Putting ochre to the test: replication studies of adhesives that may have been used for hafting tools in the Middle Stone Age

Substantial frequencies of Middle Stone Age (MSA) lithics from Rose Cottage and Sibudu Caves in South Africa have red ochre on their proximal and medial portions. Residue studies suggest that the tools were hafted and that the ochre may be part of the adhesive used for hafting the tools. Replication studies show that ochre is indeed a useful loading agent for adhesive; however, there are other potential loading agents. It is also possible to use unloaded plant resin, but this agent is brittle and difficult to work with. It appears that people living in the MSA had wide knowledge of ingredients suitable for hafting tools, and that they chose different adhesive recipes because of the required properties of the adhesive. Brittle, unloaded adhesive allows a projectile head to disengage its haft and implant itself in an animal; robust adhesive keeps a spearhead safely in its shaft.     

Lap Shear and Impact Testing of Ochre and Beeswax in Experimental Middle Stone Age Compound Adhesives

The production of compound adhesives using disparate ingredients is seen as some of the best evidence of advanced cognition outside of the use of symbolism. Previous field and laboratory testing of adhesives has shown the complexities involved in creating an effective Middle Stone Age glue using Acacia gum. However, it is currently unclear how efficient different adhesive recipes are, how much specific ingredients influence their performance, and how difficult it may have been for those ingredients to be combined to maximum effect. We conducted a series of laboratory-based lap shear and impact tests, following modern adhesion testing standards, to determine the efficacy of compound adhesives, with particular regard to the ingredient ratios. We tested rosin (colophony) and gum adhesives, containing additives of beeswax and ochre in varying ratios. During both lap shear and impact tests compound rosin adhesives performed better than single component rosin adhesives, and pure acacia gum was the strongest. The large difference in performance between each base adhesive and the significant changes in performance that occur due to relatively small changes in ingredient ratios lend further support to the notion that high levels of skill and knowledge were required to consistently produce the most effective adhesives.     

Evidence of hunting and hafting during the Middle Stone Age at Sibidu Cave, KwaZulu-Natal, South Africa: a multianalytical approach

Points and point fragments from Middle Stone Age layers (dated to between 50,000 and 60,000 years ago) from Sibudu Cave, KwaZulu-Natal, South Africa, were examined to establish whether they were used as hafted spearheads for hunting. A multi-analytical approach was followed, using macrofracture analysis, use-wear analysis, and residue analysis. In addition to the analytical processes, an experimental project tested the results of the macrofracture analysis on local raw materials. The study shows that points from Sibudu Cave were indeed hafted and used as hunting tools. It was further established that plant twine was probably the preferred binding material to attach the points to wooden hafts. Resin may have been used as an adhesive in combination with the binding material. A detailed examination of the ochre distribution on the points confirmed that ochre was also part of the hafting arrangement. The need to use a dependable methodology for the recognition of hunting and hafting traces on stone points from the southern African Middle Stone Age context is briefly discussed.     

Traditional Glue,Adhesive and Poison Used for Composite Weapons by Ju/’hoan San in Nyae Nyae,Namibia. Implications for the Evolution of Hunting Equipment in Prehistory

Ju/’hoan hunters from Nyae Nyae, near Tsumkwe in Namibia, demonstrate the manufacture of three fixative pastes made from plant extracts, and poison made from grubs and plant extracts. Ammocharis coranica and Terminalia sericea produce simple glue. Ozoroa schinzii latex mixed with carbonized Aristeda adscensionis grass is a compound adhesive. Composite poison is made from Chrysomelid grub viscera mixed with salivary extracts of Acacia mellifera inner bark and the tuber sap of Asparagus exuvialis. In order to document potential variability in the chaîne opératoire, and to eliminate inherent biases associated with unique observations, we studied manufacturing processes in three separate Nyae Nyae villages. Although there are methodological similarities in the Nyae Nyae area, we observed a few differences in contemporary traditions of poison manufacture. For example, some hunters make powder from Asparagus exuvialis tuber sap by boiling, reducing, hardening and grinding it, while others simply use heated sap. The Ju/’hoan hunting kit provides insights for archaeologists, but we must exercise caution when looking for continuity between prehistoric and historical technical systems. Some traditions have been lost to modern hunters, while others are new. We should also expect variability in the Stone Age because of geographically restricted resources. Simple glue, compound adhesive, and poison recipes identified in the Stone Age have no modern equivalents. By about 60,000 years ago at Diepkloof, simple glue was used for hafting tools, but at similarly-aged Sibudu there are recipes that combine red ochre powder with plant and/or animal ingredients. At Border Cave, novel poisons and compound adhesives were used in the Early Later Stone Age. It is possible that the complexity that we record in the manufacture of fixative pastes and poison used by Ju/’hoan hunters represents a hafting system both similar to and different from that observed at the Stone Age sites of Diepkloof, Sibudu, and Border Cave.     

古人类对赭石的利用行为在其演化中的意义

长期以来赭石利用行为被视为人类行为现代性的标志之一,受到国内外考古学界的普遍关注。本文回溯和梳理了全球背景下赭石利用的起源、发展及其与人类演化史的关系。在现代人广泛分布于全球之后,赭石利用行为更加丰富和多样化地出现在各地,然而现有考古证据表明该行为并不是解剖学意义上的现代人突变性的发明。赭石利用不能被单纯地定义为现代人行为,而应是有着长久演化积累的现代性行为之一。在长期传播与演化过程中,赭石的功能从意识形态、艺术表达等逐渐扩展到作为矿物成分被用于实际生产生活。赭石的利用历史可追溯到中更新世中期,但其广泛分布仍与晚更新世以来现代人的广泛扩散直接相关,对于理解现代人的意识形态、社会组织方式以及艺术表达、精神文化发展都具有重要的意义。国内目前所发表的相关考古学证据相对较少,以下马碑遗址为代表的材料,也恰处于现代人在全球广泛扩散的窗口期,并伴有进步的细小石器镶嵌使用的证据,成为认识东亚现代人行为的关键性考古证据。     

Middle Stone Age Ochre Processing and Behavioural Complexity in the Horn of Africa: Evidence from Porc-Epic Cave,Dire Dawa,Ethiopia

Ochre is a common feature at Middle Stone Age (MSA) sites and has often been interpreted as a proxy for the origin of modern behaviour. However, few ochre processing tools, ochre containers, and ochre-stained artefacts from MSA contexts have been studied in detail within a theoretical framework aimed at inferring the technical steps involved in the acquisition, production and use of these artefacts. Here we analyse 21 ochre processing tools, i.e. upper and lower grindstones, and two ochre-stained artefacts from the MSA layers of Porc-Epic Cave, Dire Dawa, Ethiopia, dated to ca. 40 cal kyr BP. These tools, and a large proportion of the 4213 ochre fragments found at the site, were concentrated in an area devoted to ochre processing. Lower grindstones are made of a variety of raw materials, some of which are not locally available. Traces of use indicate that different techniques were employed to process ochre. Optical microscopy, XRD, μ-Raman spectroscopy, and SEM-EDS analyses of residues preserved on worn areas of artefacts show that different types of ferruginous rocks were processed in order to produce ochre powder of different coarseness and shades. A round stone bearing no traces of having been used to process ochre is half covered with residues as if it had been dipped in a liquid ochered medium to paint the object or to use it as a stamp to apply pigment to a soft material. We argue that the ochre reduction sequences identified at Porc-Epic Cave reflect a high degree of behavioural complexity, and represent ochre use, which was probably devoted to a variety of functions.     

An Experimental Study of Hafting Adhesives and the Implications for Compound Tool Technology

Experimental studies of hafting adhesives and modifications to compound tool components can demonstrate the extent to which human ancestors understood and exploited material properties only formally defined by science within the last century. Discoveries of Stone Age hafting adhesives at archaeological sites in Europe, the Middle East, and Africa have spurred experiments that sought to replicate or create models of such adhesives. Most of these studies, however, have been actualistic in design, focusing on replicating ancient applications of adhesive technology. In contrast, this study tested several glues based on Acacia resin within a materials science framework to better understand the effect of each adhesive ingredient on compound tool durability. Using an overlap joint as a model for a compound tool, adhesives formulated with loading agents from a range of particle sizes and mineral compositions were tested for toughness on smooth and rough substrates. Our results indicated that overlap joint toughness is significantly increased by using a roughened joint surface. Contrary to some previous studies, there was no evidence that particle size diversity in a loading agent improved adhesive effectiveness. Generally, glues containing quartz or ochre loading agents in the silt and clay-sized particle class yielded the toughest overlap joints, with the effect of particle size found to be more significant for rough rather than smooth substrate joints. Additionally, no particular ochre mineral or mineral mixture was found to be a clearly superior loading agent. These two points taken together suggest that Paleolithic use of ochre-loaded adhesives and the criteria used to select ochres for this purpose may have been mediated by visual and symbolic considerations rather than purely functional concerns.     

A Milk and Ochre Paint Mixture Used 49,000 Years Ago at Sibudu,South Africa

Gas chromatography/mass spectrometry, proteomic and scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM/EDS) analyses of residue on a stone flake from a 49,000 year-old layer of Sibudu (South Africa) indicate a mixture of ochre and casein from milk, likely obtained by killing a lactating wild bovid. Ochre powder production and use are documented in Middle Stone Age South African sites but until now there has been no evidence of the use of milk as a binder. Our analyses show that this ochre-based mixture was neither a hafting adhesive nor a residue left after treating animal skins, but a liquid mixture consisting of a powdered pigment mixed with milk; in other words, a paint medium that could have been applied to a surface or to human skin. The significance of our finds also lies in the fact that it establishes the antiquity of the use of milk as a binder well before the introduction of domestic cattle in South Africa in the first millennium AD.     

Synthesis of an ochre suppressor tRNA gene and expression in mammalian cells.

We have used site-specific mutagenesis to change the anticodon of a Xenopus laevis tyrosine tRNA gene so that it would recognize ochre codons. This tRNA gene is expressed when amplified in monkey cells as part of a SV40 recombinant and efficiently suppresses termination at both the ochre codon separating the adenovirus 2 hexon gene from a 23-kd downstream gene and the ochre codon at the end of the NS1 gene of influenza virus A/Tex/1/68. Termination at an amber codon of a NS1 gene of another influenza virus strain was not suppressed by the (Su+) ochre gene suggesting that in mammalian cells amber codons are not recognized by ochre suppressor tRNAs. Finally, microinjection into mammalian cells of both (Su+) ochre tRNA genes and selectible genes containing ochre nonsense mutations gives rise to colonies under selective conditions. We conclude that it should be possible to isolate a wide assortment of mammalian cell lines with ochre suppressor activity.     

In vivo translation of amber and ochre codons in Escherichia coli

Summary By making certain assumptions, we have provided evidence which indicates that: amber and ochre codons function in vivo, the bulk of the E. coli mRNA is polycistronic, intercistronic space may correspond to no more than 12–24 nucleotides and ochre is used twenty five times more frequently than amber, for chain termination.     

Characterization of amber and ochre suppressors in Salmonella typhimurium.

Amber and ochre suppressor mutations in Salmonella typhimurium were selected. The amino acid insertions directed by the suppressors were inferred from suppression patterns of Escherichia coli lacI amber mutations. These amber mutations only respond to nonsense suppressors that direct the insertion of particular amino acids. Four Salmonella amber suppressors characterized insert serine, glutamine, tyrosine, and (probably) leucine. Of the three ochre suppressors characterized, two direct the insertion of tyrosine and one directs that of lysine. Of the three amber and two ochre suppressors which have been mapped by phage P22 cotransduction, all are located in the same relative position on the Salmonella map as the analogous E. coli suppressors are on the E. coli map.     

Extending the chronology of deposits at Blombos Cave, South Africa, back to 140 ka using optical dating of single and multiple grains of quartz

Optically stimulated luminescence (OSL) measurements are reported for both single aliquots (of two different sizes) and single grains of quartz from deposits within Blombos Cave. Ages have been obtained for six sediments from the Middle Stone Age (MSA) occupation levels and for two sterile sands, one underlying the archaeological sediment and one overlying the Later Stone Age occupation levels. The ages for the archaeological sediments were obtained from single-grain measurements that enabled unrepresentative grains to be rejected. The MSA occupation levels have ages that, within error limits, are in stratigraphic order and fall between the OSL age for the oldest dune sand (143.2+/-5.5 ka) and a previously published OSL age for the sterile sand ( approximately 70 ka) that separates the Middle and Later Stone Age deposits. The earliest MSA archaeological phase, M3, from where fragments of ochre were found as well as human teeth, is dated to 98.9+/-4.5 ka, coinciding with the sea-level high of oxygen isotope substage 5c. The cave then appears to be unoccupied until oxygen isotope substage 5a on the basis of four OSL ages for archaeological phase M2, ranging from 84.6+/-5.8 to 76.8+/-3.1 ka; these levels contained large hearths and bone tools. An age of 72.7+/-3.1 ka was obtained for the final MSA archaeological phase, M1, from which deliberately engraved ochre and shell beads were recovered along with bifacial stone points. We conclude that the periods of occupation were determined by changes in sea level, with abundant sources of seafood available in times of high sea level and with the cave being closed by the accumulation of large dunes during periods of low sea level, such as during oxygen isotope stages 4 and 6.     

supN ochre suppressor gene in Escherichia coli codes for tRNALys.

We describe the cloning and nucleotide sequence of a new tRNALys gene, lysV, in Escherichia coli. An ochre suppressor allele of this gene, supN, codes for a tRNALys with anticodon UUA, presumably derived by a single base change from a wild-type UUU anticodon. The sequence of the supN tRNALys is identical to the sequence of ochre suppressor tRNAs encoded by mutant alleles at the lysT locus. This locus, which contains the two previously known tRNALys genes of E. coli, is located far from the lysV locus on the chromosome.     

Site-specific mutagenesis of Escherichia coli gltT yields a weak, glutamic acid-inserting ochre suppressor

Of all the Escherichia coli tRNA genes that can give rise to an amber or an ochre suppressor by a single-nucleotide mutation, only the tRNAGlu genes have not been observed to do so. A study of the relationship between the sequences of tRNAs and the codons they translate predicts that the ochre suppressor derived from tRNAGlu would function very poorly on the ribosome. We have used site-specific mutagenesis to create the gene for such a tRNA in order to test this prediction. We cloned the tRNAGlu-Suoc gene into a high copy number plasmid, under control of the lacUV5 promoter. The mutant tRNA suppresses both amber and ochre nonsense mutations. As predicted, it is less efficient than other suppressors expressed under similar conditions.     

Evaluating the Photoprotective Effects of Ochre on Human Skin by In Vivo SPF Assessment: Implications for Human Evolution,Adaptation and Dispersal

Archaeological indicators of cognitively modern behaviour become increasingly prevalent during the African Middle Stone Age (MSA). Although the exploitation of ochre is viewed as a key feature of the emergence of modern human behaviour, the uses to which ochre and ochre-based mixtures were put remain ambiguous. Here we present the results of an experimental study exploring the efficacy of ochre as a topical photoprotective compound. This is achieved through the in vivo calculation of the sun protection factor (SPF) values of ochre samples obtained from Ovahimba women (Kunene Region, Northern Namibia) and the Palaeozoic Bokkeveld Group deposits of the Cape Supergroup (Western Cape Province, South Africa). We employ visible spectroscopy, energy-dispersive X-ray fluorescence (ED-XRF), X-ray diffraction (XRD) and granulometric analyses to characterise ochre samples. The capacity of ochre to inhibit the susceptibility of humans to the harmful effects of exposure to ultraviolet radiation (UVR) is confirmed and the mechanisms implicated in the efficacy of ochre as a sunscreen identified. It is posited that the habitual application of ochre may have represented a crucial innovation for MSA humans by limiting the adverse effects of ultraviolet exposure. This may have facilitated the colonisation of geographic regions largely unfavourable to the constitutive skin colour of newly arriving populations.     

Shells and ochre in Middle Paleolithic Qafzeh Cave,Israel: indications for modern behavior

Qafzeh Cave, the burial grounds of several anatomically modern humans, producers of Mousterian industry, yielded archaeological evidence reflecting their modern behavior. Dated to 92 ka BP, the lower layers at the site contained a series of hearths, several human graves, flint artifacts, animal bones, a collection of sea shells, lumps of red ochre, and an incised cortical flake. The marine shells were recovered from layers earlier than most of the graves except for one burial. The shells were collected and brought from the Mediterranean Sea shore some 35 km away, and are complete Glycymeris bivalves, naturally perforated. Several valves bear traces of having been strung, and a few had ochre stains on them.     

Serine substitutions caused by an ochre suppressor in yeast.

The suppressor SUQ5 in yeast can cause the production of approximately 10 to 20% of the normal amount of iso-l-cytochrome c when coupled to the ochre (UAA) mutants cyc1–2 and cyc1–72. The iso-l-cytochromes c contain residues of serine at positions that correspond to the sites of the ochre codons. SUQ5 is efficient only in strains having the non-Mendelian factor ψ⁺, although the low amount of suppressed iso-l-cytochrome c from a ψ⁻SUQ5 cyc1–72 strain was also shown to contain serine at the ochre site. Thus SUQ5 differs from the eight other characterized suppressors of UAA in yeast, which were previously shown to insert residues of tyrosine at ochre sites (Gilmore et al., 1971) and which are only effective in strains haying the non-Mendelian factor ψ⁻, since they generally cause inviability in the ψ⁺ state. Like the tyrosine-inserting suppressors, SUQ5 can also act on another ochre allele cyc1–9, but with a very low efficiency of approximately 0.4%, while it does not appear to act at all on amber (UAG) mutants. SUQ5 was found to be 6.4 cM (centiMorgans) from tyr7 on chromosome XVI. It is suggested that the gene product of SUQ5 is serine tRNA.     

Complete set of orthogonal 21st aminoacyl-tRNA synthetase-amber, ochre and opal suppressor tRNA pairs: concomitant suppression of three different termination codons in an mRNA in mammalian cells

We describe the generation of a complete set of orthogonal 21st synthetase-amber, ochre and opal suppressor tRNA pairs including the first report of a 21st synthetase-ochre suppressor tRNA pair. We show that amber, ochre and opal suppressor tRNAs, derived from Escherichia coli glutamine tRNA, suppress UAG, UAA and UGA termination codons, respectively, in a reporter mRNA in mammalian cells. Activity of each suppressor tRNA is dependent upon the expression of E.coli glutaminyl-tRNA synthetase, indicating that none of the suppressor tRNAs are aminoacylated by any of the twenty aminoacyl-tRNA synthetases in the mammalian cytoplasm. Amber, ochre and opal suppressor tRNAs with a wide range of activities in suppression (increases of up to 36, 156 and 200-fold, respectively) have been generated by introducing further mutations into the suppressor tRNA genes. The most active suppressor tRNAs have been used in combination to concomitantly suppress two or three termination codons in an mRNA. We discuss the potential use of these 21st synthetase-suppressor tRNA pairs for the site-specific incorporation of two or, possibly, even three different unnatural amino acids into proteins and for the regulated suppression of amber, ochre and opal termination codons in mammalian cells.