Effect of seminal plasma on post-thawing quality of bovine sperm recovered from the epididymis tail
Torralvo Genes, Juan Diego
Usuga Suarez, Alexandra
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AbstractThe aim of this study was to evaluate the effect of seminal plasma supplementation according to its total antioxidant capacity (TAC) on the post-thawing quality of bovine sperm recovered from the epididymis tail. Seminal plasma (SP) from five healthy and fertile bulls was collected by electroejaculation, and then it was centrifugated and pooled. SP total antioxidant capacity was measured by ABTS and ORAC assays. Epididymal tail sperm was collected by retrograde flow technique from sixteen testis-epididymal complexes (TEC) after orchiectomy of eight bulls in slaughterhouse. Each sample was separated and supplemented as follows: a control without SP supplementation, treatment 1 (SP 10) supplemented with 10% of SP v/v, treatment 2 (SP 20) supplemented with 20% of SP v/v and treatment 3 (SP 30) supplemented with 30% of SP v/v. Each aliquot was diluted in Triladyl® extender (Minitube, Germany) to reach a concentration of 60 x 106 sperm/ml. Sperm cryopreservation was performed using a standard protocol. After thawing, we evaluated motility and sperm kinetics in the SCA® CASA System, membrane functionality by HOS test, sperm vitality (SV) and morphology by eosin-nigrosin staining procedure, lipid peroxidation and mitochondrial membrane potential by spectrofluorimetry with BODIPY and JC- 1 dyes, respectively. Evaluation of ROS production was also assayed by spectrofluorimetry. Results for TAC of SP pool were 4902,5 ± 258,6 μmol Trolox Eq/L of SP measured by ABTS assay and 1205,3 ± 710,6 μmol Trolox Eq/L of SP measured by ORAC assay. SP 20 treatment showed the highest results for PROG, VAP and VSL, respecting the control treatment (p<.05). For HOS, the treatment with better results was SP10, while SP30 showed a significant reduction in this parameter (p<.05). Control and SP30 showed the highest and the lowest results for BODIPYRed, respectively (p<.05). No effect by treatment was found for mitochondrial membrane potential (p>.05). In conclusion, seminal plasma supplementation can improve the quality of bull sperm recovered from the epididymis.