The RNA-dependent RNA polymerase of the infectious pancreatic necrosis virus is linked to viral mRNA acting as a cap substitute.

The infectious pancreatic necrosis virus (IPNV) is responsible for significant economic losses in the aquaculture industry. It is an unenveloped virus with an icosahedral capsid. Its viral genome comprises two dsRNA segments, A and B. Segment A contains a small ORF, which encodes VP5, and a large ORF, which encodes a polyprotein that generates the structural proteins and the viral protease. Segment B encodes the RNA-dependent RNA polymerase (RdRp), called VP1 in this free form, or Vpg when it covalently attaches to the viral RNA. The viral genome does not have cap or poly(A). Instead, each 5' end is linked to the Vpg. Recently, we demonstrated that mRNA-A contains an internal ribosome entry site (IRES) to command polyprotein synthesis. However, the presence of Vpg on IPNV mRNAs and its impact on cellular translation has not been investigated. This research demonstrates that IPNV mRNAs are linked to Vpg and that this protein inhibits cap-dependent translation on infected cells. Also, it is demonstrated that Vpg interacts with eIF4E and that rapamycin treatment partially diminishes the viral protein synthesis. In addition, we determined that an IRES does not command translation of IPNV mRNA-B. We show that VPg serves as a cap substitute during the initiation of IPNV translation, contributing to understanding the replicative cycle of Birnaviruses. Our results indicate that the viral protein VP1/Vpg is multifunctional, having a significant role during IPNV RNA synthesis as the RdRp and the primer for IPNV RNA synthesis and translation as the viral protein genome, acting as a cap substitute.

Labelling fish diets with 15 N -Leucine for monitoring feed consumption and bio-distribution in Atlantic salmon.

Feeding represents 50-70% of the cost of production in salmon farming, higher than any other animal farm. The improvement of this percentage is challenging as the food is thrown into the fish tank, there is no quantification of the amount of food that is consumed by the fish. In consequence, it is difficult to adjust the food composition making it more nutritive or promoting food consumption by fish. In this study, to investigate food consumption, bio-distribution and food residues, leucine containing 15 N (a stable isotope of nitrogen) was used to label the fish food. Atlantic salmon (Salmo salar) weighing 100-120 g were maintained in 30 L tanks at a density of 14 kg/m3 . Fishes were fed daily at 1% of the fish weight with pellet labelled with 15 N-leucine. The 15 N incorporation was determined 14 hours after the feeding in all the fish organs. Results showed that 14 hours after the administration of a single dose of labelled food to Atlantic salmon enables the detection of the tracer in the whole organism allowing determining the food consumption. Through the analysis of nitrogen use efficiency (NUE), we showed that the trunk, pyloric caeca and head incorporate the highest level of the marker (72.7, 8.7 and 5.7%, respectively). This methodology would permit monitoring feeding to minimize food loss, improve administration methodologies or select the preferred foods for the fish, among others to reduce production costs.

Study of RNA-A Initiation Translation of The Infectious Pancreatic Necrosis Virus.

The infectious pancreatic necrosis virus (IPNV) is a salmonid pathogen that causes significant economic losses to the aquaculture industry. IPNV is a non-enveloped virus containing two uncapped and non-polyadenylated double strand RNA genomic segments, RNA-A and RNA-B. The viral protein Vpg is covalently attached to the 5' end of both segments. There is little knowledge about its viral cycle, particularly about the translation of the RNAs. Through experiments using mono and bicistronic reporters, in this work we show that the 120-nucleotide-long 5'-UTR of RNA-A contains an internal ribosome entry site (IRES) that functions efficiently both in vitro and in salmon cells. IRES activity is strongly dependent on temperature. Also, the IRES structure is confined to the 5'UTR and is not affected by the viral coding sequence. This is the first report of IRES activity in a fish virus and can give us tools to generate antivirals to attack the virus without affecting fish directly.