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.     

The Eulau eulogy: Bioarchaeological interpretation of lethal violence in Corded Ware multiple burials from Saxony-Anhalt, Germany

The Corded Ware is one of the major archaeological traditions of Late Neolithic Europe. Its burial customs are characterized by single graves but multiple burials also occur. We present a detailed study of antemortem and perimortem trauma in a group of Corded Ware skeletons from four multiple graves and give the most probable interpretation of the site, based upon all available bioarchaeological evidence. The pattern of observed injuries in male, female, and subadult skeletons, including cranial trauma, arrow wounds, and fractures of the forearm and hands points towards a violent event that resulted in the death of all individuals, most probably a raid. In contrast to comparable Neolithic raid sites, there was no complete extermination of the local population and no use of mass graves. The burials have been arranged with care and detailed knowledge about biological kinship ties [Haak, W., Brandt, G., de Jong, H.N., Meyer, C., Ganslmeier, R., Heyd, V., Hawkesworth, C., Pike, A.W.G., Meller, H., Alt, K.W., 2008. Ancient DNA, strontium isotopes, and osteological analyses shed light on social and kinship organization of the Later Stone Age. Proceedings of the National Academy of Sciences of the United States of America 105, 18226–18231]. The combination of clear causes of death and the proven biological relationships among some of the individuals, including a nuclear family, provides new and important insights into Corded Ware mortuary customs and the reasons why and how multiple graves have been utilised.     

Determination of a kinship system using ancient DNA,mortuary practice,and historic records in an upper Canadian pioneer cemetery

Recovered and amplified ancient DNA (aDNA), from a historically documented 19th century Upper Canadian pioneer cemetery produced genotypes that were used to infer a past societal kinship system. While the results from multiplex short tandem repeat (STR) amplifications showed an unreliable polymerase chain reaction (PCR) product, a single locus HUMTH01 analysis yielded reproducible data and an allelic frequency pattern not statistically different from modern populations. Mitochondrial (mt) DNA HVR II data showed that a combined cemetery database exhibited reduced haplotype diversity indicators, as well as clusters of probable maternally related burials. The chronological persistence and replacement of mtDNA clusters approximately every two generations suggests a patrilineal/patrilocal kinship structure from a virilocal burial program for the Harmony Road cemetery. Through the integration of the aDNA analysis with archaeological material culture, historic records, and other ethnohistoric sources of information, this conclusion is supported. In this study persisting patrilineally inherited surnames act as a surrogate for aDNA Y‐chromosome haplotype analysis. These results suggest that aDNA applications on aggregate skeletal collections where sparse, or no ethnological or historical documentation exists, may result in incorrect population history inferences if the presence of a kinship interment bias is not considered. Copyright © 2003 John Wiley & Sons, Ltd.     

Warfare and mortality at Koger's Island,Alabama

Koger's Island, a late prehistoric agricultural community in the southeastern USA, is the site of a cemetery containing four mass graves of individuals with perimortem trauma and scalping cutmarks. Although the site as a whole has a demographic profile typical for a society of this kind (with high infant mortality), the mass graves contain relatively few infants or children, and an abundance of adult males. It is suggested that the skeletons found in the multiple burials represent the victims of raiding or warfare. The disproportionate number of males in the mass graves may be related to the fact that relatively few males at the site are older than 50 years, presumably because they are dying at an early age from violence.     

拊葬墓与古代家庭

在以血缘为纽带的社会组织中,其成员死后的埋葬方式直接与家庭、家族和宗族相联系。“袝葬”现象揭示了以家庭为核心的生产关系和生活方式。拊葬墓是以一人为主,其他人从属,多人共用一墓的墓葬。考古学研究中墓室的多少常常与墓主人的身份、等级联系在一起。但袝葬现象直接表现在墓葬形制特征上,是墓葬类型的一种特殊形式,因袝葬需要出现的多室墓并非是墓主人身份的象征,不能作为判别墓葬等级的标准。汉代的袝葬墓一般不改变流行的墓葬形制,而是改变某些空间的功能,西晋以后袝葬墓影响到墓葬形制的改变。拊葬墓的流行,是因为社会组织中血缘关系密切的家庭的地位增强,土地所有制、生产经营和财产关系发生了变化。     

Sex Identification in Some Putative Infanticide Victims from Roman Britain Using Ancient DNA

Previous study of infant burials has suggested that infanticide was routinely practised during the Roman period in Britain. This, together with the observation that there is an adult sex imbalance in favour of males at many Romano-British cemetery sites, has raised the question of female infanticide. We attempted to investigate this possibility by identifying sex in some infant skeletons from Romano-British contexts using ancient DNA (aDNA) techniques. Of 31 individuals sampled, sex identification was successful in 13, of which nine were males and four females. These results are discussed in the light of previous work on DNA-based seeking of infant burials.     

Familienanalyse in kupferzeitlichen Kollektivgräbern aus Umm an-Nar; Abu Dhabi

Skeletal material from three collective burials from the island of Umm an-Nar (Arabian peninsula) dating to the 3rd millennium B.C. was submitted to a kinship analysis with the aid of a set of genetically determined odontologic traits. The analysis yielded the following results: a) phenotype and frequency of odontological traits in the skeletal sample suggest a relatively homogeneous population; and b) there are distinct indications for genetically determined relationships within the individual burial mounds. In spite of the poor state of preservation of the skeletons, the analysis permitted conclusions about the social structure of the local population.     

Variable nucleotide tandem repeat (VNTR) typing of two palaeopathological cases of lepromatous leprosy from Mediaeval England

Using ancient DNA methods, we have examined in detail two archaeological cases of leprosy from Mediaeval England. The first was a child skeleton with rhino-maxillary changes typical of lepromatous leprosy (LL). The second case was the skeleton of a male adult who showed both typical rhino-maxillary changes and osteitis/periostitis on the leg and foot bones. Bone powder was sampled from both cases and DNA extracts were prepared. These were subjected to a series of polymerase chain reactions (PCRs) specific for regions on the Mycobacterium leprae genome. The repetitive element RLEP was used for confirmation of M. leprae DNA and then three polymorphic regions were successfully amplified and sequenced to determine the number of variable nucleotide tandem repeats (vntr) at these loci. These were the microsatellite regions ML2344 and ML2172 and the minisatellite region ML0058. Genotyping data from the strains preserved within the skeletal remains were compared with those obtained for a reference strain of M. leprae. Variation at these three loci was found between both burials and the reference strain, indicating that vntr typing of LL cases from the archaeological record is a useful way of confirming disease and an additional means of authenticating aDNA data. This demonstrates the feasibility of targeting multiple loci for phylogenetic studies of leprosy strains from archival sources.     

Middle Bronze Age Burials in the Southern Levant: Spartan Warriors or Ordinary People?

Intermediate and Middle Bronze Age tombs with weapons (mainly daggers) in the southern Levant were often interpreted as ‘warrior graves’. Taking into consideration new data from Rishon Le‐Zion (Israel), recent work on early warfare and warriors, and a study of so‐called ‘warrior graves’ in Mesopotamia (Rehm 2003), we suggest that most of these graves are not graves of elite warriors, but typical male burials. We also discuss the assumed ‘burial kit’ and the decline in numbers of weapons per burial, which is in our view related to the shift from individual burials (in the Intermediate Bronze and Middle Bronze IIA periods) to multiple shaft burials (in the Middle Bronze IIB period).     

Archaeological Bones – The Anatomy of an Abandoned Churchyard

Scattered human bones from disturbed graves in medieval and post‐medieval churchyards have generally been considered to be of minor interest in archaeological analyses. However, the material has a large information potential provided that it is carefully documented and analysed. By treating scattered bones in the same way as other archaeological finds the material is found to have great value as source material in the interpretation of cemeteries and churchyards as well as in paleodemographic analyses. This is demonstrated by analysing the dispersed bones found in the medieval/post‐medieval cemetery layers of the abandoned churchyard at Sola in Rogaland, south‐western Norway. By analysing bones from disturbed graves and incorporating both archaeological and osteological data in the analyses, it was demonstrated that it is possible to provide answers to questions about the original number of burials and the number of individuals in the churchyard, the relative chronology of grave constructions and a more accurate demographic profile of the buried population.     

Amplification and sequencing of Trichuris trichiura ancient DNA extracted from archaeological sediments

Despite high prevalence of Trichuris trichiura infection, PCR-based analysis on T. trichiura from archaeological samples has not been established so far. In the present study, we sought to perform PCR-based amplification of T. trichiura aDNA using the sediments from medieval tomb of Korea. The presence of Trichuris eggs were first detected by microscopic observation; then confirmed by PCR-based aDNA analysis. Obtained sequence showed 100% homology to that of the small subunit ribosomal RNA (SSUrRNA) gene of T. trichiura but distinct from that of other Trichuris species. PCR-based aDNA analysis in this study can serve as effective method to confirm the presence of T. trichiura eggs in the soils or coprolites collected from archaeological sites.     

Sebbersund: isotopes and mobility in an 11th−12th c. AD Danish churchyard

The important Viking Age and early Medieval site of Sebbersund in northern Jutland, Denmark, contains a large churchyard from the 11th–12th century AD. Sebbersund was an important trading center in this period and the location of one of the first churches in Denmark, perhaps an entry point for the introduction of Christianity to the country. Excavations have exposed almost 500 graves of an estimated 700 individuals in the cemetery. Here we report on the analysis of strontium isotopes in human tooth enamel from burials in the cemetery as a signal of place of birth. Some 19 samples have been measured and at least three non-local outliers identified. Futhermore, six archaeological fauna samples had been analyzed in order to define the local bioavailable strontium isotope baseline range and these values were compared to the more general bioavailable baseline range values for Denmark. The burials are evaluated in light of the available archaeological, chronological, anthropological, and isotopic information.     

Molecular and morphological analyses of avian eggshell excavated from a late thirteenth century earth oven

Using ancient DNA (aDNA) extracted from eggshell of the extinct moa (Aves: Dinornithiformes) we determined the species composition and number of eggs found in a late thirteenth century earth oven feature at Wairau Bar (South Island, New Zealand) – one of New Zealand’s most significant archaeological sites. Mitochondrial and nuclear DNA signatures confirmed this oven feature contained fragments of at least 31 moa eggs, representing three moa genera: Emeus; Euryapteryx; Dinornis. We demonstrate through the genetic identification of 127 moa eggshell fragments that thickness is an unreliable character for species assignment. We also present a protocol for assessing the preservation likelihood of DNA in burnt eggshell. This is useful because eggshell fragments found in archaeological contexts have often been thermally modified, and heat significantly increases DNA fragmentation. Eggshell is widely used in radiocarbon dating and stable isotope research, this study showcases how aDNA can also add to our knowledge of eggshell in both archaeological and palaeoecological contexts.     

Black Death in the rural cemetery of Saint-Laurent-de-la-Cabrerisse Aude-Languedoc,southern France, 14th century: immunological evidence

An excavation carried out in 2007 in Saint-Laurent-de-la-Cabrerisse (Aude-Languedoc, southern France), revealed a medieval rural cemetery used during the 8th–14th centuries. One hundred and forty nine graves were identified. Amongst those, three burials radiocarbon dated to the 14th century contained the remains of several individuals. No paleotraumatological evidences of violence due to warfare or to a civil massacre were identified in the exhumed skeletons. Therefore, we hypothesized that the simultaneous inhumations could have occurred during the “Black Death” outbreak or during one of its resurgences. A rapid diagnostic test for the detection of Yersinia pestis F1 antigen was applied to the nine putative plague victims to authenticate the cause of their death. Seven of the nine individuals tested from the three graves were positive to Y. pestis F1 antigen. We additionally tested fourteen skeletons from single graves archaeologically dated to the 13th–14th centuries. Four of them were positive to Y. pestis F1 antigen, thus showing a higher mortality rate due to plague than originally expected. The Saint-Laurent-de-la-Cabrerisse plague victims are the oldest samples from which Y. pestis F1 antigen has been, hitherto, identified. Few rural French cemetery containing plague victims have previously been identified and amongst them, only one dates to the 14th century. In the present report, we provide suggestive evidence of a second rural cemetery from southern France where victims of Y. pestis infection were buried. Our findings provide new information about the management of the plagued corpses during the “Black Death” in small rural communities.     

BEAR-CLAWS IN GERMANIC GRAVES

Summary. The remains of bear-skins for cremation burials of the late pre-Roman and early Roman Iron Age are a long neglected 'grave-good'. This is because of their scattered occurrence in graves on the continent in Sweden, north Germany, Bohemia and in the Celto-Germanic 'contact-zone' as far west as England. Their distribution and connection with many high-status graves marks them as Germanic'prestige goods'. This raises the question of ethnicity in these graves.     

Adipocere Inside Nineteenth Century Femora: The Effect of Grave Conditions.

Adipocere has, infrequently, been reported from archaeological contexts normally on the external surface of bodies. In contrast to those cases, this study focuses on a white, powdery and greasy substance found inside two right human femora during sampling. These samples were obtained from two identified individuals buried in the late 19th century, who were exhumed from a rock‐cut and a brick‐lined grave in a steeply sloping graveyard with wet soil conditions. Both individuals were buried in coffins. Fourier transform infrared spectroscopy was used to test the composition of the substances, and both were found to conform to the spectrum of adipocere. This is likely to be a breakdown product of the fats in bone marrow in an anaerobic, moist environment mediated by bacteria. None of the other individuals (n = 6) buried in similar graves displayed evidence of adipocere; this includes those whose femora were in a similar state of preservation (n = 4). Contemporaneous data on precipitation for the month of burials do not highlight any obvious trends, but one of the individuals was found in a water‐logged grave. The similar preservation of other femora buried in brick‐lined graves highlights the interplay of multiple factors in the formation and degradation of adipocere. More importantly, it demonstrates differential preservation, which may impact on DNA and other biomolecular research. Furthermore, this adds to the limited data currently available on adipocere found in archaeological contexts. Copyright © 2013 John Wiley & Sons, Ltd.     

mtDNA and osteological analyses of an unknown historical cemetery from upstate New York

Thirteen burials located on Jackson Street in Youngstown, NY, USA were recovered from a construction site and excavated in 1997. Based on the artifact assemblage, it was suggested that the cemetery was used sometime between the late 1700s and 1840. No historical records existed, and initial assessment of the skeletal remains was not able to determine any cultural affiliation. We carried out osteological and genetic investigations in order to gain insight into ancestral affiliation and kinship of the unknown individuals from the burials. Due to poor preservation of the remains, dental traits and limited osteological observations were available for only a few individuals. We performed DNA extraction and sequenced the mitochondrial DNA (mtDNA) control region following standard ancient DNA procedures. Our results suggest that ten individuals have evidence of biological affiliation with Native Americans, and in particular, four individuals have maternal Native American ancestry. One male individual was determined to be of European ancestry, from both the mtDNA and osteological results. This burial may reflect admixture as a result of frequent contact between Native Americans and Europeans during the eighteenth and nineteenth centuries and attempts by missionaries to convert Native Americans to Christianity. Our study demonstrates the usefulness of a multifaceted approach through archaeological, osteological, and genetic analysis that provides valuable perspectives in understanding the individuals buried at the Jackson Street Burials.     

Post-mortem DNA damage hotspots in Bison (Bison bison) provide evidence for both damage and mutational hotspots in human mitochondrial DNA

Post-mortem damage-driven mutations are a phenomena associated with ancient DNA (aDNA) studies. A previous study has demonstrated that the distribution of such mutations in human mitochondrial DNA is not random, but is concentrated in ‘hotspots’ that correlate with sites of elevated mutation rate in vivo. However, as the previous study was undertaken on human samples, it is possible for a critic to argue that the results might be biased through the presence of modern contaminant DNA sequences among the ancient DNA extracts. In this study we confirm the phenomena of DNA damage hotspots using a data set that is unlikely to be affected by contamination – cloned mitochondrial control region sequences extracted from 81 ancient bison (Bison bison). Furthermore, using published data from modern bovid specimens, we confirm that the damage hotspots correlate with sites of in vivo hypermutation. In conclusion, the aDNA sequences from archaeological specimens provide evidence that structural elements of mitochondrial DNA confer a degree of in vivo and post-mortem protection from sequence modification. This in turn provides useful insights into the debate as to whether mutational hotspots or mitochondrial recombination might best explain homoplasies observed on phylogenetic trees of human mitochondrial sequences.     

SEX IDENTIFICATION OF SLAVE SACRIFICE VICTIMS FROM QIN STATE TOMBS IN THE SPRING AND AUTUMN PERIOD OF CHINA USING ANCIENT DNA

Ancient DNA (aDNA) analysis was employed to determine the sex of slave sacrifice victims from Qin State tombs during the Spring and Autumn Period of China. It is difficult to obtain sex information from fragmentary skeleton samples with the aid of skeletal morphology methods. aDNA was extracted from the dentine in selected tooth samples of sacrificial slaves using a modified traditional method, which combines together phenol/chloroform extraction, silicon dioxide adsorption and ultrafiltration concentration. Based on the sequence differences between the amelogenin homologous gene in the X and that in the Y chromosome, a pair of specific primers was designed to identify the sex of the selected samples. In the selected eight typical samples, the aDNA analytical results revealed that three were males and two were females. These findings indicate that molecular sex identification might provide more valuable information for archaeological research on the institution of slave sacrifice in the Spring and Autumn Period of China.     

Tempo and mode of formation of the Late Epigravettian necropolis of Arene Candide cave (Italy): direct radiocarbon evidence

Existing ¹⁴C dates of charcoal and bone samples from different layers of the Late Epigravettian sequence at Arene Candide (Liguria, Italy) place the formation of the deposit between 12,000 and 10,000 (uncal.) BP, but it is unclear how long this cave was used for funerary purposes. Clarifying this point has important archaeological and anthropological implications, given number and variety of inhumations. These include single and double burials of richly ornamented adults, adolescents and children and disarticulated accumulations of bones. Six new direct ¹⁴C AMS dates obtained from human skeletons place more precisely the necropolis at the Late Pleistocene-Early Holocene boundary, and appear distributed in two distinct groups that indicate two phases of funerary utilisation of the cave. Although separated by many centuries, the two phases of interment show recurring formal elements. Both groups include skeletons in anatomical connection exhibiting similar grave goods as well as chaotic accumulations of bones and, more importantly, the double burial of an adult with a child lying to its left. The reappearance of similar funerary patterns in the same site at two widely separated points in time suggests that the cave held a symbolic significance over several centuries. This, and the additional evidence of funerary use of the cave during the preceding Gravettian, points to the endurance of Upper Palaeolithic cultural traditions.