Antibiotic-free extended boar semen preserved under low temperature maintains acceptable in-vitro sperm quality and reduces bacterial load
Antibiotic-free extended boar semen preserved under low temperature maintains acceptable in-vitro sperm quality and reduces bacterial load
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DOI：
10.1016/j.theriogenology.2020.03.003
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发表时间：
2020-06-01
期刊：
影响因子：
2.8
通讯作者：
Bortolozzo, Fernando Pandolfo
中科院分区：
文献类型：
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作者：
Menezes, Tila de Alcantara;Goncalves Mellagi, Ana Paula;Bortolozzo, Fernando Pandolfo
本研究旨在评估低温储存的延长的猪精液的精子质量和集落形成单位（CFU mL(-1)）。正常精子射精（n = 34）用Androstar (R) Premium稀释，加或不加抗生素（氨苄西林和硫酸阿帕霉素）。延长剂量的精液在3种储存温度（5℃、10℃和17℃）下储存120小时。在析因设计中，使用GLIMMIX程序作为重复测量来分析变量。低温贮藏（5℃和10℃）下延长剂量的精液在整个贮藏过程中总活力均在75%以上。抗生素x温度对总运动性（P = 0.004）和进行性运动性（P = 0.005）有显著影响。在延长抗生素剂量下，随着贮藏温度的升高，总活力和进动率分别为80.2%、84.5%和89.1%，5、10和17℃时总活力和进动率分别为70.5%、76.0%和82.9%,P < 0.05)。在不含抗生素的延长剂量精液中，5℃贮藏时总运动性和进速运动性低于10℃和17℃贮藏时的81.8%、85.4%和86.6%，5℃、10℃和17℃贮藏时总运动性和进速运动性分别为71.9%、76.7%和78.9%,P < 0.05)。经热阻试验，17℃时抗生素给药总运动性和进行性运动性高于5℃（P < 0.05）；但在不使用抗生素的情况下，延长精液剂量的保存温度对其无显著影响（P > 0.05）。5℃和10℃下不加抗生素的延长剂量精液CFU mL（-1）数低于17℃下（P < 0.05）；然而，在延长抗生素剂量的精液中，没有观察到储存温度的影响（P > 0.05）。无论储存温度如何，未加抗生素的延长精液细菌负荷均高于加抗生素的延长精液（P < 0.05）。抗生素对顶体和精子膜完整性无显著影响（P < 0.05）。随着贮藏温度的升高，正常顶体所占比例较高（5℃、10℃和17℃时分别为93.6%、94.3%和96.8%,P < 0.0001）。与10℃和5℃贮藏的精液相比，17℃贮藏的精液的膜完整性更高（P < 0.0001）。在整个贮藏过程中，pH值均有所上升，但在17℃贮藏的精液中，pH值在120 h时出现下降（P < 0.05）。尽管低温对精子质量有负面影响，但由于精子在体外的生存能力可以维持5天，因此可以在5℃下储存延长剂量的猪精液，满足人工授精所需的精液质量要求。然而，使用无抗生素的延长精液剂量需要优化精液剂量处理过程中的卫生程序。(C) 2020爱思唯尔公司版权所有。
This study aimed to assess the sperm quality and number of colony-forming units (CFU mL(-1)) in extended boar semen stored at low temperatures with or without antibiotics. Normospermic ejaculates (n = 34) were diluted in split samples with Androstar (R) Premium with or without antibiotics (ampicillin and apramycin sulfate). The extended semen doses were stored for 120 h under three storage temperatures (5, 10, and 17 degrees C). Variables were analyzed as repeated measures using the GLIMMIX procedure, in a factorial design. The extended semen doses under low-temperature storage (5 and 10 degrees C) had total motility above 75% throughout the storage. The interaction antibiotic x temperature was significant for total (P = 0.004) and progressive motility (P = 0.005). In extended boar semen doses with antibiotics, the total and progressive motility increased as the storage temperature increased (80.2%, 84.5%, and 89.1%; 70.5%, 76.0%, and 82.9% for total and progressive motility at 5, 10, and 17 degrees C, respectively; P < 0.05). In extended semen doses without antibiotics, the total and progressive motility were lower when stored at 5 degrees C than at 10 degrees C and 17 degrees C (81.8%, 85.4% and 86.6% and 71.9%, 76.7%, 78.9% for total and progressive motility at 5, 10, and 17 degrees C, respectively; P < 0.05). After the thermoresistance test, total and progressive motility of doses with antibiotics were higher at 17 degrees C than 5 degrees C (P < 0.05); however, they were not affected (P > 0.05) by storage temperature in extended semen doses without antibiotics. The number of CFU mL(-1) was lower in extended semen doses without antibiotics stored at 5 and 10 degrees C than at 17 degrees C (P < 0.05); however, in extended semen doses with antibiotics, no effect of storage temperature was observed (P > 0.05). The bacterial load was greater in extended semen without antibiotics than with antibiotics, regardless of the storage temperature (P < 0.05). The acrosome and sperm membrane integrity were not influenced (P > 0.05) by using antibiotics. A higher percentage of normal acrosomes was observed as the storage temperature increased (93.6%, 94.3%, and 96.8% at 5, 10, and 17 degrees C, respectively; P < 0.0001). The membrane integrity was higher (P < 0.0001) in extended semen doses stored at 17 degrees C than at 10 or 5 degrees C. The pH rose throughout the storage in all the treatments, except in extended semen doses stored at 17 degrees C without antibiotics, in which a decrease in the pH occurred at 120 h (P < 0.05). Although the sperm quality being negatively affected by low temperatures, the storage of extended boar semen doses at 5 degrees C is possible since the sperm viability in vitro was maintained for up to 5 days, fulfilling the requirements of semen quality to be used in artificial insemination. Nevertheless, the use of extended semen doses without antibiotics requires the optimization of hygiene procedures during semen dose processing. (C) 2020 Elsevier Inc. All rights reserved.