NST Online by Roy Goh, 15 October 2019

KOTA KINABALU: In vitro fertilisation (IVF) treatment on an egg retrieved from Iman – the last Sumatran rhino in Malaysia – failed earlier this month.

Specialists attempting to save the species from extinction saw the fertilised cell degenerate within days after thawed-out sperm – harvested from Tam, the last male rhino – was injected into it.

Tam himself has since died.

It is learnt that the egg cell had failed to divide after fertilisation, and no embryo was formed.

However, a source said that while the failure is disheartening, “The team believes efforts should continue and that we should learn from the experience”.

The treatment was led by Professor Arief Boediono, a world-class IVF practitioner from Universiti Pertanian Bogor, Indonesia; as well as the Centre for Wildlife and Livestock Innovation, Faculty of Sustainable Agriculture, University Malaysia Sabah (UMS) in Sandakan.

The process of retrieving the egg from Iman was conducted by a team of experts from the Leibniz Institute of Zoo and Wildlife Research in Berlin, Germany, led by Professor Thomas Hildebrandt, at the Tabin Wildlife Reserve in Lahad Datu, together with the Borneo Rhino Alliance (Bora).

“We believe the quality of the sperm was low, and that to continue the IVF treatment effort, we need to try and get sperm from rhinos in captivity in Indonesia.

Despite the failed attempt of in vitro fertilisation (IVF) treatment, there is still hope for Iman and Sumatran rhinos.

“For now, Iman is relatively healthy, and though she has a tumour in her uterus, she is still producing oocytes, or eggs,” he said, but added that there is worry that the rhino may stop ovulating.

It was previously believed that the poor quality of Tam’s sperm was a factor in similar failures in previous in vitro attempts. Experts in Sabah, however, will continue to collaborate with Indonesia in the pioneering efforts.

Sabah Wildlife Department director Augustine Tuuga had previously expressed hope in the attempt to produce an embryo through the in vitro process for the continuation of the species.

Tam’s sperm was retrieved, frozen and kept for breeding purposes before he died in May. However, Iman can no longer get pregnant due to the uterine tumour that is plaguing her.

In August, Deputy Chief Minister Datuk Christina Liew led a delegation of state representatives and the department to Indonesia to discuss conservation efforts for the Sumatran rhinoceros.

The Star online by Stephanie Lee, 30 September 2019

Prof Hildebrandt and his team from the Leibniz Institute of Zoo and Wildlife Research in Berlin performing the delicate procedure to harvest egg cells from Iman, Malaysia’s last surviving Sumatran Rhino

KOTA KINABALU: A single egg cell – that is all that scientists managed to harvest from Malaysia’s last living female Sumatran rhino – Iman. The egg cell (oocyte) was harvested at the Tabin Wildlife Reserve in Lahad Datu on Monday (Sept 30).

Scientists hope to fertilise the oocyte using the in-vitro fertilisation (IVF) method with frozen sperm, which was extracted from Tam, the male rhino that died in May this year of renal failure.

The single oocyte has been sent to the Reproductive Innovation Centre for Wildlife and Livestock at the Faculty of Sustainable Agriculture in Universiti Malaysia Sabah (UMS), Sandakan.

Sabah Wildlife Department director Augustine Tuuga said this is great news and it would have been even better if they had managed to harvest more oocytes.

“But this will allow us to continue with the programme to try to create embryos of this critically-endangered species,” he said.

“We remain ready to collaborate with Indonesia for further attempts,” he added.

The extraction was performed by Prof Thomas Hildebrandt and his team from Leibniz Institute of Zoo and Wildlife Research, Berlin, in collaboration with the local Borneo Rhino Alliance team.

Preparation and maturation of the oocyte is being handled by Prof Arief Boediono of the Bogor Agriculture University, Indonesia.

Professor Abdul Hamid Ahmad, Dean of the Faculty of Sustainable Agriculture from UMS, said the Reproductive Innovation Centre is now open for use and he is pleased that the IVF attempt will be made there.

“There are not enough wild Sumatran rhinos left alive to save the species,” he said.

Advanced reproductive technology can ensure that the living genomes of rhinos are kept alive long after their deaths.

Read more at https://www.thestar.com.my/news/nation/2019/09/30/scientists-harvest-single-egg-cell-from-iman–malaysia039s-last-sumatran-rhino#WiemEY7G8EXRGrvQ.99

By Avila Geraldine, NST Online May 27, 2019

KOTA KINABALU: Malaysia’s last remaining male Sumatran rhinoceros, affectionately called Tam, has died today.

Tam died at about noon at Borneo Rhino Sanctuary in Tabin Wildlife Reserve, Lahad Datu.

Sabah Deputy Chief Minister cum Tourism, Culture and Environment Minister Datuk Christina Liew said the exact cause of death would be known after the autopsy.

“Invariably, everything that could have possibly been done, was done, and executed with great love and dedication.

“(Tam’s) last weeks involved the most intense palliative care as humanly possible, rendered by the Borneo Rhino Alliance (BORA) team under veterinarian Dr Zainal Zahari Zainuddin at the Borneo Rhino Sanctuary,” she said in a statement, here.

Liew said Tam’s death was related to old age and involved multiple organ failures.

It was reported that Tam’s appetite and alertness had declined significantly since the end of last month.

Urine analysis tests indicated that Tam was suffering from organ damage and poor kidney function.

It spent most of its time lying down, having received round-the-clock attention and medication from keepers as well as veterinarians.

In Aug 2010, a wildlife team captured the male rhinoceros at the Kretam oil palm plantation in Tawau.

At the time of its capture, the rhinocerous was thought to be in its mid-20s. Tam was taken to the Tabin Wildlife Reserve where it had lived ever since.

Liew said Tam’s living genome was now preserved in cell culture.

“We hope that with emerging technologies at cell and molecular levels, he may be able to contribute his genes to ensure the survival of the species,” she added.

With Tam’s death, Malaysia is now left with one female Sumatran rhinoceros, Iman. She was captured in 2014.

In 2017, another female rhinoceros Puntung was euthanised because she was suffering from a painful and incurable cancer.

Iman appears to be the last Sumatran rhinoceros that was found in the wild. Since her rescue, no other Sumatran rhinoceros has been detected in Sabah, indicating that the species may have become extinct.

IZW experts Professor Thomas Hildebrandt and Dr Robert Hermes extract eggs from Iman, assisted here by BORA staff Hassan Sani.

The 4 July 2018 announcement by Professor Thomas Hildebrandt of Leibniz Institute for Zoo and Wildlife Research and his co-workers that embryos of white rhinos have been successfully made in vitro represents one of the most exciting breakthroughs in years for rhino conservation. We at BORA, the Borneo Rhino Alliance, are ecstatic at this development, as it represents a new dawn for not just white rhinos, but also for another, even more endangered species – the Sumatran rhino.  While there are only two females of the northern white rhino subspecies left, the white rhino species stands at close to 20,000 individuals. But there are fewer than 100 Sumatran rhinos left in the world. Even this may be an optimistic estimate with some reports claiming only 30 are left in the wild.

Unlike the African and Indian rhino species, Sumatran rhino numbers have drastically plummeted not primarily by poaching. Instead, since the 1930s, the four-part threat was, and still is –

1) Not enough Sumatran rhinos in any one place to form a breeding population 2) Few or no mates in any one location, so very little to no mating 3) Inbreeding due to isolation over several generations

4) Reproductive tract pathology and infertility due to the above three syndromes

As a result, for many decades, there have not been enough Sumatran rhino births to sustain the species, whether or not poaching can be prevented. Natural breeding was seen as the solitary key solution but in the past five years, there has only been one birth in captivity, while there have been three deaths. In the absence of concerted efforts to maximize all available avenues of increasing the number of Sumatran rhino births, the oldest and smallest of all rhino species has been on a one-way fast track to extinction.

File image of Puntung, a female rhino rescued by BORA vet Dr Zainal Zainuddin. Photo by C C Azrie Alliamat

There is only one clear way now to save the species – all the last remaining Sumatran rhinos have to be brought into a single managed breeding programme, where two complementary approaches are taken: allowing the fertile ones to breed naturally, and supporting the infertile ones to allow their limited eggs and sperm to create embryos in vitro.

BORA has been desperately working for the past decade to try to breed new generations of Sumatran rhinos. But we faced incredible obstacles in the form of not enough rhinos left in Malaysia and the rhinos’ reproductive tract pathology and infertility. With the help of Professor Hildebrandt (IZW, Germany), Professor Cesare Galli (AVANTEA, Italy) and Professor Arief Boediono (IPB, Indonesia) and their colleagues, we have tried to create a Sumatran rhino embryo in vitro. It was a process that had been derided by many of our counterparts as being fantasy and foolish thinking.

But now with this success for the white rhino, the issue for the Sumatran rhino is no longer about “can in vitro fertilization be done?” but “why are we not prioritizing such work for Sumatran rhinos?”.

Only a few tens of Sumatran rhinos remain alive in Indonesia, and only two, a female and a male in Malaysia. Professor Hildebrandt and his co-workers have been aiming for in vitro production of Sumatran rhino embryos since 2011, when there were three females and the male alive in well-managed facilities in the Malaysian State of Sabah and five managed in the Sumatran Rhino Sanctuary in Indonesia. But the advanced reproductive technology group has never managed to gain interest from some of the most prominent institutions that need to collaborate or provide support to generate a single recovery programme.

The government of Malaysia, Sime Darby Foundation, and numerous interested individual donors from around the world have provided critical financial support for attempts to apply advanced reproductive technology to Sumatran rhinos, but the shocking lack of interest outside Malaysia has stymied the much-needed species recovery programme. The amount of funding going to advanced reproductive technology on Sumatran rhinos simply has been not sufficient enough to allow the scientific process to catch up to the sort of work done on the northern white rhino. The funds from international donors, which currently include a generous donation from government of USA, have gone to camera trapping, surveys, anti-poaching patrols and meetings in Indonesia and USA. This approach fails to recognize that the greatest threat doesn’t come from outside the Sumatran rhino population. It comes from within. Too few Sumatran rhinos, that are scattered and not breeding. All these efforts are akin to putting a band aid over a tumor.

For years, we’ve been trying to engage and work with prominent institutions that are able to influence or lead the charge to save the Sumatran rhino but we’ve been forced to refrain from taking them to task.

But today, we make our stand. The science of advanced reproductive technology is now beginning to deliver, and all of us must embrace it as we do everything in our power to save the most endangered of rhino species by increasing the number of births. We call on the Government of Indonesia, the International Rhino Foundation and the International Union for Conservation of Nature, to recognize the incredible and critical opportunity we have in front of us. Create a programme that prioritises Sumatran rhino births, through both natural breeding of fertile rhinos and in vitro fertilization for those that need such help. The alternative is to choose certain extinction.

Dr John Payne,

Executive Director of Borneo Rhino Alliance

View featured gallery highlighting IVF efforts to save the Sumatran rhino

The little-known and smallest member of the rhinoceros family, the Sumatran rhinoceros, is critically endangered. Today between 30 and 100 are isolated on the islands of Sumatra and Borneo in Southeast Asia. In a new study, researchers urge conservationists to translocate the two island groups–representing two subspecies of the Sumatran rhino–and to create a cell bank to preserve the genetic diversity uncovered by this work.

“It is heartbreaking as a geneticist to recommend that two subspecies, which are probably as different as humans were from the Neanderthals, should be combined into a single conservation unit,” said Al Roca, a professor of animal sciences in the College of Agricultural, Consumer and Environmental Sciences and the Carl R. Woese Institute for Genomic Biology at the University of Illinois.

The Sumatran rhino is the smallest of the remaining rhino species, but it is critically endangered — with 100 or fewer left in the wild today. Photo credit: Terri Roth

Published in the Journal of Heredity, the study analyzed 13 samples taken from zoos and the wild as well as 26 museum specimens to reveal differences in the species’ mitochondrial DNA, the small proportion of the genome that is passed down only from mothers to their offspring.

The study exposed 17 distinct mitochondrial haplotypes, a group of genes inherited from one parent, that can be used to trace a species’ migrations and distribution across thousands of years. The mitochondrial DNA also confirmed the classification of three subspecies of Sumatran rhinos: D. s. lasiotis (likely extinct), D. s. harrissoni, and D. s. sumatrensis.

In the wild, Sumatran Rhinos are solitary creatures, only coming together to breed. In such few numbers, it is increasingly difficult for them to find each other in their mountainous habitat. What’s more, if they are not able to mate, females develop reproductive diseases that prevent them from successfully breeding.

“My strongest recommendation is that they are brought into breeding centers as soon as possible because they aren’t going to survive in the wild in such low numbers,” Roca said. “A population of 10 individuals loses 5 percent of their genetic diversity each generation, which they cannot spare.”

“Unfortunately, at this point, we have to act quickly and risk losing unique genetic lineages in order to save a whole species,” said first author Jessica Brandt, now a professor of biology at Marian University.

This genetic erosion can be prevented, or slowed, by combining the remaining rhinos to create a larger population. A century of captive breeding efforts have yielded few babies, but recent successes suggest ex situ breeding facilities could help save this species from the brink of extinction–the result of poaching and habitat loss due to legal and illegal logging for desirable hardwoods.

To ensure the long-term genetic health of the species, the authors implore conservationists to preserve the genomes of every living Sumatran rhino. In the future, preserved cell lines could be used to create artificial gametes, to reverse the effects of inbreeding and harmful mutations.

“We may one day be able to use stored cells to bring back what was once lost, reversing the effects of inbreeding, drift, and our own folly,” Roca said. “Because they are at such low numbers, every single living Sumatran rhino is genetically very valuable, and preserving cells with genetic material from each surviving individual is of paramount importance.”

Read the original article here.

This work was made possible by the United State Fish and Wildlife Service Rhinoceros and Tiger Conservation Fund, the International Rhino Foundation, the World Wildlife Fund, the National Science and Engineering Research Council (Canada), and ACES Office of International Program.

The paper “Genetic structure and diversity among historic and modern populations of the Sumatran rhinoceros (Dicerorhinus sumatrensis)” is published by the Journal of Heredity(doi.org/10.1093/jhered/esy019).

Co-authors also include Peter J. Van Coeverden de Groot (Queen’s University in Canada), Kelsey E. Witt (University of Illinois), Paige K. Engelbrektsson (National Museum of Natural History), Kristofer M. Helgen (University of Adelaide), Ripan S. Malhi (University of Illinois), and Oliver A. Ryder (Institute of Conservation Research, San Diego Zoo).

The Carl R. Woese Institute for Genomic Biology research facility at the University of Illinois is dedicated to transformative research and technology in life sciences using team-based strategies to tackle grand societal challenges.