Use of antioxidants in TRIS extender for cooling and freezing epididmal sperm cells from dogs submitted to elective orchiectomy.

• Grant number: 09/12360-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: August 01, 2010
• End date: February 29, 2012
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Wilter Ricardo Russiano Vicente
• Grantee: Paula Andressa Pennacchi Savi
• Host Institution: Faculdade de Ciências Agrárias e Veterinárias (FCAV). Universidade Estadual Paulista (UNESP). Campus de Jaboticabal. Jaboticabal , SP, Brazil

Abstract

The aim of the present study is to assess sperm cells quality before and after chilling and freezing processes in modified TRIS extender supplemented with two different antioxidants (vitamin E and/ or N-acetyl-cysteine). Twenty experimental units (EU) will be used. Each EU will correspond to a spermatic "pool" recovered from the epididimus of three healthy male dogs aged between 1 to 5 years, previously submitted to elective orchiectomy. Nevertheless, three groups composed by 7 EU each will be formed. Group I (GI), Group II (GII), and Group III (GIII), will be supplemented with 0,5mM, 2,5mM e 5mM of antioxidants, respectively. In all groups, fresh semen will be evaluated on motility, progressive motility, and vital staining. Sperm morphology will be assessed by Pope's staining and membrane integrity will be evaluated by hypo-osmotic swelling test, and fluorescent probes. In addition thermo-resistance test will be performed, and reactive oxygen species (ROS) will be measured as well as mitochondrial activity. Subsequently each EU will be divided in 4 aliquots of equal volume and centrifuged. The supernatant will be discarded and sperm cells resuspended in TRIS extender described by Peña and Linde-Forsberg (2000) and modified by Martins (2005), supplemented by vitamin E, N-acetyl-cysteine, or both. Each aliquot will be divided in two samples one for chilling (for 48 hours), and another for freezing. All samples will be evaluated after chilling and thawing. One way ANOVA with Tukey test as a post hoc test will be performed to compare means (P<0.05).

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

SAVI, Paula Andressa Pennacchi. Uso de antioxidantes no meio diluente tris para refrigeração e congelação de células espermáticas provenientes de epidídimo de cães submetidos à orquiectomia eletiva. 2012. Master's Dissertation - Universidade Estadual Paulista (Unesp). Faculdade de Ciências Agrárias e Veterinárias. Jaboticabal Jaboticabal.