Additional comments on blood residue and analysis in archaeology.

Additional comments on blood residue and analysis in archaeology. In the March Antiquity, Eisele et al. (1995) presentedimmunological results from prehistoric sites, and from simulatedarchaeological contexts, which suggest that residues will not survive onprehistoric stone tools. Over the last several years, we have submittedover 200 archaeological items to a commercial laboratory forimmunological residue analysis. In an effort to evaluate the reliabilityof these archaeological results, we initiated an actualistic test usingexperimentally generated artefacts coated with blood of known animals.Here, we present results for n of these experimental samples. Ourresults also suggest that archaeologists should view immunologicalresults with caution. The 54 experimental samples reflect a variety of artefact See artifact. types andmaterials encountered archaeologically. Using 10 species of animalscollected from recent road kills and commercial butchers, we coatedfreshly knapped artefacts with blood by cutting and scrapping a specimenfor several minutes. After allowing the artefacts to dry, we removedvisible hairs from several of the specimens by wiping them with a cottoncloth or lightly rubbing them with sand. However, in almost all casesblood was still visible macroscopically mac��ro��scop��ic? also mac��ro��scop��i��caladj.1. Large enough to be perceived or examined by the unaided eye.2. Relating to observations made by the unaided eye. , and it is our opinion that thisprocedure did not remove the blood from any specimens. Several`blank' lithic lith��ic?1?adj.Consisting of or relating to stone or rock.Adj. 1. lithic - of or containing lithium2. lithic - relating to or composed of stone; "lithic sandstone" tools and sediment samples, along with sediment andfire-cracked rock (FCR FCRfeed conversion rate. ) samples from an experimental hearth used to cooka rabbit, were also submitted for analysis. All experimental samples were sent to a commercial laboratory thatuses a gel immunoelectrophoresis ImmunoelectrophoresisA combination of the techniques of electrophoresis and immunodiffusion used to separate the components of a mixture of antigens and make them visible by reaction with specific antibodies. technique, crossoverimmunoelectrophoresis (CIEP CIEP Council on International Economic PolicyCIEP Counterintelligence Equipment Program ). Complete details of this technique, andits application to archaeological materials, are presented by Newman& Julig (1989). The experimental samples were tested against aseries of polyclonal antiserum polyclonal antiserumImmunology A Gemische of antibodies with distinct epitope reactivities, produced in response to a broad antigenic stimulus, which may be harvested from a person exposed to a particular pathogen and administered to another whose response to the , including those for bovine, chicken,deer, dog, elk, guinea-pig, human, pronghorn pronghornor prongbuck,hoofed herbivorous mammal, Antilocapra americana, of the W United States and N Mexico. Although it is often called the American, or prong-horned, antelope, it does not belong to the true antelope family of Africa , rabbit, rat, turkey,beans, corn, mesquite, pinyon, squash and yucca yucca(yŭk`ə), any plant of the genus Yucca, stiff-leaved stemless or treelike succulents of the family Liliaceae (lily family), native chiefly to the tablelands of Mexico and the American Southwest but found also in the E United States . Results, reported by the laboratory (Table 1) fall into fivegroups: * the laboratory correctly identified the blood of the animal,or did not report any blood on a blank sample., * the laboratorycorrectly identified the blood of the primary animal, but also noted theblood of other animals., * the animal was correctly identified, but notas the primary blood on the item., the laboratory reported the blood ofthe wrong animal; * no blood was detected though blood was present.Table 1. Experimental animals and results. artefact experimental laboratory type residue results1 lithic tool human human2 lithic tool elk deer, elk, human3 lithic tool elk deer, elk4 lithic tool elk deer, elk, human5 lithic tool elk human, deer, elk6 lithic tool squirrel negative7 lithic tool squirrel negative8 lithic tool mouse deer, elk9 lithic tool mouse human10 lithic tool coyote negative11 lithic tool coyote negative12 lithic tool bobcat negative13 lithic tool bobcat negative14 lithic tool bobcat negative15 lithic tool deer deer, elk16 lithic tool deer deer, elk, human17 lithic tool deer deer, elk18 lithic tool blank negative19 lithic tool blank negative20 lithic tool lank human21 lithic tool blank negative22 lithic tool blank negative23 lithic tool blank negative24 lithic tool blank negative25 lithic tool cow bovine, elk26 lithic tool cow bovine27 lithic tool turkey negative28 lithic tool turkey chicken29 lithic tool turkey chicken30 lithic tool rabbit human, rabbit31 lithic tool rabbit rabbit32 lithic tool rabbit rabbit33 lithic tool rabbit rabbit34 lithic tool rabbit rabbit35 lithic tool rabbit rabbit36 lithic tool rabbit rabbit, mouse37 lithic tool rabbit turkey38 lithic tool rabbit rabbit39 lithic tool rabbit rabbit40 lithic tool rabbit rabbit41 lithic tool rabbit rabbit42 lithic tool rabbit rabbit43 lithic tool rabbit rabbit44 lithic tool(1) rabbit negative45 lithic tool(2) rabbit negative46 lithic tool(2) rabbit negative47 lithic tool(2) rabbit negative48 lithic tool(2) rabbit negative49 sediment blank negative50 sediment(3) rabbit negative51 sediment(3) rabbit negative52 sediment(3) rabbit negative53 fire-cracked rabbit negative rock(3)54 fire-cracked rabbit negative rock(3)1 Artefact heated to 250 [degrees]C for 30 minutes.2 The samples are resubmitted after a 5-month period.3 Sample recovered from an experimental hearth. Of the five groups, we consider groups 1 and 2 to be successful:blood was correctly identified as either the only (Group 1) or thestrongest (Group 2) reaction. Group 3,4 and 5 results are problematic.Group 1 results were obtained from thirteen samples and Group 2 resultsfrom seven samples. Group 3 results were obtained from five samples,Group,4 results from four samples, and Group 5 results from theremaining 25 samples. The results of these experiments are dishearteningas the laboratory correctly identified only 20 of the 54 samples (37%). Many of the misidentified samples are cross-reactions. As discussedby Nolin et al.(1994; see also Newman & Julig 1989), cross-reactionsbetween closely related species (e.g. deer, elk, moose) within a Family(e.g. Cervidae) are not unexpected with the use of commerciallypurchased polyclonal polyclonal/poly��clo��nal/ (-klon��'l)1. derived from different cells.2. pertaining to several clones.polyclonalderived from different cells; pertaining to several clones. sera. However, several of our results representcross-reactions beyond the Family level: an artefact coated with elkblood (Sample 5) reacted strongest with anti-human serum and an artefactcoated with mouse blood (Sample 8) strongly with anti-deer and anti-elkserum (see also Child & Pollard 1992). All of the samples exposed toheat failed to produce reactions to any antiserum antiserum/an��ti��se��rum/ (an��ti-se?rum) a serum containing antibody(ies), obtained from an animal immunized either by injection of antigen or by infection with microorganisms containing antigen. , supporting thesuggestion by Cattaneo et al. (1994) that excessive heat may render anyresidue immunologically undetectable. Yet, even eliminating those heatedcases from TABLE 1, the overall success, rate is still below 50%. Referees for Antiquity had many questions to ask about theimmunological method used to analyse our experimental samples. Forexample, Had some of the antisera lost their original activity? Was themethod standardized and controlled for the material being tested?'Such questions must be addressed by the commercial laboratory thatconducted the analysis. As archaeologists submitting either experimentalor prehistoric samples for analysis, we can only assume that appropriatelaboratory procedures are in place. Clearly, the results presented inTABLE 1 indicate that additional experimental work is needed, includinga focus on laboratory procedures and appropriate immunologicaltechniques. (since this paper was submitted, we have sent additionalsamples for analysis, with results that show the same patterns.) While laboratory procedures can be questioned, and inter-laboratoryprocedures vary, the discrepancies revealed by our experiment mayreflect more than laboratory or procedural problems. As Sensabaugh etal. (1971; see also Eisele et al. 1995) have pointed out, the biologicalactivity of proteins may be rapidly lost soon after death of theorganism. In our study, the time-lapse between coating of the sampleswith blood and subsequent testing was generally 1-2 months. During thistime period the artefacts were sealed in plastic bags and refrigerated.Eisele et al. (1995; see also Cattaneo et al. 1993; Gurfinkel &Franklin 1988) demonstrate that degradation of blood proteins may occurover this short length of time. As such, any dependence on immunologicalresults, especially at time-scales commonly used by archaeologists, maybe problematic.Acknowledgements. This research is funded by the Legacy ResourceProgram, Department of the Army. Ms Sue Green-Leach and three Antiquityreviewers provided useful comments on a previous version of this paper. ReferencesCattaneo, C., K. Gelsthorpe, P. Phillips g, R.J. Sokol. 1993. Bloodresidues on stone tools: indoor and outdoor experiments, WorldArchaeology 25(1): 29-43. Cattaneo, C., K. Gelsthorpe & R.J. Sokol.1994. 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