Cryopreservation Of Spermatoza And Embryo Of Pseudosciaena Crocea

In the past few decades, methods for successful cryopreservation of spermatozoa and embryo have been developed for domestic animals. In recent years more and more work for marine animals are developing . In the present paper, attempting to develop simple, reliable and efficient methods for cryopreservation of Pseudosciaena crocea are reported.In the study of sperm, the best levels of various factors have been found. Motility, fertility and hatchery rate of 92, 48.8 and 51% are obtained in cryopreservation of the spermatozoa of Pseudosciaena crocea using a cryoprotectant dilution (X5+10%GLY+0.015g/ml fucose) and freezing rate 20/min. To find out the optimal freezing plan a number of experiments have been done and the results demonstrate that method of two steps to insert into LN2 is more tolerant than the other ones. In this method samples are hold at -15 in the LN2 container for 10 min and above the surface of LN2 for 5 min before immerged into LN2. The optimal thawing temperature is 43癈 in water bath. Motility of 85. 3% can be obtained by this freezing progress. When the dilution rate is 1:3 there is no distinct different among various volumes of sample between 0. 5-1. 6ml. When volume of sample is 0.5ml freezing sample with dilution rate of 1:3 can obtain the most motility. Motility analysed by CASA is 93. 75%, half of them are move towards rapidly.In the study of embryo, among four kinds of cryoprotectants METH has the lest toxicity to embryo of Pseudosciaena crocea and GLY has the most one. Embryos loaded in 20%METH can keep 81. 8% alive after 40 min, yet those in 20%GLY only have 79% alive after 6 min. METH is able to keep modalityof embryo most intact and obtain intact modality rate of 60%, and it is PG that make frozen embryo lest intact. The optimal cryoprotectant is X3+20%METH. 0. 15g/ml acetamide can reduce some toxicity of METH and METH10 %+GLY10% can improve the freezing effect. For embryo of Pseudosciaena crocea the freezing damage occurs below ?0. The optimal freezing plan isand the optimal thawing method is holding samples at -80癈 for 2 min then thaw them in water bath at 43癈, when half of them have thawed put samples at room temperature to thaw completely. Then they are unloaded in 0. 5M sucrose solution for ten minutes before cultured in seawater. Planting an ice nucleus at - 26癈 is effective measure to decrease the freezing damage and enhance intact modality rate. In this plan 84.8% Sparus macrocephalus embryos are able to keep forms comparative normal, however, revival has not been found so far.