reproduction Archives - Rhino Review

San Diego Zoo researchers retrieve rhino’s eggs to recover critically endangered species

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Karla Rendon-Alvarez, NBC San Diego | March 11, 2020

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Researchers at the San Diego Zoo Safari Park collected a female southern white rhinoceros’ eggs as a step to genetically recover the northern white rhino.

Original photo as published by NBC San Diego.

On March 6, a team of more than 30 veterinarians, wildlife care specialists and researchers from the San Diego Zoo Global and Embryo Plus South Africa teamed up to perform an ovum pick-up on 9-year-old rhino, Nikita. The non-surgical procedure gathers a live animal’s eggs and is modeled after a similar method used on horses and cows.

Nikita the rhino was under anesthesia during the procedure as researchers located her ovaries by ultrasound. The animal’s eggs were then retrieved by a tiny needle that was inserted into each follicle. Those eggs are expected to mature in vitro and fertilize by intracytoplasmic sperm injection.

Meet the women racing to save the northern white rhino from extinction (California)

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Adeline Chen, CNN | March 8, 2020

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SAN DIEGO, CALIFORNIA: There are only two northern white rhinos left on the planet, and they’re both female. Unless scientists can make a dramatic breakthrough, the entire species will die with those two individuals.

In a nondescript building just north of San Diego, California, the fight to save the northern white rhino is coming down to the wire. However, the battleground here looks less like a scene from a wildlife documentary and more akin to something out of a science fiction novel.

At the San Diego Zoo Institute for Conservation Research, an army of scientists armed with liquid nitrogen, microscopes, and ultrasound machines is working around the clock to create an unprecedented first in the conservation world: they are looking to turn frozen rhino skin cells into baby rhinos.

It’s not just the science that is groundbreaking, but also the team looking to save this species. Composed mostly of women, the lab is a rarity in a field traditionally dominated by men.

Marlys Houck: Freezing Time

The first step in this conservation effort began more than four and a half decades ago in 1975 when scientists established the institute’s “Frozen Zoo.” In a small room measuring no more than 36 square meters the skin cells of more than 10,000 individuals across 1,100 species sit in giant steel tanks suspended in time, frozen in liquid nitrogen.

Among the collection are the skin samples of 12 northern white rhinos. These are vital to the group’s efforts because there is such a small gene pool of living northern whites.

Original photo as published by CNN: Marlys Houck, curator of the Frozen Zoo.

The population has been decimated by poachers, who target rhinos because of the belief in parts of Asia that their horns can cure various ailments. The two surviving females both live under guard at the Ol Pejeta Conservancy in Kenya. Even though embryos have been produced in an Italian lab using eggs extracted from the pair, any future descendants from this kind of embryo would carry the genes of those two females.

That may not be enough genetic diversity to maintain a stable population. The hope is that the skin samples of those 12 individuals at the Frozen Zoo contain enough diversity to sustain the northern white species long-term.

The arduous task for these scientists is to create a rhino population from those samples.

Marlys Houck is curator of the Frozen Zoo. She graduated high school in 1979, the same year the Frozen Zoo froze its very first northern white rhino skin cell. She later joined the institute to work on the rhino project.

“I was hired specifically to try to make the cells of the rhinos grow better because they were one of the most difficult to grow cell lines,” she told CNN.

Since then, she’s figured out how to successfully grow and freeze the skin cells of the northern white.

The impact of this work is not lost on her. “We’re losing species so rapidly,” she said. “One of the things we can do is save the living cells of these animals before it’s too late.”

“We’re at the forefront of science today,” she added. “If we do everything right … these cells should be here 50 years from now being used for purposes that we can’t even imagine today.”

Marisa Korody: The Science of Stem Cells

Marisa Korody is one of the four scientists tasked with turning these frozen cells into new life. They have to reprogram the frozen skin cells into pluripotent stem cells. In layman’s terms, Korody explains that “stem cells can become any cell type in the body if they’re given the right signals.”

The aim is to ultimately turn the stem cells into sperm and eggs. The ambitious feat has only been achieved in animals by Japanese scientists. While Korody and her team have looked to that research as a road map, she admits that doing the same with rhinos is uncharted territory. “We don’t really know what twists and turns we need to take in order to get from A to B,” she said.

Original photo as published by CNN: Marisa Korody.

“They haven’t even figured out how to do this in humans,” she added. “We have as much information as we possibly can about humans. We have a fraction of that for rhinos.”

Korody says being at the forefront of this kind of science has been a dream job. “This was really the first project that’s trying to apply this type of science to conservation as a whole,” she said.

She may spend most of her time at work looking through the lens of a microscope, but her mind is always on the final goal for the rhinos: “We want to be able to put them back into the wild one day and have them living free.”

Barbara Durrant: Carrying the Future

Because the remaining two female northern white rhinos can’t carry a pregnancy, even if the team can create embryos, the last obstacle is finding rhinos who can carry them to term.

The woman tasked with that job is Barbara Durrant. As the director of reproductive sciences, she’s spent four years studying the reproductive systems of six female southern white rhinos at the institute’s sister facility, the Nikita Kahn Rhino Rescue Center.

Though the rhinos at the center are a different species, Durrant says they are the closest relative to the northern white. The aim is to eventually have them be surrogates for northern white embryos.

Original photo as published by CNN: Barbara Durrant, director of reproductive sciences.

On any given day, Durrant can be found conducting ultrasounds to help her understand each rhino’s distinct reproductive cycle. In 2019, two of the center’s females gave birth to southern white babies. Both were conceived via artificial insemination, giving Durrant and the teams working on the rhino project hope for the future.

Durrant believes one reason the project works so well is because there are so many women involved. “Women are naturally collaborative with each other,” she said. “Because we have so many obstacles along the way and challenges and setbacks, we support each other and we have sympathy for each other.”

Houck says women tend to be naturally nurturing. “The cells are living little organisms that we’re growing and tending almost every day, and I think women are drawn to taking care of something and growing it into something more.”

“It’s wonderful leading a team of women, and I really think they’re changing the world,” she added. “People are going to look back and see it was this amazing group of women who quietly, unrecognized, work at this and just get better and better.”


Scientific breakthrough marks pioneer cheetah births through IVF, shines light on ongoing IVF on rhinos in Kenya

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Caroline Chebet, Standard Media | March 6, 2020

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Scientists and researchers have recorded a breakthrough following a successful birth of two cheetah cubs by a surrogate mother through In Vitro Fertilisation (IVF).

The first-ever scientific reproduction of cheetahs happened at a Columbian zoo, a breakthrough marked after more than 15 years of research.

According to a statement released by the Colombus Zoo and Aquarium, the achievement could ensure survival of cheetahs in their native ranges in Africa.

“These two cubs may be tiny, but they represent a huge accomplishment, with expert biologists and zoologists working together to create this scientific marvel,” the zoo said in a statement.

Original photo as published by Standard Media: A female cheetah and her seven cubs roam the savanna of Maasai Mara National Park in Southern Kenya. [Reuters]

Conservation scientists, the zoo said, have been doing researches to boost the number of cheetahs which, according to the International Union of Conservation and Nature, are listed as vulnerable with decreasing population trend in their native ranges. The two cubs were born on February 19.

The procedure to birthing the two cubs involved fertilisation of sperm and eggs in a laboratory and then incubated to create embryos. The embryos were implanted into the surrogate mother’s womb, where they developed into foetuses.

The breakthrough on the scientific reproductive techniques on cheetahs come in the wake of the assisted reproduction techniques being tried on the critically endangered northern white rhinos in Kenya.

On scientific milestones to assist in reproduction of northern white rhino, three embryos have been created successfully to be implanted on a surrogate mother at Ol Pejeta conservancy in Laikipia.

The ground-breaking of procedure on the northern white rhinos was marked in 2019 when a team of scientists and conservationists successfully harvested the eggs from the two remaining female, artificially inseminated them using frozen sperm from deceased males and created three viable northern white rhino embryos.

Last January the scientists noted that there was a significant increase in the chances of successfully producing offspring. The procedure also proved to be safe and reproducible, and can be performed on a regular basis before the animals become too old.

Currently, there are plans to select a group of female southern white rhinos at Ol Pejeta Conservancy, which could serve as surrogate mother for the northern white rhino embryo.

This is also expected to be a success, with the first attempt set to be crucial since it has never been achieved before. It is expected that implantation will be undertaken any time this year.

And while the incorporation of scientific technologies in conservation has been on the rise, the breakthroughs shine light in race against extinction.

Scientists estimate that the cheetah population has declined to approximately 7,500.


Can science save the last two white rhinos left on the planet? (Kenya)

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Zeenat Hansrod & Sébastien Nemeth, RFI | March 4, 2020

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Northern white rhinos no longer exist in the wild. The last two remaining female individuals are under constant surveillance in Kenya while scientists are working on groundbreaking techniques to save the species from complete extinction.

Najin and her daughter Fatu are under intense surveillance in their 700-acre (about 280-hectare) enclosure at the Ol Pejeta conservancy near the town of Nanyuki, on the equator in central Kenya. Their head caregiver, Zacharia Mutai, says his team considers them as family members.

“We know them very well and must ensure that they are healthy and well looked after.

“Having the last two is something very serious. We are trying all our best to protect and preserve them. We don’t want to face extinction any more.”

Mutai was the caregiver of Sudan, the last male northern white rhino, which died in 2018.

The two remaining females, 31-year-old Najin and 20-year-old Fatu, are clinically infertile and cannot carry a pregnancy. Furthermore, Najin has a large tumour on her right ovary and Fatin has a damaged uterus.

Original photo as published by RFI: Najin (L) and Fatu (R) are the last two northen white rhinos in the world. The two female rhinos at the Ol Pejeta conservancy in Kenya. (Ami Vitale/BioRescue)

Groundbreaking Techniques

In order to save the species from extinction, scientists are working on artificial reproduction techniques which have never been attempted with rhinoceros before.

The efforts are pioneered by the BioRescue project at the Leibniz Institute for Zoo and Wildlife Research (Leibniz-IZW) in Germany. The team of international scientists are using in vitro fertilisation (IVF) and stem cell technology for reproduction purposes.

“We have collected a lot of semen from four different northern white rhino bulls over the last 20 years,” says Professor Thomas Hildebrandt who heads the BioRescue project.

“This frozen semen allows us to do in vitro fertilisation and we hope to combine that approach together with groundbreaking stem cell technology.”

Hildebrandt, a veterinarian specialised in the reproduction of wild animals, hopes that the combined technology will help produce a viable population that could be released into the wild within 15 to 20 years.

Last year, in a unique procedure at the Ol Pejeta conservancy, his team managed to collect the eggs from Fatu and Najin, but only three embryos from Fatu’s eggs have managed to survive.

“Our goal is to produce the first offspring of the northern white rhino with IVF technique as soon as possible so that this baby can learn how to be a northern white rhino from Najin and Fatu.

“So, our timeline for that is about three years from now,” he added.

The embryos are ready for transfer into surrogate southern white rhinos mothers. Hildebrandt hopes that it will happen before the end of 2020. It will then be followed by a gestation period of 16 months.

Meanwhile in Kenya, Stephen Ngulu, the wildlife vet at the Ol Pejeta conservancy, told RFI that Najin and Fatu are scrupulously monitored.

“I have to observe their walking, their skin, check the eyes, teeth, feet or any wounds. I collect blood and we will test for various parasites, bacterial and viral diseases.”

Stem Cell Technology to Save Endangered Species

“The stem cell approach is needed because we need a gene pool large enough to create a solid, viable population of northern white rhinoceros,” said Hildebrandt, who has spent the last 20 years working with the northern white rhinos.

The technique, inspired by the work of the 2012 Nobel Prize-winning stem cell biologist Shinya Yamanaka, has only been performed on lab mice, but “nobody has been capable, so far, to do that with two-ton species like the rhino.”

The scientists are using stem cell technology to create eggs and sperm from deceased northern white rhinos.

“We have not only harvested sperms from the four different northern white rhino bulls but we also collected skin samples from 12 unrelated individuals.

“We have the best scientists on board and we hope to make significant progress in this field in the next three to five years,” Hildebrandt told RFI.

“It is very ambitious, but without dreams you can’t change the world.”

24/7 Armed Surveillance

The northern white rhino is endemic to swamp areas extending over Uganda, Sudan, Democratic Republic of Congo, Central African Republic and Chad.

Extensive poaching and civil war led to their near extinction. Najin and Fatu were born in captivity and brought to Kenya in 2009 from the Dvur Kralove Safari Park in the Czech Republic.

The world’s two remaining northern white rhinos live under the constant surveillance of 42 armed guards from the National Police Reservists.

“We have a system where we can track the walkie talkies of the patrols. We have night vision, we have thermal images,” explains Emilio Gichuki at the Ol Pejeta conservancy.

He added that poaching is still an acute problem which they are trying to resolve by involving the neighbouring communities.


Inbreeding among white rhino is the enemy within (South Africa)

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Tanya Farber, Times Live | January 17, 2020

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Poachers have killed almost 1,800 SA rhino in the past two years, but a first-of-its-kind study has uncovered a threat to the species that is just as deadly yet far more subtle.

German scientists say inbreeding is playing a major part in propelling the southern white rhino towards extinction as fenced reserves have reduced the range over which they roam.

Over 13 years, a team from the Leibniz Institute for Zoo and Wildlife Research collected genetic material from 104 offspring, 37 mothers and 13 males on a game farm near Thabazimbi in Limpopo.

They uncovered three problems that have severely reduced the gene pool:

• Many females stick to the same partner over several consecutive breeding periods, so all their offspring inherit the same genes.

• Rhinos do not avoid reproducing with family members and do not have the same innate inbreeding avoidance of many other species. “Females tended to mate more frequently with closely related males, and one daughter produced a child with her father,” said lead researcher Petra Kretzschmar.

• The reproductive success of males is very unequal, so a few males dominate the gene pool.

All of this was not problematic in the past, but now that all rhino live in fenced reserves, they cannot disperse far enough to avoid inbreeding.

This is the largest scientific study to date on the sexual preferences of white rhino and was published in the journal Evolutionary Applications. “During the colonial period, intense hunting decimated white rhinos to a few individuals. All currently living white rhinos originate from this small founder population,” say the researchers, so the gene pool was already severely compromised.

According to The Boucher Legacy, a rhino protection organisation started by new Proteas coach Mark Boucher, “in the early 1900s there were fewer than 50 white rhinos left in the wild. Today, however, the population numbers are up to about 18,000, but the poaching crisis caused the white rhino population to decline by 15% between 2012 and 2017.”

Now that inbreeding has been confirmed, it is not only poaching that will need to be tackled.

“We need to keep the white rhinos as genetically diverse as possible, if we want to give them a chance to adapt to anthropogenic challenges such as poaching, climate change or diseases,” said Kretzschmar. She and the team did not expect females to keep mating with the same male for several reproductive cycles because long-term bonds with mating partners were previously unknown for rhinos.

“They live on their own and only get together shortly before mating,” said Kretzschmar. “This is why it took us 13 years of field research to uncover the secrets of their mating behaviour.”

In the past, they would range far and wide – away from biological family members. According to the World Wildlife Fund, this behaviour is part of their complex social structures.

Groups of sometimes 14 rhinos may form, notably females with calves. Adult males defend territories of roughly 2.5km², which they mark with vigorously scraped dung piles,” said the WWF.

“The home range for adult females can be more than seven times larger, depending on habitat quality and population density. Males competing for a female may engage in serious conflict, using their horns and massive size to inflict wounds.”

But this complex social structure has been affected by changes in land use. “Today, all remaining rhinos live in modestly sized conservation areas and private game reserves surrounded by fences or human settlements. They cannot disperse far enough,” said Alexandre Courtiol, a senior author of the new study.

So where to from here?

The researchers said SA national parks had been less successful in mitigating the effect of poaching than private game reserves and that the reserves could become “the last refuges for the species”.

They hold a third of the worldwide rhino population and have higher budgets than state-owned conservation areas. According to the researchers, however, “game farmers often pay little attention to genetic diversity”. They “remove or introduce new rhinoceros into their population according to the size of the horns or the origins of the new animals”, without taking the genetic makeup of an individual into account.

The researchers, therefore, recommend permanent monitoring of offspring and their genetic relatedness to rhinos nearby and a regular exchange of unrelated animals between protected areas.

“This is the only way to preserve the long-term genetic heritage of the species,” said Kretzschmar.


Indonesia to capture 3 wild Sumatran rhinos to add to breeding population

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Rahmadi Rahmad / Translated by Basten Gokkon, Mongabay | December 10, 2019

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EAST LAMPUNG, INDONESIA: Officials in Indonesia say they hope to capture three Sumatran rhinos from the wild for a recently expanded sanctuary where experts are carrying out breeding attempts to save the species from extinction.

The Sumatran Rhino Sanctuary (SRS) in Way Kambas National Park, in Lampung province at the southern end of Sumatra Island, now spans 250 hectares (620 acres), following an expansion announced on Oct. 30.

“We are working to capture three wild rhinos in Way Kambas National Park,” Ade Kurnia Rauf, a senior adviser to Indonesia Rhino Foundation (YABI), told Mongabay. He added this was in line with Indonesia’s Emergency Action Plan on Sumatran Rhinoceros, issued on Dec. 6 last year.

Original photo as published by Mongabay.

While waiting for new rhinos to occupy the extension sometime next year, officials at the SRS have already moved one of the seven existing Sumatran rhinos (Dicerorhinus sumatrensis) at the facility into the new paddocks; Harapan, a 12-year-old male, is himself the product of an earlier successful captive-breeding program carried out at Cincinnati Zoo in the U.S.

The SRS at Way Kambas was opened in 1996, covering 100 ha (247 acres) and envisioned as a way to provide a heavily protected semi-wild habitat in which captive rhinos could breed naturally. Experts at the sanctuary are also tasked with carrying out research and programs to maintain a viable captive population that should be able to be released back into the wild eventually.

“Way Kambas National Park, where some of Sumatra’s last lowland tropical forest exists, is the last frontier for nearly extinct wildlife, such as the Sumatran rhino, [so] that we must protect its sustainability,” said Indra Exploitasia, the director of biodiversity conservation at Indonesia’s environment ministry.

Indonesia has launched a program to track and tally up Sumatran rhinos in the wild, including in Way Kambas National Park outside the SRS. Ade said five teams had been deployed to look for wild rhinos in Way Kambas and, by February, to start setting up pit traps to safely capture them alive.

The park agency estimates the rhino population in Way Kambas, which spans some 130,000 ha (321,200 acres), at some 33 individuals; some analyses give a much lower figure of around a dozen. The sanctuary itself is home to seven captive rhinos: three males and four females. Two of the rhinos were conceived and born at the sanctuary.

“We need new males and females to be relocated into the SRS. The reason is clear: to avoid inbreeding,” said Subakir, the head of Way Kambas National Park Agency.

In tandem with efforts to breed the species in captivity, conservationists are calling on the government to protect the last remaining wild habitats of the critically endangered animal so that there’s somewhere to release them back into when the situation allows.

The government of Lampung province has promised to increase protection for its forests from human pressure, especially within national parks, saying it wants the province to be the stronghold for the species.

“We must protect the Sumatran rhino, Indonesia’s treasure, from extinction,” said Arinal Djunaidi, the governor of Lampung, at a ceremony to mark the opening of the SRS extension on Oct. 30. He said he would seek an agreement with the environment ministry to boost law enforcement for national parks across Lampung.

Unlike with other rhino species, poaching isn’t the biggest threat to Sumatran rhinos. It’s the lack of natural breeding in the wild — a result of their habitats being carved up and destroyed, isolating individual rhinos and making it less likely that they’ll encounter one another to breed — that poses the greatest danger to the species.

“We’re fighting against extinction,” said Zulfi Arsan, a veterinarian at the SRS. “The population decline rate is [higher] than the birth rate.”

Despite its name, the species used to roam other parts of Asia, including India, Thailand, Vietnam, Cambodia and Malaysia. But the combination of habitat loss and poaching has left Indonesia as the final refuge for these rhinos; Malaysia’s last Sumatran rhino, a female held in captivity, died of ill health on Nov. 23.

Widodo Ramono, the executive director of YABI,said his foundation had partnered with an international group, the Sumatran Rhino Survival Alliance — which consists of Indonesia’s environment ministry, the IUCN, the International Rhino Foundation, Global Wildlife Conservation, National Geographic and WWF — to protect the species.

“The expansion of the SRS can be used to its best to increase the rhino population while paying attention to sustainability, heritability, and good management of habitat,” he said.

Experts at SRS will still prioritize natural breeding, but are also open to using advanced reproductive technology, including in vitro fertilization.

“We must be very careful in carrying out anything,” Zulfi said. “Obviously, to produce a healthy rhino needs healthy parents who don’t have any reproductive problems.”

Fewer than 80 Sumatran rhinos are believed to live in small populations scattered in the dwindling forests of Sumatra and Indonesian Borneo. In Sumatra, experts believe the rhinos survive in Gunung Leuser National Park, Bukit Barisan Selatan National Park and Way Kambas National Park. In Borneo, no more than 10 individuals are estimated to roam the forests of East Kalimantan province. Last year, a female rhino was captured from the wild there and relocated to a second SRS facility there. In 2016, another female rhino had been caught, but died a few weeks after her capture.

“We must act quickly against time to save this species that has lived on Earth since 20 million years ago,” Widodo said. “The most important effort now is to produce as many rhinos as possible at SRS in the safest setting there is.”


Indonesia plans IVF for recently captured Sumatran rhino

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Basten Gokkon, Mongabay | November 5, 2019

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EAST LAMPUNG, INDONESIA: Indonesian authorities will make their first attempt at in vitro fertilization of a Sumatran rhino, aiming to boost the critically endangered species’ gene pool in the process.

The egg for the IVF attempt will come from Pahu, a solitary female Sumatran rhino (Dicerorhinus sumatrensis) being held at the Kelian Lestari Sumatran Rhino Sanctuary (SRS) in West Kutai district, in the Bornean province of East Kalimantan. Indra Exploitasia, the director of biodiversity conservation at the environment ministry, told Mongabay that the plan was to fertilize egg cells harvested from Pahu with sperm collected from one of the males living at the Way Kambas SRS in Lampung province, on the island of Sumatra.

Pahu was captured from the wild in November 2018 as part of a captive-breeding program for the species. For a year she has been held alone in the facility leading some to question what Indonesian authorities plan for her future.

Original photo as published by Mongabay: Pahu is the sole captive rhino at the Kelian Lestari Sumatran Rhino Sanctuary (SRS) in Indonesian Borneo. (Image courtesy of the Indonesian Ministry of Environment and Forestry/Sumatran Rhino Rescue Team Kalimantan.)

Pahu is believed to be quite old as rhinos go, about 25 years old, but experts say they’ve found no obvious reproductive problems with her.

However, she weighs only around 360 kilograms (790 pounds), less than half the weight of a typical adult Sumatran rhino, and experts suspect she might be suffering from dwarfism. Her small size has raised fears that attempting to mate her naturally with a much-larger male could lead to injury or even death; Sumatran rhino mating is a violent, raucous affair. Her size has also prompted doubts that she would be able to bring a regular-sized baby to term.

Pahu’s isolation at the Kelian Lestari SRS is another potential obstacle. Previous research has indicated that female Sumatran rhinos do not ovulate naturally when males are not present. However, Dedi Candra, a veterinarian working for Indonesia’s environment ministry, says some egg cells do develop without males present, albeit at a slower pace, and that researchers have had some success artificially inducing ovulation. Widodo Ramono, executive director of the Indonesia Rhino Foundation (YABI), says that male rhino urine alone may be enough to stimulate ovulation. Conservationists in Indonesia have already made use urine from captive males, flying a liter to Kalimantan to help lure Pahu into the pit trap where she was captured.

“We are currently monitoring Pahu’s reproductive health,” Indra told reporters in Way Kambas on Oct. 30. “We must know first when she ovulates, so the egg cells can be retrieved and then fertilized in a test tube.”

If the plan goes through, it will be the first IVF attempt on captive Sumatran rhinos by Indonesia, says Widodo. Scientists here previously attempted artificial insemination — injecting sperm into the uterine cavity —with Bina, one of the captive female rhinos at Way Kambas, but it was unsuccessful. For that attempt, they used semen collected from Andalas, a rhino born in captivity at Cincinnati Zoo and now a resident at Way Kambas, where he has sired two offspring, both through natural mating.

Last month, experts in Malaysia attempted to use IVF to fertilize an egg harvested from an older female rhino using sperm collected from a now-deceased male. But that attempt was also unsuccessful, with Malaysian experts citing the low quality of the sperm, taken when the male was very old. The Malaysian conservationists have long requested a transfer of sperm from the Indonesian captive rhinos, but Indonesian authorities have repeatedly declined, citing the need to sort out a long list of paperwork.

If the IVF attempt with Pahu’s egg and sperm from Andalas or his younger brother, Harapan, is successful — or Malaysia sends egg cells retrieved from its last rhino to Indonesia and the treatment works — the new offspring would represent a new hope for the species. The populations in Sumatra, D. s. sumatrensis, and Borneo, D. s. harrissoni, are subspecies that have been genetically separated for hundreds of thousands of years. Mixing the two would give a much-needed boost to the gene pool of a species so diminished — as few as 40 are believed to remain on Earth — that inbreeding is a real risk.

The idea of mixing the Sumatran and Bornean bloodlines initially met with disapproval from conservationists. But in recent years there’s been a growing sense of urgency among researchers that the situation is so dire that it’s better to focus on preserving the species at all costs rather than trying to maintain two separate subspecies.

Indra said the planned IVF attempt would most likely use sperm from Andalas, who is a proven breeder. “Harapan has never had a chance to mate naturally,” Indra said. “So we don’t know yet the quality of his sperm, and we haven’t tried to collect samples from him.”

Meanwhile, the surrogate mother could be any captive female Sumatran rhino as long as she’s not going under a natural breeding program, Indra added.

A previous global effort to breed captive Sumatran rhinos, launched in the 1980s, fell through a decade later after more than half of the animals died without any calves being produced. But a string of successful captive births at Cincinnati Zoo, and later Way Kambas, and a growing consensus that the species will go extinct without intervention, have laid the groundwork for the latest captive-breeding effort.

The species was brought to the edge of extinction by habitat loss, with Sumatra and Borneo losing vast swaths of forest to oil palm plantations and coal mines, as well as poaching. Now, conservationists believe a low birthrate is the primary threat to Sumatran rhinos’ survival. The network of SRS breeding centers (the Indonesian government plans to open a third in Aceh, at the northern tip of Sumatra) holds a combined eight rhinos — seven at Way Kambas and one at Kelian Lestari, including two calves born in captivity. Malaysia has one in captivity, an aging and ailing female named Iman, but otherwise the species is believed to be functionally extinct there.

Strides in assisted reproductive techniques for rhinoceros: What do they signify?

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The International Rhino Foundation Blog | September 13, 2019

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There is no easy answer for ensuring the survival of critically endangered rhinoceros species. The International Rhino Foundation (IRF) believes we need to maximize options and minimize regrets, tackling the myriad of challenges facing rhinos using multi-faceted approaches. Some will fail, but some will succeed.

Beginning in mid-August, media across the world reported that an international team of reproductive scientists, including from Berlin’s Leibniz Institute for Zoo and Wildlife Research (IZW) recovered five eggs each from both remaining female northern white rhinos, Ceratotherium simum cottoni, at Ol Pejeta Conservancy in Kenya. The two females have reproductive tract issues which prevent them from carrying a pregnancy to term. Working in concert with Avantea, an Italian laboratory, the team matured seven of the eggs in vitro (in a culture dish). The eggs were then injected with frozen-thawed sperm from two deceased northern white rhino bulls.

Original photo as published by The International Rhino Foundation.

Earlier this week, the team revealed that two of the eggs grew to early stage embryos. This is a phenomenal achievement – the embryos have been frozen and later will be implanted into a southern white rhino (Ceratotherium simum simum) surrogate. Other partners include the Kenya Wildlife Service and. Dvur Králové Zoo in the Czech Republic.

We follow these developments with great interest, knowing that there is a long road ahead before these methods are readily reproduceable. In the words of Dr. Terri Roth, Director the Cincinnati Zoo & Botanical Garden’s Center for Conservation & Research of Endangered Wildlife (CREW) and IRF Vice President for Asia, “It’s a long path from developing a berry-like cluster of cells in a Petri dish to having a herd of healthy rhinos on the ground.”

The work with northern white rhinos has proceeded in a lengthy, stepwise process. Last year, in a paper published in Nature, IZW’s Dr. Thomas Hildebrandt and colleagues described successfully recovering southern white rhino eggs, maturing and fertilizing them with northern white rhino sperm, and successfully developing them to the blastocyst (early embryo) stage in vitro (in a culture dish).

The team also established embryonic stem cell lines from the blastocysts, which were later frozen. (A stem cell is a cell that has the potential to develop into many different cell types in the body during early life and growth.) Conservation geneticists at the San Diego Zoo’s Institute for Conservation Research in collaboration with the Center for Regenerative Medicine at the Scripps Research Institute also are carrying out groundbreaking research, recently turning rhino stem cells into beating cardiac tissue.

The IRF applauds the efforts of all of our colleagues working on Assisted Reproductive Technology (ART) and conservation genetics for rhinos. We also wish to note the importance of keeping these recent accomplishments in perspective, particularly for a subspecies that is functionally extinct. Attempting to use ART to try to “bring back” the northern white rhino is a complex, highly difficult endeavor, and creating early stage embryos is but the first step on a long journey.

Recovering immature eggs and maturing them in vitro is a difficult task in itself, particularly when eggs are being recovered from females with reproductive issues. The next step, injecting the eggs with sperm and allowing them to develop into early stage embryos and placing them into the uterus of surrogate is also complicated. Developing a viable pregnancy and having the surrogate carry it to term with a successful delivery is another hurdle.

Once it is reliable, ART may play a vital role in securing the future for rhino populations. There recently have been other important ART advances for rhinos.

In July, scientists at the San Diego Zoo’s Institute for Conservation Research produced a southern white rhino calf using frozen semen and artificial insemination (AI).

In May, reproductive scientists from the South East Zoo Alliance for Reproduction and Advanced Conservation (SEZARC) induced ovulation in a greater one-horned rhino (Rhinoceros unicornis) and used AI to produce a calf at Zoo Miami.

In 2014, the Cincinnati Zoo & Botanical Garden’s Center for Conservation & Research of Endangered Wildlife (CREW) and the Buffalo Zoo in New York, produced a greater one-horned rhino calf using AI with the sperm of a deceased male who never contributed to the gene pool of the species during his lifetime.

These are but a few of the more recent achievements; we encourage reading Pennington and Durrant’s historical review of rhino ART efforts in Mammal Review and the Nature article referenced above for more information.

The Northern White Rhino
The conservation community and international governments should have attacked conserving the northern white rhino more vigorously much earlier – certainly before only a handful were left in captivity. The IRF spent millions of dollars protecting the northern white rhino in Garamba National Park in the Democratic Republic of Congo but, despite protection, the species was lost when the Park became a conflict zone and we had to pull out to ensure the safety of our staff. Governments in range states are responsible for conserving their biological heritage, but if no political will is present, there is only so much that outside stakeholders can do.

If and when northern white rhinos are successfully created, inevitably there will be issues over where to locate them – the countries in which they perished are still in conflict, and invariably, there is an ongoing poaching threat, making repatriation improbable. There also are complex ethical issues to be resolved: should we “re-create” a subspecies that has gone extinct? Furthermore, the initial attempt at maintaining an ex situ breeding population failed, which begs the question, what will we do different to make sure the next effort succeeds?

Finally, ART work is exceedingly expensive. Some argue that funding at the level that has been raised for the work with northern white rhinos could be used for field conservation. However, we do not believe that the donors are in all cases the same; thus, the funding being used to support ART work is likely not competing for funding that would be available for in situ rhino conservation.

Application to other rhino species
All five rhinoceros species are in peril, with four of the five (with the exception of the white rhino) listed as threatened on the IUCN Red List of Threatened Species. The most critically endangered species, the Sumatran rhino (Dicerorhinus sumatrensis), numbers fewer than 80 individuals scattered in four fragmented locations in Indonesia.

Our hope is that critically endangered rhino species may someday benefit from this work, and that perfecting ART methods with the non-threatened white rhino may help them in the future.

But it’s also very possible that the technology, which could contribute to increasing numbers very slightly in the big scheme of things, may not be in place in time to contribute to increasing Sumatran rhino numbers either; the species is declining precipitously despite in situ protection and some success with natural breeding in captivity.

Nevertheless, efforts to continue developing ART should definitely move forward. No species has been saved by “high tech” approaches alone, but such technologies have proven valuable when integrated with natural breeding and protection of wild populations.

To-date, as described by in a paper by Howard and colleagues in Animal Conservation, the black-footed ferret (Mustela nigripes) is the only species where AI (with frozen sperm) has been used for genetic restoration. However, because there are no wild northern white rhinos left and the two still alive cannot reproduce naturally, it would be a long time, perhaps two decades, before enough rhinos could be created to form a viable population – Population Viability Analysis has shown that populations of fewer than 15 rhinos are not viable.

There is no easy answer for ensuring the survival of critically endangered rhinoceros species – but we do need to tackle the myriad of challenges they face using multi-faceted approaches. ART is one of the developing tools in the toolkit.

Scientists have successfully created embryos of the near-extinct northern white rhino

By Conservation, Science and technology No Comments
Abdi Latif Dahir, Quartz | September 11, 2019

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The race to save the northern white rhinoceros from extinction is looking very hopeful. Scientists today announced they were able to create two embryos of the rare rhino, raising hopes that new members of the endangered species could be conceived in the future. The in-vitro creation was made by a consortium of veterinarians and conservationists from Germany, Kenya, and the Czech Republic working at the Avantea laboratory in Cremona, Italy. The embryos are now stored in liquid nitrogen and will soon be transferred into a surrogate mother.

The news comes just weeks after a team of veterinarians achieved another milestone by harvesting 10 oocytes from the two last two remaining female northern white rhinos living in Kenya. Najin and Fatu, a mother and daughter duo, are the only two northern white rhinos left in the world and currently live under 24-hour armed guard in Ol Pejeta Conservancy in Kenya’s central highlands.

Previously housed in a zoo in the Czech Republic, they were transferred to Kenya in 2009 in the hopes that the climate, natural habitat, and dietary conditions would provide more favorable breeding conditions. But those hopes were repeatedly dashed—especially after the death of Sudan, the last male northern white rhino, last year.

Original photo as published by Quartz Africa: Najin (front) and her daughter Fatu, the last two northern white rhinos.

Yet scientists kept raising money to develop reproductive technologies and hoped the eggs along with preserved sperm from deceased northern white males could be used to inseminate the abundant female southern white rhino. The two current embryos were created using eggs collected from Fatu, the youngest of the two northern white rhinos, and frozen sperm from Suni, a deceased northern white rhino male. Najin’s eggs didn’t make it to a viable embryo despite the fact that one egg initiated segmentation.

“The entire team has been developing and planning these procedures for years,” says Thomas Hildebrandt from the Leibniz Institute for Zoo and Wildlife Research in Germany. “Today we achieved an important milestone on a rocky road which allows us to plan the future steps in the rescue program of the northern white rhino.”

Widespread poaching and civil wars have decimated the numbers of the rhino population, with the northern white rhino suffering the greatest losses. Kenya’s tourism minister Najib Balala said the government was encouraged by the new developments and was committed to championing the pioneering process globally. “It has been a decade of race against time and we are excited at the progress in reversing the hitherto bleak outlook for the northern white rhino.”

Scientists in Italy fertilize 7 northern white rhino eggs

By Science and technology No Comments
Andrea Foa, Phys.org | August 26, 2019

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Silvia Colleoni’s hand holding a syringe was trembling as she injected liquid into a micro pipette to facilitate the aspiration of sperm that had been removed and later frozen from one of the last then-living male northern white rhinos on Earth.

Her nervousness in the Avantea laboratory in northern Italy on Sunday was understandable. Whether the artificial fertilization of eggs taken from the last two living females succeeds could determine whether the species has a future, or is doomed to end.

“It does create a little emotion,” Colleoni said, reflecting in a phone interview the day after the fertilization procedures that could result in as many as seven embryos.

“After it was over, I was calm, but it’s a manual task, any error, any slipping, if it falls, will result in irreparable damage.”

Original photo as published by Phys.org: Researcher Paola Turini works at the Avantea laboratory during the inseminating of eggs from the last two remaining female of northern white rhinos with frozen sperm from two rhino bulls of the same species, in Cremona, Italy, Sunday, Aug. 25, 2019. The northern white rhino is on the verge of extinction but Sunday’s operation raises hopes that they’ll survive. (AP Photo/Antonio Calanni)

The Associated Press was granted exclusive access to the laboratory to film the procedure being carried out on Sunday.

Eggs that were removed last week in Kenya from the last two female northern white rhinos, Najin and Fatu, were fertilized in the lab with frozen sperm from two now-dead males. It will be about 10 days before it is known whether the eggs have become embryos, the Italian assisted-breeding company said Monday.

Wildlife experts and veterinarians are hoping that the species can reproduce via a surrogate mother rhino, since neither Najin nor Fatu can carry a pregnancy.

On Tuesday, Colleoni predicted that “there will be more tension, more emotion,” when she peers through a microscope to see if the fertilized eggs start dividing, in two, then four, then more cells.

She recounted how, using joysticks, she guided the fertilization process. “It takes so much concentration, a lot of attention,” Colleoni said. To facilitate success, an electronic impulse is also sent to the eggs.

“We expect some of them will develop into an embryo,” Cesare Galli, a founder of Avantea and an expert in animal cloning, said after the procedures were carried out.

The ultimate goal is to create a herd of at least five animals that could be returned to their natural habit in Africa. That could take decades.

Avantea said that only seven of 10 eggs extracted last week from the females in Kenya were suitable for use in the artificial insemination process Sunday.

Galli said that the sperm used came from two northern white bulls, named Suni and Saut, who had been living in a zoo in the Czech Republic. The sperm of both now-dead males were used to enhance chances of reproductive success, especially since Suni is the half-sibling of Najin. In addition, Saut’s sperm was difficult to work with, the company said.

Galli, a founder of the company, said that to improve chances for a species’ continuation, it is better not to wait to “get to the last two individuals before you use this technology.”

The last living male was a 45-year-year-old named Sudan, who gained fame in 2017 with his listing as “The Most Eligible Bachelor in the World” on the Tinder dating app in a fundraising effort. Sudan was euthanized after age-related complications.

Galli said that Sudan’s sperm was still in Kenya, raising the possibility that in any future attempts to create more embryos, scientists might seek the transfer of Sudan’s sperm for that purpose.

Decades of poaching decimated the northern white rhino’s numbers.

Sudan was the last of his kind to be born in the wild, in the country he was named after.

Other rhinos—the southern white rhino and the black rhino—are also prey for poachers, who kill them for their horns to supply illegal markets in parts of Asia.

The efforts to save the northern white rhino species have been led by the Leibniz Institute for Zoo and Wildlife Research in Germany, Avantea, the Dvur Kralove Zoo in Czech Republic, the Ol Pejeta Conservancy in Kenya and the Kenya Wildlife Service.