Refrigerated handling condition on post mortem goat testicles and its influence the quality of epididymal sperm [manuscript]
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Date
2017
Authors
Hannah Lei M. Harada
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Abstract
The goal of cryoconservation is to provide a parallel effort to conserve and utilize valuable germplasms for the envisaged genetic improvement. Selected, pedigreed and trained semen donors such as large ruminants are utilized in the Philippines for breed conservation. However, for small ruminants not trained for semen collection, an alternative option is to use epididymal sperm. In this study, slaughterhouse-derived testicle utilization (n=12) was performed to examine the influence of 5-8°C handling temperature during 4h (Group 1) and 24 h (Group 2) of transport on the quality and survivability of epididymal sperm across fresh, pre-freeze and post-thaw treatments. Epididymal sperm from dissected cauda epididymis were recovered in a buffered base media composed of Tris, citric acid monohydrate, lactose and raffinose (TCLR). After centrifugation, the average sperm pellet volume of Group 1 and 2 were 430µL and 150µL with a mean sperm concentration of 340.5x10⁷ per ml and 302.5x10⁷ per ml. The proportions of live perm were 90.5% and 84.67% with 60% initial motility that served as criteria to proceed cryopreservation. Sperms were diluted with TCLR buffer containing 20% egg yolk (v/v) and 7% glycerol (v/v) at a sperm concentration of 200x10⁶ sperm per ml. After 2-2.5 hours of equilibration in the refrigerator, microscopic evaluation of sperm under pre-freeze treatment still showed ideal acceptable results. After subjecting the samples to cryopreservation (-196°C), post thaw motility score of frozen epididymal sperm had a major decrease while fluorescence based live sperm assay SYBR/PI yielded 27.17 and 28.25%. Despite of loss of quality especially motility due to alterations of the physical environment and sperm aging, samples were qualified for IVF and ICSI. Thus, refrigerating the testicles before laboratory processing is a good option in rescuing valuable goat sperm reserves.