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

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.

Understanding Reproductive Aging in Wildlife to Improve Animal Conservation and Human Reproductive Health

“…Zoo animal populations represent an ideal model to study reproductive aging process since lifespan is frequently longer than in the wild, so repeated observations under controlled conditions are possible. Reproductive aging also represents a major issue in conservation breeding, especially for genetically valuable individuals that are too often not reproducing ( Hermes et al, 2004 ; Crosier et al, 2020 ).…”

“…Interestingly in cheetahs ( Acinonyx jubatus ), uterine environment in aging females is the real issue while oocyte quality is not affected ( Crosier et al, 2011 ). This can be circumvented by performing in vitro fertilization (IVF) on oocytes from old donors and transfer the resulting embryos into young recipient females ( Crosier et al, 2020 ).…”

Canid Reproductive Biology: Norm and Unique Aspects in Strategies and Mechanisms

“…Oocytes can be collected surgically in dogs via laparotomy and retrograde flushing of the oviduct [ 120 ]. However, less-invasive, laparoscopic oocyte aspiration techniques utilized in other taxa (such as that which allowed for the collection of oocytes for the recent successful cheetah in vitro fertilization [ 121 ], will require significant additional fine-tuning to get through the ovarian bursa without damaging the future reproductive potential of the animal. Still, laparoscopic ovariectomies are becoming increasingly common in domestic veterinary practice [ 122 ], and continued advances in this area will support future applications to endangered canid assisted reproductive technology development.…”

An Investigation of Ovarian and Adrenal Hormone Activity in Post-Ovulatory Cheetahs (Acinonyx jubatus)

“…Over almost the last four decades, research utilizing the SSP population has yielded a substantial body of work, advancing our understanding of topics, such as species’ nutritional needs [ 3 , 4 ], behavior [ 5 , 6 , 7 , 8 ], husbandry [ 9 , 10 ], disease [ 11 , 12 , 13 , 14 ], genetics [ 15 , 16 ], and reproductive physiology [ 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 ]. Despite our expanding knowledge base on cheetahs, the species still pose significant reproductive challenges when under human care that does not appear to be shared by free-ranging populations.…”