
TY  - JOUR
A1  - Rodriguez-Vilar, L. 
A1  - Hernandez, M. 
A1  - Lopez-Sanchez, C. 
A1  - Vazquez, J. M. 
A1  - Martinez, E. A. 
A1  - Roca, J. 
T1  - 130 EFFECTIVENESS OF BUTYLATED HYDROXYTOLUENE AS EGG YOLK SUBSTITUTE FOR CRYOPRESERVATION OF BOAR SPERMATOZOA
JF  - Reproduction, Fertility and Development
JO  - Reprod. Fertil. Dev.
Y1  - 2006
VL  - 19
IS  - 1
SP  - 182
EP  - 183
KW  - 
UR  - https://doi.org/10.1071/RDV19N1AB130 
N2  - Butylated hydroxytoluene (BHT) has proven to be efficient as a supplement for cryopreservation boar spermatozoa (Roca et al. 2004 J. Androl. 25, 397&#8211;405). Moreover, it has been successfully used as an egg yolk substitute to cryopreserve goat spermatozoa (Khalifa and El-Saidy 2006 Anim. Reprod. Sci. 93, 303&#8211;315). The objective of this study was to evaluate the effectiveness of BHT as an egg yolk substitute for freezing boar spermatozoa. Nine sperm-rich ejaculate fractions were collected from 3 boars (3 ejaculates per boar) using the gloved-hand method. After centrifugation (2400g for 3 min), the sperm pellet of each ejaculate was split into 5 aliquots. The aliquots were diluted (to a final concentration of 1 × 109 sperm/mL) in a Tris-citric-glucose extender with 3% glycerol and supplemented with 20% egg yolk (positive control, PC aliquot) or BHT at the final concentrations of 0 (negative control, NC aliquot), 0.2, 0.4, and 0.8 mM. Diluted semen samples were dispensed into 0.5-mL straws, and frozen in a programmable cell freezer at 20°C min. Thawing was carried out in a water bath at 70°C for 8 s. Post-thaw sperm survival was assessed according to total sperm motility (TSM, %) using a CASA system (SCA®; Microptic, Barcelona, Spain), and plasma membrane integrity (PMI, %) and acrosome membrane integrity (AMI, %) using a flow cytometric procedure (SYBR-14/propidioum iodide/FITC-phycoerythrin), at 30 and 150 min post-thawing in diluted Beltsville thawing solution with spermatozoa held in a waterbath at 37°C (3 straws per ejaculate). Data were analyzed using a ANOVA mixed model including the main effects of aliquot, boar, post-thaw assessment time, and their interactions, with ejaculate and straw as random effects. All main effects had significant influence (P &#8804; 0.01) in all post-thaw sperm assessments. However, no interactions (P &#8805; 0.05) among main effects were shown. Data were combined for the 2 post-thaw assessment times. The best (P &#8804; 0.05) post-thaw sperm quality (mean ± SEM) was achieved in PC aliquots (47.11 ± 3.10, 58.98 ± 2.78, and 51.35 ± 3.42 for TSM, PMI, and AMI, respectively). In NC aliquots, the percentage of TSM, PMI, and AMI were always below 1% (P &#8804; 0.05). BHT has a beneficial (P &#8804; 0.05) effect on post-thaw sperm assessments, and no differences (P &#8805; 0.05) among concentrations were shown. The mean post-thaw sperm quality in the BHT aliquots was 8.50 ± 0.80, 20.29 ± 0.53, and 16.03 ± 0.55 for TSM, PMI, and AMI, respectively. On the basis of these data, we can conclude that BHT has a protective effect for boar spermatozoa during the cryopreservation process. However, BHT alone is insufficient to replace the protective effect of egg yolk. This work was supported by CICYT (AGF2005-00706), Madrid, Spain. 
ER  -

