Stoops Monica A, Campbell Mark K, DeChant Christopher J, Hauser Joe, Kottwitz Jack, Pairan Randal D, Shaffstall Wendy, Volle Kurt, Roth Terri L
Center for Conservation and Research of Endangered Wildlife, 3400 Vine Street, Cincinnati Zoo & Botanical Garden, Cincinnati, OH 45220, USA.
Center for Conservation and Research of Endangered Wildlife, 3400 Vine Street, Cincinnati Zoo & Botanical Garden, Cincinnati, OH 45220, USA.
Anim Reprod Sci. 2016 Sep;172:60-75. doi: 10.1016/j.anireprosci.2016.07.003. Epub 2016 Jul 18.
The objective of this study was to design an artificial insemination (AI) protocol using cryopreserved spermatozoa to obtain pregnancies in captive Indian rhinoceroses (Rhinoceros unicornis). Four methods developed varied by timing and approach, as follows; Method 1: females (n=2) were inseminated pre- and post-ovulation under general anesthesia, Method 2: females (n=2) were inseminated pre-ovulation without anesthetic via endoscopy, Method 3: females (n=1) were inseminated pre-ovulation without anesthetic via manual insertion of an insemination catheter, Method 4: females (n=2) were inseminated same as Method 3 with the addition of standing sedation. Semen deposition site varied as a result of changes in AI technology and experience. All females conceived following intrauterine AI using three methods. Four pregnancies (n=3 females) produced via Method 3 and 4 resulted in term births (n=2 male calves, n=2 female calves) at 481.8±12.8days post-AI. Unfortunately, two early pregnancy losses were documented in a fourth female conceiving via Method 2. Pregnancy rates were 0%, 22%, 17%, and 50% for Method 1-4, respectively. Method 3 and 4 rates improved to 29% and 67%, respectively when accounting for AI's conducted only on ovulatory estrous cycles. Spermatozoa (n=5 males) were cryopreserved 0.3-9.3 y prior to successful AI procedures. The lowest dose of frozen-thawed sperm resulting in conception was 500×10(6) motile sperm. Mean time from AI to ovulation in conceptive and non-conceptive cycles was 26±11.8h and 66±80.7h, respectively.
本研究的目的是设计一种使用冷冻精子的人工授精（AI）方案，以使圈养的印度犀牛（独角犀）成功受孕。开发了四种方法，在时间安排和操作方式上各有不同，具体如下：方法1：2头雌性犀牛在全身麻醉下于排卵前后进行授精；方法2：2头雌性犀牛在排卵前通过内窥镜检查进行无麻醉授精；方法3：1头雌性犀牛在排卵前通过手动插入授精导管进行无麻醉授精；方法4：2头雌性犀牛与方法3相同，但增加了站立镇静。由于人工授精技术和经验的变化，精液沉积部位有所不同。使用三种方法进行子宫内人工授精后，所有雌性犀牛均受孕。通过方法3和4产生的4次妊娠（涉及3头雌性犀牛）在人工授精后481.8±12.8天产下足月幼崽（2头雄性幼崽，2头雌性幼崽）。不幸的是，通过方法2受孕的第四头雌性犀牛出现了两次早期妊娠丢失。方法1至4的妊娠率分别为0%、22%、17%和50%。仅考虑在排卵发情周期进行的人工授精时，方法3和4的妊娠率分别提高到29%和67%。在成功的人工授精程序之前，精子（来自5头雄性犀牛）已冷冻保存0.3 - 9.3年。导致受孕的最低冻融精子剂量为500×10⁶ 个活动精子。在受孕和未受孕周期中，从人工授精到排卵的平均时间分别为26±11.8小时和66±80.7小时。
