Ketchum (2013) Figure 5 Shows Sample 26 Hair to be from a Black Bear

Ketchum et al. (2013) exhibited as their Fig. 5 hair photomicrographs of Sample 26 (the Smeja find) and a human hair comparison.  They claimed that:  “Most   of   the   submitted   hairs  were   not microscopically consistent with any of the hairs from the reference collection of common animal hairs that included human, cat, dog, cow, horse, deer, elk, antelope, moose, sheep, fox, bear, coyote, wolf, rat, mouse, monkey, beaver, squirrel, llama and others.” (emphasis mine), and that Figs. 5B Left and 5C Right are from a sasquatch.  The hair analyst was David W. Spence of Southwest Institute of Forensic Sciences in Dallas, Texas.

In view of the controversy around this study, especially Sample 26, which was independently analyzed as black bear by three additional laboratories, these photomicrographs should have been more carefully examined long ago.  “Better late than never.”

Identification of hairs by microscopy can be subjective, because there is variation among individuals and even among different hairs from a single individual, such that forensic determination in some criminal cases has been challenged and overturned.  Never the less, it remains a good screening tool, and in many cases it is definitive for identifying or classifying wildlife, especially when the number of possibilities is small. 

We are fortunate that, in the case of Ketchum et al. Sample 26, we are only called upon to judge whether the hair in their Fig. 5 is from a primate or a bear, which have very different hair morphologies.  With a small amount of image enhancement (limited to brightness, contrast, and sometimes also color), this call is very obvious.  Enhancements of this sort can be made at the microscope level before photographing with included software or later with Photo Shop or other image software such as that included with Microsoft Word®.  We used all three approaches here.  Understand that these enhancements are not the same as the attempts to bring out of focus pictures into clarity as is done by Scott Carpenter with Blurity!.  Blurity! is for motion blurring, not out of focus blurring.  All of the photomicrographs in Figs. 1-13 are in focus pictures, some of which were initially just too dark to resolve details before enhancement

Fig. 1 is from the Ketchum et al. (2013) paper, their Figure 5B Left, and purports to show a Sample 26 sasquatch hair.  Fig. 2 is basically the same photograph, extended in length from p. 109 of Carpenter (2019), who supports her interpretation.  Both figures have been enhanced in brightness and contrast here.  By comparison to Figs. 3-7 it is very clear that the Ketchum et al. (2013) Fig. 5B Left photomicrograph matches black bear extremely well.  Shared characteristics are: continuous, nodose, corpuscular medulla of disc shaped cells, and Medullary Index (MI) of 0.25-0.50 for guard hairs.   Medullary Index = Diameter of medulla / Diameter of shaft.  Underfur in reference Figs. 5 and 7, exhibit lower MI and single ladders of the same corpuscular disc shaped cells. 

In contrast,  primate hairs usually show discontinuous, thinner medulla (MI < 0.2, often much less) or no medulla as shown in Figs. 8 for human, 10 for orangutan, and 11 for gorilla.  Fig. 9 for chimpanzee shows a more continuous thicker medulla, but because of the intense pigmentation, the real width of the medulla is somewhat uncertain.  For such samples the microscopist must increase the light intensity in transmission or go to dark field or crossed polarizers, which was not done in Fig. 9.  Crossed polarizers bring out different birefringent colors for otherwise indistinguishable features. (See Fig. 10 for gorilla).

Cuticle comparisons are also insightful.  Fig. 12. is from the Ketchum S26 (Fig. 5C Right).  Fig. 13 is a black bear cuticle.  These are extremely similar, in fact, they are indistinguishable in the context of species identification.

In summary, it is hard to imagine why Mr. David Spence did not recognize the unmistakable features of bear hair for Sample 26.  But then, it’s just one of many red flags that Ketchum et al. did not recognize in their quest to prove their hypothesis.  

Note: All photomicrographs are for research and education only, "Fair Use."

  

Fig. 1.  "Sasquatch" Ketchum et al. (2013). Fig. 5B (Left)  Contrast and brightness adjusted.

Fig. 2. "Sasquatch." Carpenter (2019, p. 109).  Contrast and brightness adjusted. 

Fig.3  Black bear. Hart private collection.

Fig. 4.  Black bear.  Hart private collection.

 Fig. 5.  Bear underfur. Deedrick (2004).  

Fig. 6  Black bear guard hair. Alaska Fur ID Project.  Contrast, brightness, and color adjusted.

Fig. 7.  Black bear underfur.  Alaska Fur ID Project.

Fig. 8.  Human.  Public Domain.  Variety of medulla: none, discontinuous, nearly continuous.

Fig. 9.  Chimpanzee.  Chris Murphy.  Left original.  Right brightness, contrast, and color enhanced.

Fig. 10.  Orangutan.  McCrone Associates.  Brightness and contrast adjusted.  Arrow points to thin medulla.

Fig. 11. Gorilla.  Crossed polarizers.  McCrone Associates.  Brightness and contrast adjusted.  Red arrows point to thin medulla (yellow).

Fig.  12.  "Sasquatch."  Ketchum et al. (2013) Sample 26. Fig. 5 C Right, cuticle.

Fig. 13.  Black bear cuticle.  Hart private collection.

References

Alaska Fur ID Project.  https://alaskafurid.wordpress.com/

Carpenter S. (2019) Truth Denied: The Sasquatch DNA Study.   Self-Published.

Deedrick D. W. (2004)  Microscopy of Hair Part II: A Practical Guide and Manual for Animal Hairs.  Forensic Science Communications.  July 2004 - Volume 6 - Number 3. 
https://archives.fbi.gov/archives/about-us/lab/forensic-science-communications/fsc/july2004/research/2004_03_research02.htm 

Ketchum M. S. et al. (2013) Novel North American hominins: next generation sequencing of three whole genomes and associated studies. DeNovo  1:1.  

http://sasquatchgenomeproject.org/sasquatch_genome_project_002.htm

Sample 26 mtDNA Electropherograms Show Degradation/Contamination

Figure 1.  Ketchum S26 Electropherograms from SGP website.  Green bars above each peak proportional to quality of read.  Peak color indicates base:  A G  T  C.  For research and education only, "Fair Use." 

Melba Ketchum has consistently claimed in her paper [1], on radio, on TV, on the Sasquatch Genome Project (SGP) website [2], on YouTube [3],  and at every other opportunity,  that her samples are not contaminated or degraded, in spite of very anomalous results, many of which are easily explained by such.  She posted Sample 26 (the Smeja find) raw data mtDNA electropherogram files on the 
SGP website.  I downloaded all 272 such files, which require special software to open, examine and assemble into a complete mtDNA sequence.  I purchased DNABaser software from Heracle Biosoft to do just that.  It cost $159 and runs on my PC.  The above Fig. 1 is a screen grab of  six randomly selected, representative electropherograms of the 272.

This data was obtained by the Sanger Dideoxy Sequencing Method, outlined in Fig. 2.

Figure 2.  Sanger Dideoxy Method.  After Wikipedia.  For research and education only, "Fair Use."   

In this method, segments are elongated (as complementary strands) and terminated with "fluorochrom" (i.e. fluoroprobe, or fluorescent tag) nucleotides with different fluorescent wavelengths for each of the four nucleotides (A, G, T, C).  The successive elongated segments are separated by electrophoresis according to their length, then detected and identified by their characteristic fluorescent wavelength (stimulated by a laser).  Software makes the plots as represented schematically in Fig. 2 lower right or actually in Fig. 1 and generates a sequence (Fig. 1 below each electropherogram, Fig. 2 lower right).

All but the shortest (few hundred bp) sequences must be sequenced in segments, and then assembled by computer, based on overlaps, as shown in Fig. 3.

Figure 3.  Assembly.  Overlaps underlined.  After Wikipedia.  For research and education only, "Fair Use."


Mitochondrial DNA assembly is handily done on a PC.  Only very large data sets (e.g. whole nuclear genomes) from next generation sequencing, require a "super computer" or a network of linked servers to assemble.

Sanger sequencing and subsequent assembly does not work for severely degraded DNA, whether from a single or multiple sources.  This is because of the presence of sequence fragments which are self-primed and which are never the less elongated and tagged with a fluorescent probe.  These fragments coelute with the fragments of interest as shown in Fig. 1 ("Poor").  The "Excellent" electropherogram has no such coelution:  each peak stands alone and is separated from its neighbors so that the fluorescent detector can make an unambiguous base call (A, G, T, or C). The "Good" electropherogram can also be used because coeluting peaks are much smaller in most cases and can be ignored.  The few ambiguous, double peaks will hopefully be clarified by another overlapping sequence fragment.  Otherwise these few remain unknown bases (or possibly polymorphisms).  The "Poor" electropherograms are totally uninterpretable.  They have multiple overlapping peaks of comparable intensity.  Those "?" electropherograms might have some salvageable sequence runs, but should be considered dubious at best.   So, two usable e-grams out of these six.  A well collected and preserved single species DNA sample would produce only usable e-grams.

Unfortunately, the 272 electropherograms contained only 47 that could be assembled into a sequence of 5260 bp out of an expected 16,568 bp human mitochondrial genome.  The remaining 225 electropherograms contained only 11 which could be assembled into a 1923 bp sequence.  Other assemblies were shorter still.

The first assembly (47 e-grams) aligned with the published S26 mtDNA sequence [4], base positions 11401-16551, only 96% with 74 gaps.   The second best assembly (11 e-grams) aligned with published [4] S26 base positions 13718 - 15435 only 97% with 27 gaps.  This is rather poor agreement between these raw data and the published S26 sequence [4].   The 47 e-gram sequence aligned with the 11 e-gram sequence only 93% with 1643 identities and 69 gaps.

Thus, only 32% of the mtDNA genome could be sequenced with these data, and the sequence agreed very poorly with the published S26 sequence produced by Family Tree DNA [4].  FTDNA assumes a human sequence and uses human primers, so DNA of any other species present would not be sequenced. 

These data call into question the mtDNA sequencing and haplogroup designation of Sample 26, and hence also the Ketchum claim that this sample represents a human-unknown primate hybrid.  Most likely, the sample is a black bear contaminated by one or more humans and severely degraded, as shown in my previous blogs. 

References

[1]  Ketchum M. S. et al. (2013) Novel North American hominins: next generation sequencing of three whole genomes and associated studies. DeNovo 1:1.  Online only: http://sasquatchgenomeproject.org/ 

[2] http://sasquatchgenomeproject.org/sasquatch_genome_project_002.htm

[3]  https://youtu.be/yW4yPPXPfSw?t=1

[4]  http://sasquatchgenomeproject.org/linked/supplemental-data-2-mito-mutation-chart-complete-final.pdf