In vitro production of female bovine embryos by means of semen sexing associated techniques confirmed by PCR

• Grant number: 12/01357-7
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2012
• End date: April 30, 2013
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Vera Fernanda Martins Hossepian de Lima
• Grantee: Cibeli Martins Chioda
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

Studies have indicated that it's possible to divert the sexual proportion favoring female bovine embryos produced in vitro, given by semen centrifugation frozen in a gradient of such density that sediments X spermatozoon. This diverting has 65% accuracy. It has been shown that many factors could influence the gender proportion, including 1 - the insemination timing, 2 - the maturation state of the oocyte in the very moment of in vitro fecundation., 3- in vitro gametes co-incubation timing, and 4 - in vitro post-incubation cultivation conditions. The ability to produce as many males as females would be of great importance in the animal production industry as well as the biotechnology programs directed to animal reproduction face many problems, among them, the male production being higher than the female, the purposes of this project are: to try some modifications in all the steps of in vitro production (maturation, fecundation, and cultivation) of embryos in order to favor the development of female embryos. The modification made in the fecundation timing and the spermatozoon incubation associated with gender diverting by Percoll TM before the fecundation in order to direct the sexual proportion to females. The quality control will be done in every step of the procedure evaluating the motility parameters, vigor, and spermatozoon concentration as well as the cleavage rate and in vitro production of the embryos. The sexual proportion will be evaluated by the Polymerase Chain Reaction (PCR), such procedure allows for verifying the accuracy of the techniques mentioned above.(AU)