Cryopreservation of ram semen with incorporation of cholesterol through cyclodextrin

• Grant number: 12/09927-7
• Support Opportunities: Scholarships in Brazil - Master
• Start date: November 01, 2012
• End date: July 31, 2014
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Eneiva Carla Carvalho Celeghini
• Grantee: Leonardo Batissaco
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The concern with the quality of frozen ram semen has been the subject of many researches, particularly through the trouble of cervical transposition during artificial insemination. However, intra-cervical insemination in sheep is often used, resulting in reduced fertility with the use of frozen semen. In this experiment we tested the hypothesis that the influx of cholesterol incorporated by cyclodextrin improves sperm parameters. Five ejaculates from six White Dorper rams (n = 30) were collected, each ejaculate divided into three treatments: control, only extender (CON); extender plus cholesterol incorporated cyclodextrin (CLC + CHO); and extender plus pure cyclodextrin (CLCP).The samples were previously analyzed for motility parameters (computed and subjective evaluation), integrity of the plasma and acrosomal membranes, mitochondrial membrane potential, morphology, and production of free radicals under microscopy. After, samples were packaged in straws at a concentration of 50x106 spermatozoa / ml, identified, and cryopreserved by an automated system. After, semen was thawed (37 ° C for 30 seconds) and analyzed again for the motility parameters, mitochondrial membrane potential, morphology, and production of free radicals under microscopy, moreover, it has been added analyzes by flow cytometry on the integrity of plasma and acrosomal membrane, lipid peroxidation and cell membrane permeability, and by fluorimetry, the quantification of cholesterol and cholesterol esters. Data were assessed for normality of residuals (Shapiro-Wilk) and homogeneity of variances. When p <0.05, the data were transformed and reassessed. Comparisons between groups were analyzed by the mixed procedure (PROC MIXED) of SAS. When the group effect was significant, differences between groups were found using the Tukey test. Data that, even after transformation, continue to be significant in the normality and / or homogeneity tests were analyzed by non-parametric statistics of order (Kruskal-Wallis). The CLC + CHO treatment was more effective in preserving the parameters of motility, membrane integrity, and mitochondrial potential compared to other treatments. The CLCP group showed a decrease in the parameters of motility, membrane integrity, mitochondrial potential, showing thereby a decrease in sperm preservation. It can be concluded with this study that treatment with cyclodextrin plus cholesterol contributes to better preserve the sperm parameters of ram semen.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

BATISSACO, LEONARDO; DE ARRUDA, RUBENS PAES; RODRIGUES ALVES, MAIRA BIANCHI; TORRES, MARIANA ANDRADE; LEMES, KLEBER MENEGON; PRADO-FILHO, ROBERTO ROMANO; DE ALMEIDA, TAMIE GUIBU; CESAR DE ANDRADE, ANDRE FURUGEN; CARVALHO CELEGHINI, ENEIVA CARLA. Cholesterol-loaded cyclodextrin is efficient in preserving sperm quality of cryopreserved ram semen with low freezability. REPRODUCTIVE BIOLOGY, v. 20, n. 1, p. 14-24, MAR 2020. (12/00040-0, 12/09927-7, 13/13438-4)

Academic Publications

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

BATISSACO, Leonardo. Criopreservation of ram semen with colesterol loaded cyclodextrin. 2014. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Medicina Veterinária e Zootecnia (FMVZ/SBD) Pirassununga.