Biofilm growth in human skeletal material from ancient Mesopotamia

Investigators have long recognised the effects of microbial activity on archaeological bone. These investigators, however, have focused on single or groups of microbes rather than on complex microbial aggregates such as biofilms, a focus that has affected our understanding of archaeological bone biodeterioration. In this paper, we report on the investigation of a biofilm in archaeological human bone from the site of Tell Leilan, Syria (2900–1900 BCE). Scanning electron microscopy indicated that the biofilm is characterised by single cells and microcolonies of bacteria and fungi, as well as calcite crystals that were all embedded within extracellular polymeric substances. Using culture techniques and DNA sequencing, we isolated and identified several microbes from the biofilm including Amycolatopsis sp., Penicillium chrysogenum, Aspergillus sp., Chaetomium sp., and Cladosporium sp. Having characterised the Leilan biofilm, we are now closer to understanding the complex process of bone biodeterioration in archaeological bone collections.     

State of preservation of polymorphic plasma proteins recovered from ancient human bones

Protected by the mineral matrix, bone proteins are capable of surviving inhumation periods of several hundreds or thousands of years in soil. While the preservation of the bone matrix protein, collagen I, is the prerequisite for a variety of archaeometric approaches, such as radiocarbon dating and the reconstruction of palaeodiet by stable isotope analysis, little is known about both the rate and state of preservation of non‐collagenous proteins. We succeeded in the isolation, electrophoretic separation (SDS‐PAGE, IEF) and immunological detection (radial immunodiffusion, IEF immunoblotting and ELISA) of plasma proteins preserved in archaeological human bones. However, sample preparation and electrophoretic methods had to be adapted to the specific demands of these aged proteins, since they are not only degraded and fragmented but also cross‐linked to other organic components, either indigenous to the bone or to contaminants from the burial environment. Complex decomposition phenomena are responsible for the altered mode of migration of aged proteins through a gel. After isoelectric focusing, the ancient proteins mainly concentrate below pH 4.45 in the pH‐gradient. Thus, highly negatively charged protein components have a better chance of preservation in bone after death. Isoelectric focusing with subsequent immunoblotting of ancient protein samples revealed protein patterns which showed marked charge‐modifications in comparison with those of modern human plasma proteins due to protein degradation (e.g. α₂‐HS‐glycoprotein and α₁‐antitrypsin). Nevertheless, in combination with different immunological analyses, previous results concerning the selective enrichment of α₂‐HS‐glycoprotein in bone compared with other plasma glycoproteins could be confirmed. Copyright © 1999 John Wiley & Sons, Ltd.     

Bone preservation in human remains from the Terme del Sarno at Pompeii using light microscopy and scanning electron microscopy

Archaeological bone can show marked and complex alterations depending on the environment in which it was buried. In this study, the state of preservation of 27 femurs recovered from the archaeological site of Pompeii was evaluated by light microscopy and scanning electron microscopy. Most of the bone samples, prepared by the grinding method, showed good histological preservation, although they were characterized by microfissures (microcracking). Nine bone samples showed different states of histological preservation, including worst preservation (two femurs), due to diagenetic processes. Cryostat bone sections stained with thionin or 4′,6′-diamidino-phenylindole (DAPI) revealed the persistence of DNA within some osteocyte lacunae. Scanning electron microscopy analysis showed that ultrastructural characters, such as lamellae and collagen fibres, are recognizable only in unaltered bone. Our results reveal that most Pompeian samples are well preserved since they have a bone microstructure virtually indistinguishable from that of fresh bone. In methodological terms, although each of the various morphological methods used contributes information, histological and histochemical analyses are the most informative for studying the preservation state of bone and allow for rapid essential screening of archaeological bone.     

Bone collagen preservation in the tropics: a case study from ancient Puerto Rico

Bone collagen is a well-characterized and generally robust protein that plays a key role in both radiometric dating and stable isotopic reconstruction of paleodiet. To be useful for such purposes, however, analyzed collagen must be relatively taphonomically unaltered. To date, little research has been conducted to document the taphonomic fate of bone collagen from archaeological sites in the tropics. In the present work, a large (n = 298) dataset of archaeological bone samples from sites on the island of Puerto Rico is examined by means of radiometric, chemical, and elemental analyses. The ultimate conclusion of this work is that while collagen loss may be accelerated in the tropics versus that seen in samples from higher latitudes, what collagen remains is typically sufficiently well-preserved and taphonomically unaltered as to make radiocarbon dating and/or stable isotopic analysis worthwhile.     

Fragmentation and preservation of bird bones in uneaten food remains of the Gyrfalcon Falco rusticolus

This paper presents fragmentation patterns of bird bones in uneaten food remains of the gyrfalcon Falco rusticolus. The victims' bones show a relatively low degree of fragmentation. Elements of the pectoral girdle and wing predominate while head and leg elements are poorly represented.     

A possible case of renal osteodystrophy in a skeleton from medieval Wharram Percy,England

The skeletal remains of a child aged 13–15 years, recovered from Wharram Percy, England, and dating from AD 960–1700, were examined using gross observation, radiography and scanning electron microscopy. Lesions suggestive of renal osteodystrophy were found. Palaeopathological recognition of renal disease has hitherto relied upon the identification of soft tissue lesions or the presence of urinary calculi. This appears to be the first palaeopathological identification of renal insufficiency on the basis of skeletal changes alone. Copyright © 2007 John Wiley & Sons, Ltd.     

Microchemical and crystallographic analysis of human bones from Nasca,Peru. A possible method of direct dating of archaeological skeletal material

This paper provides two archaeometric equations obtained by crystal–chemical parameters for dating human bone samples from the archaeological site of Nasca, Peru. Based on radiocarbon dating, the burials span from 500 BC to AD 1000. Crystal–chemical modifications in bone hydroxylapatite over time were compared with a twentieth century skeleton from Italy. Chemical analysis was carried out by means of electron microprobe along a profile from the periosteal to the endosteal border of the midshaft of four human femurs. The results indicate that the Ca/P ratio is linearly correlated with time according to the equation t=7880.68Ca/P−12805.90 (r=0.97; r²=0.95). Bone apatite X-ray powder diffraction analysis showed a systematic increase in the (002) reflection intensity with time, according to the equation t=250.49h/a−1961.86 (r=0.98; r²=0.97). The two independent archaeometric equations permit good accuracy in dating osteoarchaeological remains from the south coast area of Peru. © 1998 John Wiley & Sons, Ltd.     

上海松江区广富林遗址良渚时期人骨微量元素的初步研究

近几十年来,古代人群的食谱研究已经成为现代科技考古学的一个重要组成部分,也是当前国际科技考古学研究领域的一项前沿性课题。人类骨骼的化学元素分析为重建古代居民的食谱提供了大量信息。采用电感耦合等离子发射光谱仪(ICP-AES)对上海松江区广富林遗址良渚时期出土人骨中的Ba、Ca、Mg、Mn、P、Sr、Zn等7种化学元素测定的结果显示:发达的稻作业为该组居民提供了充足的植物性食物;而水网密集的周边环境同时又为该时期居民提供了丰富水产品,从而极大地补充了其蛋白质的摄入。     

Calculus,periodontal disease and tooth decay among the prehispanic population from Gran Canaria

In the prehispanic Gran Canaria there are some anthropological differences between the coastal inhabitants who buried their dead mainly in tumuli, and those from the central mountains, mainly buried in caves. Some data, as the prevalence of auricular exostoses, and a different bone Ba/Sr ratio support the view that there were differences in economy and diet between both groups of islanders. Moreover, the proportion of carious teeth was significantly higher among the population buried in caves. In the present study we analysed the prevalence of dental calculus, periodontal disease and antemortem tooth loss in remains of 791 individuals belonging to the anthropological collection of the Museo Canario (Las Palmas). Calculus deposition was very frequent (88.51%), no differences existing between men and women or between those interred in tumuli or in caves. Age at death was the only parameter independently related to calculus deposition by stepwise multivariate analysis. Periodontal disease was observed in 66.78% of the population, significantly more in men (χ² = 4.88, P = 0.027). No differences existed between individuals buried in tumuli and in caves. Antemortem teeth loss was observed in 64.73% of individuals, no differences existing between men and women or between those interred in tumuli or in caves. There was a significant association between calculus and periodontal disease (χ² = 18.07, P < 0.0001). Both caries (χ² = 8.40, P = 0.004) and periodontal disease (χ² = 44.96, P < 0.0001) were associated with tooth decay. However, the proportion of teeth with calculus deposition (in relation to observed teeth) was significantly higher among the population buried in tumuli (Z = 3.18, P = 0.001), although no differences were observed when the proportion of antemortem lost teeth and alveoli with periodontal disease were compared among people buried in tumuli and in caves, but women showed significantly lower proportions of alveoli with periodontal disease and antemortem tooth decay. These data suggest that the population buried in caves had a different dietary pattern to that of those buried in tumuli, since calculus deposition -more frequent in the latter-may be related to the consumption of proteins. The results also point to the existence of differences in diet between men and women.     

An investigation of climatic change in the holocene epoch using archaeological charcoal from Swaziland, Southern Africa

A methodology is described for the analysis of Holocene charcoals excavated from a rock shelter in the Lubombo Mountains of northeast Swaziland. Scanning electron microscopy was used to compare these with modern reference woods. Of the ancient material 96·6% could be identified, in some cases at specific level. It is in such a subtropical area, where the woody flora is so rich, that assemblages of local taxa can be used in palaeoclimatic reconstructions. The changing taxa indicated by the charcoal fragments from the rock shelter clearly reflect minor shifts in Holocene climate, from moist to dry and back to moist in recent times. This is of relevance to the fluctuations in Stone Age populations in southern Africa. The wider use of such evidence to complement other palaeoenvironmental and archaeological data is advocated.     

‘Rid grasse of bones’: A taphonomic study of the bones from midden deposits at the Neolithic and Bronze age site of Runnymede,Surrey, England

Runnymede has large samples of Neolithic and Late Bronze Age animal bones, with contrasting preservation conditions in both periods. The bone evidence has been used to interpret the formation of the site deposits. There are few articulated bones, and no joins were found in butchered bone, indicating that the area studied did not contain primary refuse. Various aspects of bone alteration have been analysed: (i) the proportion of bones with very good surface preservation was high in the in situ Neolithic excavation units and the basal Bronze Age midden, but bones in the upper units were mostly eroded. These units are reworked flood deposits. The greater degree of fragmentation of the bone in the reworked units has been quantified, using a system of recording the ‘zones’ present on each bone, which allows calculation of the fraction present. It is also demonstrated that the reworked units contain a lower proportion of identified bones and a higher proportion of teeth and iaws than the units with well-preserved bone, (ii) Quantification of canid gnawing shows, unexpectedly, that more was recorded on well-preserved bone. Thus recognition of gnawing depends on bone condition. This also confirms that most of the erosion of the bone surface is a post-depositional phenomenon. The sequence of activities is therefore interpreted as follows: meat was cooked and consumed, and the bones discarded for the dogs. At a later stage, larger bones were picked up and thrown away in the river or midden. Some ethnographic examples of periodic cleaning of farming settlements are cited.     

Experimental study of cut marks made with rocks unmodified by human flaking and its bearing on claims of ∼3.4-million-year-old butchery evidence from Dikika, Ethiopia

In order to assess further the recent claims of ∼3.4 Ma butchery marks on two fossil bones from the site of Dikika (Ethiopia), we broadened the actualistic-interpretive zooarchaeological framework by conducting butchery experiments that utilized naïve butchers and rocks unmodified by human flaking to deflesh chicken and sheep long limb bones. It is claimed that the purported Dikika cut marks present their unexpectedly atypical morphologies because they were produced by early hominins utilizing just such rocks. The composition of the cut mark sample produced in our experiments is quite dissimilar to the sample of linear bone surface modifications preserved on the Dikika fossils. This finding substantiates and expands our earlier conclusion that—considering the morphologies and patterns of the Dikika bone surface modifications and the inferred coarse-grained depositional context of the fossils on which they occur—the Dikika bone damage was caused incidentally by the movement of the fossils on and/or within their depositional substrate(s), and not by early hominin butchery. Thus, contrary to initial claims, the Dikika evidence does not warrant a major shift in our understanding of early hominin behavioral evolution with regard to carcass foraging and meat-eating.     

Tracking changes in bone fracture morphology over time: environment, taphonomy, and the archaeological record

Zooarchaeologists have often employed studies of bone fracture morphology as a means of understanding past human cultural activity, and various methodological approaches have been developed for analyzing archaeological broken bone assemblages. It is widely understood that bones degrade over time, however, few studies have attempted to define and quantify the rate at which bones degrade and fracture morphologies change. This study examines degradation in frozen bones (−20 °C) and bones exposed to hot (40 °C) dry conditions. These two simulated environmental conditions represent extreme real-world climates, and allow for an actualistic understanding of the rates of degradation that bones experience in nature. When frozen, bones degrade slowly but significantly, and demonstrate measurable differences in samples frozen for 1, 10, 20, 40, and 60 weeks. In hot, dry conditions, bones degrade very quickly, and demonstrate measurable differences after 1, 3, 7, 14, and 21 days. These data allow for a more detailed understanding of the relationship between the cultural and natural processes that result in bone fracture, and the time period during which bones can be expected to maintain fresh fracture characteristics. This research also has implications for understanding human subsistence and survival strategies and for interpreting the archaeological record.     

Detection of blood proteins in ancient human bone using ELISA: A comparative study of the survival of IgG and albumin

ABSTRACT The immunological identification of species-specific blood proteins in skeletal remains has an important role in the reconstruction of ancient dietary, ritual and domestic behaviour. However, which protein provides the most suitable target for such work has not been considered previously, and the present investigation was carried out on human bone to assess the relative merits of IgG and albumin. Extracts of bone from 31 individuals (from the English Civil War, medieval, Early Saxon, Roman, Iron Age and Bronze Age periods) were tested by an enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies. Albumin was detected in 23 of the 31 skeletons, including those from the Iron and Bronze Ages, whereas IgG was identified in only one; this difference was very highly significant (P < 0.0005, χ² 30.0). The better detection rate for albumin was thought to be due to its higher original blood level, inherent differences in survival pattern being considered unlikely. Testing the same extract for both proteins in the same assay system ensured that any effects due to soil factors, burial conditions, physical integrity of the bone, chronological age, amount of original specimen, method of analysis and type of reagent were the same for each part of the study, thus permitting a valid comparison of antigen survival to be made. Control samples, including fresh and ancient animal bone extracts and human and animal sera, confirmed that the results were consistent and specific, with no cross-reactivity between human and animal material, and that as little as 10 ng of protein was detectable. In summary, the investigation compared the suitability of IgG and albumin for osteoarchaeological studies using a highly specific, sensitive and versatile ELISA; the results showed that albumin was a far better target molecule for such work and that it can survive in ancient bone for long periods of time.     

Growth curves of immature bones from a Scottish island population of sixteenth to mid-nineteenth century: Shoulder girdle,ilium, pubis and ischium

Growth curves were prepared for length and various other dimensions, including some innovative ones, of the scapula, clavicle, first rib, ilium, pubis and ischium of ages up to 20 years including several immature and many full-term fetuses. Most curves showed, like the previously reported diaphyses of the limb bones, what appears to be a consistent adolescent growth spurt at about 17 years, but the number of specimens supporting these parts of the curves is too small for the validity of this growth spurt to be beyond doubt. The fetal dimensions were similar to those of Fazekas and Kósa and of other recent populations. For the post-natal stages of growth, no data are available for comparison but there was close correspondence with the growth curves of some ancient peoples, Amerindian, caucasoid and negroid, whose limb-bone growth rates had also been shown previously to be well below those of contemporary caucasoids. Thus it is argued that the shoulder-girdle and hip-bone data provided further evidence that this Scottish island population was affected by stunting factors. Although there is no skeletal evidence of malnutrition, there is plenty of evidence that the way of life was full of hardship in which the children shared. They took a full part in the arduous work involved in farming marginal-quality land; the houses were damp, cold, smoky and dark; they were overcrowded although, mainly because of the high infantmortality rate, families were not large.     

Ancient DNA in human bones from Neolithic and Bronze Age sites in Greece and Crete

Attempts were made to detect ancient DNA (aDNA) in samples of 88 human skeletons from eight Neolithic and Bronze Age sites in Greece and Crete. Ancient DNA was absent in specimens from Nea Nikomedia, Lerna, Karaviádena (Zakro), Antron Grave Circle A and Mycenae Grave Circle A. For each of three skeletons from Antron Grave Circle B that were sampled, polymerase chain reactions (PCRs) gave products for nuclear but not mitochondrial DNA, but amplicon yield was low and inconsistent with replicate PCRs failing to give reproducible results. With specimens from Mycenae Grave Circle B, evidence for mitochondrial aDNA was obtained for four of the 22 skeletons that were studied, and at Kouphovouno evidence for mitochondrial and/or nuclear aDNA was obtained with eight of the 20 skeletons that were examined. We conclude that, although aDNA might be present in some Eastern Mediterranean skeletons from later centuries of the Bronze Age, it is not commonly found in material from this period and is likely to be absent from older material.     

Relative and absolute dating of the human skull and skeleton from Galley Hill,Kent

Over a period of some 100 years it has gradually been accepted that the human skeletal remains found in the late nineteenth century in ancient gravels at Galley Hill, Kent are not those of early man, but are of much more recent date. This conclusion was finally verified by means of relative (N, F) and absolute (¹⁴C) dating. Here we present the results of further analysis of the remains from Galley Hill for their uranium content by a method having general application to the relative dating of fossil human and other mammalian bone.     

Bone changes in the human skull probably resulting from scurvy in infancy and childhood

Scurvy in children is poorly known in the palaeopathological literature despite biomedical references indicating that bone changes do occur in some cases and should be observable in human skeletal remains. Research on infants' and children's skulls in museum and anatomical collections in the USA and Europe reveals a complex of features on the external surface of the greater wing of the sphenoid and adjacent bone tissue that may be caused by scurvy. The lesions are bilateral and are characterized by porosity and, in some cases, hypertrophic bone formation. These abnormal bone changes are probably the result of a reaction to chronic haemorrhage in the deep vasculature associated with temporalis muscle function. While these lesions resemble those seen in anaemia and infection, their distinctive anatomical location and association with chewing should differentiate them, in most cases, from other disease conditions. These features are also associated with porous, hypertrophic lesions of the orbital roof and provide additional evidence that caution is needed in attributing porous lesions of the orbital roof solely to anaemia. © 1997 John Wiley & Sons, Ltd.     

Egyptian Blue cakes from the Ayanis fortress (Eastern Anatolia,Turkey): micro-chemical and -structural investigations for the identification of manufacturing process and provenance

The microstructure and chemical composition of blue cakes, found during the archaeological excavation of the Ayanis fortress (Eastern Anatolia, Turkey), have been investigated by means of scanning electron microscopy combined with energy dispersive spectrometry (SEM–EDS), X-ray diffraction (XRD), micro-Raman (μ-Raman) and optical microscopy (OM) techniques. The analysis of the Ayanis cakes has shown the presence of Egyptian blue (i.e. CuCaOSi₄O₁₀, cuprorivaite), as the major component, intermixed with minor amount of other phases such as partially reacted quartz grains, an adherent glass phase and copper oxides. Since the finding of Egyptian Blue in Turkey has been never reported so far, great attention has been paid to its characterisation. The micro-chemical and micro-structural investigations of the Ayanis cakes have allowed a further insight into the manufacturing process and into the sources of the starting materials. The results of the characterisation have revealed some significant differences with respect to Egyptian blue cakes found in Egypt and Mesopotamia, as for instance the absence of tin excluded the use of bronze scraps or filings in their preparation differently from those produced in Egypt and Mesopotamia. Furthermore, some peculiarities of Egyptian blue found in Ayanis, as the detection of zinc in the cakes, allow to put forward the hypothesis of a local production considering that a large part of the bronze artefacts found at the Ayanis fortress is characterised by the presence of zinc as minor alloying element.