Siwicki A. K., M. Morand, F. Pozet, B. Kazun: Anti-Birnavirus Activity of Methisoprinol -in vitro Study with Infectious Pancreatic Necrosis Virus (IPNV). Acta Vet. Brno 2002, 71: 543-547. This study was conducted to evaluate the influence of methisoprinol, synthetic anti-viral product, on the IPNV replication in vitro by measuring viral RNA synthesis. The monolayers of RTG-2 cells in tissue culture plates (Multiwell, 24 wells, Becton Dickinson, USA) was cultivated with different concentrations of methisoprinol (Polfa, Poland) 0, 100, 200, 400, 500 and 1 000 µg/ml of medium and were followed by infection with 100 µl of IPN virus suspension containing 10 7 TCID 50 /ml, in triplicate. At 24, 48 and 72 h after infection, the IPN virusinfected and methisoprinol-applicated RTG-2 cell cultures were submitted to one hour starvation and after two hours incubation with 10 µCi/ml of [ 3 H]-uridine. Culture homogenates of each isolate were incubated with phenol/chloroform to extract RNA and followed by slab polyacrylamide gel electrophoresis for 2 h. The gel strips were dissolved and the counts per minute (cpm) evaluated in a scintillation counter. The replicative cycle of IPN virus in RTG-2 cell culture was rapid. In control group (only infected by IPNV), the incorporation of [ 3 H]-uridine was 45 000 ± 1 500 cpm at 24 h, 186 000 ± 2 450 cpm at 48 h and 554 500 ± 4 550 cpm at 72 h. The percent of inhibition of IPN viral RNA labelling under methisoprinol application ranged from 5% at 24 h to 85% at 72 h depending on concentration of tested product and time when cultures were harvested. The highest percent of inhibition at 72 h after infection was observed at the dose 1000 µg/ml. The results of these experimental studies show the inhibition of incorporation (cpm) of [ 3 H]-uridine into IPN viral RNA in cell cultures exposed with methisoprinol at various concentrations.
Fish, IPNV, methisoprinol, replicationThe family Birnaviridae presents the smallest RNA viruses and consists of three genera: Aquabirnavirus, Avibirnavirus and Entomobirnavirus. The aquatic birnaviruses are the largest group of viruses including several strains from different fish species and invertebrates. The Birnaviridae are the most extensively studied fish viruses, because they are ubiquitous in aquatic organisms, since they have been isolated all over the world from both freshwater and marine fish of different species, and are responsible for severe losses in aquaculture.Infectious pancreatic necrosis virus (IPNV), the genus Aquabirnavirus, is the prototype of the birnavirus infecting fish, molluscs and crustaceans. The singleshelled, naked, icosahedral viral particles of birnaviruses containing 2 segments (A and B) have double-stranded RNA. Their dsRNA genome is easily purified and is resistant to common RNase. Very little is known about the replication strategy of birnaviruses. A viral protein 1 (VP1) is found in the viral particle both as a free form and bound to the RNA genome-linked protein (VPg) and is thought to be the viral