Sperm And Embyo Cryopreservation Of Red Sea Bream(Pagrosomus Major) Fu-Hong, DING

Cryopreservation is one of the most important technologies to realize the long-term stability of species preservation, which can provide technological support to protect the diversity of genes. Meanwhile, it can also provide great convenience to scientific research and application. In this paper, some cryopreservation experiments about sperm and embryo of red sea bream are reported and a series of conclusions are got.For the cryopreservation of red sea bream's sperm, a set of integral cryopreservation methods and quality controlling methods are obtained. By filtering different cryopreservation methods and cryoprotectants, we have found the optimal scheme for cryopreservation; DMSO 15% is the best cryopreservation to get highest revival rate (89.29±2.77%); the corresponding average path velocity (VAP) and fertilization are 63.5and 96.8% respectively. The correlation between fertilization rate and revival rate, average velocity, the percentage of high velocity sperms are analyzed respectively, and found the correlation coefficient is extremely small between fertilization rate and revival rate, however better with the percentage of high speed sperms, which suggests the sperm can not fertilize unless it has a certain high velocity. The test of DNA damage shows no significant difference between the cryopreservated sperms (DMSO5%, DMSO10%) and fresh sperms.Program cooling method and vitrification method are applied in the study of cryopreservation of embryos and 41 revived embryos are got, which is a scientific breakthrough for the revival of embryos cryopreservated in liquid nitrogen. For the revived embryos, the longest cultured life-span is 15 days, the maximum revival rate is 33.3%. By program cooling method, the embryo's surviving rate reaches 80.989.36 % when the temperature is decreased to -30oC. Adding unpenertrative cryopreservant into vitrification liquid will significantly increase embryo's revival rate in -20oC (P>95%), the maximum revival rate can get to 87.69%. Toxicity experiments show PG has the least toxicity to embryos, and DMSO, GLY, EG take the successive turns, METH has the maximum toxicity to embryos.