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Validation of a new method to remove the seminal plasma of goat semen

Grant number: 14/05983-5
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): May 01, 2014
Effective date (End): April 30, 2015
Field of knowledge:Agronomical Sciences - Veterinary Medicine - Animal Reproduction
Principal Investigator:André Maciel Crespilho
Grantee:Nicole Flavio
Host Institution: Universidade de Santo Amaro (UNISA). São Paulo , SP, Brazil

Abstract

The preservation of semen for use in artificial insemination programs depends on the reversible reduction of metabolic activity and motility of the sperm, which can be achieved by cooling or freezing sperm. To ensure full preservation of spermatozoa during processing, is necessary to add diluents that promote sperm protection. However, the presence of phospholipases present in seminal plasma of goats limits the use of diluents, especially those having a source of egg yolk lipoprotein, and lipid degradation causing the formation of toxic compounds to sperm. For this reason, most studies focused on the preservation of goat semen recommends the use of centrifugation to remove the plasma, assuming, however, that such processing leads to injury to sperm ultrastructure. In this sense, the goal of this research proposal is to evaluate a new filtering system originally developed for separating sperm from seminal plasma of horses SpermFilter® (patent 0700624-1, U.S. 2010/0099075), for the processing of goat semen. Ejaculates from six breeding adults Anglonubiana breed (n = 5 ejaculations per goat) will be collected in an artificial vagina and fractionated 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 in 5mls. An aliquot will be used to centrifugation at room temperature for 10 minutes at 600xg (control group). The second fraction is diluted in the same solution and subjected to separation by SpermFilter® (group challenge). The two groups of samples are evaluated at 0 (after dilution), 1 (immediately after separation) and 2 (3 hours after removal of the plasma) and the kinetics of sperm (sperm evaluation technique computed, ISAS V.1.2, Valencia, Spain), plasma membrane integrity in epifluorescence microscopy count and sperm concentration using a Neubauer chamber hemocitométrica. The data generated are evaluated using the general linear model (GLM®, SAS Institute, Cary, USA) ,testing the hypothesis that the separation of seminal plasma using SpermFilter® can provide a greater number of intact and motile spermatozoa after processing.

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