILLER, T.A., Rapp, J., Wastlhuber, U., Hoffmann, R.W., Enzmann, P.J., (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 33.MORTENSEN, H.F., Heuer, O.F., Lorenzen, N., Otte, L., Olesen, N.J., (1999). Isolation of viral haemorrhagic septicaemia virus (VHSV) from wild marine species in the Baltic Sea, Kattegat, Skagerrak and North Sea. Virus Res , 63, 95-106 34.MOURTON, C., Bearzotti, M., Piechaczyk, M., Paolucci, F., Pau, B., Bastide, J.M., de Kinkelin, P., (1990). Antigen-capture ELISA for viral haemorrhagic septicaemia virus serotype I. J Virol Methods , 29(3), 325-33 35.MUNRO, A.L.S., (1996). Report on the first recorded outbreak of viral haemorrhagic septicaemia (VHS) in GB and subsequent actions to contain, eradicate and investigate the origins of infections. Scott Aquacult Res Rep ,3, 1-12 36.NISHIZAWA, T., Iida, H., Takano, R., Isshiki, T., Nakajima, K., Muroga, K., (2002). Genetic relatedness among Japanese, American and European isolates of viral hemorrhagic septicemia virus (VHSV) based on partial G and P genes. Dis Aquat Org , 48, 143-8 ������������������������������������������������������������������������������������������� Genotyping and Pathogenicity of Viral Hemorrhagic Septicemia Virus from Free-Living Turbot Psetta maxima in a Turkish Coastal Area of the Black Sea. ����������������������, 72 (4), 2373-8 38.OIE (Office International des Epizooties) (2000,2006) Manual of diagnostic for aquatic animal diseases, 3rd edn.OIE,Paris 341
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