Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, 712100, China.
Theriogenology. 2019 Dec;140:124-135. doi: 10.1016/j.theriogenology.2019.08.020. Epub 2019 Aug 21.
Bacteriospermia is a documented risk to sperm quality when boar semen is stored at 17 °C. The objective of this study was to evaluate the effects of kojic acid (KA) on sperm quality and anti-bacterial effect during liquid storage boar semen at 17 °C, as well as to explore sperm-oocyte binding and embryonic development in vitro. Boar semen was diluted with Beltsville thawing solution (BTS), and it contained KA at different concentrations (0, 0.02, 0.04, 0.06, 0.08, and 0.10 g/L). Bacterial concentrations and sperm quality parameters (motility, mitochondrial membrane potential, acrosome integrity, and plasma membrane integrity) were evaluated on each experimental day. Differences in microbial compositions were compared using 16S rDNA sequencing among the control group, 0.04 g/L KA, and 0.25 g/L gentamycin groups on experimental day 5, and the effects of KA on sperm capacitation, Western blot, total anti-oxidant capacity (T-AOC), reactive oxygen species (ROS) content, malondialdehyde (MDA) content, in vitro fertilization (IVF) parameters, sperm-oocyte binding, cleavage rates, and blastocyst rates were evaluated. The results showed that KA at the optimum concentration of 0.04 g/L significantly improved sperm quality parameters and sperm capacitation, increased T-AOC ability, enhanced IVF parameters and sperm-oocyte binding, increased cleavage and blastocyst rates, inhibited bacterial concentrations, reduced ROS and MDA content, and altered bacterial compositions (P < 0.05). Moreover, KA also increased the expression of anti-oxidant-related proteins, SOD1, SOD2 and CAT, and anti-apoptosis-related protein, Bcl 2, and decreased the expression of apoptosis-related proteins, caspase 3 and Bax in sperm (P < 0.05). These findings demonstrated that supplementation of antibiotic-free extenders for boar semen with 0.04 g/L KA has beneficial effects on liquid boar sperm preservation.
细菌污染是猪精液在 17°C 保存时导致精子质量下降的一个已知风险。本研究的目的是评估曲酸（KA）对猪精液在 17°C 液态储存时的精子质量和抗菌效果的影响，以及探索精子-卵结合和体外胚胎发育。猪精液用贝尔茨维尔解冻液（BTS）稀释，并在不同浓度（0、0.02、0.04、0.06、0.08 和 0.10 g/L）下添加 KA。在每个实验日评估细菌浓度和精子质量参数（活力、线粒体膜电位、顶体完整性和质膜完整性）。在第 5 天的实验中，通过 16S rDNA 测序比较了对照组、0.04 g/L KA 组和 0.25 g/L 庆大霉素组之间的微生物组成差异，并评估了 KA 对精子获能、Western blot、总抗氧化能力（T-AOC）、活性氧（ROS）含量、丙二醛（MDA）含量、体外受精（IVF）参数、精子-卵结合、卵裂率和囊胚率的影响。结果表明，最佳浓度 0.04 g/L 的 KA 显著改善了精子质量参数和精子获能，提高了 T-AOC 能力，增强了 IVF 参数和精子-卵结合，提高了卵裂率和囊胚率，抑制了细菌浓度，降低了 ROS 和 MDA 含量，改变了细菌组成（P < 0.05）。此外，KA 还增加了抗氧化相关蛋白 SOD1、SOD2 和 CAT，以及抗凋亡相关蛋白 Bcl-2 的表达，降低了凋亡相关蛋白 caspase 3 和 Bax 的表达（P < 0.05）。这些发现表明，在无抗生素的猪精液稀释液中添加 0.04 g/L KA 对猪精液的液态保存具有有益效果。
