Photograph of one example of hemorrhages in the fins of VHSV-infected zebrafish (A) and time course of mortalities of VHSV-infected zebrafish (B). Adult zebrafish (n = 10) were infected with 2 × 10⁶ ffu/ml of supernatant from VHSV-07.71 infected EPC cells or virus-free cell cultured medium at 14°C in 50 ml of water. Two hours later they were released into 2 liters aquaria. External hemorrhages were present in all the fish that die. Depending on the fish, hemorrhages first appeared distributed among mouth, lateral or ventral skin and bases of the fins. Figure 1A shows one example which appears in 10-40% of the VHSV-infected zebrafish. Mortalities (Figure 1B) were recorded during the following 30 days in 6 different experiments (different symbols). The vertical arrow indicates the 2-day time at which samples of fins and organs (pooled spleen, head kidney and liver) were harvested for further studies.

Representative 2D-DIGE of fin proteins from VHSV- infected (red) and non-infected (green) zebrafish. Protein extracts from zebrafish fins were analyzed by 2D-DIGE. First dimension was performed on an immobilized pH gradient gel (pH 4 to 9). Second dimension was performed on a 10% SDS-PAGE to separate proteins by their molecular weight (10 to 150 KDa). The circled numbers correspond to the spots which were analyzed by MS. About 70% of the spots analyzed could be identified depending on the experiment (Table 1). Left, molecular weight markers. Bottom, pH gradient from 4 to 9.