180 SPERM SELECTION OF STALLION PONIES THROUGH GLASS WOOL
G. A. Pessoa A B , J. M. Trentin A B , A. P. Martini B , D. R. Dotto B , L. A. M. Centeno B , M. L. Jardim B , K. V. Aires B and M. I. B. Rubin A BA Universidade Federal do Rio Grande do Sul - Programa de Pos-Graduacao em Medicina Animal: Equinos, Porto Alegre, Rio Grande do Sul, Brazil;
B Universidade Federal de Santa Maria - Embryolab, Departamento de Clinica de Grandes Animais, Centro de Ciências Rurais, Santa Maria, Rio Grande do Sul, Brazil
Reproduction, Fertility and Development 26(1) 204-205 https://doi.org/10.1071/RDv26n1Ab180
Published: 5 December 2013
Abstract
Two techniques of sperm concentration (centrifugation or filtering) and sperm separation technique with glass wool were applied to the sperm samples collected from 3 pony stallions (6 ejaculates; 2 from each stallion). Ejaculates were extended to a final concentration of 50 × 106 spermatozoa mL–1 using a nonfat dry milk-based extender and evaluations occurred at 24, 48, and 72 h after immediate ejaculate dilution and cooling. Each stallion was considered as a block, and semen from each stallion was assigned to 4 treatments: Group A (control): extended semen alone; Group B: extended-centrifuged semen; Group C: extended-sperm filtered semen; Group D: extended-glass wool-separated semen. All groups were tested for pH, osmolarity, motility, morphology, membrane functionality (hyposmotic), and cell viability (MTT assay). The experimental design was performed using a split-plot model. Data analysis at the level of 5% was performed using ANOVA and Bonferroni as post-hoc test. Data are presented as mean ± standard error. Group D had the highest rate of viable cells (P < 0.05) after the separation procedure (Table 1). Group B had a higher percentage of cells with tail defects after processing compared with the controls and Groups A, C, and D (P < 0.05). More than 60% of the cells retained on the filter showed defects (P < 0.001). Progressive motility was greater in group D at 0, 24, and 48 h (P < 0.05). Seventy-two hours after cooling, motility in groups A and B was lower than in Group D (P < 0.01). Group D showed a higher number of cells with mitochondrial activity during the cooling period. In conclusion, the technique of sperm selection by gravity using a glass wool filter resulted in an increased number of viable sperms after cooling pony semen for 24, 48, and 72 h.
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