Immune Responses of LEAPs in Fish

To detect the immune responses of LEAPs during infection, rainbow trout and grass carp were intraperitoneally injected with 100 µL Aeromonas salmonicida BG1 (29 (link)) suspension culture (1×10⁷ CFU/mL) and 200 µL Aeromonas hydrophila XS91-4-1 (30 (link)) suspension culture (8×10⁶ CFU/mL) respectively, while the control fish were intraperitoneally injected with PBS instead after anesthetized with MS222 (1:10000). At 12 h, 1 d, 3 d, 5 d, and 7 d post-injection, the liver and gut of four individuals were sampled from each group. After the RNA extraction and cDNA synthesis, the expression levels of LEAPs in infected and control fish were determined by qPCR. The expression changes of LEAPs after infection were calculated using the 2^−ΔΔCt method, with β-actin as the internal reference.