Co-infection of RGNNV and Vibrio harveyi in grouper (Epinephelus spp.) increases mortality by affecting host immune and metabolism

Five treatment groups were established, Grouper were injected with PBS only (200 μL/individual), RGNNV (105 TCID50/mL, 100 μL/individual and 100 μL PBS to make up 200 μL/individual), V. harveyi (1.6 x 106 CFU/mL, 100 μL/individual and 100 μL PBS to make up 200 μL/individual), RGNNV+V. harveyi [(105 TCID50/mL, 100 μL/individual) for RGNNV first and then (1.6 x 106 CFU/mL, 100 μL/individual) for V. harveyi], or V. harveyi+RGNNV [(1.6 x 106 CFU/mL, 100 μL/individual) for V. harveyi first and then (105 TCID50/mL, 100 μL/individual) for RGNNV].Brain tissue and liver & spleen tissues of 3 fish from each group were collected for RNA-seq. Samples from 3 individual fish were collected to prepare each sample, with three replicates of each sample.