PROPAGATION OF INFECTIOUS PANCREATIC NECROSIS VIRUS IN FISH CELL CULTURES
M. Maistrenko1, Yu. Rud2, L. Buchatsky1,2
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1Taras Shevchenko Kyiv National University, 60, Volodymyrska str., 01033, Kyiv, Ukraine
2Institute of Fisheries of NAAS, 135, Obukhivska str., 03164, Kyiv, Ukraine
The reproduction of the Ukrainian isolate of infectious pancreatic necrosis virus (IPNV) «Carpathians» in fish continuous cell cultures of RTG-2, FHM and EPC was investigated. All three cell lines were sensitive to virus. IPNV caused morphological changes, such as vacuole enlargements and cells rounding. Subsequently cells scaled from a surface and characteristic cytopathic effect (CPE) of virus on cells was visible. For cell lines of RTG-2 and FHM the complete destruction of monolayer was noted on 7–8 day after infection (d.a.i.). For culture of EPC characteristic CPE and complete destruction of cell monolayer were marked on 10-12 d.a.i. Infectious titer of IPNV «Carpathians» in studied cell lines was following for EPC 105,5-5,8 TCID50/ml,and for the lines of FHM and RTG-2 — 106,2-6,5 and 106,9-7,4 TCID50/ml respectively. The low infectious titer of virus in cells of EPC can be related to its slow reproduction in this culture. The greatest infectious titer was observed for the culture of RTG-2, that is fully appropriately, as this cell line was derived from a rainbow trout — natural IPNV reservoir. That is why the RTG-2 is the most appropriate cell lines for accumulation of the Ukrainian isolate IPNV «Carpathians». But for diagnostics of the Ukrainian isolate IPNV «Carpathians» all three cell cultures of RTG-2, FHM and EPC can be used. Results of our electronic-microscopy researches of purified viral particles revealed basic for birnaviruses morphology and ultrastructure characteristics. Virions of the Ukrainian isolate of IPNV «Carpathians» had a hexagonal form, their diameter was 70±5 nm. For subsequent research of isolate «Carpathians» the methods of molecular diagnostics are needed, which will allow to determine the genotype of this IPNV strain and his possible origin.
Keywords:INFECTIOUS PANCREATIC NECROSIS VIRUS, ISOLATE “KARPATY”, CELL CULTURE, REPRODUCTION, INFECTIVE TITER, CPE
