Article / artikel in: Der Mann im Eis. Neue Funde und Ergebnisse. K. Spindler, E. Rastbichler-Zissernig, H. Wilfing, D. zur Nedden, H. Nothdurfter (eds.), 275-278. Wien/ New York 1995

The decorated body of the man from Hauslabjoch. Preliminary results

Eddy van der Velden(1), Leo den Dulk(1), Henk Leenders(2), Koert Dingemans(3), Marius van der Bergh Weerman(3) , Sebastiaan van der Putte(4), Vojislav Vuzevski(5), and Bemard Naafs(1)

1. Department of Dermato-venereology, Academic Hospital Dijkzigt, Erasmus University, Rotterdam
2. Laboratory for Otobiology & Biocompatibility, Biomaterials Research Group, University Hospital, Leiden
3. Department of Pathology, Electron-Microscopy, Academic Medical Centre, University of Amsterdam
4. Department of Pathology, Academic Hospital, University of Utrecht
5. Department of Pathology, Erasmus University, Rotterdam, The Netherlands

Introduction

The oldest tattoos known until the discovery of the man from Hauslabjoch were those found on the bodies of the Egyptian mummies of the priestesses of Hathor (Middle Kingdom, 2040-1780 B.C.)(1,2). The new find makes the tattooing technique some 1200 years older than was assumed. In historical respect, the finds of Pasyryk (Altai mountains, 440-420 B.C.)(3,4) and Qilakitsoq (Greenland, 1475 A.D.)(5) are equally important in showing that tattooing techniques have been used over a very long period and with a wide geographical distribution.
The body of the man from Hauslabjoch shows three tattoos. The first is situated on the left lower back and consists of one group of four lines positioned above two groups of three lines. The second tattoo is located on the inside of the right knee and has the shape of a cross. The third is formed by a group of three lines on the outside of the right ankle.

Material

On December 21, 1992, punch biopsies of 4 mm diameter were taken from all three locations.
Biopsy no. 1 was taken from the lower side of the medial line of the group of four in tattoo no. 1. The biopsy was full thickness into the subcutaneous fat and was divided in two. One half, specimen no. 1A, was fixed in buffered formalin, the other half; specimen no. I B, in liquid nitrogen after saturating it in PBS.
Biopsy no. 2 was taken from the medial side of the horizontal arm of the cross of tattoo no. 2. This biopsy was also divided in two. One half; specimen no. 2A, was fixed in glutaraldehyde, the other half; specimen no. 2B, was frozen.
Biopsy no. 3 was taken from the top of the middle line of tattoo no. 3. The biopsy was divided in two. One half, specimen no. 3A, was fixed in glutaraldehyde, the other half, specimen no. 3B, was frozen.
The frozen samples were stored at -70 °c until use.
The samples were examined in order to answer three important questions: Where in the dermis is the pigment located, what material does it consist of and how has it been introduced into the skin?

Methods

• Sp(ecimen) no. 1A, fixed in buffered formalin, was carefully processed and embedded in paraffin. Slides were cut and stained with H & E and examined.
• Sp. 1 B has not yet been examined.
• Sp. 2A was embedded in Epon, after Osmium-postfixation, for TEM (Transmission Electron Microscopy), which was performed on coupes of 60 nm on copper grids with a Zeiss EM 902.
• Sp. 2B has not yet been examined.
• Sp. 3A was divided in two halves, no. 3A1 and no. 3A2.
  After Osmium fixation and a prolonged embedding procedure in Epon, Sp. 3A1 was used forTEM which was performed on coupes of 60 nm on copper grids with a Philips CM 10.
• Sp. 3A2 has been examined using TEM, SEM (Scanning Electron Microscopy), freeze-fracturing and X-ray microanalysis. TEM was performed on coupes of 80-100 nm on copper grids with a Philips EM 400, fitted with a Tracor Northern Microanalyser. SEM was performed according to the procedure described by Hayat(6). After Critical Point Drying (CPD) with a Balzer CPD-030 the specimen was glued with a carbon glue onto an aluminium stub. It was then shadowed with carbon and examined using a Philips SEM 525M, equipped with a Voyager Tracor Northern Microanalyser. The results were reproduced on slides. After X-raf microanalysis the specimen was sputtered with gold (Balzer MED 010) to get a still better resolution for SEM. Another specimen was freeze-fractured according to the procedure described by Hayat(7).
• Sp. 3B has not yet been examined.

Results

What was noted first in all three of the specimina was the absence of details of the epidermis. Electron microscopy at a low magnification shows that it has completely necrotized (Fig. 1).

Light-microscopic examination of sp. 1A did not reveal any recognizable structure.

TEM of sp. 2A showed abundant, well-preserved collagen fibres in between which pigment particles were seen. These were located in the dermis through the whole of the sp. and some could be seen in cells which had the features of fibroblasts. Distribution and size of the pigment particles showed a remarkable likeness with those of contemporary tattoos (Fig. 2).

TEM of sp. 3A1: The material was found to be very hard to cut, even after a prolonged embedding procedure. As a result, the coupes are folded and fall apart easily, which makes them difficult to evaluate. Collagen bundles are recognizable, in between which crumbled material is seen. The rest of the material is structureless and homogeneous. In TEM the collagen bundies are made up of closely packed, very regularly structured fibrils. The pattern of the transverse bands is extremely weIl-preserved (Fig. 3). The diameter of the fibrils is approx. 60 nm. The crumbled material also turned out to be collagen, but this was non-structured. The homogeneous material consists of criss-cross structures, often filament-like, which are difficult to define but which could be degenerated elastine. In between the components of the matrix structure mentioned above two more structures can be seen:
A. Homogeneous, electron-dense rounded bodies of 100-400 nm diameter. These could weIl be the expected pigment particles.
B. Bodies of 600-900 nm diameter with a clear wall. These could be micro- organisms.

SEM of sp. 3A2: During examination a number of structures was seen, which were found during x-ray microanalysis to consist of CaP (calcium phosphate). The calcium phosphate depositions are present throughout the specimen. During one of the mappings a high concentration of silicium was detected in the upper layer of the specimen. This could be sand particles (Fig. 4a, b). Diffusely spread traces of sulphur and phosphate were also detected.

TEM of sp. 3A2: Using x-ray microanalysis, different trace elements were detected. In some areas, amorphous black deposits were seen in which silicium could be detected. In other areas remnants of the post-fixation osmium and uranyl were found. Furthermore, iron, phosphate and sodium were found among others. Many aggregates of pigment particles were also seen.
Freeze-fracturing was also performed on sp. 3A2, followed by SEM. The procedure for the preparation of the specimen took a considerably longer period of time than for normal skin. Due to the age of the specimen the release of the replica took 6 weeks. The replicas showed a very good skin morphology in which the normal skin structure was visible. The replicas also showed the corneocytes located in layers showing a stepped aspect. Within this normal morphology, in some areas aggregates are found consisting of rounded pigment particles 100- 400 nm in diameter (Fig. 5). The particles are found in the same locations, and have the same shape and size as the electron-dense aggregates seen in TEM.

Discussion

At present no definite statement can be made about the method of application of the tattoos. It is likely that a method similar to well-known primitive methods was used. This may have involved the use of soot. This would either have been rubbed on the skin, after which it was punctured, or vice versa.
In the upper layer of specimen 3A2 particles were present which could be sand particles, since x-ray microanalysis showed the major constituent to be silicates. Moreover, cutting these particles presented major problems, even though a diamond knife was used.
The presence of sand particles on the skin of the Man from Hauslabjoch may support the theory that after his death hot föhn-winds desiccated his body.
Remarkable is the excellent condition of the collagen fibres (Fig. 2). Among them, a few fibroblasts are preserved. In some of these, pigment particles of the same size and morphology as those found in contemporary tattoos are present. At the moment, it is not as yet clear whether these particles are soot or another substance.
The significance of the tattoo is still a subject of speculation. One thing which may be safely assumed however is that the tattoos did not serve any decorative purposes, since they are located on parts of the body which were normally not exposed. However, in primitive societies tattooing is often used as a therapeutic, or a preventive measure. Both localisation and shape show a remarkable resemblance to contemporary therapeutic tattoos as seen in India and Africa.

Summary

Under normal light conditions, the body of the man from Hauslabjoch shows tattoos on three locations. Of each location, a 4-mm punch biopsy was taken. Different electron microscopic examination techniques showed the presence of sand particles, supporting the theory that the body was desiccated by hot winds. Size and morphology of the pigment aggregates found indicate that the method used to apply the tattoos was much the same as contemporary methods. Macroscopic evaluation shows that a remarkable resemblance exists with contemporary therapeutic tattoos found in India and Africa. Examination of the remaining specimina should give additional clues as to what materials were used.

Zusammenfassung

Unter normalen Lichtbedingungen zeigt der Körper des Mannes vom Hauslabjoch drei tätowierte Stellen. Jeder Stelle wurde ein 4-mm Biopt entnommen. Unterschiedliche elektronenmikroskopische Untersuchungstechniken zeigten die Anwesenheit van Sandpartikeln, die die Theorie bestätigt, dass der Körper von Föhnwinden ausgetrocknet worden ist. Grösse und Morphologie der gefundenen Pigmentaggregaten deuten daraufhin, dass die damals zur Aufbringung der Tätowierungen angewendeten Techniken den heutigen sehr ähnlich sind. Die Makroskopie ergibt eine bemerkenswerte Übereinstimmung mit modernen therapeutischen Tätowierungen wie diese in Afrika und Indien angewandt werden. Untersucnung der noch vorliegenden Spezimina sollte zusätzliche Ergebnisse über die verwendeten Grundstoffe ergeben.

Résumé

L:examen du corps de I'homme de Hauslabjoch, exécuté sous des conditions d' éclairage normales a revélé trois lieux tatoués. On a pris un 4 mm-biopte de chacun de ces lieux. l'Examen exécuté à l'aide du microscope électronique, a montré la présence de particles de sable, ce qui confirme l'hypothèse que des tempêtes, causées par le föhn, ont desséché le corps. l'Etendue et le caractère morpholo- gique des agrégats de piment justifient la conclusion que les méthodes employées pour appliquer les tatouages ont été quasi-identiques aux méthodes actuelles. l'Evaluation macroscopique révèle une ressemblance remarquable avec les tatouages thérapeutiques en usage aux lndes et en Afrique. l'Examen des autres spécimen devrait résulter dans des conclusions additionelles en ce qui concerne la nature des matériaux en usage en ce temps-là.

Riassunto

In una luce normale il corpo dell' uomo di Hauslabjoch dimostra tatuaggi in tre posti differenti. In ogni posto è stata fatta una biopsia di 4 mm. Esami elettromicroscopici di tecniche differenti hanno indicato la presenza di granelli di sabbia il chè sorregge la teoria che il corpo è stato disseccato da venti caldi e secchi. Le misure e la morfologia degli aggregati di pigmento dimostrano che la tecnica di tatuaggi di allora non è certamente molto differente da quella di oggi. La macroscopia fa vedere una notevole assomoglianza con i tatuaggi terapeutici moderni come li troviamo in India e in Africa. Ulteriori esami delle rimanenti specimina potranno informarci sul materiale allora usato.

1. Keimer, L., Remarques sur le tatouage dans I'Égypte ancienne, le Caire/Paris: Imprimerie de I'Institut Français d'Archéologie Orientale, 1948.
2. Helck, W, Westendorf, W, Lexicon of Aegyptology 1985; 6: 145.
3. Rudenko, S. I., Tätowierungen asiatischer Eskimos. Sovjetskaja Etnografija 1949; 1: 149-154.
4. Rudenko, S. I., Die älteste "skythische" Tätowierung. Sovjetskaja Etnografija 1949; 3: 133-143.
5. Hansen, J. P. H., Meldgaard, J., Nordquist, J., Qilakitsoq. De Grönlandske Mumier frå 1400-tallet. Kopenhagen 1985.
6. Hayat, M. A., Principles and Techniques of Scanning Electron Microscopy, v: 2, New York 1974.
7. Hayat, M. A., Principles and Techniques of Electron Microscopy, London 1984

Fig. 1. Sp. 3A, necrotized epidermis (top), TEM 65.5x

Fig.2. Sp. 2A, pigment particles in fibroblast, TEM 21,000x

Fig.3. Sp.3A1, well-preserved collagen bundles and unidentified structure (bacterium?). TEM, 69,000x

Fig. 4a. Sp. 3A2, silicate particle, SEM 4580x.

Fig. 4b. Sp. 3A2, x-ray microanalysis with Si-peak

Fig. 5. Sp. 3A2, freeze-fractured showing pigment particles, SEM 21,000x