Vongpralub Thevin, Chinchiyanond Wittaya, Hongkuntod Pornchai, Sanchaisuriya Pitcharat, Liangpaiboon Sanan, Thongprayoon Areeya, Somphol Noppadon
Department of Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand.
Wildlife Conservation Bureau, National Park, Wildlife and Plant Conservation Department, Bangkok, Thailand.
Zoo Biol. 2015 Jul-Aug;34(4):335-44. doi: 10.1002/zoo.21214. Epub 2015 Apr 23.
Little is known of the different freezing and thawing techniques for post-thaw survival of spermatozoa in Sambar deer. So, this study determined the effect of seminal plasma, egg yolk and glycerol extenders and their concentrations, plus cooling, freezing, and thawing protocols on the post-thaw quality of their semen. Semen samples were collected by electro-ejaculation from four Thai Sambar deer stags (Cervus unicolor equinus). As evaluated by post-thaw progressive motility and acrosome integrity removal of seminal plasma was beneficial; Tris-egg yolk was the most efficient extender; a 20% egg yolk concentration was better than the 0%, 10%, or 30%; and a 3% glycerol concentration was better than 5%, 7%, or 9%. Using the optimum dilution techniques, semen was loaded in 0.5 ml plastic straws. Cooling times from ambient temperature to 5°C in 3 hr resulted in higher post-thaw progressive motility and acrosome integrity than 1, 2, or 4 hr. Suspending the straws 4 cm above the surface for 15 min before plunging into liquid nitrogen was better than suspending at 2 or 6 cm. For thawing frozen semen, an intermediate thawing (50°C, 8 sec) protocol was more effective than the slower (37°C, 10 sec) or faster (70°C, 5 sec) thawing rates. Timed insemination following estrus synchronization of 10 hinds resulted in six confirmed pregnancies at 60 days. Five hinds delivered live fawns. This study provides an effective approach for semen cryopreservation and artificial insemination (AI), which should be valuable to scientists for genetics and reproductive management of Sambar deer in developing countries.
关于水鹿精子解冻后存活的不同冷冻和解冻技术，人们了解甚少。因此，本研究确定了精浆、蛋黄和甘油稀释液及其浓度，以及冷却、冷冻和解冻方案对其精液解冻后质量的影响。通过电刺激射精从四只泰国水鹿雄鹿（Cervus unicolor equinus）采集精液样本。经解冻后精子的前向运动能力和顶体完整性评估，去除精浆是有益的；Tris-蛋黄是最有效的稀释液；20%的蛋黄浓度优于0%、10%或30%；3%的甘油浓度优于5%、7%或9%。采用最佳稀释技术，将精液装入0.5毫升塑料细管中。从室温在3小时内冷却至5°C，解冻后的前向运动能力和顶体完整性高于1、2或4小时。在投入液氮之前，将细管悬于液氮表面上方4厘米处15分钟，优于悬于2或6厘米处。对于解冻冷冻精液，中等解冻（50°C，8秒）方案比慢速（37°C，10秒）或快速（70°C，5秒）解冻速率更有效。对10只母鹿进行发情同步后定时输精，60天时确认有6只怀孕。5只母鹿产下活仔。本研究为精液冷冻保存和人工授精（AI）提供了一种有效方法，这对发展中国家水鹿遗传学和繁殖管理方面的科学家应具有重要价值。
