Replication of Infectious Pancreatic Necrosis Virus in Different Cell Lines and in Rainbow Trout (Oncorhynchus Mykiss) Fingerlings

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Abstract

The results of a study of Infectious Pancreatic Necrosis Virus (IPNV) isolated in natural reservoirs in Ukraine are presented. The pathogenicity of isolates was investigated in vitro on cell cultures and in vivo on rainbow trout, Oncorhynchus mykiss (Walbaum), fingerlings. Experimental indications were that the Ukrainian IPNV isolates have affinity with reference European strains. During the reproduction of these isolates in cell cultures of FHM (fat head minnow), RTG-2 (rainbow trout gonads), and BF-2 (bluegill caudal peduncle), complicated degenerative changes were visible that finally led to the full destruction of cell monolayers. The experimental infection of rainbow trout fingerlings resulted in typical disease symptoms that were systemic. However, obvious evidence of viral infection was noted in single individuals only, and the majority of experimental fish died without visible disease symptoms. During the study of physicochemical properties, it was noted that Ukrainian isolates completely lost their infectivity with chloroform treatment and heating to 60°C. This proved that IPNV isolates are resistant to Ion concentrations in the range of pH 3.0 to 12.0.

Abstract

The results of a study of Infectious Pancreatic Necrosis Virus (IPNV) isolated in natural reservoirs in Ukraine are presented. The pathogenicity of isolates was investigated in vitro on cell cultures and in vivo on rainbow trout, Oncorhynchus mykiss (Walbaum), fingerlings. Experimental indications were that the Ukrainian IPNV isolates have affinity with reference European strains. During the reproduction of these isolates in cell cultures of FHM (fat head minnow), RTG-2 (rainbow trout gonads), and BF-2 (bluegill caudal peduncle), complicated degenerative changes were visible that finally led to the full destruction of cell monolayers. The experimental infection of rainbow trout fingerlings resulted in typical disease symptoms that were systemic. However, obvious evidence of viral infection was noted in single individuals only, and the majority of experimental fish died without visible disease symptoms. During the study of physicochemical properties, it was noted that Ukrainian isolates completely lost their infectivity with chloroform treatment and heating to 60°C. This proved that IPNV isolates are resistant to Ion concentrations in the range of pH 3.0 to 12.0.

References

  • Ahne W., Negele R.D. 1985 - Studies on the transmission of infectious pancreatic necrosis virus via eyed eggs and sexual products of salmonid fish - In: Fish and Shellfish Pathology (Ed.) A.E. Ellis, Academic Press, London: 262-270.

  • Alonso M.C., Cano I., Castro D., Perez-Prieto S.I., Borrego J.J. 2004 - Development of an in situ hybridization procedure for the detection of sole aquabirnavirus in infected fish cell cultures - J. Virol. Methods 116:133-138.

  • Ball H.J., Munro A.L.S., Ellia A., Elson K.G.R., Hodgkiss W., Farlane L.S. 1971 - Infectious pancreatic necrosis in rainbow trout in Scotland - Nature 234: 417-418.

  • Bootland L.M., Dobos P., Stevenson R.M. 1991 - The IPNV carrier state and demonstration of vertical transmission in experimentally infected brook trout - Dis. Aquat. Org. 10: 13-21.

  • Davies K.R., McColl K.A., Wang L.F., Yu M., Williams L.M., Crane M.St.J. 2010 - Molecular characterisation of Australasian isolates of aquabirnavirus - Dis. Aquat. Org. 93: 1-15.

  • Dixon P.F., Ngoh G.H., Stone D.M., Chang S.F., Way K., Kueh S.L.F. 2008 - Proposal for a fourth aquabirnavirus serogroup - Arch. Virol. 153:1937-1941.

  • Dorson M. 1985 - Experimental transmission of infectious pancreatic necrosis virus via the sexual products - In: Fish and Shellfish Pathology (Ed.) A.E. Ellis, Academic Press, London: 251-260.

  • Golovina N.A, Bauer O.N. 2007 - Ichthyopathology - Mir, Moscow, R448 p. (in Russian).

  • Gravell M., Malsberger R.G. 1965 - A permanent cell line from the fathead minnow (Pirnephales prornelas) - Ann. NY Acad. Sci. 126: 555-565.

  • Jorgensen P.E., Bregnballe F. 1969 - Infectious pancreatic necrosis in rainbow trout (Salmo gairdneri) in Denmark -Nordisk Veterinaermedicin 21: 142-148.

  • Mahy B.W. 1988 - Virology a practical approach - Mir, Moscow, Russia, 344p.

  • Mahy B.W., Kangro H.O.1996 - Virology Methods Manual -London, Academic Press, UK, 452 p.

  • Matvienko N., Buchatsky L., Deryabin O. 2011 - Isolation of IPN virus from rainbow trout in the Ukraine - 15th International Conference on Diseases of Fish and Shellfish, Split, Croatia, 12–16.09.11, Abstract book: 363.

  • OIE 2003 - Manual of diagnostic tests for aquatic animals -Fourth Edition, Paris, Office International Des Epizooties.

  • Osadchaya E.F. 1984 - Developing ichthyovirusology in the USSR and abroad - In: Biological foundations of fish farming: parasites and fish diseases (Eds) O.N. Bauer, V.A. Muselius, E.S. Scriabin, Moscow: 28-46 (in Russian).

  • Reed L., Muench H. 1938 - A simple method of estimating fifty per cent endpoints - Am. J. Hyg. 27: 493-497.

  • Santi N., Sandtro A., Sindre H., Song H., Hong J.R., Thu B., Wu J.L., Vakharia V.N., Evensen O. 2005 - Infectious pancreatic necrosis virus induces apoptosis in vitro and in vivo independent of VP5 expression - Virology 342: 13-25.

  • Sano M., Okamoto N., Fukuda H., Saneyoshi M., Sano T. 1992 - Virulence of infectious pancreatic necrosis virus is associated with the larger RNA segment (RNA segment A) - J. Fish Dis. 15: 283-293.

  • Wolf K., Quimby M.C. 1962 - Established eurythermic line of fish cells in vitro - Science 135: 1065-1066.

  • Wolf K., Quimby M.C. 1971 - Salmonid viruses: Infectious pancreatic necrosis virus morphology, pathology, and serology of first European isolations - Arch. Gesamte Virus-Forsch. 34: 144-156.

  • Wood E.M., Snieszko S.F., Yasutake W.T. 1955 - Infectious pancreatic necrosis of brook trout - AMA Arch. Pathol. 60: 26-38.

  • Zhang C.X., Suzuki S. 2004 - Aquabirnaviruses isolated from marine organisms form a distinct genogroup from other aquabirnaviruses - J. Fish Dis. 27:633-643.

Ahne W., Negele R.D. 1985 - Studies on the transmission of infectious pancreatic necrosis virus via eyed eggs and sexual products of salmonid fish - In: Fish and Shellfish Pathology (Ed.) A.E. Ellis, Academic Press, London: 262-270.

Alonso M.C., Cano I., Castro D., Perez-Prieto S.I., Borrego J.J. 2004 - Development of an in situ hybridization procedure for the detection of sole aquabirnavirus in infected fish cell cultures - J. Virol. Methods 116:133-138.

Ball H.J., Munro A.L.S., Ellia A., Elson K.G.R., Hodgkiss W., Farlane L.S. 1971 - Infectious pancreatic necrosis in rainbow trout in Scotland - Nature 234: 417-418.

Bootland L.M., Dobos P., Stevenson R.M. 1991 - The IPNV carrier state and demonstration of vertical transmission in experimentally infected brook trout - Dis. Aquat. Org. 10: 13-21.

Davies K.R., McColl K.A., Wang L.F., Yu M., Williams L.M., Crane M.St.J. 2010 - Molecular characterisation of Australasian isolates of aquabirnavirus - Dis. Aquat. Org. 93: 1-15.

Dixon P.F., Ngoh G.H., Stone D.M., Chang S.F., Way K., Kueh S.L.F. 2008 - Proposal for a fourth aquabirnavirus serogroup - Arch. Virol. 153:1937-1941.

Dorson M. 1985 - Experimental transmission of infectious pancreatic necrosis virus via the sexual products - In: Fish and Shellfish Pathology (Ed.) A.E. Ellis, Academic Press, London: 251-260.

Golovina N.A, Bauer O.N. 2007 - Ichthyopathology - Mir, Moscow, R448 p. (in Russian).

Gravell M., Malsberger R.G. 1965 - A permanent cell line from the fathead minnow (Pirnephales prornelas) - Ann. NY Acad. Sci. 126: 555-565.

Jorgensen P.E., Bregnballe F. 1969 - Infectious pancreatic necrosis in rainbow trout (Salmo gairdneri) in Denmark -Nordisk Veterinaermedicin 21: 142-148.

Mahy B.W. 1988 - Virology a practical approach - Mir, Moscow, Russia, 344p.

Mahy B.W., Kangro H.O.1996 - Virology Methods Manual -London, Academic Press, UK, 452 p.

Matvienko N., Buchatsky L., Deryabin O. 2011 - Isolation of IPN virus from rainbow trout in the Ukraine - 15th International Conference on Diseases of Fish and Shellfish, Split, Croatia, 12–16.09.11, Abstract book: 363.

OIE 2003 - Manual of diagnostic tests for aquatic animals -Fourth Edition, Paris, Office International Des Epizooties.

Osadchaya E.F. 1984 - Developing ichthyovirusology in the USSR and abroad - In: Biological foundations of fish farming: parasites and fish diseases (Eds) O.N. Bauer, V.A. Muselius, E.S. Scriabin, Moscow: 28-46 (in Russian).

Reed L., Muench H. 1938 - A simple method of estimating fifty per cent endpoints - Am. J. Hyg. 27: 493-497.

Santi N., Sandtro A., Sindre H., Song H., Hong J.R., Thu B., Wu J.L., Vakharia V.N., Evensen O. 2005 - Infectious pancreatic necrosis virus induces apoptosis in vitro and in vivo independent of VP5 expression - Virology 342: 13-25.

Sano M., Okamoto N., Fukuda H., Saneyoshi M., Sano T. 1992 - Virulence of infectious pancreatic necrosis virus is associated with the larger RNA segment (RNA segment A) - J. Fish Dis. 15: 283-293.

Wolf K., Quimby M.C. 1962 - Established eurythermic line of fish cells in vitro - Science 135: 1065-1066.

Wolf K., Quimby M.C. 1971 - Salmonid viruses: Infectious pancreatic necrosis virus morphology, pathology, and serology of first European isolations - Arch. Gesamte Virus-Forsch. 34: 144-156.

Wood E.M., Snieszko S.F., Yasutake W.T. 1955 - Infectious pancreatic necrosis of brook trout - AMA Arch. Pathol. 60: 26-38.

Zhang C.X., Suzuki S. 2004 - Aquabirnaviruses isolated from marine organisms form a distinct genogroup from other aquabirnaviruses - J. Fish Dis. 27:633-643.

Fisheries & Aquatic Life

The Journal of Stanislaw Sakowicz Inland Fisheries Institute

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