Infrared and Isotopic Evidence for Diagenesis of Bone Apatite at Dos Pilas, Guatemala: Palaeodietary Implications

The use of stable carbon isotopic analysis of bone apatite to reconstruct prehistoric diets is hindered by the possibility of diagenetic alteration of carbonate during burial. We examine apatite preservation in Classic Period Maya skeletal remains from Dos Pilas, Guatemala, using Fourier transform infrared spectroscopy (FTIR). We use weight % CO₂evolved from apatite, FTIR carbonate/phosphate absorbance ratios, phosphate peak splitting crystallinity indices, fluoride peaks, and stable oxygen isotopic ratios to identify diagenetic change in apatite chemistry. Isotopically light carbon taken up from burial soil is adequately removed from most Dos Pilas bone by treatment with dilute acetic acid, but more severe alteration cannot be reversed by standard preparation methods. Infrared criteria identify recrystallized apatite in a subset of Dos Pilas burials, that is accompanied by isotopic exchange, and that no longer preserves biogenic δ¹³C. These results illustrate that comparatively recent bone may be diagenetically altered and demonstrate a need for systematic evaluation of mineral integrity in all archaeological bone prior to interpreting paleodiets with apatite δ¹³C.     

How Did Mississippians Prepare Maize? The Application of Compound‐Specific Carbon Isotope Analysis to Absorbed Pottery Residues From Several Mississippi Valley Sites

A newly developed compound‐specific stable carbon isotope technique allows the detection of maize in absorbed organic pottery residues. This method was applied to absorbed organic residues from a variety of Mississippi Valley potsherds, and successfully identified maize components. Maize was cooked in sampled vessels less often than expected, but otherwise fitted expected patterns of maize use derived from stable carbon isotope analysis of human bone from the region. Absorbed organic pottery residue analysis is useful in determining pottery use, particularly in concert with other analytical methods.     

TESTING AN ALTERNATIVE HIGH‐THROUGHPUT TOOL FOR INVESTIGATING BONE DIAGENESIS: FTIR IN ATTENUATED TOTAL REFLECTION (ATR) MODE*

Archaeological bone undergoes alterations after burial (diagenesis) that constitute a problem for the survival of archaeological information. A common method to assess this alteration is Fourier transform infrared spectrometry (FTIR). However, the commonly applied method (FTIR–KBr) is destructive and sample preparation may influence the results. This paper tests the suitability of FTIR attenuated total reflection (FTIR–ATR), a method not commonly used to investigate bone diagenesis. FTIR–ATR requires less sample preparation and can be non‐destructive, allowing analysis of bone cross‐sections. Modern and archaeological bones were analysed using both methods and different sample preparation methods were tested. The results show that FTIR–ATR has advantages for the rapid assessment of bone diagenesis.     

PROVENANCE DETERMINATION OF NEOLITHIC TO CLASSICAL MEDITERRANEAN MARBLES BY STABLE ISOTOPES*

Two stable isotopic data bases for ?¹³C and ?¹⁸O values of marble have been accumulated: (1) 590 analyses from 42 Classical quarries of Greece, western Turkey, Tunisia, and Carrara, Italy, and (2) potential Early Bronze Age (EBA) marble sources in the central Cyclades, comprising 192 analyses from 16 sites on the islands of Paros, Naxos, Ios, and Keros. The data bases include the most important marble occurrences in the eastern Mediterranean and so can be used to source artefacts of any age. Neolithic artefacts found at Franchthi Cave, where no marble occurs locally, have been attributed to the Peloponnese and the Cyclades, direct evidence for early trade. In the Cycladic EBA, Naxos and Keros were the principal marble sources for the abundant artefacts found at Keros. The commercial marble trade in Archaic Greece began in the central Cyclades where the tradition apparently went back some 5000 years, to the Neolithic. Classical Greece and especially Rome expanded the number and locations of commercial marble sources throughout the Mediterranean.     

THE EFFICIENT COMPUTATION OF UNBIASED MAHALANOBIS DISTANCES AND THEIR INTERPRETATION IN ARCHAEOMETRY*

This paper argues for cross-validation to reduce bias in estimating Mahalanobis distances of individuals to groups, a particular problem with small sample sizes. Formulae for the efficient computation of cross-validated distances, which avoid excessive matrix inversion, are given. The use of Mahalanobis distances as measures of consistency, or as indications of outlying values, is described. The little-known technique of gamma plotting is outlined and discussed as an aid to interpreting distances in these terms. Both cross-validation and gamma plotting are illustrated in an example on marble composition.     

CATHODOLUMINESCENCE OF WHITE MARBLES: AN OVERVIEW*

Identifying the source area of white marbles has long been a problem in archaeology. To address this problem, about 1000 samples from known Classical quarrying areas in Greece, Italy and Turkey were analysed using cathodoluminescence. Twenty-one cathodomicrofacies were recognized, each of which generally characterizes a given area. In cases where the cathodomicrofacies is common to more than one area, δ¹³C and δ¹⁸O signatures provide an effective complementary dataset which allows additional distinctions. Determination keys for the 21 cathodomicrofacies observed are proposed based upon their principal cathodoluminescence colour, and their stable isotopic signature.     

IN VITRO DECOMPOSITION OF BONE COLLAGEN BY SOIL BACTERIA: THE IMPLICATIONS FOR STABLE ISOTOPE ANALYSIS IN ARCHAEOMETRY*

To understand biogenic collagen type I decomposition and to establish how diagenesis may bias archaeometric data, modern mammalian bone was inoculated with a selection of ubiquitous soil bacteria. The presence of exogenous microbial biomass in the inoculated specimens was then checked microscopically prior to collagen extraction. The experimentally degraded bone collagen showed altered amino acid compositions, attributable to the selective breakdown of certain amino acids by the bacteria. While both the bulk collagen extract and the single amino acids exhibited shifts to more negative δ¹³ C-values, enrichment was recorded for general δ¹³ N, and a depletion trend relative to unaltered collagen was observed for individual amino acid δ¹³ N. One explanation for the enrichment of the global δ¹⁵N-values is cleavage of peptide bonds, which leaves ¹⁵ N within the substrate, while the change of ¹³C is mostly due to the altered amino acid composition. On the other hand, possible repolymerization of cleavage products under experimental conditions may also be responsible for the depletion trend of individual amino acid δ¹³C- and δ¹⁵ N-values. This paper discusses the results as a basis for the development of a method for the reconstruction of the isotopic abundance of the original collagen, using the amino acid composition of the degraded product, the contribution of individual amino acids to its global δ-values and of isotope discriminations implied in the microbial decomposition.     

STABLE CARBON ISOTOPE MEASUREMENTS OF THE CARBOXYL CARBONS IN BONE COLLAGEN*

In order to explore the possibility that intramolecular differences in the δ¹³C value of bone collagen can provide useful infomation for isotopic dietary analysis, a method for reliably measuring the δ¹³C value of the peptide-bonded carboxyl carbons in collagen was developed and applied to a wide range of modem and ancient species, including humans. Carboxyl carbons were selectively removed as CO₂ by decarboxylation of collagen hydrolysates with ninhydrin (2,2-dihydroxy-l,3-indanedione). Carboxyl carbons were usually isotopically heavier than the total carbon in the collagen, with differences ranging from -0.9 to 4.0%c. Animals at higher trophic levels and with more positive δ¹⁵N values usually had smaller differences than the lower animals while humans had the largest range, reflecting their larger range of diets.     

Patterns of Diagenesis in Bone II: Effects of Acetic Acid Treatment and the Removal of Diagenetic CO3

Archaeological bones of varying preservation have been treated with 0·1 M acetic acid in order to investigate the effect on structural and chemical alterations caused by diagenesis. Acetic acid is commonly used as a “cleaning agent” for removing diagenetic carbonate from bone and enamel, in an attempt to recover original, biogenic signals for use in dietary and¹⁴C dating studies.Diagenetic parameters were measured before and after treatment on a range of archaeological bones with good and bad preservation. Histological preservation defined the behaviour of the correlating parameters, where correlation coefficients between carbonate content and crystallinity, microporosity and macroporosity increased significantly after treatment. For histologically well preserved material, acetic acid is effective at returning carbonate content to around that of modern bone. Where bone is extensively damaged by micro-organisms, “loose” diagenetic material can be removed, but a fraction largely composed of hypermineralized bioapatite remains, which, we believe, cannot be reliably used to obtain accurate biological signals.     

ON ISOTOPES AND OLD BONES*

This review charts the developments and progress made in the application of stable light isotope tools to palaeodietary adaptations from the 1970s onwards. It begins with an outline of the main principles governing the distribution of stable light isotopes in foodwebs and the quality control issues specific to the calcified tissues used in these analyses, and then proceeds to describe the historical landmark studies that have marked major progress, either in their archaeological applications or in enhancing our understanding of the tools. They include the adoption of maize agriculture, marine‐focused diets amongst coastal hunter–gatherers, trophic level amongst Glacial‐period modern humans and Neanderthals, and the use of savannah resources by early hominins in Africa. Particular attention is given to the progress made in addressing the challenges that have arisen out of these studies, including issues related to the routing of dietary nutrients. I conclude with some firm, and some more speculative, pointers about where the field may be heading in the next decade or so.     

HUMAN MOBILITY AT THE EARLY NEOLITHIC SETTLEMENT OF VAIHINGEN,GERMANY: EVIDENCE FROM STRONTIUM ISOTOPE ANALYSIS*

As evidence concerning human mobility during the transition to agriculture in central Europe, we present the results of strontium isotope analysis of human skeletons from the Neolithic village of Vaihingen, Germany. We find significantly more ‘non‐local’⁸⁷Sr/⁸⁶Sr values from humans buried in a Neolithic ditch surrounding Vaihingen than from those buried within the settlement. These results fit with previous studies showing a correlation between burial circumstances and strontium isotope signatures from LBK cemeteries of southwestern Germany ( Price et al. 2001 ; Bentley et al. 2002 ). A pilot study of Neolithic animal teeth from Vaihingen suggests that either ‘local’⁸⁷Sr/⁸⁶Sr signatures were more variable than the analysed human bones suggest, or that these domestic animals themselves were mobile, perhaps ranged by mobile pastoralists.     

AUTOMATION OF SOLUBILITY PROFILES USING A FLOW CELL

The solubility profile technique is a means of characterizing the mineral phase of bones and fossils from archaeological and palaeontological sites. The method is based on the differential dissolution of variously substituted apatites in acetate buffer at pH 4.5. The original solubility profile protocol, which involves repetitive washing and centrifuging, is extremely cumbersome and time-consuming. Here we describe an alternative, automatic method in which the fossil bone preparations are dissolved in a perspex cell. The behaviour of fossil preparations in the manual and automatic protocols are compared. We conclude that the dissolution cell is satisfactory for routine applications, but that the original, manual protocol is preferable for preliminary investigation of untested assemblages.     

MULTI‐METHOD MARBLE PROVENANCE DETERMINATIONS: THE CARRARA MARBLES AS A CASE STUDY FOR THE COMBINED USE OF ISOTOPIC,ELECTRON SPIN RESONANCE AND PETROGRAPHIC DATA*

A multi‐method approach, based on petrography, stable isotope data and electron spin resonance spectroscopy for assigning the provenance of white marbles, is presented. A total of 11 experimental variables has been used, including the quantitative measurement of the sample colour and its homogeneity. On this basis we attempt to distinguish the three main quarrying basins of the Carrara district, that is, Torano, Miseglia and Colonnata. Such discrimination, extremely uncertain on the basis of the single techniques, may be performed with acceptable confidence’ using a properly selected subset of six petrographic, isotopic and spectroscopic variables. The final resubstitution and jackknifed re‐assignment rates are, respectively, 85% and 81%. Inclusion of the remaining five variables into the classification rule does not improve the results, but simply increases data overinterpretation. The practical possibility of using such an approach for more general provenancing problems is considered.     

THE FUNDAMENTAL RELATIONSHIP BETWEEN ANCIENT DIET AND THE INORGANIC CONSTITUENTS OF BONE AS DERIVED FROM FEEDING EXPERIMENTS

The levels of ten elements (phosphorus, calcium, magnesium, strontium, barium, potassium, sodium, zinc, iron, and aluminium) have been measured in the femurs and humeri of laboratory rats grown on test diets as a model for palaeodietary studies of excavated human skeletons. High levels of dietary fibre correlate with lower Ca and higher Sr and Ba in the bone. High values of the dietary Sr/Ca ratio correlate with high Sr levels in the bone. High levels of dietary protein correlate with lower Sr and higher Zn. High values of the ratio Ba/Ca and lower levels of Fe correlate with higher levels of Ba. Because bone levels typically are controlled by several dietary components, variation of the levels of a given element in the diet does not necessarily translate into analogous variations in bone. Only K and Fe in bone correlate highly with their levels in diet. The failure of bone Sr, Ba, and Zn to correlate positively with dietary levels is fully explicable in terms of their interdependence on other dietary components, such as Ca, P, Fe, protein, and fibre.