Influence of seminal plasma derived from the rich fraction of the ejaculate on the structural characteristics and the kinetics of the swine sperm stored under refrigeration at 17°C for 72 hours

Abstract

This project aims to evaluate the effects of seminal plasma, derived from the rich fraction of the ejaculate, on sperm kinetics, integrity of plasma an acrossomal membranes and lipid peroxidation of porcine sperm stored under refrigeration at 17°C for 72 hours. Thus, six ejaculates from six boars will be used. The semen in natura will be evaluated as to the characteristics of motility by the computerized system for semen analysis, sperm morphology by differential interference contrast and sperm concentration. Shortly after this initial assessment, the ejaculate will be placed into 50 mL conical tubes in order to be divided into three treatments, as follows: Not Centrifuged (NC), centrifuged with the seminal plasma removed After Centrifugation (CS) Centrifuged and Suspended (CR). To separate the seminal plasma from the ejaculate, semen will be centrifuged at 600xg for 20 minutes. All treatments will be extended in BTS medium in order to obtain a concentration of 30 X 106 sperm per milliliter (mL) to reach a final volume of 50 ml DI. The semen doses will be hold for 90 minutes at room temperature and protected from light before they are stored under refrigeration. Semen from the different treatments will be stored at a temperature of 17°C to be evaluated at 0 (90 min post-dilution), 24, 48 and 72 hours for the following parameters: sperm motility characteristics (CASA) plasma membrane integrity and acrosomal membranes and peroxidation of sperm by flow cytometry. The treatment will be subjected to analysis of variance (PROC GLM), using the SAS program (1998). When the main effect is significant, the averages will be compared by the Tukey test. (AU)