1.In order to understand practicablity cryopreserved sperm of small abalone (Haliotis diversicolor )sperm, which have established in 2016 and modified frozen semen treatment program , using this sperm that has been cryopreserved in liquid nitrogen (LN) for 20 months under the same thawing conditions fertilized test at concentration of 2.646 109 mL at 23- 25 °C, the survival rate of subsequent fertilized eggs, embryos and juveniles was observed after fertilization. The results showed that about 20% of the fertilized eggs had begun to divide after about one hour after fertilization. In addition, Abalone juvenils can be observed by a digital microscope 27 days after fertilization.
2. In study the cryopreservation of Epinephelus tukula and Paralichthys olivaceus sperm , mature E.tukula semen and P.olivaceus semen were used as experimental material, and concentration of various semen diluents, and concentration of cryoprotectant were screened.We used liquid nitrogen and 0.5 ml cryo-straws to preserve sperms by step cooling procedure. The result showed the cryopreservation of E.tukula semen that Hank’s solution、Marine Fish Ringer and EM1-2 semen duluents was better , which preserved 90.00%±1.00、89.67%±1.53及85.67%±4.04 sperm motility after unfrozen. Using Hank’s solution as the liquid foundation to prepare cryoprotectant, no significant difference was observed in sperm motility by 10%、15% and 20% DMSO (P>0.05) . The best of all is 15% DMSO, which preserved 87.67%±1.53 sperm motility respectively after unfrozen.
The cryopreservation of P.olivaceus semen that MPRS and Hank’s solution semen diluents was better in sperm motility, which preserved 56.67%±2.08及53.67%±3.21% sperm motility after unfrozen. Using MPRS as the liquid foundation to prepare cryoprotectant, was better , which preserved 90.00%±1.00、89.67%±1.53及85.67%±4.04 sperm motility after unfrozen. It was the best in sperm motilityby 15% DMSO cryopreservation, which preserved 57.00%±2.65 sperm motility respectively after unfrozen. We established a sperm cryopreservation protocol for E.tukula and P.olivaceus on this study, and it will provide a basis for the artificial breeding and cross breeding.
3.The purpose of this program is to provide the tilapia industry with new techniques that were developed by the Germplasm Bank of Freshwater Fish, Fisheries Research Institute. These techniques will be promoted to the industry with following goals to enhance the competitive strength. (1) The technology transfer of genetically male tilapia. (2) The supply of 500,000 fries of Nile tilapia with the feature of high growth rate. (3) The application of molecular protocols to analysis the reason of low male-ratio of tilapia fry.
The technology genetically male tilapia has licensed for two companies this year, and it is expected that the fry will be stably produced after 2 years. In addition, we try to analyze the sexdetermining locus of genetically female and found GM047, GM212, UNH898, UNH216 and GM631 loci were correlated with sex determination. 4.Qualified individuals of Tridacna maxima with target colorations and patterns were selected. Spermiation of 3-year-old clams were successfully induced and fertilized with eggs from mature wild-caught adult. Through aquaculture of artificially produced T. maxima was proved completed. Serotonin injection (chemically induced method) in the field was carried out and spermiation and spawn were successfully induced. Fertilization and further larval development of T. maxima from chemically induced sperm and eggs were also observed.