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RESOLVING THE FISHER-FARMER CONUNDRUM IN PREHISTORIC EUROPE USING BIOMOLECULAR APPROACHES (AquaNeo)

Lead Research Organisation: University of Bristol
Department Name: Chemistry

Abstract

For most of the modern human evolution, humans were depending on food resources they hunted, gathered and fished. The transition to the Neolithic saw a major change in subsistence, with the introduction of domesticated plants and animals in human diet. Many coastal populations of early farmers turned their back from the sea and were not consuming marine foodstuffs. Little is known about early farmers from inland locations and their use of aquatic resources from rivers and lakes. Together with milk, aquatic resources are the only foodstuff available to farmers that are a source of vitamin D3, crucial for bone health in most of Europe where exposition to UVB are insufficient. Aquatic foods may thus have played a key nutritional role in farmers' diet, particularly in regions where milk was not greatly exploited. However, fishing and trapping skills are very much part of the hunter-gatherer life-ways. Aquatic resource use in farming communities is thus likely to show that these skills were transmitted from residual hunter-gatherer populations to early farmers (cultural hypothesis). Beyond the mere ease-of-access to productive rivers and streams, the use of aquatic resources by early farmers is hypothesised to have been shaped by nutritional necessity and access to cultural skills.

Aquatic skeletal remains are very small and fragile, and traditional methods for the detection of consumption of freshwater resources using human bone collagen is extremely challenging. We are targeting pottery sherds to detect the use of freshwater resources at early (inland) farming sites. Indeed lipids (or fats) trapped in pottery pores provide evidence for what was cooked in ceramic vessels. By characterising lipids using state-of-the-art analytical methods, we will be able to detect well-known compounds (or biomarkers) characteristic of aquatic resource processing. We will develop a novel method using a highly sensitive instrument to detect extremely low amounts of such biomarkers in the lipid assemblage of pottery extracts. We will also use the aquatic biomarkers and the carbon isotope signature of ubiquitous compounds found in animal fats (C16:0 and C18:0 fatty acids) to build a mixing model to quantify the amount of freshwater-derived animal fats in each pottery vessel.

There is very much uncertainty into whether proteins from foodstuffs would survive on archaeological pottery sherds - if so, they would provide very complementary insights to lipids as they are species-specific (while lipids are not). We will test the stability of foodstuff proteins by cooking diverse foodstuffs (including different types of fish) in replica pottery. We will then sample parts of the vessels and bury them in compost under two different conditions for 18 months to mimic archaeological degradation. We will then analyse the sherds using palaeo-proteomics methods to assess whether foodstuff proteins have survived and how well.

We have >1,500 archaeological lipid extracts and sherds curated in Bristol that were obtained from our European-funded NeoMilk project. We will select extracts and sherds from various farming sites across mainland Europe to investigate aquatic lipid biomarkers and proteins. That will enable us to assess the level and type of freshwater resource use at those sites. This novel data will be extremely valuable to detect sherds that were not used for cooking freshwater resources - they will be used to demonstrate that lipids preserved within can be 14C-dated and be used to build robust chronologies as they are not affected by an "old-carbon" effect.

Finally, we will use models to study the link between the use of freshwater resources by early farmers and ecological (accessibility to water bodies), nutritional (milk use, exploitation of hunted game) and cultural (contact with hunter-gatherer populations) parameters. This project will highlight the importance of freshwater ecosystems to human populations in the past, present and future.

Publications

10 25 50
 
Description To date, the main significant achievements of the award are (i) the development of a new analytical method for the detection of aquatic biomarkers in archaeological pottery (manuscript in preparation) and (ii) the sampling and analysis of 100s of pottery sherds and lipid extracts from across Europe for the detection of such aquatic residues.
More specifically, the objectives as described in the grant proposal and their status are described below.

O1 - Develop high sensitivity methods for the detection of aquatic biomarkers in lipid extracts from pottery sherds using GC-qTOFMS.
This objective was fully met as part of Izzy Davis' PhD studentship (funded through the NERC GW4+ DTP; but associated to this project).
O4 - Record aquatic (particularly freshwater) resource processing in pottery vessels from early farming communities at inland locations of Central Europe using the methods developed and validated in O1.
This objective is work in progress. To date, we have analysed ca. 600 pottery sherds from early farming communities in Central Europe. Diachronic and geographical trends in aquatic resource processing are currently investigated. Specific studies have been undertaker on selected regions (with >3 sites studied) or specific sites (with multiple phases).
In addition to this, we have been created a photographic atlas for the identification of aquatic remains in the faunal assemblage (Davis et al. 2024, International Journal of Osteoarchaeology).
O6 - Show that secure chronologies can be built using compound-specific 14C dating of pottery lipids devoid from aquatic biomarkers.
This objective is work in progress. Few sites and specific pottery sherds have been earmarked to carry out the study.
O5 - Test the correlation between aquatic resource use and ecological / nutritional / cultural factors using a formal model comparison framework.
The appointment of a PDRA mentored by Prof Mark Thomas is currently on hold, but this work package will be tackled in the next few months.
O2 - Test the stability of food proteins on ceramics, using cooking and degradation experiments with foodstuffs and mixtures of foodstuffs common in prehistory; O3 - Develop new methods for the quantification of the relative contribution of different foodstuffs to lipid residues using a Bayesian approach.
O2 and O3 are currently on hold, partly because other groups have been targeting those research questions.
Exploitation Route On of the main significant achievement of the award will be providing a high-resolution method for others to detect aquatic biomarkers in archaeological pottery.
Sectors Environment

 
Description Centre For Chemical Characterisation In Heritage Science (C3HS)
Amount £1,000,000 (GBP)
Funding ID AH/Z505808/1 
Organisation Arts & Humanities Research Council (AHRC) 
Sector Public
Country United Kingdom
Start 08/2024 
End 08/2026
 
Description Royal Society Dorothy Hodgkin Renewal
Amount £674,937 (GBP)
Funding ID DHF\R\241006 
Organisation The Royal Society 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2024 
End 03/2028
 
Title New photographic guide for the identification of European freshwater fish 
Description We developed a photographic guide for the bones commonly used in taxonomic identification, covering 32 species of freshwater and migratory fish present in central Europe. The guide aims to act as an accessible and user-friendly resource to assist the identification process. 
Type Of Material Biological samples 
Year Produced 2024 
Provided To Others? Yes  
Impact While a photographic guide atlas cannot replace the nuances of a physical reference collection or fully illustrate morphological variability within species, it provides an accessible and practical resource for species identification in archaeological context. . Davis I, Sykes N, Hochmuth M, Outram A, Roffet-Salque M. (2024). A photographic atlas for European freshwater and migratory fish remains and key considerations for their analysis. International Journal of Osteoarchaeology, 2024, e3284. 
URL https://onlinelibrary.wiley.com/doi/10.1002/oa.3284
 
Title Novel high-resolution mass spectrometric method for the identification of aquatic biomarkers in archaeological pottery sherds 
Description We exploit the full-spectrum, high-resolution and accurate mass features of the GC-qTOFMS (Agilent Technologies) enabling the simultaneous, highly selective and highly sensitive detection of suites of compounds in complex mixtures using a non-targeted approach. We use the deep coverage of GC-qTOFMS for the detection of aquatic biomarkers in archaeological lipid extracts, which has never been been done before. In contrast to single quadrupole GC-MS requiring several individual runs using full-scan and SIM mode, the GC-qTOFMS will allow the detection and verification of peak identities of all established aquatic biomarkers (IFAs / DHYAs / APAAs) with full sensitivity and mass spectra within one analysis. 
Type Of Material Biological samples 
Year Produced 2023 
Provided To Others? No  
Impact This new method is high-throughput, delivering higher data quality than traditional methods based on low resolution GC-MS operated in SIM mode, limiting the number of false negatives and false positives. Manuscript is in preparation. 
 
Description Collaboration on 14C dating 
Organisation University of Bristol
Department Bristol Radiocarbon Accelerator Mass Spectrometry Facility
Country United Kingdom 
Sector Academic/University 
PI Contribution Provided research questions for the application of compound-specific 14C dating.
Collaborator Contribution Provided expert knowledge in compound-specific 14C dating.
Impact Submission of an AHRC-DFG grant early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with archaeozoologists for access to reference collection 
Organisation Deutsches Archäologisches Institut
Country Egypt 
Sector Public 
PI Contribution (Note that the institution was registered as based in Egypt but is in Germany). Photographing fish bones for their collection for publication in as an Atlas.
Collaborator Contribution Access to the modern fish reference collection at the Deutsches Archäologisches Instituts, Berlin, Germany.
Impact Davis I, Sykes N, Hochmuth M, Outram A, Roffet-Salque M. (2024). A photographic atlas for European freshwater and migratory fish remains and key considerations for their analysis. International Journal of Osteoarchaeology, 2024. e3284.
Start Year 2022
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation Archäologisches Landesmuseum Baden-Württemberg
Country Germany 
Sector Public 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation Deutsches Archäologisches Institut
Country Egypt 
Sector Public 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation Landesamt für Archäologie Sachsen
Country Germany 
Sector Public 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation National Center for Scientific Research (Centre National de la Recherche Scientifique CNRS)
Country France 
Sector Academic/University 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation University of Gdansk
Country Poland 
Sector Academic/University 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation University of Las Palmas de Gran Canaria
Country Spain 
Sector Academic/University 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on archaeological pottery sherds 
Organisation University of Wroclaw
Country Poland 
Sector Academic/University 
PI Contribution Analyses of lipid residues preserved in pottery sherds from various regions in mainland Europe to investigate spatial and diachronic changes in aquatic resource use in Prehistory.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with archaeological information (e.g. contexts, dating).
Impact Collaboration were initiated before the award start date but were reinforced with this award. Some of the collaborations started with this award. Manuscripts in preparation. Multi-disciplinary collaboration: archaeology, chemistry. AHRC-DFG grant submitted with one of the partners early 2024 (unsuccessful).
Start Year 2023
 
Description Collaboration with foreign archaeologists for lipid residue analyses on pottery sherds (cooking experiments) 
Organisation University of Kiel
Country Germany 
Sector Academic/University 
PI Contribution Analyses of lipid residues preserved in pottery sherds from experimental cooking experiments.
Collaborator Contribution Provided material (pottery) for lipid residue analyses with experimental information.
Impact Manuscript in preparation. Multi-disciplinary collaboration: archaeology, chemistry.
Start Year 2022
 
Description Collaboration with specialist in data processing 
Organisation Friedrich-Alexander University Erlangen-Nuremberg
Country Germany 
Sector Academic/University 
PI Contribution n/a
Collaborator Contribution Bespoke training in data processing (2022), support in data processing using MZMine (since 2022).
Impact n/a Multi-disciplinary collaboration: chemistry, mass spectrometry, data science.
Start Year 2022
 
Description After-school activity at Bristol Grammar School 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Ca. 20 pupils attended a 1h-workshop designed and delivered by us.
Year(s) Of Engagement Activity 2024
 
Description Bristol Innovation Partnership - Inspiring Innovation in STEM day 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Local 14/15-year-olds visiting from ca. 12 local schools.
The event was followed by an invitation from Bristol Grammar School to propose an after-school activity for their pupils.
Year(s) Of Engagement Activity 2024
 
Description FUTURES Festival - Schools' Research Fair at the SS Great Britain 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact Senior School Groups, years 7-8, ages 11-13 attended the event on 20.10.2023. Learning outcomes for schools: experience hands-on research exhibits, knowledge in different research disciplines, inspiration about careers in reserach.
Year(s) Of Engagement Activity 2023
URL https://futuresnight.co.uk/
 
Description Festival of Nature 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Display at the Festival of Nature in Bristol (10th, 17th-18th June 2023), the UK's largest celebration of the natural world.
Year(s) Of Engagement Activity 2023
URL https://www.bnhc.org.uk/festival-of-nature/
 
Description Radio programme broadcasted on France Culture 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Guest interviewee in a 29-minute Radio programme 'Carbone 14, le magazine de l'archéologie' broadcasted on the public radio channel France Culture in October 2022 (with 200k podcasts).
Triggered an invitation to a conference as keynote speaker.
Year(s) Of Engagement Activity 2022
URL https://www.radiofrance.fr/franceculture/podcasts/carbone-14-le-magazine-de-l-archeologie/l-epopee-d...