Description of Research Projects

1 PhD and 1 postdoc position: Palaeogenomic / Biostatistic analysis of ancient DNA data from Mesolithic and Neolithic skeletal remains

Institute of Anthropology, Johannes Gutenberg University Mainz, Germany; Joachim Burger

Apply to: BEAN.MAINZ at googlemail.com 

Data garnered from the extraction and analysis of ancient DNA from human skeletal remains can be used to reconstruct the demography of past populations; parameters such as population size and timing of genetic bottlenecks can be inferred from genomic data. Through the use of next generation ancient DNA sequencing, the early-stage researcher (ESR) in Mainz will gather genetic data from relevant skeletal collections originating from western Anatolia and south-eastern Europe (particularly the Balkan region). Approximately 200 Mesolithic and Neolithic skeletons from Turkey, Serbia, Greece, Bulgaria, Romania, and Hungary will be analysed. Many of these specimens are readily available from the research collections of BEAN project participants and associates Sofija Stefanović (UoB), Necmi Karul (UoI), and Christina Papageorgopoulou (JGU Mainz), allowing the ESR to begin analysis immediately.

The state of the art of human genetic diversity analysis is moving from typing a few known SNPs to resequencing of large amounts of sequence using next generation sequencing (NGS). This technology is particularly promising for ancient DNA analysis as it uses short fragments of size typical in fossil DNA, it produces several orders of magnitude more data than PCR and gives a representative sample of all DNA in the sample which is an aid to distinguishing endogamous ancient and contaminating modern DNA.

The aim of this sub-programme is to investigate demographically relevant markers from each skeleton for further population genetic analysis and computer simulations. To this end, the ESR will use a capture/enrichment approach to target i) full mitochondrial genomes ii) the majority of the exome on the human X-chromosome iii) 1,000 ancestry informative neutral autosomal SNPs iv) 200 loci that are candidates for being under selection during the last 10,000 years. Initial NGS data analysis will be carried out by the ER in Mainz. DNA sequence data will be used for standard population genetic analysis in Mainz and forwarded to the projects led by Mark Thomas at UCL and Mathias Currat of the University of Geneva for further statistical inference methods. DNA libraries from skeletons with excellent DNA preservation will be provided to the project led by Dan Bradley of Trinity College Dublin for a deeper Y-chromosomal sequencing approach.



1 PhD position: Orienting Y-chrmosome lineages in space and time

Trinity College Dublin, Ireland, Smurfit Institute; Dan Bradley

http://www.gen.tcd.ie/molpopgen/dan.php

Apply to: dbradley at tcd.ie

There is a huge wealth of Y chromosome data from modern surveys using a standard reference panel of SNPs that are known to show strong geographical structure. Patterns observed using thousands of samples from Europe and the Near East have been subject to very strong interpretations where, for example the spread of farmers into Europe have been cited as the origin of alternately, i) virtually all, and ii) only a minority of modern European Y chromosome lineages. The postulation of an association between specific SNP lineages and the Near Eastern Neolithic also has led to strong inference about migrations into North Africa at the dawn of agriculture. There is a clear need to produce time-stamped Y chromosome lineages to test and distinguish between these hypotheses.

Simply typing SNPs that have been ascertained using modern sampling gives a risk of missing vital patterns of variants in the past. One cannot assume that all past variants, or even major families of variants from thousands of years ago are represented today; indeed there are indications from other systems that sharp discontinuity from ancient to modern is possible. For this reason we propose to re-sequence the major non-repetitive regions of the Y chromosome in ancient samples. This will cover positions where SNPs have already been ascertained from modern samples, thus embedding these specimens in existing phylogenetic patterns. However, this will also uncover relationships that may be unobtainable from modern study but which may be critical in interpreting relationships among ancient groups. 

The ESR of this sub-programme will investigate these anthropological and methodological questions using the DNA libraries generated by the ESR in Mainz, which derive from approximately 200 Mesolithic and Neolithic skeletons from western Anatolia and southeastern Europe. Indexed next generation sequencing libraries will be generated by ligation and PCR. We will use RNA bait capture and Illumina GA next generation sequencing in a range of carefully selected samples to produce ~1 Mb of sequence per individual. The calling of authentic SNPs from ancient material will be non-trivial, given the error rates associated with ancient DNA and NGS. However, methods exist for this and the testing for known SNPs will provide a training data set, allowing a calibration of quality control parameters. Whereas NGS has opened up the prospects of large amounts of data being harvested from ancient samples, the analysis of whole genomes is not yet feasible as a population screening tool. However, the minority of the Y chromosome sequence which is comprised neither of large repeats or simple sequence represents a reasonable target size for the economical screening of multiples samples. The data obtained from this Y-chromosome re-sequencing sub-programme will be compared to modern European Y-chromosome reference data to examine the contribution of these ancient populations to modern European Y-chromosome diversity. These results will be forwarded to the ESRs supervised by Mathias Currat (UNIGE), Mark Thomas (UCL), and Jean-Pierre Bocquet-Appel (CNRS) for incorporation into demographic and genetic models.



1 PhD position: Assessing the transition to agriculture in Western Anatolia and the Balkans using a spatially-explicit computer simulation approach

University of Geneva Switzerland, Mathias Currat

Apply to: mathias.currat at unige.ch

This sub-programme will investigate the processes by which the Neolithic culture diffused from Western Anatolia to the Balkans by using an inter-disciplinary model-based computer simulation approach. The basic principles of this method in the context of human evolution are: i) model possible alternative evolutionary (e.g. demographic) scenarios in a specific region (e.g. demographic diffusion model (DDM) versus cultural diffusion model (CDM) in Anatolia and the Balkans); ii) implement these models into a computer program; iii) simulate numerous independent instances of each alternative scenario by varying the input parameters within a range of plausible values, in order to explore the space of parameters; iv) compute output statistics (e.g. genetic) for each simulation; v) compare the distributions of the statistics obtained under each alternative model to the values estimated from real data in real populations, in order to evaluate formally which scenario (and which combination of parameter values) is the most probable, or at least to discard scenarios that do not fit the observed data at all.

The computer simulation approach offers the advantage of merging information from different fields in the model, possibly archaeology, zooarchaeology, landscape archaeology, palaeogeography, linguistics and genetics. The goal of this sub-programme is to develop hypothetical demographic scenarios for the Neolithic transition in Western Anatolia and the Balkans based on archaeological data and to evaluate these scenarios using genetic data sampled in present and past populations from the region. The simulation of alternative scenarios will be performed using a modified version of the program SPLATCHE2 accounting for aDNA. This coalescent-based program has been designed to simulate the molecular diversity of samples of genes in a heterogeneous and dynamic environment. Its goal is to translate environmental information into genetic information in order to study the impact of demographic factors on the genetic structure of populations. This approach has proved invaluable to the study of patterns of genetic variation in spatially-explicit contexts, notably in the context of human range expansions and more specifically to study the genetic consequences of the Neolithic transition in Europe at a continental level. Ancient DNA samples will be an invaluable information source as they provide snapshots of the genetic structure of populations at different time points in the past, and thus anchor the models with real data from distinct time-periods. SPLATCHE2 can be used in concert with the ABC toolbox package, a powerful statistical tool which implements the Approximate Bayesian Computation Approach (ABC) to evaluate evolutionary scenarios.

This research plan will be strengthened by being embedded within the BEAN ITN. The definition of the alternative models and the choice of parameter values will be carried on in close interaction with field archaeologists and palaeogeography experts (BEAN participant Necmi Karul (UoI), as well as associates Çiler Çilingiroğlu (EgeU) and Barbara Horejs (ÖAI)). This is a critical point as there is often a lot of uncertainty on the values of the parameters when modelling evolutionary scenarios including palaeodemographic and palaeoenvironmental information. A close collaboration is thus necessary to ensure the development of realistic models. Second, the genetic samples from Anatolia and south-eastern Europe typed by the ESRs working in ancient DNA laboratories belonging to the BEAN ITN (JGU Mainz and TCD) will be used for the statistical evaluation of scenarios (in addition to published data). A fine coverage of the area under study, in terms of genetic samples, is essential to infer past population demography from genetic data. Joachim Burger (JGU Mainz) and Dan Bradley’s (TCD) teams are among the few laboratories in the world that produce ancient DNA (aDNA) data for the Neolithic period in Europe, and the inclusion of those data will dramatically increase the statistical power of the method of discrimination among alternative scenarios.



1 PhD position: Tracing genes and culture through the Neolithic

University College London, United Kingdom, Mark Thomas

http://www.ucl.ac.uk/mace-lab/people/mark

Apply to: http://www.ucl.ac.uk/mace-lab/opportunities.

Demographic processes, including migration, population growth and decline, and admixture have been shown not only to affect patterns of genetic variation, but also the accumulation and distribution of culturally inherited traits. It is therefore to be expected that both genetic and cultural data contain information on the shared population histories and processes that have shaped them. Recent advances in modelling cultural evolutionary processes have drawn heavily on the “neutral model” in population genetics, and have been widely applied to explain typological variation in many archaeological and cultural datasets. Furthermore, through the application of newly developed statistical inference methods such as approximate Bayesian computation (ABC), which rely on comparisons of simulated and observed datasets, and models that incorporate both genetic and cultural processes, it is possible to infer population histories and evolutionary trajectories, and to explicitly compare different competing hypotheses. Critically, this latter approach allows the formal integration of both cultural and biological data on an equal footing in a coherent statistical framework for the first time.

By considering genetic (Bradley and Burger) and cultural (Shennan and Karul) data generated by network partners, the ESR in this project will test a range of different hypotheses on the spread of Neolithic people and lifeways from the Near East to Europe around 7,000 years ago. Genetic data will include both ancient and modern human DNA, and cultural data will include information on the frequencies of different pottery and other artefact styles through time. Genetic variation will be analysed using serial coalescent approaches, which allows the incorporation of ancient DNA data from different time periods with modern genetic data. The distribution of culturally inherited traits through time will be studied using a newly developed neutral model. This approach uses a generation time-independent Moran process to overcome many of the unrealistic assumptions of previous neutral models of cultural change and incorporates factors such as population size and structure. In addition to looking at changes in the frequencies of specific artefact types through time, we will also consider archaeological proxies for overall accumulation of cultural complexity using the cultural transmission model described by Powell et al.. We will generate expectations of genetic and cultural data under the same range of demographic scenarios and compare these different hypotheses using ABC, which rests on the ability to generate simulated data from models that describe realistic demographic scenarios. The model will incorporate genetic and cultural data generated by partner ESRs and ERs, and the predictions of the model will be used to test the strength of the correlation between the genetic and material culture correlates of the Neolithic expansion.


1 Postdoc position in biotech industry: Next generation genomics protocols for studies of migration using aDNA

GATC BioTech, Konstanz Germany, Janet Kenlies

Apply to: j.kenklies at gatc-biotech.com

The isolation of extremely small amounts of ancient DNA from archaeological material must be performed under clean room conditions to prevent contamination by modern human mitochondrial or nuclear DNA or contemporary microbe DNA (prokaryote or eukaryote). This process begins at the excavation site and continues with the isolation of the DNA at JGU Mainz. The small amounts of material isolated represent additional challenges to further processing in a standard molecular biology laboratory using manufacturer NGS protocols, because it is not possible to characterize the material to estimate concentration, fragment size distribution or degradation. Sequencing library preparation requires the ligation of adaptors to the sample DNA that may also contain a barcode to allow multiplexing of samples and aid in identification of contaminating DNA sequences. The DNA fragments must also be size selected to isolate fragments 80bp to 200bp in length (including adaptors) prior to sequencing. DNA from the target organism must also be enriched using commercially available methods (e.g. Agilent SureSelect). The enrichment oligo sequences will have to be optimized to perform well for the desired genomic target to ensure coverage of loci necessary for population genetic analysis. Difficulties in working with such small amounts of material will be identified and a protocol will be developed that ensures sample purity and minimizes the work required in a clean room laboratory. Sample preparation will be optimized to allow processing in a standard molecular biology laboratory as early in the processing protocol as feasible. Sequencing is performed using Next Generation Sequencing (NGS) technology to generate the raw data for the bioinformatic analysis (SNP discovery or comparative genomics).

The bioinformatic analysis can be divided into two phases: Phase 1 is processing of the raw sequence data. The HiSeq 2000 quality metrics are used to call the bases in the DNA sequence. This can be done using the Illumina software or external software that can be more flexibly matched to the characteristics of the samples sequenced (e.g. Ibis http://bioinf.eva.mpg.de/Ibis/). After base calling, sequence reads that do not belong to the target species can be identified and removed. Correction of misidentified bases due to chemical degradation of ancient DNA accumulated over millennia before excavation (e.g. the deamination of cytosine to uracil) is performed. It will then be possible to recalculate the quality scores of affected bases and, in conjunction with mapping to the reference human genome, improve the identification of SNPs. Phase 2 analysis compares the sequence data to the reference human genome by mapping the reads using one of the standard algorithms (e.g. BWA http://bio-bwa.sourceforge.net/). In a second step SNPs and Indels are identified and characterized. The base quality scores can be used to define a SNP or Indel quality score that can be used to improve the confidence of the analysis. The data and preliminary analysis are then sent to BEAN partners for further analysis.

Completion of this project will provide GATC Biotech with valuable laboratory protocols, operating procedures and processes, analysis pipelines and experience sequencing and analysing large genomes in complex samples containing DNA from many different organisms including possible contamination from persons who collected and processed the samples. This represents a special case of metagenome sequencing that will give GATC a competitive advantage in the palaeogenetics field. This project will allow GATC to develop products to meet the needs of palaeogenetecists and researchers working with metagenomics of complex samples and contaminated samples, environmental samples having many organisms, but few target organisms, and agricultural samples containing symbiotic and syntropic organisms in complex metabolic interdependencies.


1 PhD position: Ceramic assemblages as evidence of social interaction in the Early Neolithic Balkans

University College London, United Kingdom, Stephen Shennan

Apply to: ioa-director at ucl.ac.uk

Research on the scientific characterisation of Early Neolithic Starčevo-Criş pottery in the Balkans, carried out under Shennan’s supervision in 2002–2005 and supported by the Leverhulme Trust, provided a new picture of the Neolithic of the central Balkans, emphasising the similarities in pottery production techniques at Starčevo-Criş sites over a wide geographical area and throughout a period of ca. 800 years; the same technology and raw materials were used regardless of vessel shape, surface treatment, or decoration. Additionally, radiocarbon results indicate that changes in in pottery forms, surface treatments and motifs, appear to have occurred more or less simultaneously throughout the study area. However, it is clear from the results of the petrographic, SEM-EDS and XRD analyses that the pottery was always manufactured locally. What was transmitted from one community to another was not pottery itself, but the idea of making pottery in a certain style and by a particular method. This coherence in Starčevo-Criş pottery production stands in contrast to the differences in early Neolithic pottery production between the central Balkans and the adjoining Impressed Ware region on the Adriatic coast, which suggest the existence of a persistent cultural boundary.

Given the similarities between some of the artefacts and symbols found at early Neolithic sites in Anatolia and in the central Balkans (the Körös-Starčevo-Criş Cultures), many authors have assumed that the Neolithic of the Balkans originated in Anatolia, and that the space between these two areas represents the bridge between Asia and Europe. However, it has been pointed out by several authors that there exist significant differences between Early Neolithic traditions in Greece, the Balkans, and Anatolia that are largely incompatible with this hypothesis. In the light of these results and the ongoing debates about the processes involved in the spread of farming from southwest Asia to Europe, this research sub-programme will have two main aims: to characterise the spatial and temporal continuity in ceramic traditions, and to interpret these results in terms of the economic and social context of the early Neolithic.

Given the uniformity in production methods found in the central Balkans, we plan to use established ceramic characterisation methods to determine whether Early Neolithic ceramics in Bulgaria, Thrace, and western Anatolia, which are similar in surface appearance, were produced by the same methods. The ESR on this project will carry out a systematic analysis of the similarities and differences in the painted and impressed decoration and surface treatment used in the pottery assemblages at different sites, assessing the extent to which they relate to patterns in social interaction dependent on proximity in space and time. In turn these can be compared to independent evidence of interaction based on the exchange of lithic raw materials and other goods. We plan to incorporate the results of these analyses into a framework for modelling the demographic correlates of the patterning in ceramic technology and style observed in these regions. Traditional archaeology in the Balkans has described and discussed changing decorative patterns across time and space, but there has been no systematic recording and comparison of assemblages to develop and test models, only long-established forms of interpretation, for example in terms of successive invasions. While the nature of previous data collection means that a quantitative description of ceramic assemblage variation would be suspect, a detailed presence-absence trait analysis would be both valid and practicable and would provide the basis for model-testing.

The results of BEAN network participant Necmi Karul’s trade network analysis will be compared with these results in order to make broader inferences about the mobility of material culture and technology in the Anatolian and Balkan Neolithic. BEAN network associate Çiler Çilingiroğlu will expand the scope of this analysis to include material from additional contemporaneous sites in western Anatolia.



1 PhD position: Lithics and raw materials as source for mobility and migration in Neolithic and Chalcolithic periods: a case study from western Anatolia

Department of Prehistory, University of Istanbul, Turkey, necmi Karul

Apply to: karul at istanbul.edu.tr

Preliminary analyses of the lithic assemblages from Çukuriçi Höyük, a tell site in Turkey dating from 6,2002,600 BC, suggest that the site is a rich repository of information concerning interregional exchange, communication and mobility in the western Anatolian Neolithic. This research sub-programme intends to investigate the nature and extent of trade networks in western Anatolia using Çukuriçi Höyük as a focal point. The primary goals of this research sub-programme are to evaluate different models of raw material circulation and to determine possible technological influences between the regions of interest.

The majority of the knapped stone artefacts at Çukuriçi Höyük are made of obsidian, although there is also a significant quantity of lithics made from flint at the site. Evidence from neutron activation analysis (NAA) and surveys of material availability in the region surrounding Çukuriçi Höyük suggest that certain raw materials, particularly flint, were acquired over long distances.. These preliminary results point toward differences in exchange networks and mobility within the central Anatolian coastal group, possibly a distinction between inland and coastal sites. Building upon these results, the ESR in this sub-programme will combine new data from excavations of late Neolithic levels (mid-7th millennium BC) at Çukuriçi Höyük planned in 201112 and funded by an ERC project directed by BEAN network associate Barbara Horejs (ERC Starting Grant project “From Sedentism to Proto-Urban Societies in Western Anatolia”, Project no. 26339).The ESR will also investigate the knapping technology in use at the site in order to compare the artefacts and technology at Çukuriçi Höyük with those at other yet-unpublished sites in the region, such as Ulucak, Aktopraklık, and Ege Gübre. The results of these comparisons will allow inferences to be made about regional mobility and exchange, including technology transfer and the frequency of transport of readymade tools.

The second focus of this project will be on the raw materials traded and exploited throughout the exchange network. Obsidian samples from Çukuriçi Höyük and other sites in the region will be analysed using NAA. With new data from Çukuriçi Höyük and other yet-unpublished sites in the region, a more detailed picture of the region’s prehistoric networks can be expected. Because obsidian is a rather scarce resource with readily-characterized source-specific chemical properties it provides an excellent material basis for the definition of possible circles of exchange and influence. In addition to the obsidian analyses, a new method for geochemical analyses on non-obsidian artefacts will be tested within this project. We propose to compare flint and chert artefacts with samples from known primary sources both micropalaeontologically (microscopic analysis of the fossils in the sample) and geochemically in order to correlate artefacts with source material locations. Combining these new data with results from other settlement sites in western Anatolia (inland and coast), the Marmara region, Greece and the Balkans will clarify questions concerning the movement of raw material and resources, as well as delineate regional networks of communication, exchange, and mobility. The comparison of raw material exchange networks on the Greek mainland versus those in western Anatolia should be particularly informative. Additionally, an inter-temporal comparison of raw material and lithic assemblages from the 3rd versus the 67th millennium BC will allow us to evaluate changes in the circulation of raw materials and technology, and therefore of human mobility. BEAN associate Barbara Horejs will help to organize the raw material characterization component of this project, and BEAN associate Çiler Çilingiroğlu will provide complementary archaeological data from comparable sites in the region. The results of this analysis will be shared with the groups led by Stephan Shennan, Mark Thomas, and Mathias Currat for inclusion in models of cultural trait and material culture distribution.


1 PhD position: Testing the assumptions of the Neolithic demographic transition (NDT) through direct analysis of skeletal remains

Faculty of Archaeology, University of Belgrade, Serbia, Sofija Stefanović

http://www.f.bg.ac.rs/en2

Apply to: smstefan at f.bg.ac.rs

The Neolithic Demographic Transition (NDT) is usually explained as a consequence of two primary factors: changes in nutrition that led to the gradual reduction of low-calorie food and to increase of high-calorie food; and the adoption of a sedentary way of life that led to the reduced female mobility. A possible tertiary factor is a shortened period of lactation, which could allow for a greater number of pregnancies and deliveries. At present, very little work has been done toward investigating the causes and correlates of the Neolithic Demographic Transition (NDT) through the examination of direct skeletal evidence. Here we propose to investigate a suite of physical correlates associated with dietary intake, mobility, and age at weaning in an assemblage of over 200 well-preserved Mesolithic and Neolithic skeletons from the Danube Gorges.

Although a number of cross-cultural studies suggest that agricultural products increase fertility, it remains uncertain whether access to domesticates is a necessary precondition for increased fertility. Here we test whether the incursion of the Neolithic (and therefore agriculture) into the Danube gorges is correlated with an increase in the availability of calorie-dense foods, and whether there is a relationship between nutritional changes and changes in fertility in this region. We propose to analyse the strontium: calcium ratios of the skeletons in our assemblage, and to compare the results from Mesolithic skeletons with those of Neolithic skeletons in order to assess the relative predominance of grains in the diets of the two populations. The principal source of strontium in the diets of the two populations can be inferred from Sr/Ca ratios, if we assume that grains are a novel introduction to the Danube Gorges diet not available to Mesolithic populations.

There are also indicators of interdependence between the length of lactation and fertility. However, we have very little direct evidence about the duration of breastfeeding in prehistory at present. The investigation of Sr/Ca ratios across the neonatal line in human dentition has been developed as an indicator of age at weaning , and this project intends to use Sr/Ca to examine decrease in breast-feeding began and provide evidence of regional differences in the onset of weaning. We intend to examine 100 juvenile Mesolithic and Neolithic skeletons from the Danube Gorges collection, as well as 10 juvenile skeletons from the Serbian Neolithic sites of Jaričište and Gomolova. Histological sections of the relevant dentition will be prepared and measured through laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

Reduced mobility of females in the Neolithic is considered as an important prerequisite for the NDT, as a mobile foraging lifestyle is incompatible with the provisioning of multiple dependent offspring. Previous studies of the mobility of Mesolithic and Neolithic populations have not included direct skeletal evidence from the Balkan region. A preliminary study of musculo-skeletal stress and of femoral morphology in the Danube Gorges indicated that the Mesolithic females in the area were not highly mobile, suggesting that in some regions, Mesolithic females were already at least semi-sedentary. By analysing female mobility during the Mesolithic we can test whether a reduction in female mobility, considered an essential prerequisite for the NDT, began before the Neolithic. To this end, we propose to conduct a macroscopic and radiological examination of musculo-skeletal stress markers of the lower limbs and analysis of femoral cross sectional geometry in the Danube Gorges collection. Results from these analyses will be incorporated into the palaeodemographic models produced by the ESRs in the CNRS group, as well as the models of ESRs at UCL and UNIGE in work package 1.


1 PhD position: Model of the mode, tempo, and demography of the Neolithic expansion in Greece and Bulgaria from the Levant, using archaeological and bioarchaeological data

CNRS, Paris, France,  Jean-Pierre Bocquet-Appel

Apply to: jean-pierre.bocquet-appel at evolhum.cnrs.fr

The emergence of the European Neolithic is rooted in the Levant with the simultaneous emergence of the agricultural production system and a major qualitative demographic shift between 11,500–9,000 calBP. This unprecedented demographic shift in human history, designated the Neolithic Demographic Transition (NDT), was characterized by an increase in the average individual female fertility of almost 50% in 3,000 years, resulting in an increase in the population growth-rate and size of the Levantine area. This population explosion has left many archaeological traces. But at the end of the Pre-Pottery Neolithic B, a demographic collapse seems to occur in this geographical Levantine area, with a reduction in the sizes of the sociological units (villages, hamlets), an internal reorganization of their structures toward a reduction in domestic surfaces, and an increase in the geographical dispersion of these agricultural sociological units. This apparent demographic collapse in the east coincides chronologically with the European westward expansion of the farming system in Greece and Bulgaria. A convenient geographical device could be formed in subdividing the surface of the westward expansion into a Levantine core area (containing the origins of agriculture and the NDT) and a western area of colonization in Greece and Bulgaria.

This sub-programme plans to use archaeological, palaeoanthropological and palaeogenetic data from Neolithic humans, animals and cultigens to trace the expansion of Neolithic settlements westward from the Levant to Greece and Bulgaria. More specifically, we plan to create a database-informed demographic reconstruction of the populations of key settlements along the expansion corridor by estimating the space-time densities of radiocarbon-dated Mesolithic and Neolithic sites in the region, analysing the age distributions and stature markers present in burial assemblages to understand population dynamics, plotting the phylogeographic distribution of cultural artefacts, and incorporating palaeogenetic data on humans and their domesticates to infer population structure. This database will be incorporated into a GIS platform to investigate spatial correlates of the demographic phenomena of interest. Our goal is to estimate the timing and expansion rate of the movement of the Neolithic into Greece and Bulgaria, and to describe the genetic structure and demographic parameters of the populations involved. We also plan to investigate whether the demographic, biological, and cultural characteristics of this population expansion can be correlated with the 8,200 calBP Holocene cooling event. By analogy with the genetic drift model, we plan to investigate whether the modes of colonization from the core area of the Levant toward Greece and Bulgaria (demic, leapfrog of clan, sub-clan, family, individuals) can be inferred using these data. Anthropological data produced by BEAN network contributing scientist Christina Papageorgopoulou and BEAN network participant Sofija Stefanović will be incorporated into the database used in this project.