Briefly, 1000 μl of hemolymph–anticoagulant mixture (1:9) was centrifuged at 800 ×g and 4 °C for 20 min. Plasma was removed, and the pellet was re-suspended with 1ml of 0.85% sodium chloride solution, and centrifuged again. The supernatant was discarded, and the pellet was re-suspended in 100 μl of distilled water at 4 °C for 15 min