In a groundbreaking scientific breakthrough, two cheetah cubs have been born through an in vitro fertilization process with the embryo transferred to a surrogate mother.

The births took place this week at the Columbus Zoo and Aquarium after years of careful planning. With in vitro fertilization, or IVF, sperm and eggs are fertilized in a laboratory and then incubated to create embryos. The embryos are implanted into a female's womb, where they may develop into fetuses. IVF has become a more common process with humans and some other species, but it previously has been unsuccessful in large cats, including cheetahs and lions.

In this case, however, the two cubs were delivered by an older cheetah named Isabelle (Izzy) on February 19th—although the cubs' biological mother is Kibibi.

The cheetahs' care team observed the births through a remote camera and continue to monitor Izzy and her cubs closely. Izzy, a first-time mom, has been providing great care to her cubs at this time. The care team performed a well check on the cubs on Friday and determined that Izzy gave birth to a male cub and a female cub, both of which have been observed nursing.

"These two cubs may be tiny, but they represent a huge accomplishment, with expert biologists and zoologists working together to create this scientific marvel," said Dr. Randy Junge, the Columbus Zoo's Vice President of Animal Health. "This achievement expands scientific knowledge of cheetah reproduction, and may become an important part of the species' population management in the future."

Kibibi first received hormone injections on November 14th, 2019 to stimulate follicle development. Eggs (oocytes) were taken from the 6-year-old big cat a few days later, as her genes are considered to be valuable in maintaining a strong lineage of cheetahs in human care. Izzy's bloodline is already well-represented in the genetic registry, so she was selected as a surrogate. After the age of 8, cheetahs' ability to reproduce declines significantly, and because Izzy is 3 years old, she has a better chance of safely delivering healthy, full-term cubs.

Once Kibibi's eggs were extracted, the eggs were then fertilized on November 19th in a Columbus Zoo laboratory using thawed semen originally collected in February 2019 from two cheetahs: a male from Fossil Rim Wildlife Center in Glen Rose, Texas and another from Smithsonian's National Zoo and Conservation Biology Institute (SCBI) in Front Royal, Virginia.

On November 21st, the early-stage embryos from Kibibi were then implanted into Izzy. It was only the third time scientists had ever attempted this procedure.

On December 23rd, an ultrasound revealed the remarkable news: two fetuses were growing in Izzy, the father of which is 3-year-old Slash from Fossil Rim Wildlife Center.

"I am very proud of the team for this accomplishment," said Jason Ahistus, Fossil Rim Wildlife Center Carnivore Curator. "It gives the cheetah conservation community another tool to use in cheetah management, both in situ and ex situ. It really opens the door to many new opportunities that can help the global cheetah population. This is a big win for the cheetah."

According to the International Union for Conservation of Nature (IUCN), cheetahs have a population classification of "Vulnerable" and a decreasing population trend in their native range of Africa. Due to threats including habitat loss and fragmentation, conflict with livestock and game farmers, as well as unregulated tourism, cheetahs now inhabit just 10% of their historic range. This geographic separation has left the species genetically "bottlenecked," creating the potential for inbreeding. Scientists estimate that the cheetah population has declined to only approximately 7,500 individuals.

The achievement at the Columbus Zoo brings the potential to help ensure the survival of cheetahs in their native range. Conservation scientists have long sought ways to boost the numbers and help maintain genetic diversity of the species. Yet, attempts to artificially inseminate cheetahs often have not been successful, with the last one occurring in 2003.

IVF previously had been somewhat successful in small domestic cats and African wildcats, but it is still rare in larger cat species, with only the birth of three tiger cubs reported back in 1990. These breakthrough births mark a significant advancement in the field, and may provide valuable information to boost future conservation efforts in cat species.

"The first thing we had to do is show that this technique works," said Dr. Junge. "Then we have to become proficient in it, so we can do it efficiently and reliably. With experience, we may be able to freeze embryos and transfer them to Africa."

The Columbus Zoo's animal care team is limiting access to the cheetahs' private birthing area. As a cheetah's gestation is typically 93 days and her due date was estimated to be February 22nd, Izzy's care team put her on a 24-hour birth watch beginning Sunday, February 16, and the Zoo's veterinary team stood ready with an incubator warmed up in case the cubs arrived early. Because complications during a birth can occur, the team also was prepared to perform an immediate Caesarean section if Izzy or the cubs became distressed.

The Columbus Zoo has extensive experience with cheetahs, having raised many cubs. Izzy and Kibibi are two of the Zoo's ambassador cheetahs, most of whom arrived at the Zoo to be raised by hand when their mothers were unable to care for them. As a result, the cheetah ambassadors are accustomed to humans and have formed extremely close bonds with their care providers. The cheetahs are trained to voluntarily allow ultrasounds, X-rays, blood draws and other medical procedures, so the risks of anesthesia often can be avoided. Their training also allowed Zoo staff to be near Izzy during the delivery to assist, if needed.

"In the 19 years that I've worked with cheetahs, one of the big challenges is that we have no idea if a female is pregnant until at least 60 days following a procedure or breeding. Working with the Columbus Zoo and Aquarium was a game-changer because their females are highly cooperative. We knew that Izzy was pregnant at five weeks by ultrasound and we continued to collect ultrasound data throughout her entire pregnancy. It was a remarkable opportunity and we learned so much," said Adrienne Crosier, cheetah biologist at the Smithsonian Conservation Biology Institute, one of the scientists who performed the embryo transfer.

Through conservation projects in Africa, the Columbus Zoo works with communities in cheetahs' natural range, reducing farmers' livestock losses to cheetahs by introducing Anatolian shepherd guard dogs; assisting with cheetah health exams; and learning more about cheetah populations through camera traps, scat analysis and habitat monitoring.

Reprinted from the Columbus Zoo and Aquarium

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