Recovery of Protein and DNA Trapped in Stone Tool Microcracks

Microcracks produced during the manufacture of stone tools may harbour ancient biological residues. Our studies test this hypothesis using light, scanning electron, fluorescence, and confocal microscopy to characterize microcracks produced in obsidian microblades by pressure flaking. Cell-sized fluorescent latex beads penetrated microcracks to depths exceeding 50 μm below the microblade surface. Fluorescently labelled blood protein and DNA were trapped in microcracks within 5 min of exposure. Washing artifacts with water did not remove blood residues from microcracks. However, 60–80% of trapped blood protein and DNA were released by an 18-hour incubation in 4M guanidine hydrochloride or by sonication in 5% ammonium hydroxide. These experiments suggest that residues trapped in artifact microcracks represent an important and often overlooked source of ancient DNA and protein.     

Analysis of DNA from Ethnoarchaeological Stone Scrapers

Analysis of DNA residues on stone tools provides a direct method of determining what the tools were used on. However, little is known about the taphonomy of DNA on tools. The discovery of present-day, stone-tool using hide workers in Ethiopia therefore provides a unique opportunity to study the survival and authenticity of DNA residues on stone tools. We collected stone scrapers from contexts ranging from excavated to confirmed use, as well as unused scrapers to test for the presence of DNA residues. We amplified segments of the mitochondrial genome with PCR to determine which animal species a tool was used on. We were able to recover authentic DNA from scrapers of known use and from a subset of excavated scrapers. We did, however, also obtain DNA unrelated to the use of the tools. Thus, caution must be used when interpreting results of DNA analysis of stone tools.     

Evaluating protein residues on Gainey phase Paleoindian stone tools

Blood protein analysis provides a method for acquiring and interpreting archaeological data bearing on human–animal relationships. The present study makes use of cross-over immunoelectrophoresis (CIEP) and a large sample of stone tools (N = 130) from an early Paleoindian site pertaining to the Gainey phase (ca. 11,200 BP) of the Midwestern USA. Results are used to interpret toolkit organization and site structure, and indicate that hafted tools with longer use-life potentialities are more likely to be associated with preserved residues than are tools of a more ad hoc or situational character. Protein residues derived from cervids are the largest category of identified samples, a result consistent with an interpretation that these animals were important first-line resources of Gainey phase populations. The identification of caribou (Rangifer sp.) is notable. Interpretations of subsistence importance, however, must be tempered by an appreciation of the possible confounding effects of tool manufacture and maintenance in early Paleoindian contexts.     

Small things in perspective: the contribution of our blind tests to micro-residue studies on archaeological stone tools

Our blind tests are distinctive for they were conducted on replicated stone tools used for a variety of tasks that included the processing of animal remains and plants. The analyst was required to differentiate an array of residues from microscopic morphological characteristics, using light microscopy. The original aim of our first tests was to assess the analyst's ability to identify a variety of plant and animal residues, but issues and problems that arose during the testing process made it clear that greater value might be gained from the lessons that we learnt about methodology and the direction for future micro-residue research. We show that problems identified during our first tests stimulated research. Amongst other things, we learnt to distinguish plant and animal remains more confidently than previously. Our residue analyses are firmly embedded in wider archaeological research and our tests help to explain why there are sometimes contradictions between the evidence from archaeologically recovered remains and residues on stone tools. A further outcome of the tests is that we have adopted a multi-stranded approach that provides a cautious, but secure strategy for identifying and interpreting use-residues. Our studies of contaminants through replications have also been invaluable for distinguishing incidental residues from use-related residues.     

DNA from ancient stone tools and bones excavated at Bugas-Holding,Wyoming

DNA residues may preserve on ancient stone tools used to process animals. We studied 24 stone tools recovered from the Bugas-Holding site in northwestern Wyoming. Nine tools that yielded DNA included five bifaces, two side scrapers, one end scraper, and one utilized flake. The excavators did not handle three of these tools with bare hands, and they were sealed in separate plastic bags to minimize contact with modern DNA. Five sediment samples did not yield mammalian DNA. One tool contained DNA from three species, and these templates competed during polymerase chain reaction (PCR) amplification. DNA identifications suggested a butchery pattern indicative of a human population under resource stress or a broader range of industrial activities at Bugas-Holding. We describe several improvements in DNA residue analysis, including a more effective DNA recovery protocol, methods to measure sensitivity and inhibition of PCR in each sample, and strategies to surmount competition between templates during PCR.     

Raman spectroscopy and scanning electron microscopy confirm ochre residues on 71 000‐year‐old bifacial tools from Sibudu,South Africa

Micro‐residue analysis of stone tools is generally performed with optical light microscopy and the visual observations are then compared with experimental, replicated pieces. This paper complements such archaeological research by providing physico‐chemical evidence. Raman spectroscopy and scanning electron microscopy have been used to confirm the presence of hematite on red‐stained medial and proximal parts of 71 000‐year‐old Still Bay bifacial tools from Sibudu Cave. Our results confirm the conclusion from optical light microscopy that the tools were hafted with an ochre‐loaded adhesive. However, we point to some instances when hematite residues are incidental or may be inclusions in the rock used to make the stone tools.     

Developing FTIR Microspectroscopy for the Analysis of Animal-Tissue Residues on Stone Tools

The analysis of microscopic residues on stone tools provides one of the most direct ways to reconstruct the functions of such artifacts. However, new methods are needed to strengthen residue identifications based upon visible-light microscopy. In this work, we establish that reflectance Fourier-transform infrared microspectroscopy (FTIRM) can be used to document IR spectra of animal-tissue residues on experimental stone tools. First, we present a set of reflectance FTIRM standards for the most commonly identified animal-tissue residues on stone tools: skin, meat, fat, hair, blood, feather barbules, fish scales, and bone. We provide spectral peak assignments for each residue and demonstrate that high-quality reflectance FTIRM spectra can be generated under ideal circumstances. Second, we document the spectra for these residues when they are located on a stone substrate such as flint or obsidian. We discuss procedures for correcting spectra that are affected by specular reflection and explain the effects of spectral interference from the stone. Our results show that reflectance FTIRM is sensitive to small intra-sample differences in composition. This means that it will record the effects of decomposition in ancient residues. The methodological developments we present here will help lithic residue analysts incorporate in situ reflectance FTIRM into their analysis protocols to strengthen identifications.     

The morphological identification of micro-residues on stone tools using light microscopy: progress and difficulties based on blind tests

Fifty-three stone flakes were knapped for a series of four blind tests on replicated flakes with residues derived from the processing of plant and animal products. Some flakes were hafted before use. Tests 1 and 2 were pioneering efforts published in 2004; lessons learned from these early studies shaped the new research reported here and lead to improved methodology and interpretive skills. A high level of accuracy was obtained for test 4. Test 3 showed that the rock type of a tool could influence the ability of the analyst to recognize and interpret residues. Test 4 in the series resulted in the most accurate interpretations because, prior to Test 4, identification difficulties experienced during the first three blind test sessions were addressed by examining many stone tools that had been used for various replicated tasks. The preparatory exercise was particularly useful for resolving issues that had previously caused problems for correctly identifying animal residues. The new work reported here highlights some of the difficulties that can be experienced in the morphological identification of microscopic organic residues, particularly the distinction between animal and plant residues. Some solutions for these problems are suggested.     

COLD NEUTRON PROMPT GAMMA ACTIVATION ANALYSIS—A NON‐DESTRUCTIVE METHOD FOR CHARACTERIZATION OF HIGH SILICA CONTENT CHIPPED STONE TOOLS AND RAW MATERIALS*

Recently, several archaeometrical projects have been started on the prehistoric collection of the Hungarian National Museum. Among the analytical methods applied, non‐destructive prompt gamma activation analysis has a special importance. We have also tested the potential of this method on chipped stone tools, with the aim of determining their exact provenance. On the basis of major and trace element components, characterizations of stone tools and their raw materials—silicites (flint, chert, radiolarite and hornstone) as well as volcanites (felsitic porphyry and obsidian)—were performed. We discuss some important results concerning each group, as case studies. Compiling the data set of different PGAA analysis series, compositions of 110 samples are reported, including 76 archaeological pieces. In the future, we plan to extend the number of investigated objects in each class.     

UNDER THE HAMMER: RESIDUES RESULTING FROM PRODUCTION AND MICROWEAR ON EXPERIMENTAL STONE TOOLS*

This paper presents the results of an experimental programme investigating the effects of production processes on stone tools and their interference with use‐related features. In the first part of the experimental programme, a series of flint flakes was retouched with different kinds of hammers in order to assess the extent and nature of percussion residues. After careful environmental scanning electron microscope (ESEM) observation and energy‐dispersive X‐ray (EDX) analysis, the retouched flakes were thoroughly cleaned and the underlying flint deformation was recorded. The second part of the experimental programme involved selecting several uncleaned retouched pieces for use in short woodworking, hide‐processing and butchery experiments. These pieces were observed and analysed before, during and after use. Some of the percussion features proved to be surprisingly resistant to use, and in some cases turned out to be strikingly similar in appearance to use‐generated features.     

Identifying stone tools used in mining,smelting, and casting in Middle Bronze Age Cyprus

Abstract

Metallurgical production sites are often difficult to identify in the archaeological record because ore beneficiation and slag processing in the past involved the use of ground stone tools that were similar to those used in other contexts to prepare cereals and foods. Analysis of the ground stone assemblage from a Middle Bronze Age copper mining and production site at Ambelikou Aletri in Cyprus provided an opportunity to distinguish industrial and domestic ground stone tools and to identify the types of tools used in different stages of metal production. A comparison of tool morphologies, raw materials, and wear and breakage patterns from Ambelikou Aletri with those from contemporary domestic contexts, suggests that distinctions in the nature and structure of industrial and domestic tool kits do exist and those distinctions have an important role to play in identifying mining, smelting, and casting sites in the future.     

CHARACTERIZATION OF NATURAL PRODUCTS ON NATIVE AMERICAN ARCHAEOLOGICAL AND ETHNOGRAPHIC MATERIALS FROM THE GREAT BASIN REGION,U.S.A.: A PRELIMINARY STUDY

Although documentary evidence provided by ethnographers makes reference to the source of adhesives, sealants, dyestuffs and medicines used by the communities of the Great Basin, much of it is sporadic and unspecified. However, observations of amorphous deposits surviving on stone tools, ceramics, basketry and wooden artefacts have prompted an investigation of their identity, procurement and use. The aim initially is to study samples from a number of archaeological and ethnographic contexts from the region. Comparative material includes higher plant and insect resins. The study provides a unique opportunity to document use patterns in plant and animal resources in the Great Basin region.     

A COMPARISON OF METHODS FOR ESTABLISHING FATTY ACID CONCENTRATION GRADIENTS ACROSS POTSHERDS: A CASE STUDY USING LATE BRONZE AGE CANAANITE AMPHORAE*

Canaanite amphorae fragments in fabrics believed to be associated with the bulk transport of vegetable oils from the fourteenth‐century BC site of Amarna, Egypt, have been examined. A comparison is made between solvent extraction, saponification and the use of TMTFTH (m‐trifluoromethylphenyl)trimethylammomum hydroxide, used here for the combined extraction and derivatization of ceramic‐absorbed organic residues. Extracts were studied using gas chromatography and gas chromatography‐mass spectrometry. The extraction of fatty acids from small ceramic samples has established concentration gradients of absorbed organic matter from the inner to the outer surfaces of the sherds.     

NEOLITHIC ‘GREENSTONE’ AXE BLADES FROM NORTHWESTERN ITALY ACROSS EUROPE: A FIRST PETROGRAPHIC COMPARISON*

Neolithic polished stone axe blades, manufactured with uncommon lithologies such as Alpine eclogites, jades and other HP metaophiolites, were exploited from primary and secondary occurrences in Piemonte and Liguria and dominate the north Italian and south‐east French polished stone blades used as functional tools (for deforestation and woodworking). In other European countries the same lithologies are found less frequently or only occasionally as axe blades; in north‐west Europe they were frequently used for manufacturing ceremonial axes that have shapes that are not present in the Italian Neolithic tradition. This paper undertakes a preliminary examination of the case, prevalently from a petrographic point of view, comparing the well‐established Italian petroarchaeometric knowledge with the rather poorer petrographic basis of the European eclogite/jade axes. A provisional picture of the distribution of these axes in Europe is provided and some interim interpretations and open problems are discussed.     

A Case of Ancient Bladder Stones from Oluz Höyük,Amasya, Turkey

A biological object was recovered from the pelvic region of an adult woman coded to SK13 unearthed from Oluz Höyük in Amasya, Turkey. The skeleton was dated to the medieval period on the basis of the burial customs and the calibrated ¹⁴C dates. The find was identified as a bladder stone on morphological, radiographic and chemo‐analytical grounds with the help of X‐ray diffraction, polarised energy dispersive X‐ray fluorescence spectroscopy, confocal Raman spectroscopy, and microscopic techniques. The mineralogical composition of urinary stone was found to be as calcium phosphate (apatite). Bladder stone disease is endemic in poor agricultural regions where the typical diet is mostly based on grain carbohydrate consumption with scarce intake of animal protein. Copyright © 2013 John Wiley & Sons, Ltd.     

METALLURGY OF BRONZE USED IN TOOLS FROM MACHU PICCHU,PERU*

Although the tin‐bronze artisans’ tools found at Machu Picchu appear to have been cast to their final shape, examination of their microstructures shows that they were forged and annealed. As found, the tools were broken or cracked due to poor ductility of the bronze. Rolling and annealing of samples of bronze formulated with sulphur and iron additions to replicate the metal used at Machu Picchu show that the minimum amount of deformation needed to induce the observed recrystallization is a 12% reduction in thickness. The sulphur and iron impurities retard nucleation of new grains during annealing, but do not inhibit subsequent grain growth. Tensile tests show that while up to 10% porosity in cast bronze has no effect on hardness or initial strain‐hardening rate, it reduces the tensile strength and ductility enough to embrittle the metal. Hardness is an inadequate indicator of the strength properties of cast bronze intended for use in tools or weapons. The results show that when the metalsmiths at Machu Picchu started making bronze tools for use by stone and woodworkers, they had not mastered the art of making metal with adequate strength properties for this service. The forging and annealing procedure they used did not enhance the mechanical properties of their tools, and may reflect an inherited metallurgical tradition.     

Experiments with Lithic Tools: Understanding Starch Residues from Crop Harvesting

Stone knives were used widely in Neolithic East Asia, presumably in the harvesting of grain crops, but their function has not been clearly understood due to the lack of study of residues from these tools. To address this issue, starch grain analysis was employed to study the residues on the surface of ancient stone knives and large amounts of starches were recovered. The sources of these starches, however, were not well understood, because harvesting of crops involves the cutting of stems rather than direct contact with starchy seeds. To determine whether harvesting could deposit these types of residues, we designed a simulation experiment using stone flakes to harvest ears of wheat, rice and foxtail millet, then analysed the residues on both the flakes and in the plant tissues. A large number of starch grains were found in the stems, including both typical morphotypes from seeds and newly described types that occur only in stems, which can be used as indicators of harvesting. Our study demonstrates that starch grains from residues on the surfaces of archaeological stone knives can indicate not only that the tools were used to harvest ears, but also the type of crops harvested.     

Slicing Cut Marks on Animal Bones: Diagnostics for Identifying Stone Tool Type and Raw Material

Abstract

Butchering implements leave identifying signatures on bones. From these signatures, it is possible to distinguish the different raw materials and types of chipped stone butchering tools. The results of recent experiments enable us to distinguish the different types of raw materials and tools used in the butchering process, in particular those implements that produce slicing cut marks. Three types of chipped stone tools (blades,flakes, and side scrapers) and raw materials (flint, obsidian, and quartzite), as well as issues relevant to lithic production and use were examined and tested. Silicone molds of cut marks produced by each of the instruments were made and subjected to analysis in light optical and scanning electron microscopes under various levels of magnification. The criteria used for distinguishing tool type, raw material, type of production and use characteristics of the respective cutting instruments are presented, as well as a discussion of the application of the experimental results to the Early Bronze Age I site of Afridar, Israel. The data from Afridar indicate that almost all of the butchering marks on animal bones from the site were made by stone tools, in particular haphazardly-made flakes.     

The Archaeological Site of Persepolis (Iran): Study of the Finishing Technique of the Bas‐Reliefs and Architectural Surfaces

One of the aims of the 5‐year Iranian/Italian project for Persepolis, called ‘From Palace to Town’ was to contribute to the conservation of the stone monuments of the imperial site. As part of the activities dedicated to this purpose, a diagnostic study was carried out. Various aspects were considered: petrographic characterization of the stone, forms and factors of decay, and in situ testing of suitable conservation treatments. The present paper reports on the unexpected results of the study on the finishing of architectural surfaces. The results obtained on a limited, but nevertheless significant, number of samples collected from the monuments of the imperial Terrace, allow us to state that the dark grey limestone used for several (or many?) monuments was covered on purpose with a thin, fine whitish layer containing fluorapatite, as major component, and calcite. It is highly probable that the fluorapatite was obtained from calcined animal bones and that slaked lime was used as a binder. Further evidence for this is the discovery of a kiln with the remains of calcined bones and, nearby, a waste pit with animal bones containing fluorapatite. A second white layer, obtained with barium sulphate, was detected in one of the samples beneath the external, earthy encrustation. It could be perhaps interpreted as the remnants of a polychrome finishing.