Abstract

This study was performed to find out the optimal kind of additive and concentration of ethylene glycol (EG) as the cryoprotectant on the D-shaped larvae of arkshell, Scapharca broughtonii. The larvae in straws was carried in the programed freezer set uo at 0$^{\circ}C$, frozen to -12$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min, held for 10 minutes, seeded at -12$^{\circ}C$, finally refrozen to -35$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min and then directly plunged into liquid nitrogen. The survival of larvae frozen in 1.0 M and 2.0 M ethylene glycol added with 0.5 M sucrose were high (50.5 ${\pm}$ 1.3% and 51.9 ${\pm}$ 1.7%, respectively) in the early D-shaped larvae cryopreserved in 1 M and 2 M ethylene glycol diluted with 0.2 M and 0.5 M fructose, glucose and sucrose. In the late D-shaped larvae were cryopreserved according to five concentrations of ethylene glycol added with 0.5 M sucrose, the survival of larvae frozen in 2.0 M ethylene glycol was the highest as 51.9 ${\pm}$ 1.7%. The morphological differences in cells between unfrozen and frozen-thawed D-shaped larvae were not found by the scanning and transmission electron microscopy.