Carbon and nitrogen isotopic variability in foxtail millet (Setaria italica) with watering regime.

RATIONALE: Carbonised plant remains are analysed for reconstruction of past climates and agricultural regimes. Several recent studies have used C4 plants to address related questions, and correlations between modern C4 plant δ13 C values and rainfall have been found. The millets were important food crops in prehistoric Eurasia, yet little is known about causes of isotopic variation within millet species. Previous research has shown there to be significant isotopic variation between millet accessions. Here we compare isotope ratios from plants grown under different watering regimes. This allows for a consideration of whether or not Setaria italica is a good proxy for environmental reconstruction. METHODS: We compare stable isotope ratios of Setaria italica plants grown in a controlled environment chamber with different watering regimes. We compare the carbon isotope ratios of leaves and grains, and the nitrogen isotope ratios of grains, from 12 accessions of Setaria italica. RESULTS: We find significant isotopic variability between watering regimes. Carbon isotope ratios are positively correlated with water availability, and on average vary by 1.9‰ and 1.7‰ for leaves and grains, respectively. Grain nitrogen isotope ratios also vary with watering regime; however, the highest isotope ratios are found with the 130-mL watering regime. CONCLUSIONS: The carbon isotope ratios of Setaria italica are strongly correlated with water availability. However, the correlation is the opposite to that seen in studies of C3 plants. The difference in isotopic ratio due to watering regime is comparable with that seen between different accessions; thus distinguishing between changing varieties of Setaria italica and changing climate is problematic. In terms of grain nitrogen isotope ratios, the highest δ15 N values were not associated with the lowest watering regime. Again, δ15 N variation is comparable with that which would be expected from an aridity effect or a manuring effect, and thus distinguishing between these factors is probably problematic.

Carbon and nitrogen isotopic signatures of hair, nail, and breath from tropical African human populations.

RATIONALE: Stable isotopic analyses are increasingly used to study the diets of past and present human populations. Yet, the carbon and nitrogen isotopic data of modern human diets collected so far are biased towards Europe and North America. Here, we address this gap by reporting on the dietary isotopic signatures of six tropical African communities: El Molo, Turkana (Kerio), Luhya (Webuye), Luhya (Port Victoria), and Luo (Port Victoria) from Kenya, and Baka from Cameroon; representing four subsistence strategies: fishing, pastoralism, agriculturalism, and hunter-gatherer. METHODS: We used an elemental analyser coupled in continuous-flow mode to an isotope ratio mass spectrometer to measure the carbon and nitrogen isotopic ratios of hair (n = 134) and nail (n = 80) and the carbon isotopic ratios of breath (n = 184) from these communities, as well as the carbon and nitrogen isotopic ratios of some food samples from the Kenyan communities. RESULTS: We expand on the known range of δ13 C values in human hair through the hunter-gatherer Baka, with a diet based on C3 plants, and through the agriculturalist Luhya (Webuye), with a diet based on C4 plants. In addition, we found that the consumption of fish from East African lakes is difficult to detect isotopically due to the combined effects of high nitrogen isotopic ratios of plants and the low nitrogen isotopic ratios of fish. Finally, we found that some of the communities studied are markedly changing their diets through increasing sedentism and urbanisation. CONCLUSIONS: Our findings contribute substantially to the understanding of the environmental, demographic, and economic dynamics that affect the dietary landscape of different tropical populations of Africa. These results highlight the importance of studying a broader sample of human populations and their diet, with a focus on their precise context - from both isotopic and more general anthropological perspectives.

East Anglian early Neolithic monument burial linked to contemporary Megaliths.

In the fourth millennium BCE a cultural phenomenon of monumental burial structures spread along the Atlantic façade. Megalithic burials have been targeted for aDNA analyses, but a gap remains in East Anglia, where Neolithic structures were generally earthen or timber. An early Neolithic (3762-3648 cal. BCE) burial monument at the site of Trumpington Meadows, Cambridgeshire, UK, contained the partially articulated remains of at least three individuals. To determine whether this monument fits a pattern present in megalithic burials regarding sex bias, kinship, diet and relationship to modern populations, teeth and ribs were analysed for DNA and carbon and nitrogen isotopic values, respectively. Whole ancient genomes were sequenced from two individuals to a mean genomic coverage of 1.6 and 1.2X and genotypes imputed. Results show that they were brothers from a small population genetically and isotopically similar to previously published British Neolithic individuals, with a level of genome-wide homozygosity consistent with a small island population sourced from continental Europe, but bearing no signs of recent inbreeding. The first Neolithic whole genomes from a monumental burial in East Anglia confirm that this region was connected with the larger pattern of Neolithic megaliths in the British Isles and the Atlantic façade.

New chronology for Ksâr 'Akil (Lebanon) supports Levantine route of modern human dispersal into Europe.

Modern human dispersal into Europe is thought to have occurred with the start of the Upper Paleolithic around 50,000-40,000 y ago. The Levantine corridor hypothesis suggests that modern humans from Africa spread into Europe via the Levant. Ksâr 'Akil (Lebanon), with its deeply stratified Initial (IUP) and Early (EUP) Upper Paleolithic sequence containing modern human remains, has played an important part in the debate. The latest chronology for the site, based on AMS radiocarbon dates of shell ornaments, suggests that the appearance of the Levantine IUP is later than the start of the first Upper Paleolithic in Europe, thus questioning the Levantine corridor hypothesis. Here we report a series of AMS radiocarbon dates on the marine gastropod Phorcus turbinatus associated with modern human remains and IUP and EUP stone tools from Ksâr 'Akil. Our results, supported by an evaluation of individual sample integrity, place the EUP layer containing the skeleton known as "Egbert" between 43,200 and 42,900 cal B.P. and the IUP-associated modern human maxilla known as "Ethelruda" before ∼ 45,900 cal B.P. This chronology is in line with those of other Levantine IUP and EUP sites and demonstrates that the presence of modern humans associated with Upper Paleolithic toolkits in the Levant predates all modern human fossils from Europe. The age of the IUP-associated Ethelruda fossil is significant for the spread of modern humans carrying the IUP into Europe and suggests a rapid initial colonization of Europe by our species.

Red deer bone and antler collagen are not isotopically equivalent in carbon and nitrogen.

RATIONALE: Bone and antler collagen δ(13) C and δ(15) N values are often assumed to be equivalent when measured in palaeodietary, palaeoclimate and palaeocological studies. Although compositionally similar, bone grows slowly and is remodelled whereas antler growth is rapid and remodelling does not occur. These different patterns of growth could result in isotopic difference within antler and between the two tissue types. Here we test whether red deer (Cervus elaphus) bone and antler δ(13) C and δ(15) N values are equivalent, and whether intra-antler isotopic values are uniform. METHODS: Bone and antler were isotopically analysed from six stags that lived in a temperate maritime climate on the Isle of Rum, Scotland. Multiple antlers from different years were sampled per individual, together with a single bone sample per individual. Up to 12 samples were taken along the length of each antler (total of 25 antlers, 259 samples) so that a chronological record of the isotopic composition during antler growth could be obtained. Collagen was extracted and its δ(13) C and δ(15) N values were measured by continuous-flow isotope ratio mass spectrometry. RESULTS: Intra-antler collagen isotope signatures vary, and show that not all antlers from an individual or a growth year are equivalent in carbon and nitrogen isotopic ratios. δ(15) N values typically increase with distance along antler length, but no overall trend is observed in δ(13) C values. An isotopic offset is visible between bone and antler, with bone δ(13) C and δ(15) N values being higher in most cases. CONCLUSIONS: Bone and antler collagen δ(13) C and δ(15) N values are not isotopically equivalent and are therefore not directly comparable in palaeodietary, palaeoclimate and palaeocological studies. Bone and antler collagen isotopic differences probably relate to differential metabolic processes during the formation of the two tissues. Intra- and inter-antler isotopic variations probably reflect the isotopic composition of an individual's diet rather than physiological parameters, and may have the potential to provide high-resolution individual-specific information in modern and ancient cervid populations. Copyright © 2016 John Wiley & Sons, Ltd.

Intraspecific carbon and nitrogen isotopic variability in foxtail millet (Setaria italica).

RATIONALE: Isotopic palaeodietary studies generally focus on bone collagen from human and/or animal remains. While plant remains are rarely analysed, it is known that plant isotope values can vary as a result of numerous factors, including soil conditions, the environment and type of plant. The millets were important food crops in prehistoric Eurasia, yet little is known about the isotopic differences within millet species. METHODS: Here we compare the stable isotope ratios within and between Setaria italica plants grown in a controlled environment chamber. Using homogenised samples, we compare carbon isotope ratios of leaves and grains, and nitrogen isotope ratios of grains, from 29 accessions of Setaria italica. RESULTS: We find significant isotopic variability within single leaves and panicles, and between leaves and panicles within the same plant, which must be considered when undertaking plant isotope studies. We find that the leaves and grains from the different accessions have a ca 2‰ range in δ(13) C values, while the nitrogen isotope values in the grains have a ca 6‰ range. We also find an average offset of 0.9‰ between leaves and grains in their δ(13) C values. CONCLUSIONS: The variation found is large enough to have archaeological implications and within- and between-plant isotope variability should be considered in isotope studies. The range in δ(15) N values is particularly significant as it is larger than the typical values quoted for a trophic level enrichment, and as such may lead to erroneous interpretations of the amount of animal protein in human or animal diets. It is therefore necessary to account for the variability in plant stable isotope values during palaeodietary reconstructions. Copyright © 2016 John Wiley & Sons, Ltd.

Fecal carbon and nitrogen isotopic analysis as an indicator of diet in Kanyawara chimpanzees, Kibale National Park, Uganda.

OBJECTIVES: The shorter-term overview from feces provides scope to investigate dietary fluctuations. We assess the correlation of stable isotopic fecal values with recorded seasonal diet of 10 adult chimpanzees (P. t. schweinfurthii) of the Kanyawara community (Kibale National Park, Uganda) and whether fecal nitrogen levels (%N) indicate a change in crude protein intake. MATERIALS AND METHODS: We recorded food eaten by each ape and collected both concurrent fecal samples (N = 115) and plant foods eaten by this community (N = 64). We compared fecal δ13 C and δ15 N values (also %N) with: (a) plant values; (b) feeding data; and (c) food-items found macroscopically in the fecal samples. Interspecies and intraspecies differences in plant and fecal isotope values (and %N) as well as seasonality in diet were determined using parametric and nonparametric tests. RESULTS: No difference in plant δ13 C and δ15 N values was found at intraspecies or interspecies level. Fecal isotope values reflected a diet of C3 plants from evergreen forest vegetation. Seasonal differences in δ13 C and δ15 N corresponded with aspects of feeding and fecal macroscopic data, but only at community level. A change in crude protein intake was not indicated from %N content. DISCUSSION: This study further validates the use of staple isotope analyses of primate feces to provide a dietary overview, revealing seasonal differences at community level; however, conclusive results may be limited for individuals when using short sampling periods. Further study of variables that influence fecal %N content is also suggested to interpret crude protein intake.

Genetic history of Cambridgeshire before and after the Black Death.

The extent of the devastation of the Black Death pandemic (1346-1353) on European populations is known from documentary sources and its bacterial source illuminated by studies of ancient pathogen DNA. What has remained less understood is the effect of the pandemic on human mobility and genetic diversity at the local scale. Here, we report 275 ancient genomes, including 109 with coverage >0.1×, from later medieval and postmedieval Cambridgeshire of individuals buried before and after the Black Death. Consistent with the function of the institutions, we found a lack of close relatives among the friars and the inmates of the hospital in contrast to their abundance in general urban and rural parish communities. While we detect long-term shifts in local genetic ancestry in Cambridgeshire, we find no evidence of major changes in genetic ancestry nor higher differentiation of immune loci between cohorts living before and after the Black Death.

Carbon and nitrogen isotopic ratios of urine and faeces as novel nutritional biomarkers of meat and fish intake.

PURPOSE: Meat and fish consumption are associated with changes in the risk of chronic diseases. Intake is mainly assessed using self-reporting, as no true quantitative nutritional biomarker is available. The measurement of plasma fatty acids, often used as an alternative, is expensive and time-consuming. As meat and fish differ in their stable isotope ratios, δ(13)C and δ(15)N have been proposed as biomarkers. However, they have never been investigated in controlled human dietary intervention studies. OBJECTIVE: In a short-term feeding study, we investigated the suitability of δ(13)C and δ(15)N in blood, urine and faeces as biomarkers of meat and fish intake. METHODS: The dietary intervention study (n = 14) followed a randomised cross-over design with three eight-day dietary periods (meat, fish and half-meat-half-fish). In addition, 4 participants completed a vegetarian control period. At the end of each period, 24-h urine, fasting venous blood and faeces were collected and their δ(13)C and δ(15)N analysed. RESULTS: There was a significant difference between diets in isotope ratios in faeces and urine samples, but not in blood samples (Kruskal-Wallis test, p < 0.0001). In pairwise comparisons, δ(13)C and δ(15)N were significantly higher in urine and faecal samples following a fish diet when compared with all other diets, and significantly lower following a vegetarian diet. There was no significant difference in isotope ratio between meat and half-meat-half-fish diets for blood, urine or faecal samples. CONCLUSIONS: The results of this study show that urinary and faecal δ(13)C and δ(15)N are suitable candidate biomarkers for short-term meat and fish intake.

The earliest evidence of millet as a staple crop: New light on neolithic foodways in North China.

There is a growing body of archaeobotanical evidence for the harvesting of millet in Eurasia prior to 5,000 cal. BC. Yet direct evidence for the extent of millet consumption in this time period is rare. This contradiction may be due to millet crops making only a minor contribution to the diet before 5,000 BC. In this article, drawing from recent excavations in North China, we present evidence for millet crops making a substantial contribution to human and animal diets in periods, which correspond chronologically with the time depth of the archaeobotanical record. We infer that in eastern Inner Mongolia, human adoption of millets, which may or may be not related to substantial agriculture, happened at the Early Neolithic, with direct dates between 5,800 and 5,300 cal. BC.

Were there royal herds? Understanding herd management and mobility using isotopic characterizations of cattle tooth enamel from Early Dynastic Ur.

During the third millennium BC, Mesopotamia (the land between the Tigris and Euphrates Rivers, in modern Iraq-Syria), was dominated by the world's earliest cities and states, which were ruled by powerful elites. Ur, in present-day southern Iraq, was one of the largest and most important of these cities, and irrigation-based agriculture and large herds of domesticated animals were the twin mainstays of the economy and diet. Texts suggest that the societies of the Mesopotamian city-states were extremely hierarchical and underpinned by institutionalised and heavily-managed farming systems. Prevailing narratives suggest that the animal management strategies within these farming systems in the third millennium BC were homogenous. There have been few systematic science-based studies of human and animal diets, mobility, or other forms of human-animal interaction in Mesopotamia, but such approaches can inform understanding of past economies, including animal management, social hierarchies, diet and migration. Oxygen, carbon and strontium isotopic analysis of animal tooth enamel from both royal and private/non-royal burial contexts at Early Dynastic Ur (2900-2350 BC) indicate that a variety of herd management strategies and habitats were exploited. These data also suggest that there is no correlation between animal-management practices and the cattle found in royal or private/non-royal burial contexts. The results demonstrate considerable divergence between agro-pastoral models promoted by the state and the realities of day-to-day management practices. The data from Ur suggest that the animals exploited different plant and water sources, and that animals reared in similar ways ended up in different depositional contexts.

Isotope data from amino acids indicate Darwin's ground sloth was not an herbivore.

Fossil sloths are regarded as obligate herbivores for reasons including peculiarities of their craniodental morphology and that all living sloths feed exclusively on plants. We challenge this view based on isotopic analyses of nitrogen of specific amino acids, which show that Darwin's ground sloth Mylodon darwinii was an opportunistic omnivore. This direct evidence of omnivory in an ancient sloth requires reevaluation of the ecological structure of South American Cenozoic mammalian communities, as sloths represented a major component of these ecosystems across the past 34 Myr. Furthermore, by analyzing modern mammals with known diets, we provide a basis for reliable interpretation of nitrogen isotopes of amino acids of fossils. We argue that a widely used equation to determine trophic position is unnecessary, and that the relative isotopic values of the amino acids glutamate and phenylalanine alone permit reliable reconstructions of trophic positions of extant and extinct mammals.

Lipid residues in pottery from the Indus Civilisation in northwest India.

This paper presents novel insights into the archaeology of food in ancient South Asia by using lipid residue analysis to investigate what kinds of foodstuffs were used in ceramic vessels by populations of the Indus Civilisation in northwest India. It examines how vessels were used in urban and rural Indus settlements during the Mature Harappan period (c.2600/2500-1900 BC), the relationship between vessels and the products within them, and identifies whether changes in vessel use occurred from the Mature Harappan to Late Harappan periods, particularly during climatic instability after 4.2 ka BP (c.2100 BC). Despite low lipid concentrations, which highlight challenges with conducting residue analysis in arid, seasonally-wet and alkaline environments, 71% of the vessels yielded appreciable quantities of lipid. Lipid profiles revealed the use of animal fats in vessels, and contradictory to faunal evidence, a dominance of non-ruminant fats, with limited evidence of dairy processing. The absence of local modern reference fats makes this dataset challenging to interpret, and it is possible that plant products or mixtures of plant and animal products have led to ambiguous fatty acid-specific isotopic values. At the same time, it appears that urban and rural populations processed similar types of products in vessels, with limited evidence for change in vessel use from the urban to the post-urban period. This study is a systematic investigation into pot lipid residues from multiple sites, demonstrating the potential of the method for examining ancient Indus foodways and the need for the development of further research in ancient organic residues in South Asia.

Water-related occupations and diet in two Roman coastal communities (Italy, first to third century AD): correlation between stable carbon and nitrogen isotope values and auricular exostosis prevalence.

The reconstruction of dietary patterns in the two Roman imperial age coastal communities of Portus and Velia (I-III AD) by means of stable isotope analysis of bone remains has exposed a certain degree of heterogeneity between and within the two samples. Results do not correlate with any discernible mortuary practices at either site, which might have pointed to differential social status. The present study tests the hypothesis of a possible connection between dietary habits and occupational activities in the two communities. Among skeletal markers of occupation, external auricular exostosis (EAE) has proved to be very informative. Clinical and retrospective epidemiological surveys have revealed a strong positive correlation between EAE development and habitual exposure to cold water. In this study, we show that there is a high rate of occurrence of EAE among adult males in both skeletal samples (21.1% in Portus and 35.3% in Velia). Further, there is a statistically significant higher prevalence of EAE among those individuals at Velia with very high nitrogen isotopic values. This points to fishing (coastal, low-water fishing) as the sea-related occupation most responsible for the onset of the ear pathology. For Portus, where the consumption of foods from sea and river seems to be more widespread through the population, and where the scenario of seaport and fluvial activities was much more complex than in Velia, a close correlation between EAE and fish consumption by fishermen is less easy to establish.

Radiocarbon and stable isotope investigations at the Central Rhineland sites of Gönnersdorf and Andernach-Martinsberg, Germany.

The late glacial open-air sites of Gönnersdorf and Andernach-Martinsberg in the German Central Rhineland are well known for their Magdalenian occupation and activities. The latter site also produced evidence for a younger, Final Palaeolithic occupation of the locality by people of the Federmessergruppen. Both sites are particularly well preserved, largely due to their burial beneath volcanic deposits of the late glacial Laacher See eruption. We conducted a program of AMS radiocarbon dating and stable isotope analyses with the aim of improving understanding of the chronological history and ecological setting of the two sites. Previously published radiocarbon dates appeared to indicate that the earliest Magdalenian occupation at Gönnersdorf fell around 12,900 uncalibrated (14)C yr BP, while the earliest occupation at Andernach may have been more than 500 radiocarbon years earlier. The AMS determinations presented here revise this impression and suggest that the onset of occupation at the two sites was in fact simultaneous and prior to the warming of Greenland Interstadial GI 1e. At Gönnersdorf, a chronological hiatus exists between the main Magdalenian faunal assemblage and mega-faunal remains interpreted as collected sub-fossil material. At Andernach-Martinsberg, there is a clear chronological hiatus between the Magdalenian occupation and subsequent Federmessergruppen activities at the site. However, an intermediate radiocarbon date on an atypically preserved horse bone is suggestive of ephemeral human visits to the site between these well demonstrated phases. A date of similar age on an elk bone from Gönnersdorf may indicate broadly contemporaneous human presence at Gönnersdorf too. Stable isotope analysis of faunal remains from Gönnersdorf and Andernach-Martinsberg was conducted with the aim of both reconstructing and comparing local environmental conditions at the two sites, and also potentially identifying subtle variations in the chronological development of the two sites not detectable at the level of precision of current radiocarbon dating techniques. No spatial trends in the faunal isotope signatures were observed within each site. In the case of samples with both radiocarbon and isotope data, no chronological pattern was observed for the isotope results. The Magdalenian faunal isotope signatures at the two sites resembled each other, suggesting comparable climatic and environmental conditions. The faunal delta(13)C signatures at Gönnersdorf and Andernach-Martinsberg were similar to those at contemporary European sites. While the faunal delta(15)N values were similar to those at contemporary sites in Germany, the UK, and Belgium, they were lower than those from the South of France. This difference in delta(15)N values is thought to relate to regional differences in the timing of changes in soil and plant nitrogen cycling in response to ameliorating climatic conditions.

Trans-Atlantic slavery: isotopic evidence for forced migration to Barbados.

The question of the ultimate origin of African slaves is one of the most perplexing in the history of trans-Atlantic slavery. Here we present the results of a small, preliminary isotopic study that was conducted in order to determine the geographical origin of 25 enslaved Africans who were buried at the Newton plantation, Barbados, sometime between the late 17th and early 19th century. In order to gain a more nuanced understanding of the slaves' origin, we used a combination of carbon, nitrogen, oxygen, and strontium isotope analyses. Carbon and nitrogen isotope ratios were determined in bone and dentinal collagen; oxygen and strontium isotopes were measured in tooth enamel. Results suggest that the majority of individuals were born on the island, if not the estate itself. Seven individuals, however, yielded enamel oxygen and strontium ratios that are inconsistent with a Barbadian origin, which strongly suggests that we are dealing with first-generation captives who were brought to the island with the slave trade. This idea is also supported by the fact that their carbon and nitrogen stable isotope values differ markedly between their teeth and bones. These intra-skeletal shifts reflect major dietary changes that probably coincided with their enslavement and forced migration to Barbados. While it is impossible to determine their exact origins, the results clearly demonstrate that the slaves did not all grow up in the same part of Africa. Instead, the data seem to suggest that they originated from at least three different areas, possibly including the Gold Coast and the Senegambia.

Stable isotopic evidence for diet at the Imperial Roman coastal site of Velia (1st and 2nd centuries AD) in Southern Italy.

Here we report on a stable isotope palaeodietary study of a Imperial Roman population interred near the port of Velia in Southern Italy during the 1st and 2nd centuries AD. Carbon and nitrogen stable isotope analyses were performed on collagen extracted from 117 adult humans as well as a range of fauna to reconstruct individual dietary histories. For the majority of individuals, we found that stable isotope data were consistent with a diet high in cereals, with relatively modest contributions of meat and only minor contributions of marine fish. However, substantial isotopic variation was found within the population, indicating that diets were not uniform. We suggest that a number of individuals, mainly but not exclusively males, had greater access to marine resources, especially high trophic level fish. However, the observed dietary variation did not correlate with burial type, number of grave goods, nor age at death. Also, individuals buried at the necropolis at Velia ate much less fish overall compared with the contemporaneous population from the necropolis of Portus at Isola Sacra, located on the coast close to Rome. Marine and riverine transport and commerce dominated the economy of Portus, and its people were in a position to supplement their own stocks of fish with imported goods in transit to Rome, whereas at Velia marine exploitation existed side-by-side with land-based economic activities.