In vitro buffalo calf brings hope to endangered species

Pumelelo and his surrogate mother, Suri.

The first Cape buffalo calf to be conceived through in vitro fertilisation (IVF) was born on 28 June 2016, on the farm Lekkerleef Buffalo Ranch of Frans Stapelberg, near Marble Hall in the Limpopo Province of South Africa.

The news was announced by Dr Morné de la Rey, managing director of Embryo Plus, at a press conference held at Lekkerleef, where members of the press were introduced to the healthy three-month old buffalo bull calf, Pumelelo.

Three-month old in vitro buffalo bull calf, Pumelelo.

The assisted reproductive technology used in the production of this calf was developed by Embryo Plus from Brits in the Northwest Province, a world-leader in the embryo industry. The Embryo Plus team, led by Dr Morné de la Rey (BVSc), is developing techniques that can be used to save critically endangered wildlife species.

“The use of assisted reproductive techniques (ARTs) in wildlife management, although still in its infancy, is becoming more of a reality. This success is of major importance for the prospective breeding of endangered species, and this is the reason why we are undertaking this work,” Dr De la Rey said.

Vasti, the biological mother and egg donor of Pumelelo.
Vasti, the biological mother and egg donor of Pumelelo.

In a process similar to ovum recovery in women undergoing in vitro fertilisation (IVF), the oocytes of a buffalo cow were collected by a technique called ovum pick-up (OPU), which took place with the cow under full anaesthesia. A needle was guided trans-vaginally and ova (eggs) were aspirated from the ovaries of the cow, using ultrasound to visualise the process. “The sedation went very well and the buffalo cow incurred very little stress,” Dr De la Rey said.

The eggs were matured and fertilised in vitro with frozen-thawed Cape buffalo semen, then grown in a laboratory incubator in a process known as in vitro production (IVP). After seven days of growth, the embryo was transferred into a surrogate buffalo cow, which carried the foetus to term for eleven months. She gave birth to a perfect, healthy buffalo calf on 28 June. DNA samples taken from the calf confirmed the parentage of the cow and bull used in the IVF/IVP process.

Goliat, the biological father and sperm donor of Pumelelo.
Goliat, the biological father and sperm donor of Pumelelo.

“Extensive preliminary research was necessary to mature and fertilise the eggs, and to incubate the embryos to an advanced stage of development, as all species have different requirements for growth and utilise different nutrients during the laboratory phase of the largely uncharted path of IVF/IVP in African game species,” Dr De la Rey said. “This breakthrough is of great significance as it is the first of its kind in the world and holds great promise for the continued survival of endangered species.”

The buffalo used for this procedure are part of a herd that has been maintained at the Stapelberg farm for more than 17 years. The facility was chosen in part because they have worked extensively with the animals and trained them to participate in routine veterinary procedures without the need for anaesthesia.

“The superior ranch management implemented by Stapelberg and his team was a significant factor in the success of the IVF project,” Dr de la Rey said. “These facilities make the handling of the buffalo easier and a large percentage of the vaccinations and routine treatments is accomplished without anaesthetising the animals.”

Northern White Rhino Project

Dr De la Rey’s work is considered to be among the first steps in an international collaborative effort to save endangered wildlife species such as the northern white rhino. There are only three northern white rhino left in the world – a rhino bull named Sudan, and his two cows, Najin and Fatu, who live in the Ol Pejeta Conservancy in Kenya. The last northern white rhino in the United States died at the San Diego Zoo Safari Park in 2015.

The San Diego Zoo Global has pledged to work with a team of scientists, including Embryo Plus, to save this rhino species and has created a herd of southern white rhinos that have been trained for non-invasive procedures that are providing unprecedented information about the reproductive physiology of rhinos.

“We were delighted to hear about the success of this technique with Cape buffalo,” Dr Barbara Durrant, reproductive physiologist for San Diego Zoo Global, said. “We look forward to working with Embryo Plus in an effort to bring back the northern white rhino for the future.”

In 2003, Dr Morné de la Rey also received world-wide acclaim for his involvement in the cloning of the first animal on the African continent, a healthy Holstein heifer named Futhi, born in April 2003 at the Embryo Plus Centre at Brits. No semen or bull was involved in the birth of this calf. Futhi was derived from a single cell taken by biopsy from the ear of a donor cow, which was then inserted into an empty cow egg and later implanted in a recipient cow.

According to Dr De la Rey, every success achieved with assisted reproductive technology in wild animals such as the Cape buffalo is a step closer to understanding the reproductive requirements of other wildlife species. “The first steps of wildlife conservation will always remain habitat protection followed by animal protection. In addition to these basic steps, however, there is a real need for improved breeding practices to ensure an increase in numbers of these endangered species.”

Dr De la Rey and the team of San Diego Zoo will start an exciting research project at the Rhino Pride Foundation Sanctuary, which currently houses more than 40 southern white rhinos, with the goal of obtaining success in IVF in rhino.

Rhinos are considerably more difficult to work with than most other species, due to the size and sensitive nature of the animals. Specialized equipment will have to be designed, tested and manufactured for effective use on rhino. Once these procedures have been successfully implemented with southern white rhino, the techniques could be applied on the northern white rhino to try and save them from extinction.

This will be Dr De La Rey and the research team’s next goal, and parallel to this there is the possibility of doing similar work with the extremely endangered mountain bongo of Kenya. There are only 30 to 50 bongo left in three areas in Kenya. The planned research for this project will be conducted at the envisaged endangered species breeding centre to be established at Ol Pejeta Conservancy, Kenya. – Embryo Plus