Experts Studying The 14,000-Year-Old Remains Of A Frozen Siberian Puppy Made A Genetic Breakthrough

Image: YouTube/The Siberian Times

Scientists are poring over the body of a puppy; but this is no ordinary animal. The young pup has lain frozen in the Siberian permafrost for 14,300 years. Painstakingly, the researchers extract samples of tissue from cartilage, liver and muscle. And once they’ve analyzed the samples in the lab they make a groundbreaking discovery – and it may have huge implications for science.

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But is this young dog a wild wolf or a domesticated animal once owned by a human, or perhaps a hybrid of the two? It’s impossible to be sure, but at the site where the puppy was found, there are signs of human activity. Researchers found animal bones with the tell-tale signs of butchering, as well as traces of fire.

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So the young animal might indeed have belonged to someone as a pet, or was perhaps a working animal. The remains of the puppy were found some 25 miles from the Siberian village of Tumat in 2015 on a sheer bank of the Syallakh River. Tumat is in Yakutia’s Ust-Yana region, which is also known as the Sakha Republic. And the village has now given its name to the young animal which has become known as the Tumat puppy.

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When they found this young dog, scientists were excavating at that steep river bank because of a previous find in 2011; that year, they had discovered another puppy. They’d been hoping to find evidence of human activity, which they did. But they were also lucky enough to come across this second puppy, perhaps the sibling of the first one that was found. Apparently, the researchers believe that the two dogs may have died together in a mudslide.

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The expedition leader, head of exhibitions at Yakutsk’s Mammoth Museum Sergei Fedorov, described the discovery of the second puppy to The Siberian Times in 2015. He said that it had actually been a matter of luck and that they come across the young dog’s body after not much more than an hour of digging.

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So it turns out that there are not one, but two Tumat puppies. But before we get into more detail about the second one found in 2015, let’s rewind and find out about that first discovery in 2011. It was villagers from Tumat who’d made that earlier find, almost at the same spot as where the 2015 discovery was to come.

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The Tumat villagers who found the first puppy were brothers Igor and Yury Gorokhov and their pal Aiaal Tomsky. They were actually scouring the banks of the Syallakh River in search of mammoth bones when they happened across the remains of the puppy. Fortunately, they’d had the presence of mind to contact scientists at the North-Eastern Federal University in the city of Yakutsk, the regional capital.

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Scientists reckoned that the puppy found by Tomsky and the Gorokhovs was around three months old when it died and that it was a female. Amazingly, the three mammoth tusk hunters had stumbled across the oldest mummified dog ever to have been discovered. Apparently, older canine remains have previously emerged, but never in such a well-preserved condition.

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In 2014 Dr. Mietje Germonpre, of the Royal Belgian Institute of Natural Sciences’ paleontology department, went to Yakutsk to see the puppy’s remains for herself. Speaking to The Siberian Times in 2015, she said, “After studying the mummy and looking at the measurements of the skulls belonging to ancient dogs and wolves, I can say this find is unique.”

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“It’s amazing,” Germonpre continued. “In other museums around the world you will only find the remains of adult dogs, but this is a puppy. Also all external signs and scan results indicate that it is a primitive dog, and at the moment it is the most ancient one found in northern Siberia.”

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“The oldest dog remains were found in the Goyet [Caves] in Belgium, and were 36,500 years old, and there are many finds dating to about 26,000 years ago – but they are not so well preserved,” Germonpre explained. “Here we see the skin and wool and even the internal organs survived.”

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Germonpre went on to describe two theories about how the relationship between humans and dogs had developed. She continued to The Siberian Times, “The first [theory] is that dogs arrived near sites where humans lived and picked up the scraps and gradually they co-existed. The second version talks about the active involvement of man, where the people themselves were the initiator of the relationship, and brought the puppies to their home and trained them.”

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And Germonpre was hopeful that this particular mummified puppy would help to settle the debate about how dogs had become domesticated. She pointed out that her own studies had led her to believe that the second theory she had outlined was the most plausible. In the light of the 2011 find, she added, “Now we can get more arguments.”

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It was four years after the discovery of the first Tumat dog before a post mortem was carried out on the animal at the North-Eastern Federal University in April 2015. One of the scientists involved, Dr. Darima Garmaeva, described the challenges the researchers faced to The Siberian Times. She said, “Our task is to estimate the preservation of the ancient animal tissues at the macro and micro level.”

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“What is of real interest is the fact the animal has a completely preserved carcass, which is unique by itself, with nothing like it in the world,” Dr. Garmaeva pointed out. “Although the tissues are mummified, they have no post-mortem decomposition, as it usually happens with biological material.”

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And back in June 2015, Fedorov was clearly keen to get back to the Syallakh River where the first puppy had been found four years previously. He said, “Near the place where the dog was found local people often find things that obviously belonged to ancient humans, such as stone implements and bone arrowheads. We plan to go to the site this summer together with the archaeologists to find any traces left by humans – and possibly the owners of the dog.”

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Fedorov and his colleagues returned to the site in the summer of 2015, where they found traces of human settlement. As we’ve seen, the team discovered a number of fascinating things. These included signs of fire, animal remains that had been butchered by human hand and tools fashioned from bone. But they also made another extraordinary discovery; a second mummified puppy was found, just six feet or so from the 2011 one.

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Speaking to The Siberian Times, Fedorov described the animal they’d found. He said, “The condition of our new find is perfect. It is preserved from nose to tail, including the hair. You can see the hair on the paw on the picture.” This extraordinary level of preservation in a 14,300 year-old body was thanks to the properties of the permafrost.

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“The find of the new remains was pure luck,” Fedorov added. “We had no inkling we would get it. But having worked there for a full hour, at the beginning of the second hour, some two meters away from the first find, and down the rock, we saw the front paws and the head.” Fedorov went on to describe how the muddy bank had moved since their last visit four years earlier.

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The movement of the bank meant that the second Tumat puppy was found in almost the same spot as the first one. The proximity of the two puppy bodies could indicate that the two were related, perhaps even from the same litter. However, one important question remained – were these the remains of young wolves or of domesticated dogs owned by humans?

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Initial DNA tests on the 2011 Tumat puppy had shown that it was probably a dog rather than a wolf, but there was still some doubt around those findings. The genetic make-up of domesticated dogs and wolves is close, so reaching a completely definitive answer is problematic. Therefore, the scientists planned to examine both the mummified puppies more closely.

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The results of the detailed autopsy of the Tumat puppies were reported in March 2016; and the second puppy turned out to be especially interesting. It had something unique – a well-preserved brain. A video of the autopsy shows the scientists carefully handling the body and then gently washing it to remove some of the mud and debris.

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The video starts off with the scientists carefully collecting some of the material that encrusts the puppy’s body, before they start to wash it. The rather lumpy cadaver already has recognizable features including the paws and the small tail. You can also just about make out the shape of the animal’s head.

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After the carcass has been washed, one of the scientists pulls back the puppy’s lips. And there, quite clearly we can see its teeth, obviously canine in character. The shape of the paws is also quite clear once the corpse has been cleaned. Although still matted, the animal’s fur is also recognizable.

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Sergei Fedorov, the scientist we met earlier, told The Siberian Times, “The carcass is preserved really very well. And one of the most important things is that the brain is preserved. The degree of preservation is about 70 to 80 percent. We will be able to say more precisely after it is extracted.”

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“For now we can see it on MRI scans,” Fedorov continued. “Of course, it has dried out somewhat, but the parencephalon, cerebellum and pituitary gland are visible. We can say that this is the first time we have obtained the brain of a Pleistocene canid.” The parencephalon and cerebellum are parts of the brain while the term canid refers to the group of animals that includes wolves, coyotes, dogs and others.

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And what the autopsy film also shows is the presence of South Korean professor Hwang Woo-suk. He has hit the headlines more than once for his groundbreaking and sometimes controversial cloning experiments. In 2006 Hwang faced scientific fraud charges and was eventually sentenced to an 18-month suspended jail sentence in 2010.

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Speaking to The Siberian Times, Fedorov described the South Korean’s reactions when he saw the second Tumat puppy. He said, “Professor Hwang Woo-suk was also satisfied with the degree of preservation. He was very excited. We examined the carcass thoroughly, palpated the soft tissues, searching for the areas preserved best of all. As a result, he took the samples from the skin, muscles and ear cartilage.”

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Despite his distinctly shady history, Hwang had remained active in the field of cloning. And in 2012 he teamed up with Russian scientists in order to clone a woolly mammoth using DNA extracted from mummified specimens found in Siberia. However, this project never came to fruition.

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But there Hwang was at the 2016 autopsy of the Tumat puppies. And according to a report in The Siberian Times, the professor was interested in attempting to clone a dog from those Tumat puppy remains. He said, “It is my dream to restore not only the woolly mammoth, but also some of the ancient (extinct) animals, such as ancient dogs, ancient deer, ancient bison, and ancient tigers.”

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However, none of Hwang’s dreams have come to pass and nor are they anywhere near doing so, as far as we know. Of course, no resurrected woolly mammals or ancient puppies are currently wandering the Siberian steppe. But scientists had more realistic hopes of knowledge that could be gained from not only the body of the puppy, but the dirt it was entombed in.

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Dr. Artemiy Goncharov, head of research at St. Petersburg’s North-Western State Medical University Department of Epidemiology, Parasitology and Desinfectology told The Siberian Times, “We took samples of the ground which surrounded the carcass to find out the bacteria there. Later we will compare them with the bacteria from the puppy’s intestines.”

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“We hope to find ancient bacteria among them. Also we took samples to find the parasites – ticks, fleas,” Goncharov continued. “We hope to find the parasites which were characteristic for this exact species.” And there has been a significant scientific breakthrough related to the Tumat puppy, although not actually from Goncharov’s work.

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In July 2019 scientists announced a highly significant development in their work on the second Tumat puppy. They had managed to extract genetic material called RNA from the animal’s tissues. Ribonucleic acid (RNA) is distinct from the more familiar DNA; it serves a different purpose within a living organism, which we’ll explain in a moment.

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The University of Copenhagen’s Dr. Oliver Smith led the team which made the new discovery of RNA, which they were able to sequence. Amazingly, this had never been done before from such an ancient animal. While DNA is the permanent record of genetic material contained in living things, RNA, as the EurekAlert! website puts it, is the “the short-lived working copy of a gene.”

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While DNA can be preserved for long periods of time, RNA is much less stable and prone to far quicker deterioration. So scientists have previously thought that the chances of sequencing ancient RNA from archaeological finds was slim to non-existent. But now Smith and his team had done just that; they had extracted the material from the puppy’s liver.

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In fact, scientists have occasionally extracted RNA from ancient plant samples, but it has never been sequenced before from animal remains anything like as ancient as the Tumat puppy. At 14,300 years old, these remains are some 13,000 years older than any animal that has previously yielded a viable RNA sample.

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Smith pointed out another characteristic of RNA; it acts as an “intermediary” between DNA and proteins in living things. And the doctor went on to describe why this discovery of ancient RNA could be so significant. He told EurekAlert!, “… We think the future of ancient RNA has great potential.”

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“For example, many of the most clinically relevant viruses around today have RNA genomes, and the RNA stage is often crucial to understanding the intricacies and complexities of gene regulation,” Smith continued. “This might have repercussions when discussing the environmental stresses and strains that drive evolution.”

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So that second Tumat puppy, probably killed in a mudslide 14,300 years ago, has now made a unique contribution to modern science. Its genetic material, preserved in the frozen Siberian soil, may help us to understand the evolutionary story of not only dogs and wolves, but possibly of the humans who came to domesticate them.

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