2.2.2. Juvenile oysters
To test the effects of emersion, juvenile oysters were exposed to
air for 3 h. The treated oysters were then returned to the water and sampled 24 and 96 h post treatment (Fig. 1B). Control oysters were
removed from the water and sampled immediately at 0, 24, and 96 h.
2.3. Hemolymph collection
Oysterswere shucked open by severing the adductormuscle. Hemolymph
was collected by penetrating the pericardial cavity located near
the adductor muscle, using a 21-gauge needle fitted to a 1 mL syringe.
Whole hemolymph was then transferred into sterilized 1.5 mL
microcentrifuge tubes and stored in liquid nitrogen at −80 °C. Hemocyte
lysates (HL) were produced by freeze-thawing the hemolymph
(−80 °C/room temperature) three times. The lysed samples were
centrifuged at 5000×g for 30 min (4 °C) to remove cellular debris.