Residues on stone artefacts: state of a scientific art.

Residues on stone artefacts: state of a scientific art. At the 1996 Society for American Archaeology The Society for American Archaeology (SAA) is the largest organization of professional archaeologists of the Americas in the world. The Society was founded in 1934 and today has over 7000 members. meeting in fabulous NewOrleans, residues and functional analysis of stone artefacts were thespecific focus of offerings scattered through the programme in posters(e.g.D. Lee, T. Murphy, L. Hooper and R. Donahue), workshop (N. Tuross,M. Wachowiak, R. Evershed, C. Kolman), at least two symposia (B. Hardy,B. Kimura & B. Hardy, T. Loy, D. Hyland, Charters et. al., Cummingset al., Tuross et al., C. Heron, H. Ceri and M. Newman) and severalgeneral sessions (E. Lohse, J. Furby & R. Fullagar, B. Williamson,M. Newman). We could not see them all, and we know some presenterslisted in the programme did not show. A symposium (sponsored by theConservation Analytical Laboratory at the Smithsonian Institution) and aworkshop (sponsored by the Conservation Analytical Laboratory at theSmithsonian Institution and the National Center for Preservation andTraining, National Park Service, Natchitoches, Louisiana) were dedicatedto residue analysis. Both, organized by Noreen Tuross, were a successwith lively debate. In dealing with much broader issues, thepresentations seemed to portray conflicting views on the veracity ofscientific techniques, and on the achievements of integrated functionalanalysis of stone artefacts. While papers presented in the generalsessions assumed the validity of the methodologies being used, some inthe sponsored workshop and symposium cast doubt on: * the distinctiveness of microscopic residues; * the integrity/reactivity of ancient bio-molecules; * the viability of the current methodologies; * and consequently on the reliability of taxonomic identificationson ancient stone artefacts.Archaeologists could be forgiven for being both confused whetherresidues survive on stone tools, or whose technique is best. Asarchaeologists with one foot in the lab and one in the field, we wereboth stimulated and depressed by the workshop and sponsored symposium sponsored symposiumA collection of papers published as a separate issue or as a special section–eg, an appendix in a regular issue of a peer-reviewed medical or scientific journal; symposia are usually underwritten by a drug company with a marketing 'agenda'. .We offer here our post-conference views on residues, on functionalanalysis of stone tools and on where consensus seems to lie.Stone artefact See artifact. functionStone artefact function has been investigated by diverse lines ofevidence in many theoretical contexts for over a century. Many articlessummarize these notions and review lines of evidence, which include tooldesign, raw material characteristics, use-wear, residues, context,ethnography and the study of long-term trends (e.g. Hayden 1979;Kamminga 1982; van Gijn 1990; Yerkes & Kardulias 1994). In truth,artefact function rarely, if ever, is determined from a single line ofevidence, although many claim use-wear or residues provide sufficientevidence of tool function. The rise and rise of residue studies can becorrelated with the increased availability of specialized techniques inbiochemistry for analysing low concentration samples. We thinktime-consuming techniques of observing and recording visible residueshave been neglected, a casualty of the high-tech methods now availablefor targeting the particular species of animal or plant contactmaterial.Detailed microscopic observations and recording are essentialpre-requisites to any residue analysis. In contrast to the impressionsoffered at the workshop, microscopically visible organic structures arecommon under optimal preservation conditions. Many residue analysts do'whole tool' extractions in the process of analysis, aprocedure which is often inappropriate for archaeological material as itis both destructive and does not target particular residues with theirvisible structures and associations with utilized edges.Survival, detection and taxonomic identification of animal relatedresiduesBlood, hair, bone and cartilage are the residues most often preservedon the surfaces of stone artefacts used in butchering animals. These andother traces have been documented by micro-wear analysts and in specificresidue studies (Briuer 1976; Anderson 1980; Keeley 1980; Kamminga 1982;Loy 1983; Fullagar 1986; van Gijn 1991; Furby 1995). For example, bloodforms characteristic films which are often visible at low magnificationand clearly identifiable at high magnifications (Brown 1988; Furby1995). In contrast, the sponsored workshop at the 1996 SAA (Systems Application Architecture) A set of interfaces designed to cross all IBM platforms from PC to mainframe. Introduced by IBM in 1987, SAA includes the Common User Access (CUA), the Common Programming Interface for Communications (CPI-C) and Common Communications sent themessage that 'we can't separate residues from other stuff withany certainty' (Tuross & Wachowiak 1996).Handling of the artefacts during excavation was another point raisedat the sponsored symposium and workshop. There was a clear dichotomybetween the recommended handling and storage methods of those examiningpottery and those studying blood residues. It was recommended (byTuross) that excavators wear plastic gloves during the recovery ofartefacts, which may be a reasonable precaution if submitting materialfor DNA analysis DNA analysisAny technique used to analyze genes and DNA. See Chromosome walking, DNA fingerprinting, Footprinting, In situ hybridization, Jeffries' probe, Jumping libraries, PCR, RFLP analysis, Southern blot hybridization. . However, it is a complicating procedure for the levelof analysis that most people are pursuing. Recent excavations at CuddieSprings in central northern New South Wales New South Wales,state (1991 pop. 5,164,549), 309,443 sq mi (801,457 sq km), SE Australia. It is bounded on the E by the Pacific Ocean. Sydney is the capital. The other principal urban centers are Newcastle, Wagga Wagga, Lismore, Wollongong, and Broken Hill. were undertaken without thebenefit of plastic gloves, yet the subsequent analysis of the stoneartefact assemblage was successful, both in the identification andcharacterization of blood and in DNA analysis of the material (Furby1995; Loy et al. submitted). The excavator's hands are covered inthe enclosing sediments and the artefacts themselves are usually cloakedin sediment. With at least two layers of sediment between the excavator ex��ca��va��torn.An instrument, such as a sharp spoon or curette, used in scraping out pathological tissue.excavator (eks´k and the stone there is, in our experience, little or no detectablecontamination.Most interest in identifying residues associated with butcheringanimals has been in blood residue analysis, particularly in thedetermination of species of origin (Loy 1983; Newman & Julig 1989;Tuross & Dillehay 1995; Petraglia et al. 1996). It is difficult toreconcile the range of studies currently being reported which assumeblood to be present in the absence of careful microscopic observations.The issue at stake appears to be whether residues can be characterizedto determine species of origin. As blood residues are preserved on thesurface of stone only under a limited number of depositional conditionsand environments, it is imperative that the range of residues on thestone artefacts are documented before the artefact is tested. In ourexperience, where blood does survive, it is usually found in stepfractures, at the edge of undetached flakes or in other protected areas,and in quantities significantly less than must have been depositedduring use.And what are the real costs of microscopy? While the sponsoredworkshop implied that tens of thousands of dollars are required for thevisual identification, this is simply not true. Archaeologists caneasily examine their assemblage and sample it for artefacts with visiblesigns of use and organic residues. While compound metallographic andbiological microscopes can cost in excess of $10,000, they are commonlyavailable in many university departments; more importantly, a hand lensor simple stereo-microscope can be used for examination of stoneartefacts before they are submitted for analysis. Description andrecording of the residues present can then be undertaken by a residuespecialist.A range of methods is available to characterize purported residues asblood (Newman & Julig 1989; Cattaneo et al. 1990; Hyland et al.1990; Tuross & Dillehay 1995; Loy & Hardy 1992; Loy & Wood1989). Constructing a hierarchy of evidence hierarchy of evidence,n the sequence of scientific evidence; a means of judging evidence presented in medical literature. Criteria for judging include how the clinical subjects were selected, the nature of the control group, the means by which the data enables different lines ofevidence to be addressed, as in the forensic sciences, to provide aconfident level of interpretation. Loy & Hardy (1992) presented arange of techniques for identifying blood residues including the use ofmicroscopy, Ames Hemastix and immunoassay ImmunoassayAn assay that quantifies antigen or antibody by immunochemical means. The antigen can be a relatively simple substance such as a drug, or a complex one such as a protein or a virus. using the 'dot-blot'method. At the SAA sessions, Loy indicated how false positives for AmesHemastix can effectively be eliminated by the addition of 0.5M, pH 8di-sodium EDTA EDTA:see chelating agents. which complexes metal ions in solution. These three stepshave been demonstrated to provide a viable and reproducible method forcharacterizing blood residues (Furby et al. submitted).An impression current at the residue workshop and symposium is thatit will be at least another five years before blood residue analysiswill be able to produce any meaningful results. Yet it is widelyaccepted that blood residues can be identified using relatively simpleassays in series coupled with microscopic observation. On the otherhand, species of origin determinations are problematic, under themethods described by Tuross, Newman and other workers; in many casesthey may provide no additional useful information beyond that obtainedfrom simple assays for Haem haemsee heme. or Immunoglobulins. Haemoglobin haemoglobinor US hemoglobinNouna protein in red blood cells that carries oxygen from the lungs to the tissues [Greek haima blood + Latin globus ball]Noun 1. crystallization CrystallizationThe formation of a solid from a solution, melt, vapor, or a different solid phase. Crystallization from solution is an important industrial operation because of the large number of materials marketed as crystalline particles. , a simple method proposed by Loy (1983), has beenreplicated in the Australian Museum laboratory. The results areequivocal and this does not appear to be a technique that can yet beexploited by archaeologists in identifying species of origin (Garling1994).It is important to document the environmental conditions from whichany stone assemblage is recovered the soil pH, particle size analysis,environmental setting etc the information that will eventually allow usto predict the most likely circumstances under which residues willsurvive. Loy (1990) has suggested that greater than 20% clay must bepresent for organic residues to be preserved, a figure related to thepresence of certain clay minerals, e.g. montmorillonite Montmorillonite is a very soft phyllosilicate mineral that typically forms in microscopic crystals, forming a clay. It is named after Montmorillon in France. Montmorillonite, a member of the smectite family, is a 2:1 clay, meaning that it has 2 tetrahedral sheets sandwiching a .As argued by many conference presenters, blood residue analysis needsto be critically evaluated by archaeologists themselves and questionsasked as to what information they really need. In many cases, it wouldbe sufficient to identify and characterize blood residues without theadditional complication of species of origin analyses. Species of originmay not be critical to the research question, a point also made byTuross et al. (1996). There was tacit criticism in New Orleans that a'residue industry' is emerging prematurely, driven bycommercial interests, with many analyses obtained just because themethodology is available. Presentations in New Orleans implied that somestudies are not part of an integrated approach to the analysis of stonetool function. If these sentiments are true, study of residues is not tobe split between laboratory technicians and archaeologists, but belongsin the context of a collaborative and integrated approach to the site asa whole.Previously, Fiedel (1996) critically reviewed a range of bloodresidue analyses, arguing that results were frequently irreconcilablewith contextual archaeological data. The same issues were raised in NewOrleans. We emphasize three points. First, the criticisms are largelydirected to the problem of cross-reactivity with CIEP CIEP Council on International Economic PolicyCIEP Counterintelligence Equipment Program (cross-overimmuno-electrophoresis), and the lack of consistency between CIEP andother techniques of taxonomic identification. Newman's defence ofCIEP at the conference cannot deny problems of cross-reactivity, thoughthe ability to control for this and other variables does not appear tobe unattainable. Second, as Fiedel points out, there is a need forfurther blind tests to assess more accurately the reliability,specificity and sensitivity of different techniques. Ceri, Newman andtheir colleagues seem keen to participate, as they have co-operatedpreviously in print. Third, there seems little doubt about the survivalof blood residues on utilized tool edges, based on presence ofmicroscopically visible cracked plaques, erythrocytes, mammalian hairand consistent biochemical, immunological and DNA DNA:see nucleic acid. DNAor deoxyribonucleic acidOne of two types of nucleic acid (the other is RNA); a complex organic compound found in all living cells and many viruses. It is the chemical substance of genes. tests as demonstratedby Loy for the Toad River Canyon site in British Columbia (seereferences in Fiedel and presentations at the New Orleans conference).The lowest common denominator low��est common denominatorn.1. See least common denominator.2. a. The most basic, least sophisticated level of taste, sensibility, or opinion among a group of people.b. appears to be that blood residues dosurvive. Taxonomic identifications are limited by reference collections,cross-reactivity, protein concentrations and sensitivity of thetechnique used. Which technique is most reliable has yet to beascertained, though DNA tests appear most promising (see below).Plant residuesPlant residues were hardly mentioned in the workshop, but fared wellin the symposium (with an excellent paper by Carl Heron) and in thegeneral sessions where papers dealt with resins, starch grains,phytoliths and other microscopically visible structures. Plantprocessing has also been associated with highly visible polishes onstone tool surfaces (e.g. silica gloss), with some attempts to determinetaxonomic identifications in conjunction with residues (Anderson 1980;Fullagar 1993; Loy et al. 1992). Given the presence of visiblestructures of likely taxonomic significance, it seems easier forarchaeologists to accept the survival of plant residues on stoneartefacts and their direct association with utilized tool edges.Nevertheless, in order to interpret residues like starch grains andphytoliths, it is essential to have both extensive reference collectionsof different parts of many plant species and also extractions from localsediments.In addition to visible structures, much research has been undertakento identify plants through biochemical and other means. At thesymposium, Heron reported on long-term research to identify lipids usinggas chromatography and mass spectrometry.DNA: a final solution?The recognition of morphologically identifiable residues on stonetools has naturally led to the application of DNA analysis to residuestudies. The technique which has made this possible is the PolymeraseChain Reaction (PCR PCRpolymerase chain reaction. PCRabbr.polymerase chain reactionPolymerase chain reaction (PCR)). From a minute amount of target DNA (theoreticallyas little as one molecule), PCR can create millions of copies, enoughfor analysis through cloning or sequencing. Determination of thebase-pair sequence of the DNA can allow phylogenetic phy��lo��ge��net��icadj.1. Of or relating to phylogeny or phylogenetics.2. Relating to or based on evolutionary development or history. analysis and, insome cases, determination of the species of origin. Sequence analysis ofDNA is a potentially more accurate way to determine species of origin ofresidues than the immunological techniques previously used. However,because PCR is capable of amplifying minute amounts of DNA,contamination by modern DNA is a significant hazard. In order to ensurethat putative ancient DNAs are authentic, a number of informalprecautions have been proposed including: extractions carried out inlabs where the DNA of interest has not previously been analysed, no DNAcontrol PCR reactions, dedicated pipettes, aerosol resistant or positivedisplacement pipettes, periodic UV irradiation of solutions andpipettes, etc. Strict adherence to these controls greatly reduces thepossibility of contaminants. Because ancient DNA is highly degraded,fragments are usually less than 500 base pairs in length (e.g. Paabo1989; Lindahl 1993). This general rule has been used to test theauthenticity of putative ancient DNA fragments recovered, as thoselarger than 500 base pairs are suspect as modern contaminants. AncientDNA studies in archaeological contexts have most often been applied tobone (e.g. Parr et al. 1996; Stone et al. 1996; Hoss et al. 1996) butDNA has also been recovered from preserved soft tissue (e.g. Hoss et al.1996; Handt et al. 1994; Paabo 1989). As researchers improve theirprecautions against contamination, the results of ancient DNA studieshave become more believable and valuable.Residues on stone tools have recently been analysed for ancient DNAwith the goal of attempting accurately to determine the species oforigin of a residue (Hardy et al. in press; Loy, this symposium). Hardyet al. have recovered DNA from Middle Palaeolithic stone tools in excessof 35,000 years old from the site of La Quina, France, and haveidentified some to species. In the New Orleans symposium, Loy reportedthe recovery of DNA from stone tools of similar age and used sequenceanalysis to identify the DNA as diprotodont (an extinct Australianmarsupial marsupial(märs`pēəl), member of the order Marsupialia, or pouched mammals. ) in origin. Despite objections raised at the New Orleanssymposium, use residues can clearly survive on stone tools and mayretain recoverable DNA. DNA analysis can potentially provide speciesidentification and phylogenetic information about the origin of the DNA.Furthermore, DNA analysis of residues is not limited to residues ofanimal origin, but can be expanded to include investigations of plantremains on stone tools as well. DNA analysis, not necessarily a finalsolution in residue investigations, is capable of providing preciseidentification of the species with which a stone tool was associated -when performed with the proper controls and precautions. By combiningDNA analysis with other techniques, including use-wear and microscopicresidue analysis, archaeologists can construct a much more completepicture of prehistoric tool function than is otherwise available.ConclusionNoreen Tuross is thanked for organizing the successful sponsoredsessions, and we agree with her call not to accept residue analyseswithout scepticism, particularly the taxonomic identifications usingCIEP. But we express surprise at how residues have been analysed withsuch a low level of basic microscopic observation and such little regardfor other traces of use. In our experience, structurally distinct tracesof plant and animal tissue, visible microscopically, undoubtedly surviveover tens and probably hundreds of thousands years. Much can be done, atlittle expense, with careful observations.The consensus at the workshop and symposium is that residues dosurvive on the surfaces of stone artefacts for considerable timeperiods. There is debate over the reliability and viability of varioustechniques for characterizing these residues. Few of the protagonistsmeet head-on in disputing the best techniques, and samples and artefactsare rarely treated in parallel. Blind testing is still a problem due tothese variations in treatments; no one technique can yet be ruled out(see Fiedel 1996). In our experimental collections produced over 10years ago, residues are still present, reactive and with structuralmaterial still clearly visible. A key issue, not discussed in the SAAsessions, is the process of recording and characterizing the visibleresidues.A back-to-basics approach may be useful for archaeologists wishing tosample their collections for more sophisticated analyses. The debateover survival, reactivity and the most viable techniques has not yetbeen resolved. On the other hand, and in contrast to the impressionsfrom the SAA Workshop, we believe that residue analysis is within therealm of the archaeologist, at least in the initial identification andobservation stage. A positive point made at the SAA conference is thatresidue analysis needs to be planned in the context of meaningful andappropriate archaeological research questions which are achievable giventhe resources available. Archaeologists can continue, as they have forover a hundred years, examining artefacts and pursuing the differentlines of evidence; an integrated approach to the identification of toolfunction. There is no magic bullet to determine artefact function, andno substitute for careful microscopic observations.ReferencesANDERSON, P. 1980. A testimony of prehistoric tasks: diagnosticresidues on stone tool working edges, World Archaeology 12: 181-94.BRIUER, F.L. 1976. New clues to stone tool function: plant and animalresidues, American Antiquity 41: 478-84.BROWN, P.D. 1988. 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