Wojtusik Jessye, Stoops Monica A, Roth Terri L
Cincinnati Zoo and Botanical Garden, Center for Conservation and Research of Endangered Wildlife, 3400 Vine St., Cincinnati, OH, 45220, USA.
Cincinnati Zoo and Botanical Garden, Center for Conservation and Research of Endangered Wildlife, 3400 Vine St., Cincinnati, OH, 45220, USA.
Theriogenology. 2018 Nov;121:72-77. doi: 10.1016/j.theriogenology.2018.07.042. Epub 2018 Aug 13.
Semen cryopreservation for the black rhinoceros (Diceros bicornis) and Indian rhinoceros (Rhinoceros unicornis) relies on extenders containing egg-yolk (EY). Use of such media is not ideal as inter-batch composition varies and there is risk of pathogenic contamination. The goal of this study was to test animal protein-free extenders. Semen collected via electroejaculation from 10 rhinoceros (6 black, 4 Indian) was diluted with extender containing EY, 1% or 2% soy lecithin (1%SL; 2%SL), coconut water (CW), or coconut milk (CM), cryopreserved and evaluated for sperm motility, viability, morphology, progression, and acrosomal integrity at 0, 1, 3, 6 and 24 h post-thaw. Mean ± SD fresh ejaculate motility was 84.5 ± 7.6%, progression: 3.6 ± 0.6 (scale 0-5), viability: 83.4 ± 7.1%, intact acrosomes: 71.3 ± 6.9%, and morphologically normal: 78.8 ± 13.6%. Motility and progression decreased in all groups post-thaw, were greatest in EY, and decreased over time (P ≤ 0.05). Motility and progression did not differ (P > 0.05) between 1%SL and 2%SL, but were lower (P ≤ 0.05) in CM and CW, and acrosomal integrity was higher (P ≤ 0.05) in EY, 1%SL and 2%SL than in CM and CW. Post-thaw viability was greatest in EY and 2%SL followed by 1%SL, then CM and CW (P ≤ 0.05). Morphology did not differ among treatments (P > 0.05). Morphology, acrosomal integrity, and viability were maintained over time (P > 0.05). Although some rhinoceros sperm survived cryopreservation in SL treatments, reduced post-thaw motility rendered all treatments inadequate substitutes for EY-based extenders.
黑犀牛（双角犀属）和印度犀牛（独角犀属）的精液冷冻保存依赖于含有蛋黄（EY）的稀释液。使用此类培养基并不理想，因为不同批次的成分存在差异，且存在病原污染风险。本研究的目的是测试无动物蛋白的稀释液。通过电刺激从10头犀牛（6头黑犀牛、4头印度犀牛）采集精液，并用含有EY、1%或2%大豆卵磷脂（1%SL；2%SL）、椰子水（CW）或椰奶（CM）的稀释液进行稀释，冷冻保存，并在解冻后0、1、3、6和24小时评估精子活力、生存力、形态、前进运动能力和顶体完整性。新鲜射精的平均活力±标准差为84.5±7.6%，前进运动能力：3.6±0.6（0 - 5级），生存力：83.4±7.1%，完整顶体：71.3±6.9%，形态正常：78.8±13.6%。解冻后所有组的活力和前进运动能力均下降，在EY组中最高，并随时间下降（P≤0.05）。1%SL和2%SL之间的活力和前进运动能力无差异（P>0.05），但在CM和CW组中较低（P≤0.05），EY、1%SL和2%SL组的顶体完整性高于CM和CW组（P≤0.05）。解冻后的生存力在EY和2%SL组中最高，其次是1%SL组，然后是CM和CW组（P≤0.05）。各处理间形态无差异（P>0.05）。形态、顶体完整性和生存力随时间保持稳定（P>0.05）。尽管在SL处理中一些犀牛精子在冷冻保存后存活，但解冻后活力降低使得所有处理都不足以替代基于EY的稀释液。
