First Birth of Cheetah Cubs from In Vitro Fertilization and Embryo Transfer

SIMPLE SUMMARY: Although the cheetah is the most studied of all wild felid species, reproduction of cheetahs in zoological settings has never been self-sustaining. A large proportion (~30%) of the Association of Zoos and Aquariums population is excluded from breeding due to advanced age, health, behavior, or management issues. Development of assisted reproductive technologies (ARTs) to enable the genetic contribution of sub-fertile individuals is now a priority research and management focus. We have previously demonstrated that aging females produce eggs that have the same developmental competence as young females. The objective of the present study was to produce embryos in vitro from older donor oocytes and transfer them into younger recipients to obtain pregnancies and live births. Good quality oocytes were collected from three synchronized donors, fertilized in vitro with frozen-thawed semen, and cultured for two days. Resulting embryos were transferred into the oviduct of three synchronized recipients. Pregnancies were monitored via fecal levels of progestogens, ultrasonography, and radiography. Two cubs were born naturally after 90 days of gestation, representing the first cheetah births resulting from the transfer of embryos produced in vitro. ABSTRACT: Approximately 30% of the Association of Zoos and Aquariums cheetah population (~350 total animals) is unlikely to breed naturally due to advanced age, health, or behavioral issues. Aging cheetah females (≥9 y old) are unlikely to become pregnant via natural breeding if they are nulliparous. We previously demonstrated that oocytes recovered from aged females were of similar quality compared with those recovered from younger females (2–8 y old). We hypothesize that transfer of 4–8 cell embryos produced by in vitro fertilization with oocytes from old donors could result in pregnancy after transfer into younger recipients. Female cheetahs (n = 3 aging donors and n = 3 young recipients) received 300 IU equine Chorionic Gonadotropin (eCG) and 3000 IU Luteinizing Hormone (LH) while fecal metabolites of estrogens and progestogens were closely monitored. At 28 h post-LH injection, oocytes were aspirated laparoscopically from donors and inseminated in vitro with cryopreserved sperm. After 48 h of in vitro culture, resulting embryos (4–8 cells) were transferred into the oviducts of recipient females. Pregnancy was confirmed in one recipient via ultrasound 32 days after transfer and by radiograph 62 days after transfer. Two cubs were born naturally after 90 days of gestation, representing the first cheetah births resulting from transfer of embryos produced in vitro.