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Durrington Walls was a large Neolithic settlement in Britain dating around 2500 BCE, located very close to Stonehenge and likely to be the campsite where its builders lived during its main stage of construction. Nineteen coprolites recovered from a midden and associated pits at Durrington Walls were analysed for intestinal parasite eggs using digital light microscopy. Five (26%) contained helminth eggs, 1 with those of fish tapeworm (likely Dibothriocephalus dendriticus) and 4 with those of capillariid nematodes. Analyses of bile acid and sterol from these 5 coprolites show 1 to be of likely human origin and the other 4 to likely derive from dogs. The presence of fish tapeworm reveals that the Neolithic people who gathered to feast at Durrington Walls were at risk of infection from eating raw or undercooked freshwater fish. When the eggs of capillariids are found in the feces of humans or dogs it normally indicates that the internal organs (liver, lung or intestines) of animals with capillariasis have been eaten, and eggs passed through the gut without causing disease. Their presence in multiple coprolites provides new evidence that internal organs of animals were consumed. These novel findings improve our understanding of both parasitic infection and dietary habits associated with this key Neolithic ceremonial site.

The cycling of soil organic matter (SOM) and carbon (C) within the soil is governed by the presence of key macronutrients, particularly nitrogen (N) and phosphorus (P). The relative ratio of these nutrients has a direct effect on the potential rates of microbial growth and nutrient processing in soil and thus is fundamental to ecosystem functioning. However, the effect of changing soil nutrient stoichiometry on the small organic molecule (i.e., metabolite) composition and cycling by the microbial community remains poorly understood. Here, we aimed to disentangle the effect of stoichiometrically balanced nutrient addition on the soil metabolomic profile and apparent microbial carbon use efficiency (CUE) by adding a labile C source (glucose) in combination with N and/or P. After incorporation of the added glucose into the microbial biomass (48 h), metabolite profiling was undertaken by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). 494 metaboliteswere identified across all treatments mainly consisting of lipids (n = 199), amino acids (n = 118) and carbohydrates (n = 43), >97% of which showed significant changes in concentration between at least one treatment. Overall, glucose-C addition generally increased the synthesis of other carbohydrates in soil, whileaddition of C and N together increased peptide synthesis, indicative of protein formation and turnover. The combination of C and P significantly increased the number of fatty acids synthesised. There was no significant change in the PLFA-derived microbial community structure or microbial biomass following C, N and P addition. Further, N addition led to an increase in glucose-C partitioning into anabolic processes (i.e., increased CUE), suggesting the microbial community was N, but not P limited. Based on the metabolomic profiles observed here, we conclude that inorganic nutrient enrichment causes substantial shifts in both primary and secondary metabolism within the microbial community, leading to changes in resource flow and thus soil functioning, however, the microbial community illustrated significant metabolic flexibility.

Food is often one of the most distinctive expressions of social, religious, cultural or ethnic groups. However, the archaeological identification of specific religious dietary practices, including the Jewish tradition of keeping kosher, associated with ritual food practices and taboos, is very rare. This is arguably one of the oldest known diets across the world and, for an observant Jew, maintaining dietary laws (known as Kashruth) is a fundamental part of everyday life. Recent excavations in the early medieval Oxford Jewish quarter yielded a remarkable assemblage of animal bones, marked by a complete absence of pig specimens and a dominance of kosher (permitted) birds, domestic fowl and goose. To our knowledge, this is the first identification of a Jewish dietary signature in British zooarchaeology, which contrasted markedly with the previous Saxon phase where pig bones were present in quantity and bird bones were barely seen. Lipid residue analysis of pottery from St Aldates showed that vessels from the possible Jewish houses were solely used to process ruminant carcass products, with an avoidance of pig product processing, correlating well with the faunal data. In contrast, lipid analysis of pottery from comparative assemblages from the previous Saxon phase at the site and a contemporaneous site in the city, The Queen’s College, shows that the majority of these vessels appear to have been used to process mixtures of both ruminant and non-ruminant (pig) products. Here, the combination of organic residue analysis, site excavation and animal and fish bone evidence was consistent with the presence of Jewish houses in eleventh- and twelfth-century St Aldates, Oxford, hitherto only suspected through documentary information. This is the first identification of specific religious dietary practices using lipid residue analysis, verifying that, at least 800 years ago, medieval Jewish Oxford communities practised dietary laws known as Kashruth.

In European and many African, Middle Eastern and southern Asian populations, lactase persistence (LP) is the most strongly selected monogenic trait to have evolved over the past 10,000 years1. Although the selection of LP and the consumption of prehistoric milk must be linked, considerable uncertainty remains concerning their spatiotemporal configuration and specific interactions2,3. Here we provide detailed distributions of milk exploitation across Europe over the past 9,000 years using around 7,000 pottery fat residues from more than 550 archaeological sites. European milk use was widespread from the Neolithic period onwards but varied spatially and temporally in intensity. Notably, LP selection varying with levels of prehistoric milk exploitation is no better at explaining LP allele frequency trajectories than uniform selection since the Neolithic period. In the UK Biobank4,5 cohort of 500,000 contemporary Europeans, LP genotype was only weakly associated with milk consumption and did not show consistent associations with improved fitness or health indicators. This suggests that other reasons for the beneficial effects of LP should be considered for its rapid frequency increase. We propose that lactase non-persistent individuals consumed milk when it became available but, under conditions of famine and/or increased pathogen exposure, this was disadvantageous, driving LP selection in prehistoric Europe. Comparison of model likelihoods indicates that population fluctuations, settlement density and wild animal exploitation—proxies for these drivers—provide better explanations of LP selection than the extent of milk exploitation. These findings offer new perspectives on prehistoric milk exploitation and LP evolution.

Chemical analyses of carbonized and absorbed organic residues from archaeological ceramic cooking vessels can provide a unique window into the culinary cultures of ancient people, resource use, and environmental effects by identifying ingredients used in ancient meals. However, it remains uncertain whether recovered organic residues represent only the final foodstuffs prepared or are the accumulation of various cooking events within the same vessel. To assess this, we cooked seven mixtures of C3 and C4 foodstuffs in unglazed pots once per week for one year, then changed recipes between pots for the final cooking events. We conducted bulk stable-isotope analysis and lipid residue analysis on the charred food macro-remains, carbonized thin layer organic patina residues and absorbed lipids over the course of the experiment. Our results indicate that: (1) the composition of charred macro-remains represent the final foodstuffs cooked within vessels, (2) thin-layer patina residues represent a mixture of previous cooking events with bias towards the final product(s) cooked in the pot, and (3) absorbed lipid residues are developed over a number of cooking events and are replaced slowly over time, with little evidence of the final recipe ingredients.

Archaeology has yet to capitalise on the opportunities offered by bioarchaeological approaches to examine the impact of the 11th-century AD Norman Conquest of England. This study utilises an integrated multiproxy analytical approach to identify and explain changes and continuities in diet and foodways between the 10th and 13th centuries in the city of Oxford, UK. The integration of organic residue analysis of ceramics, carbon (δ13C) and nitrogen (δ15N) isotope analysis of human and animal bones, incremental analysis of δ13C and δ15N from human tooth dentine and palaeopathological analysis of human skeletal remains has revealed a broad pattern of increasing intensification and marketisation across various areas of economic practice, with a much lesser and more short-term impact of the Conquest on everyday lifestyles than is suggested by documentary sources. Nonetheless, isotope data indicate short-term periods of instability, particularly food insecurity, did impact individuals. Evidence of preferences for certain foodstuffs and cooking techniques documented among the elite classes were also observed among lower-status townspeople, suggesting that Anglo-Norman fashions could be adopted across the social spectrum. This study demonstrates the potential for future archaeological research to generate more nuanced understanding of the cultural impact of the Norman Conquest of England, while showcasing a method which can be used to elucidate the undocumented, everyday implications of other large-scale political events on non-elites.

Cereal cultivation in Britain dates back to ca. 4000 BCE, probably introduced by migrant farmers from continental Europe. Widespread evidence for livestock appears in the archaeozoological record, also reflected by ubiquitous dairy lipids in pottery organic residues. However, despite archaeobotanical evidence for domesticated plants (such as cereals), organic residue evidence has been near-absent. Our approach, targeting low-abundance cereal-specific markers, has now revealed evidence for cereals (indicating wheat) in Neolithic pottery from Scottish ‘crannogs’, dating to ca. 3600 – 3300 BCE. Their association with dairy products suggests cereals may have been regularly prepared together as a milk-based gruel. We also observed a strong association between the occurrence of dairy products and smaller-mouthed vessels. Here, we demonstrate that cereal-specific markers can survive in cooking pots for millennia, revealing the consumption of specific cereals (wheat) that are virtually absent from the archaeobotanical record for this region and illuminating culinary traditions among early farming communities.

The development of pastoralism transformed human diets and societies in grasslands worldwide. The long-term success of cattle herding in Africa has been sustained by dynamic food systems, consumption of a broad range of primary and secondary livestock products, and the evolution of lactase persistence (LP), which allows digestion of lactose into adulthood and enables the milk-based, high-protein, low-calorie diets characteristic of contemporary pastoralists. Despite the presence of multiple alleles associated with LP in ancient and present-day eastern African populations, the contexts for selection for LP and the long-term development of pastoralist foodways in this region remain unclear. Pastoral Neolithic (c. 5000 to 1200 BP) faunas indicate that herders relied on cattle, sheep, and goats and some hunting, but direct information on milk consumption, plant use, and broader culinary patterns is rare. Combined chemical and isotopic analysis of ceramic sherds (n = 125) from Pastoral Neolithic archaeological contexts in Kenya and Tanzania, using compound-specific δ(13)C and Δ(13)C values of the major fatty acids, provides chemical evidence for milk, meat, and plant processing by ancient herding societies in eastern Africa. These data provide the earliest direct evidence for milk product consumption and reveal a history of reliance on animal products and other nutrients, likely extracted through soups or stews, and plant foods. They document a 5,000-y temporal framework for eastern Africa pastoralist cuisines and cultural contexts for selection for alleles distinctive of LP in eastern Africa.

The emergence of agriculture in Greece denotes the start of the Neolithic in Europe, however, little is known about dietary practices in the region. Archaeobotanical and zooarchaeological remains indicate reliance on cereals and pulses, together with meat-based subsistence practices, including sheep/goat and pig husbandry. Preliminary investigations of dietary practices obtained through lipid residue analysis of pottery of a small number of sites in the region have confirmed primarily carcass products were processed. The weak evidence for dairy products contrasts with finding of dairy-based subsistence strategies in NW Anatolia, which is surprising given its close proximity. This paper aims to build on this earlier work to provide a more detailed model for the dietary changes throughout the region, both chronologically and spatially. To achieve this >900 potsherds from 11 sites spanning the Early (EN) to Late Neolithic (LN) periods from the north of Greece have been investigated using the lipid biomarker approach involving high temperature-gas chromatography (HT-GC), GC-mass spectrometry (GC-MS) and GC-combustion-isotope ratio MS (GC-C-IRMS) to determine the nature and origins of organic residues preserved in the fabric of pottery vessels. Lipid residue analysis of pottery vessels revealed ruminant and non-ruminant carcass fats comprise the majority of animal fat types identified, reflecting the high abundance of sheep/goat and pig in faunal assemblages. The emergence of dairying in northern Greece can now be dated to the site of EN/Middle Neolithic (MN) Ritini (5900/5700 - 5500 cal. B.C.E.), however, the frequency of dairy fat residues was low, overall, indicating that dairying was not intensively practised. The δ13C values of the fatty acids extracted from potsherds reflect a predominately C3 diet, however, in the EN and MN there is greater variation with some lipids exhibiting enriched δ13C values indicating a significant abundance of C4 plants in the ecosystem(s) covered by the study. Significantly, plant-derived n-alkanes (C22 to C34) detected in pottery vessels provide the first evidence for plant processing identified in lipid residues from ceramic vessels in Neolithic northern Greece, supporting the abundant archaeobotanical evidence for the processing of cereals and pulses.

Organic residue analyses of archaeological ceramics can provide important insights into ancient foodways. To date, however, there has been little critical reflection on how lipid residues might (or might not) reflect dietary practices or subsistence strategies more generally. A combination of ethnoarchaeological research and chemical and isotopic analyses of lipid residues from pottery made and used by modern Samburu pastoralists in northern Kenya was undertaken to supplement the interpretive framework used in archaeological investigations. A total of 63 potsherds were collected from various contexts, including settlement sites and rockshelters, and analysed using gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS) and gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The results showed that the free fatty acids, palmitic (C16:0) and stearic (C18:0), dominated the lipid profiles, with extremely broad ranges of δ13C values. The majority of the Δ13C values from Samburu pots suggest that vessels were intensively used to process ruminant carcass products, yet the Samburu economy is not, in fact, meat-based at all. Despite an overall reliance on dairy products, milk is rarely processed in ceramic vessels, largely due to cultural prohibitions. Surprisingly, a number of vessels from one site, Naiborkeju Hill, were used to process dairy products. Compound-specific radiocarbon dating of lipids from these sherds suggests that this pottery originated from an earlier period, demonstrating a possible shift in ceramic use by pastoralist communities in this region over time. The overall conclusion is that lipid residues may not necessarily reflect, in a simple way, the day-to-day consumption or the perceived relative importance of different foodstuffs. In the Samburu case, lipid residues reflect the functional and ideological suitability of ceramics for processing only certain types of food (meat/fat/bones), despite an overall reliance on milk. These conclusions are important when considering the origins and development of African pastoralism, for example, as interpreted from the archaeological record.

The analysis of lipids and other biomolecules preserved in archaeological artefacts, using chromatographic and mass spectrometric techniques, is a powerful approach, which has provided unprecedented insights into the diet and cultural practices of past populations. In more recent years, the now-mature field of organic residue analysis (ORA) has entered a new phase, undertaking large scale analyses and providing broader perspectives on the uses of pottery over a range of both temporal and spatial scales. However, it has become apparent from the literature that there are significant pitfalls in applying the technique, often due to inexperience or lack of knowledge, that can lead to the production of data which is essentially worthless, because it either lacks analytical rigour or a valid archaeological interpretation. This is partly because ORA sits between chemistry and archaeology and projects are sometimes conducted by researchers not sufficiently familiar in the respective other discipline. Consequently, there have been numerous examples in recent years that showed a lack of understanding of critical points in ORA. This has two serious consequences: 1. the often-wide perpetuation of false knowledge, and 2. the use of a method that is both costly and destructive of irreplaceable archaeological material, which is not compensated for with useful information-gain. Here, we address some common errors encountered either in published literature or during peer-review and discuss the most important errors and misconceptions around ORA. In particular, we discuss use of unsuitable biomarkers, contamination, poor analytical data quality, the limitations of gas chromatography (GC), GC-mass spectrometry (GC-MS) and MS databases, and problems with the interpretation of ORA data. Here, we offer best practice advice and we hope that this will help, and encourage, all researchers looking to apply ORA in their studies, as we are optimistic that these pitfalls can be avoided, and the quality of published research consequently raised.

We unequivocally confirmed pre-Clovis human occupation of the western Great Basin using fecal lipid biomarkers and a new 14C date., When and how people first settled in the Americas is an ongoing area of research and debate. The earliest sites typically only contain lithic artifacts that cannot be directly dated. The lack of human skeletal remains in these early contexts means that alternative sources of evidence are needed. Coprolites, and the DNA contained within them, are one such source, but unresolved issues concerning ancient DNA taphonomy and potential for contamination make this approach problematic. Here, we use fecal lipid biomarkers to demonstrate unequivocally that three coprolites dated to pre-Clovis are human, raise questions over the reliance on DNA methods, and present a new radiocarbon date on basketry further supporting pre-Clovis human occupation.

Little is known about the types of intestinal parasites that infected people living in prehistoric Britain. The Late Bronze Age archaeological site of Must Farm was a pile-dwelling settlement located in a wetland, consisting of stilted timber structures constructed over a slow-moving freshwater channel. At excavation, sediment samples were collected from occupation deposits around the timber structures. Fifteen coprolites were also hand-recovered from the occupation deposits; four were identified as human and seven as canine, using fecal lipid biomarkers. Digital light microscopy was used to identify preserved helminth eggs in the sediment and coprolites. Eggs of fish tapeworm (Diphyllobothrium latum and Diphyllobothrium dendriticum), Echinostoma sp., giant kidney worm (Dioctophyma renale), probable pig whipworm (Trichuris suis) and Capillaria sp. were found. This is the earliest evidence for fish tapeworm, Echinostoma worm, Capillaria worm and the giant kidney worm so far identified in Britain. It appears that the wetland environment of the settlement contributed to establishing parasite diversity and put the inhabitants at risk of infection by helminth species spread by eating raw fish, frogs or molluscs that flourish in freshwater aquatic environments, conversely the wetland may also have protected them from infection by certain geohelminths.

The spread of early farming across Europe from its origins in Southwest Asia was a culturally transformative process which took place over millennia. Within regions, the pace of the transition was probably related to the particular climatic and environmental conditions encountered, as well as the nature of localized hunter–gatherer and farmer interactions. The establishment of farming in the interior of the Balkans represents the first movement of Southwest Asian livestock beyond their natural climatic range, and widespread evidence now exists for early pottery being used extensively for dairying. However, pottery lipid residues from sites in the Iron Gates region of the Danube in the northern Balkans show that here, Neolithic pottery was being used predominantly for processing aquatic resources. This stands out not only within the surrounding region but also contrasts markedly with Neolithic pottery use across wider Europe. These findings provide evidence for the strategic diversity within the wider cultural and economic practices during the Neolithic, with this exceptional environmental and cultural setting offering alternative opportunities despite the dominance of farming in the wider region.

Strontium isotope ratios are widely used in archaeology to differentiate between local and non-local populations. Herein, strontium isotope ratios of 36 human tooth enamels from seven archaeological sites spanning the Early to Late Neolithic of northern Greece (7th–5th millennia B.C.E.) were analysed with the aim of providing new information relating to the movement of humans across the region. Local bioavailable 87Sr/86Sr signals were established using tooth enamel from 26 domestic animals from the same Neolithic sites. 87Sr/86Sr values of faunal enamel correlate well with predicted strontium isotope ratios of the local geology. This is consistent with animal management occurring at a local level, although at Late Neolithic sites strontium isotope values became more varied, potentially indicating changing herding practices. The strontium isotope analysis of human tooth enamel likewise suggests limited population movement within the Neolithic of northern Greece. Almost all individuals sampled exhibited 87Sr/86Sr values consistent with having spent their early life (during the period of tooth mineralisation) in the local area, although movement could have occurred between isotopically homogeneous areas. The strontium isotope ratios of only three individuals lay outside of the local bioavailable 87Sr/86Sr range and these individuals are interpreted as having spent their early lives in a region with a more radiogenic biologically available 87Sr/86Sr. Mobility patterns determined using Sr isotope analysis supports the current evidence for movement and exchange observed through studies of pottery circulation. Suggesting limited movement in the Early and Middle Neolithic and greater movement in the Late Neolithic.

Paisley Caves in Oregon has become well known due to early dates, and human presence in the form of coprolites, found to contain ancient human DNA. Questions remain over whether the coprolites themselves are human, or whether the DNA is mobile in the sediments. This brief introduces new research applying an integrated analytical approach combining sediment micromorphology and lipid biomarker analysis, which aims to resolve these problems.

In the absence of any direct evidence, the relative importance of meat and dairy productions to Neolithic prehistoric Mediterranean communities has been extensively debated. Here, we combine lipid residue analysis of ceramic vessels with osteo-archaeological age-at-death analysis from 82 northern Mediterranean and Near Eastern sites dating from the seventh to fifth millennia BC to address this question. The findings show variable intensities in dairy and nondairy activities in the Mediterranean region with the slaughter profiles of domesticated ruminants mirroring the results of the organic residue analyses. The finding of milk residues in very early Neolithic pottery (seventh millennium BC) from both the east and west of the region contrasts with much lower intensities in sites of northern Greece, where pig bones are present in higher frequencies compared with other locations. In this region, the slaughter profiles of all domesticated ruminants suggest meat production predominated. Overall, it appears that milk or the by-products of milk was an important foodstuff, which may have contributed significantly to the spread of these cultural groups by providing a nourishing and sustainable product for early farming communities.

In the absence of any direct evidence, the relative importance of meat and dairy productions to Neolithic prehistoric Mediterranean communities has been extensively debated. Here, we combine lipid residue analysis of ceramic vessels with osteo-archaeological age-at-death analysis from 82 northern Mediterranean and Near Eastern sites dating from the seventh to fifth millennia BC to address this question. The findings show variable intensities in dairy and nondairy activities in the Mediterranean region with the slaughter profiles of domesticated ruminants mirroring the results of the organic residue analyses. The finding of milk residues in very early Neolithic pottery (seventh millennium BC) from both the east and west of the region contrasts with much lower intensities in sites of northern Greece, where pig bones are present in higher frequencies compared with other locations. In this region, the slaughter profiles of all domesticated ruminants suggest meat production predominated. Overall, it appears that milk or the by-products of milk was an important foodstuff, which may have contributed significantly to the spread of these cultural groups by providing a nourishing and sustainable product for early farming communities.

The investigation of organic residues associated with archaeological pottery using modern analytical chemical methods began in the 1970s. It was recognised early on that the analysis of lipids (i.e. fats, waxes and resins) preserved in surface residues or the fabric of single potsherds, representative of single vessels, was a powerful method for ascertaining pottery use, with a high degree of specificity. Subsequent developments saw a significant change in scale, with studies often involving lipid analyses of tens to hundreds of potsherds per archaeological assemblage, providing information that extended beyond pottery use. The identification of animal and plant foodstuffs processed in pots provides insights into herding and farming, and can also detect trade in exotic organic goods. Information about the environment and climate can be extrapolated from the isotopic composition of compounds detected in potsherds, potentially providing novel avenues of investigation. The direct dating of lipids in potsherds is opening up new opportunities for building archaeological chronologies, while the integration of lipid residue analyses with other environmental and cultural proxies within interdisciplinary projects is already providing unprecedented insights into past lifestyles, from site to regional scales.

This paper presents the findings from an investigation of organic residues extracted from pottery sherds from Late Iron Age and Norse phases from Bornais, South Uist, and the Late Norse period from Jarlshof on Shetland. These data confirm intensive and/or specialized processing of marine products in pottery on Shetland, either for consumption or other uses, such as rendering of oil from fish livers. In contrast, at Bornais, little increase in the intensity of marine product exploitation can be identified between the residues from the Later Iron Age and Norse phases; however, an emphasis on dairy products is identifiable throughout all phases and pottery types. While the findings from these two sites clearly cannot be extrapolated as entirely representative of the wider respective regions, what emerges is further evidence for diverse economic or cultural patterns at different locations within Scandinavian Scotland.