Cryopreservation of Canine Spermatozoa with Blueberry Leaf Extract (Vaccinium corymbosum L.)

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Mariana Deppe; Felipe Pezo; Marjorie Reyes-Díaz & Jennie Risopatrón


During cryopreservation, oxidative stress damage leads to a deterioration of the quality of thawed semen, which could be reduced by the addition of antioxidants to freezing extender. This study was designed to determine the effect of the addition of blueberry leaf extract (EHA) to freezing extender, on quality of cryopreserved canine sperms. Sperm devoid from seminal plasma were frozen with different concentrations of EHA (0 %, control; 1 %, EHA1; 2 %, EHA2; 4 %, EHA4 y 6 %, EHA6) added to freezing extender. Post-thawing progressive motility was evaluated; the viability and plasma membrane integrity (SYBR-14/PI) and acrosomal membrane integrity (FITC-PNA/PI) were assessed by flow cytometry. Progressive motility was similar to the control with concentrations of EHA1 and EHA4 (P >0.05); at concentrations of EHA2 and EHA6 a significant decrease of this parameter compared to control (P <0.01and P <0.001, respectively) was observed. The addition of EHA1, EHA2 and EHA4 to the freezing extender showed no significant differences with respect to the control on viability and plasma membrane integrity (P >0.05); however with the addition of EHA6 values significantly lower (P <0.001) were exhibited. The concentrations of EHA used showed no significant differences with respect to the control on acrosome membrane integrity. In conclusion, the results of this study revealed that none of the concentrations of EHA used were effective in improving canine thawed semen quality.

KEY WORDS: Canine; Spermatozoa; Cryo- preservation; Antioxidants; Blueberry.

How to cite this article

DEPPE, M; PEZO, F.; REYES-DÍAZ, M. & RISOPATRÓN, J. Cryopreservation of canine spermatozoa with blueberry leaf extract (Vaccinium corymbosum L.). Int. J. Morphol., 34(2):653- 659, 2016.