Research On Separation Of X, Y Spermatozoa In Boar Semen

It has very important mean on improving husbandry efficiency and promoting basic research development in controlling the mammal offspring sex so get more required animals. Controlling the sex-determine gene, separating the X, Y spermatozoa, identifying the embryos and selective transfer, selective pregnancy and controlling the environment facts were used to realize the aim. Separation the X, Y spermatozoa is the most direct and efficient method among them, so most researches attached great importance to do it.More researches were done to separate X, Y spermatozoa in bovine, goat and rabbit, including swim-up, electrophoresis, density-grads centrifuge, Percoll method, immune method, layer-out. However, their veracity and repetition can not be validate. Since the flow cytometry was invented, much manpower and resource were delivered to it by government and society to get sexed sperm, though its separate veracity reached to 80-90% and be the most veracious method now. Disadvantages such as expensive price, high technique need, low efficiency and harm to sperm were restrict its use in husbandry. So finding a cheap, practical, harmless and efficient way to separate the X, Y spermatozoa become urgent need in research and production.In this study, boar X, Y spermatozoa was separated with Wang's tube under acid inducement, identified, froze, in vitro fertilized and embryos sexed. Boar semen was separated with Wang's tube under acid inducement, identified with Quarine dying and observed the results under fluorescence microscope. In order to explore separated sperm conservation method and test its anti-froze capability, liquid nitrogen was used to froze separated sperm, sperm quality was tested after be thawed. Thawed sperm in vitro fertilized with mature ovum, observe the effect of separated sperm on fertilization and oosperm development. Embryos were identified with PCR technique and appraise separate efficiency again. After the above experimentation, the follow results had been obtained.1. When pH is 5.5, the result of separate X, Y spermatozoa with Wang's tube under acid inducement is better. the motility improved 11.9%(95 % VS 85 %,P<0.01), X spermatozoa is 72.6 %,Y spermatozoa is 27.4 %,there is significant difference between separated and original(P<0.01). Wang's tube separate method under acid inducement is a easy but efficient way.2. After the separated sperm thawed, the motility, acrosome integrity, membrane integrity,...