Chapter 4: Dima chemically analyzed

The deep-frozen body of the woolly mammoth calf Dima, found in 1977 in NE Siberia, is one of the best-preserved bodies ever found, if not the best one. It lay in a lens of pond ice. - How well has its body been preserved? Why has it died? Has it stayed frozen since it died thousands of years ago? American scientists have analyzed now a piece of frozen muscle from Dima’s body. What have they found out?

Ellen M. Prager, Department of Molecular and Cell Biology, Division of Biochemistry and Molecular Biology, University of California, Berkeley, was so kind to send me on December 5, 1994 a copy of her paper on "Protein survival of mammoth muscle and evidence for leaching". It has come out in a Russian book in 1981. She sent me a copy of this article in Russian and a complete typescript of this article in English. I shall quote now briefly from this work, Protein Survival in Mammoth Muscle and Evidence for Leaching, by Ellen M. Prager, D. Alice Taylor, Vincent M. Sarich, and Allan C. Wilson, Department of Biochemistry, University of California. N. K. Vereshchagin and V. M. Mikhelson, eds. Nauka Press, Soviet Academy of Science, Leningrad, 1981:

"Intact frozen carcasses of the Siberian mammoth would be expected to offer an excellent opportunity to examine fossil proteins (and perhaps even nucleic acids) in a better state of preservation than one finds in ordinary fossils. Consequently, we were delighted to receive samples of tissue from the mammoth discovered frozen and intact near Magadan in 1977. Joy soon turned into disappointment, however, as we found that the samples lacked nucleic acids and contained proteins that were largely insoluble. Eventually, though, we succeeded in showing with an immunological approach that mammoth albumin has survived in a nearly native state in a muscle sample. In this sample, we also detected collagen and perhaps actin and myosin by use of the electron microscope." (1981:4).

Experimental procedure, materials, tissues

"A frozen sample of muscle (19.5 grams) from the right hind leg of the baby woolly mammoth (Mammuthus primigenius) known as Dima was sent to Berkeley from the U.S.S.R. in March 1978. The tissue was dark brown and appeared rather dry. Sera and tissues from the following species were obtained fresh and stored at –15°C: Indian elephant, Elephas maximus, African elephant, Loxodonta africana.

Summary: With the electron microscope, evidence was obtained for the survival of collagen and for structures suggestive of actin and myosin filaments in muscles. More than 90% of the dry weight of mammoth muscle was recovered as amino acids after acid hydrolysis. All 17 of the amino acids normally found in acid hydrolysates of proteins were present.

"Despite the high content of amino acid residues in mammoth muscle, these residues were not soluble in solvent for normal amino acids and proteins. It appears that the original proteins have undergone extensive modification, including cross-linking, following death of the mammoth. Elemental analysis reveals major differences between mammoth and elephant muscle in the content of minor elements. Most remarkable is the absence of phosphorus in mammoth muscle. We attribute the difference in elemental composition to limited autolysis and extensive leaching of the mammoth carcass by soil water." - Prager, E. M. et al. (1981:2).

Leaching of Muscle

The deep-frozen body of the mammoth calf Dima has been leached by soil-water. – How does one know that? What is proving that?

Ellen M. Prager et al.: "The near absence of phosphorus is indicative of post-mortem changes, including hydrolysis of cellular organic phosphates to inorganic phosphate and the loss of inorganic phosphates by diffusion into water surrounding the carcass. The leaching hypothesis explains not only the dearth of phosphorus but also the low content of sodium, potassium, calcium, magnesium, and chlorine in mammoth muscle.

"Leaching of the carcass is expected to have occurred according to the Tolmachoff’s hypothesis as to the conditions of death, burial, and preservation of mammoth carcasses (Tolmachoff, 1929; Farrand, 1961). His hypothesis states that most of the preserved mammoths died suddenly by accidentally being buried in a cold mud stream or drowned and then silted over in cold river. Weeks or months later, the carcass is thought to have frozen. In this period, some autolysis and extensive leaching of small molecules could go on. Under these conditions, one would expect nucleic acids to by hydrolyzed enzymically, whereas proteins like serum albumin probably would survive. ... Leaching might also have occurred during those occasional periods when the frozen carcass thawed, for example, during unusually warm spells in summer.

"If ions diffused out of the carcass, small molecules and ions from the surrounding water would inevitably have diffused into the carcass. It would be valuable, therefore, to analyze the soil in which Dima was found. Knowledge of the soil-pH and the concentrations of soluble inorganic and organic compounds should make it easier to understand the elemental composition of surviving mammoth tissue. The organic compounds in soil water would be of special interest because of their possible involvement in the cross-linking that is presumably responsible for the insolubility of most amino compounds in mammoth muscle.

"The high iron content of mammoth muscle might be explained by diffusion from soil. This is only likely, however, if the soil water was distinctly acidic and anaerobic so that iron would be soluble. ... For their part in providing mammoth tissues, we thank Dr. Andrew S. Antonov, Academician, Dr. Yuri A. Ovchinnikov, and Professor Nikolai K. Vereshchagin." - Prager, E. M. et. al. (1981:1).

Elemental Composition: Results

What is the elemental composition of Dima’s muscle?

Ellen M. Prager: "The phosphorus results are most notable. Mammoth muscle contains essentially no phosphorus. The sodium and potassium levels are also conspicuously low in mammoth muscle. In contrast, one of the minor elements, iron, is present in mammoth muscle at an unusually high concentration. In muscle, iron is normally present as heme, associated with proteins such as myoglobin, cytochromes, and hemoglobin. However, in mammoth muscle, only about 1% of the iron was in the form of heme. This was apparent from the peroxidase test. The results of the pyridine hemochromogen test were also consistent with a low heme level in mammoth muscle, but quantification was difficult owing to interference by a yellow pigment seen in all extracts of mammoth muscle." (1981:9, 10).  

Table 1. Elemental Composition of Mammoth and Elephant Muscle

Elemental composition of muscle from young mammoth Dima. After: Prager, E. M. et al. (1981) Table 1.

Result

A frozen sample of muscle (19.5 g) from the right hind leg of the woolly mammoth Dima was sent from the Zoological Institute of the Soviet Academy of Sciences in Leningrad (now St. Petersburg) to The University of Berkeley, Department of Molecular and Cell Biology, to Ellen M. Prager and co-workers, Division of Biochemistry and Molecular Biology. Their research report was published in Russian under the title Magadan Baby Mammoth. It was edited by N. K. Vereshchagin and V. M. Mikhelson, and published by Nauka Press, Soviet Academy of Sciences, Leningrad, in 1981. They found out:

The samples lacked nucleic acids. The proteins were largely insoluble. The original proteins have undergone extensive leaching, including cross-linking, after the mammoth died. Most remarkable: Phosphorus in the mammoth muscle is absent; due to limited autolysis and extensive leaching of the mammoth carcass by soil water. And the near absence of phosphorus in the mammoth muscle indicates that it was changed after death. Cellular organic phosphates were hydrolyzed into inorganic phosphate. And the inorganic phosphate was lost from the muscle, when it diffused into the water, surrounding the carcass. The muscle of the mammoth calf Dima contains more than 620 times less phosphorus (P), than that of the Indian elephant.

The mammoth muscle also contains only little sodium, potassium, calcium, magnesium, and chlorine. This also proves that the body has been leached. The mammoth muscle contains 11.3 times less sodium (Na) than that of the Indian elephant. It contains 25.3 times less potassium (K), than the muscle of the Indian elephant. It contains 3 times less organic and inorganic chlorine, and 2.6 times less organic sulfur (S), than the Indian elephant. The mammoth muscle contains essentially no phosphorus. The sodium and potassium levels are conspicuously low in the mammoth muscle.

But one of the minor elements, iron, is present in the mammoth muscle at an unusually high concentration. In muscle, iron is normally present as heme. However, in the mammoth muscle, only about 1% of the iron is in the form of heme. The mammoth muscle contains so much iron, because it has seeped in from the surrounding soil-water. This soil-water had to be distinctly acidic and anaerobic, so that the iron would be soluble. The mammoth muscle contains 3.7 times more iron, than that of the Indian elephant. But only about 1% of this iron is in the form of heme. Normally, the iron in the body is nearly all heme-iron. The mammoth muscle contains, therefore, 7.2 times less heme-iron, than that of the Indian elephant.

And the muscle from Dima’s right hind leg contains 8.6% more carbon, than that of the Indian elephant. Hence, much foreign carbon (C) from the surrounding soil-water must have seeped into the body.

The American scientists in southern California do conclude: The mammoth calf Dima was buried in a cold mud stream or has drowned. Then it was silted over in a cold river. Weeks or months later, the carcass was frozen. In this time, some autolysis and extensive leaching of small molecules could go on. The body of the young woolly mammoth might also have been leached sometimes, when the summer was very warm, when the carcass thawed. If ions diffused out of the carcass, small molecules and ions from the surrounding ground water would inevitably have diffused into the carcass!

This means: The body of the mammoth calf has lost much of its original carbon. It was leached out into the water of the surrounding watery, acidic, anaerobic silt. Foreign carbon from the surrounding soil-water then seeped into the body. This foreign carbon then combined itself with the original tissue, still remaining in the body. This contaminated protein is "not amenable to purification by current methods" (Gillespie, R. 1980:241).

That is: The foreign carbon atoms, oozing into the body from the outside, cannot be separated from the body’s original carbon atoms in the laboratory. Some of these foreign carbon atoms, seeping into the carcass from outside, may be younger than the mammoth, some may be just as old, and some may be very much older, containing much "dead" carbon.

What does that mean? – It means: The radiocarbon date will only able to tell us, how much radioactive (¹⁴C) and how much stable carbon (¹²C, ¹³C) the tested sample contains. And these carbon atoms one is able to count now very accurately. But this radiocarbon date certainly does not tell us, when this elephant has really lived. That is why these radiocarbon dates, done on leached-out carcasses, are wrong.

All of these (or most of these) animals of the mammoth fauna, preserved in the flesh, have died and have been buried at the same time and by the same cause. They perished suddenly in the global Flood of Noah’s days in the year 2370 B.C.E., according to Bible chronology. This agrees fully with all the scientific facts, known now.

Only the radiocarbon dates done on samples, which have not been leached by soil-water, should be reliable. Because their original carbon has not been leached out and has not been replaced by foreign carbon from the surrounding soil-water. I am thinking here especially about the Scrolls from the Dead Sea. That are rolled-up parchments. They have usually been wrapped up in a linen-cloth. Then they were stored in a waterproof, closed jar. And this jar, covered with a lid, was stored then in a dry, cool cave. There the scrolls were protected from water and from the burning summer heat at the ground’s surface. This dry heat is destroying the proteins in the sample just as much, as the soil-water, surrounding the carcass.

Baby mammoth Dima. Its feet are still covered by its hair-cover. At other parts of its body the hair has fallen out. Drawing from: R. D. Guthrie, Frozen Fauna of the Mammoth Steppe (1990:8) Fig. 1.5. Dima was a male, 6 to 8 months, or about 1 year old. His body was well preserved, was completely frozen. His gastrointestinal (stomach-intestines) tract contained silt, clay, and gravel, and the remains of plants. The seeds from the gastrointestinal tract are from late summer to early winter. Mineral particles were found in Dima’s gastrointestinal tract, and also throughout the trachea, bronchi, and alveoli of the lungs. The widening of the pulmonary alveoli indicates death by asphyxia (suffocation, drowning). Heart muscle hemorrhaging and other damage was found, suggesting great exertion just before death. Il’inskii interpreted this as running. He died in autumn. R. D. Guthrie (1990:9)