Trzcińska Monika, Bryła Magdalena, Gajda Barbara, Gogol Piotr
National Research Institute of Animal Production, Department of Animal Reproduction Biotechnology, Balice, Kraków, Poland.
National Research Institute of Animal Production, Department of Animal Reproduction Biotechnology, Balice, Kraków, Poland.
Theriogenology. 2015 Feb;83(3):307-13. doi: 10.1016/j.theriogenology.2014.07.045. Epub 2014 Oct 30.
The present study was to determine the effect of butylated hydroxytoluene (BHT) on quality and fertilizing ability of frozen-thawed boar semen. In the first experiment, five crossbreds of Polish Landrace and Large White boars (five ejaculates per boar) were frozen in 0.5 mL straws after dilution with lactose-egg yolk-glycerol extender supplemented with 0 (control), 0.5, 1.0, and 2.0 mM BHT. The sperm quality was verified based on the motility (computer-assisted sperm analysis; total motility, %; progressive motility, %), membrane integrity (YO-PRO-1/propidium iodide [PI] assay), acrosome integrity (fluorescein isothiocyanate-conjugated with peanut agglutinin/PI), and lipid peroxidation (chemiluminescence method) at 15 minutes postthaw. In the second experiment, the semen cryopreserved in extender supplemented with 1.0 and 2.0 mM BHT were selected for intrauterine artificial insemination of synchronized gilts. An intrauterine artificial insemination with low numbers of spermatozoa (500 × 10(6)) was surgically infused into each uterine horn. The highest (P < 0.001) progressive motility (%), membrane integrity, and acrosomal integrity were noted by the addition of 1.0 and 2.0 mM BHT to the freezing extender. Moreover, the various concentrations (0.5-2.0 mM) of BHT caused a considerable decrease in lipid peroxidation in relation to the control extender (P < 0.001). The highest reproductive performance of inseminated gilts (farrowing rate, 86.7%; litter size, 10.8 ± 1.6) was observed when semen was cryopreserved in extender supplemented with 1.0 mM BHT. These findings demonstrate that the addition of 1.0 mM BHT to the freezing extender efficiently improves the fertilizing ability of postthaw boar spermatozoa.
本研究旨在确定丁基羟基甲苯（BHT）对冻融公猪精液质量和受精能力的影响。在第一个实验中，将五头波兰长白杂交公猪（每头公猪采集五次射精）的精液用补充了0（对照）、0.5、1.0和2.0 mM BHT的乳糖-蛋黄-甘油稀释液稀释后，分装于0.5 mL细管中冷冻。解冻后15分钟，基于精子活力（计算机辅助精子分析；总活力，%；直线运动活力，%）、膜完整性（YO-PRO-1/碘化丙啶[PI]检测）、顶体完整性（异硫氰酸荧光素标记花生凝集素/PI）和脂质过氧化（化学发光法）来验证精子质量。在第二个实验中, 选择保存在补充了1.0和2.0 mM BHT的稀释液中的精液，对同期发情的后备母猪进行子宫内人工授精。通过手术将少量精子（500×10⁶）注入每个子宫角进行子宫内人工授精。在冷冻稀释液中添加1.0和2.0 mM BHT时，精子的直线运动活力（%）、膜完整性和顶体完整性最高（P<0.001）。此外，与对照稀释液相比，不同浓度（0.5 - 2.0 mM）的BHT均能显著降低脂质过氧化水平（P<0.001）。当精液保存在补充了1.0 mM BHT的稀释液中冷冻时，授精后备母猪的繁殖性能最高（产仔率86.7%；窝产仔数10.8±1.6）。这些结果表明，在冷冻稀释液中添加1.0 mM BHT可有效提高解冻后公猪精子的受精能力。
