Evaluation of a new method of removal of seminal plasma for cryopreservation of goat semen

• Grant number: 14/05984-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: May 01, 2014
• End date: April 30, 2015
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: André Maciel Crespilho
• Grantee: Daniele Campos do Nascimento
• Host Institution: Universidade de Santo Amaro (UNISA). São Paulo , SP, Brazil

Abstract

Artificial insemination (AI) using the cryopreserved semen is the main and the most important technique reproductive management for livestock production. Although many aspects related to the freezing of goat semen have been successfully adapted from the acquired knowledge in biotechnology bovine semen, some particularities presented by semen of goats, and the presence of phospholipase A, require differential processing for these samples. The removal of seminal plasma is a prerequisite for the cryopreservation of goat semen step is usually performed by centrifugation. However, previous studies have shown that centrifugation deleterious effect on the viability and integrity of spermatozoa, justifying the search for alternative technologies for removal of seminal plasma goat. In this sense, the Sperm Filter® (patent 0700624-1, U.S. 2010/0099075) system, a new method of separation of seminal plasma has recently been proposed for equine semen, showing advantages over centrifugation. The purpose of this research proposal is reviewed Sperm Filter® system for removal of seminal plasma prior to cryopreservation of goat semen. Ejaculates from 6 adult breeding Anglo Nubian (n = 5 ejaculations per goat) will be collected by artificial vagina and separated into two equal aliquots. Each sample is diluted in Lactated Ringer's solution Sodium preheated to 37° C in order to fix the final volume of 5 mls. An aliquot is intended to centrifugation at room temperature at 600xg for 10 minutes (control group). The second fraction is diluted in the same solution and subjected to separation by the Sperm Filter® (Challenge Group). Two groups of samples are cryopreserved in medium Tris egg yolk fructose (TRIS) containing 6.4% of glycerol cryoprotectant and evaluated at 0 (after dilution into TRIS), 1 (immediately after freezing/thawing) and 2 (3 hours after thawing) and sperm kinetics of plasma membrane integrity and count of sperm concentration. The generated data are evaluated by a general linear model (GLM® , SAS Institute, Cary, USA), testing the hypothesis that the Seminal plasma separation using Sperm Filter ® may provide a more intact and motile sperm post- thawing.