The present study examined the possibility of long term storage, by cryopreservation in liquid nitrogen, of the sperm of Filefish (Thamnaconus septentrionalis), and the changes in motility, survival rate and ultrastructure of the sperm after freezing and thawing. The sperm was collected by stripping and stored on ice until experiments. For selection of the immobilizing solution, diluted artificial seawater (ASW) of 20, 30 and 40% were tested. The sperm motility was significantly inhibited in 30% ASW, and restored entirely after 100% ASW was added again. Two cryoprotectants, dimethyl sulfoxide (SO) and glycerol, were added to 30% ASW to formulate the extenders at the concentrations between 5 to 20% by volume for freezing. The sperm was diluted at the ratio of 1 :6 with the extenders, inserted into 0.5ml plastic straws and frozen at a freezing rate of /min to after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The straws were thawed in a water bath for 15 sec. The highest post-thawed sperm motility and survival rate were obtained with 5% glycerol Afterward, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of /min to showed the best result Some ultrastructural changes of sperm, such as the detachment of plasmatic and nuclear membranes, destruction of mitochondria, were observed after cryopreservation. Morphological normality of the sperm in 5% glycerol frozen at the ratio of 1/min to was better than that of others.

The present study examined the possibility of cryopreservation of the D-shaped and umbo larvae of arkshell (Scapharca broughtonii), in terms of the survival rates after freezing and thawing. D-shaped and umbo larvae of arkshells were obtained from a shellfish farming on Yosu city. The average shell lengths were m and , respectively. Five cryoprotectants (CPAs), dimethyl sulfoxide (DMSO), glycerol, ethylene glycol (EG), propylene glycol (PG), and methanol, were tested at the concentrations of 1.5, 2.0 and 2.5 M. After larvae suspended in CPAs, cryoprotectants were loaded in 0.5 ml straws at a larval density of 50-100 larvae per straw, and epuilibrated for 10 and 20 minute at room temperature (), repectively. Straws were cooled at a rate of /min from to , held for 5 min at , and then cooled at /min to and equilibrated for 5 min followed by plunging in liquid nitrogen. After storage in liquid nitrogen for 1 day, straws were thawed in a water. As soon as straws were observed to melt, larvae were diluted with an equal volume of ASW and then washed twice with a large volume of ASW at an interval of 2 min to unload the CPAs. The results showed that after equilibration for 10 and 20 minute at room temperature, no larvae survived using methanol as CPAs, and it was observed that larval shells all open slightly, and larval flesh broke down and slopped over the shells. The highest survival rates (D-shaped larvae: 77.6%, umbo larvae: 59.3%) were obtained with 2M DMSO, and 1.5M glycerol yielded survival rates of 53.8% for D-shaped larvae and 37.5% for umbo larvae. The surviving D-shaped larvae showed active rotary motion and perfect membrane integrity and cytoplasmic normality, and the vigorous movement of veliger cilia was observed inside the closed shells. The breakdown of tissue occurred in the abnormal larvae, and the isolated cell often run out of shells.