咖啡因补充以及酶促和机械精液液化协同作用对单峰驼精子冷冻保存能力的影响。
The influence of caffeine supplementation and concerted utilization of enzymatic and mechanical semen liquefaction on freezability of dromedary camel spermatozoa.
作者信息
El-Bahrawy K A
机构信息
Animal & Poultry Production Division, Desert Research Center, 1 Mathaf El Matariya Street, B.O.P. 11753, Cairo, Egypt.
出版信息
Int J Vet Sci Med. 2017 Oct 9;5(2):121-127. doi: 10.1016/j.ijvsm.2017.09.005. eCollection 2017 Dec.
Assisted reproductive technologies have been reported to improve reproductive efficiency and genetic potential in camelids. Two experiments were carried out to determine efficiency of centrifugation in the presence of a mucolytic agent for liquefaction of dromedary semen. In the first experiment, three groups, namely: I. Tris lactose (TL, control), II. Tris lactose supplemented with amylase (TL_A) and III. Tris lactose supplemented with amylase followed by seminal plasma removal via centrifugation (TL_A_) and re-suspension into enzyme free TL media. After equilibration, control group recorded 62.85 ± 4.28% motility and 1.3 ± 0.13 viscosity score, while TL_A group values were 72.88 ± 3.30% and 0.83 ± 0.07%, respectively. TL_A_ group showed significant viscosity reduction (0.33 ± 0.05) and motility decline 47.85 ± 3.04% with increment in abnormalities and detached acrosome. The second experiment investigated the effect of caffeine addition to tolerate enzymatic and mechanical stress. Using 4 mM caffeine in amylase-treated semen (TL_AC) improved post-thaw motility 50.0 ± 1.29% and recovery rate 77.8 ± 3.83% compared to the control (40.17 ± 2.79% and 62.55 ± 8.39%), respectively. Caffeine supplemented centrifuged samples (TL_AC_) showed superiority ( < .05) in post-thaw motility and recovery rate (38.33 ± 6.41%, 62.76 ± 8.10%) compared to centrifuged samples TL_A_ without caffeine addition (25.00 ± 2.88% and 40.47 ± 3.48%), respectively. Sperm kinetics showed that TL_A exhibited high (p < .05) values for mostly all sperm kinetics. Caffeine treatments showed superiority in velocity curved line (VCL, µm/s) 94.24 ± 8.44 for TL_AC and 104.25 ± 8.72 for the TL_AC_ group compared to 86.8 ± 5.54 for TL_A, and 85.73 ± 5.99 for the TL_A_ groups. In conclusion, preforming a combined enzymatic-mechanical protocol in the presence of an antioxidant may be crucial for refinement of camel semen cryopreservation.
据报道，辅助生殖技术可提高骆驼科动物的繁殖效率和遗传潜力。进行了两项实验，以确定在存在黏液溶解剂的情况下离心对单峰驼精液液化的效果。在第一个实验中，分为三组，即：I. 三羟甲基氨基甲烷乳糖（TL，对照组）；II. 添加淀粉酶的三羟甲基氨基甲烷乳糖（TL_A）；III. 添加淀粉酶后通过离心去除精浆（TL_A_）并重新悬浮于无酶的TL培养基中。平衡后，对照组的活力为62.85±4.28%，黏度评分为1.3±0.13，而TL_A组的值分别为72.88±3.30%和0.83±0.07%。TL_A_组显示黏度显著降低（0.33±0.05），活力下降至47.85±3.04%，同时异常和顶体脱落增加。第二个实验研究了添加咖啡因以耐受酶促和机械应激的效果。与对照组（分别为40.17±2.79%和62.55±8.39%）相比，在经淀粉酶处理的精液（TL_AC）中使用4 mM咖啡因可使解冻后活力提高至50.0±1.29%，回收率提高至77.8±3.83%。与未添加咖啡因的离心样本TL_A_（分别为25.00±2.88%和40.47±3.48%）相比，添加咖啡因的离心样本（TL_AC_）在解冻后活力和回收率方面具有优势（p<0.05）（分别为38.33±6.41%，62.76±8.10%）。精子动力学表明，TL_A组几乎所有精子动力学参数的值都很高（p<0.05）。咖啡因处理组在速度曲线（VCL，μm/s）方面表现出优势，TL_AC组为94.24±8.44，TL_AC_组为104.25±8.72，而TL_A组为86.8±5.54，TL_A_组为85.73±5.99。总之，在抗氧化剂存在的情况下进行酶促 - 机械联合方案可能对优化骆驼精液冷冻保存至关重要。
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