Stable isotope evidence for the consumption of millet and other plants in Bronze Age Italy

Stable carbon and nitrogen isotope analysis was carried out on human and animal bones from four inland Early and Middle Bronze Age sites in Northern and Southern Italy. The main aims of the investigation were to explore the contribution of plant foods to the human diet and to examine any dietary differences between and within each of the sites. At two of the sites in Northern Italy, human and animal bones were significantly enriched in ¹³C. This finding was attributed to the consumption of domestic millets (Panicum miliaceum and/or Setaria italica), which are C₄ pathway plants. Conversely, individuals from the two Bronze Age sites in Southern Italy were significantly depleted in ¹³C compared to those from the north. Here, millet was absent from the diet, and protein from C₃ plants made a much greater dietary contribution than animal protein. This finding highlights the importance of cereal cultivation, most likely of wheat and barley, in the south of Italy during the Bronze Age. Overall, our results support the idea that the widespread cultivation of millet first occurred in Northern Italy, following its introduction from across the Alps in Central Europe. Finally, we found no significant differences in the stable isotope values between individuals at each site, when grouped by their sex or presence of grave goods. This leads to the conclusion that any status difference that may have existed is not reflected in the long‐term dietary record, or at least not as measurable by stable isotope analysis. Am J Phys Anthropol, 2009. © 2008 Wiley‐Liss, Inc.     

Multivariate carbon and nitrogen stable isotope model for the reconstruction of prehistoric human diet

Using a sample of published archaeological data, we expand on an earlier bivariate carbon model for diet reconstruction by adding bone collagen nitrogen stable isotope values (δ(15) N), which provide information on trophic level and consumption of terrestrial vs. marine protein. The bivariate carbon model (δ(13) C(apatite) vs. δ(13) C(collagen) ) provides detailed information on the isotopic signatures of whole diet and dietary protein, but is limited in its ability to distinguish between C(4) and marine protein. Here, using cluster analysis and discriminant function analysis, we generate a multivariate diet reconstruction model that incorporates δ(13) C(apatite) , δ(13) C(collagen) , and δ(15) N holistically. Inclusion of the δ(15) N data proves useful in resolving protein-related limitations of the bivariate carbon model, and splits the sample into five distinct dietary clusters. Two significant discriminant functions account for 98.8% of the sample variance, providing a multivariate model for diet reconstruction. Both carbon variables dominate the first function, while δ(15) N most strongly influences the second. Independent support for the functions' ability to accurately classify individuals according to diet comes from a small sample of experimental rats, which cluster as expected from their diets. The new model also provides a statistical basis for distinguishing between food sources with similar isotopic signatures, as in a previously analyzed archaeological population from Saipan (see Ambrose et al.: AJPA 104(1997) 343-361). Our model suggests that the Saipan islanders' (13) C-enriched signal derives mainly from sugarcane, not seaweed. Further development and application of this model can similarly improve dietary reconstructions in archaeological, paleontological, and primatological contexts.     

A stable carbon isotope study of dissolved inorganic carbon cycling in a softwater lake

The dissolved inorganic carbon (DIC) cycle in a softwater lake was studied using natural variations of the stable isotopes of carbon,¹²C and¹³C. During summer stratification there was a progressive decrease in epilimnion DIC concentration with a concomitant increase in ¹³C_DIC), due to preferential uptake of¹²C by phytoplankton and a change in the dominant CO₂ source from inflow andin situ oxidation to invasion from the atmosphere. There was an increase in hypolimnion DIC concentration throughout summer with a concomitant general decrease in ¹³C_DIC from oxidation of the isotopically light particulate organic carbon that sank down through the thermocline from the epilimnion.Mass balance calculations of DI¹²C and DI¹³C in the epilimnion for the summer (June 23–September 25) yield a mean rate of net conversion of DIC to organic carbon (C_org) of 430 ± 150 moles d^-1 (6.5 ± 1.8 m moles m^-2 d^-1. Net CO₂ invasion from the atmosphere was 420 ± 120 moles d^-1 (6.2 ± 1.8 m moles m^-2 d^-1) with an exchange coefficient of 0.6 ± 0.3m d^-1. These results imply that at least for the summer months the phytoplankton obtained about 90% of their carbon from atmosphere CO₂. About 50% of CO₂ invasion and conversion to C_org for the summer occurred during a two week interval in mid-summer.DIC concentration increased in the hypolimnion at a rate of 350 ± 70 moles DIC d^-1 during summer stratification. The amount of DIC added to the hypolimnion was equivalent to 75 ± 20% of net conversion of DIC to C_org in the euphotic zone over spring and summer implying rapid degradation of POC in the hypolimnion. The ¹³C of DIC added to the deep water (-22.) was too heavy to have been derived from oxidation of particulate organic carbon alone. About 20% of the added DIC must have diffused from hypolimnetic sediments where relatively heavy CO₂ (-7) was produced by a combination of POC oxidation and as a by-product of methanogenesis.     

Lapita diet and subsistence strategies on Watom Island,Papua New Guinea: New stable isotope evidence from humans and animals

Stable isotope ratios (δ¹³C and δ¹⁵N) were analyzed from the bone collagen of individuals (n = 8) from a Lapita burial ground (ca. 2800–2350 BP) on Watom Island, located off northeast New Britain in the Bismarck Archipelago. The aim of this study was to assess the diet and subsistence strategies of humans that lived during the later Lapita period in Near Oceania. To aid in the interpretation of the human diet we analyzed the stable isotope ratios of faunal material from the site (n = 27). We also aim to assess methods of animal husbandry at the site over time from an analysis of the stable isotope ratios (δ¹³C and δ¹⁵N) of pig bones (n = 22) from different temporal periods (Lapita, post‐Lapita, and late prehistoric). The protein diet of the humans consisted of marine organisms from the inshore environment and some deep‐water species, most likely marine turtle, in addition to higher trophic level terrestrial foods, likely pig and native animals (e.g., fruit bat, Cuscus and bandicoot). Although the sample sizes were small, females (n = 4) displayed more variable δ¹³C and δ¹⁵N values compared with males (n = 4), which may be associated with the movement of adult females to the island. The stable isotope analysis of the pig bones indicated that there were few differences between the diets of the pigs from the Lapita and post‐Lapita layers, suggesting that the method of pig husbandry was similar between these two periods and was likely relatively free‐range. Am J Phys Anthropol 157:30–41, 2015. © 2015 Wiley Periodicals, Inc.     

Sources and fates of dissolved organic carbon in lakes as determined by whole-lake carbon isotope additions

Four whole-lake inorganic ¹³C addition experiments were conducted in lakes of differing trophic status. Inorganic ¹³C addition enriched algal carbon in ¹³C and changed the C-DOC by +1.5‰ to +9.5‰, depending on the specific lake. This change in C-DOC represented a significant input of algal DOC that was not completely consumed by bacteria. We modeled the dynamics in C-DOC to estimate the fluxes of algal and terrestrial carbon to and from the DOC pool, and determine the composition of the standing stock. Two experiments in lightly stained, oligotrophic lakes indicated that algal production was the source of about 20% of the DOC pool. In the following year, the experiment was repeated in one of these lakes under conditions of nutrient enrichment, and in a third, more humic lake. Algal contributions to the DOC pool were 40% in the nutrient enriched lake and 5% in the more humic lake. Spectroscopic and elemental analyses corroborated the presence of increased algal DOC in the nutrient enriched lake. Natural abundance measurements of the C of DOC in 32 lakes also revealed the dual contributions of both terrestrial and algal carbon to DOC. From these results, we suggest an approach for inferring the contribution of algal and terrestrial DOC using easily measurable parameters.     

Origin and fate of organic carbon in the freshwater part of the Scheldt Estuary as traced by stable carbon isotope composition

We investigated the seasonal and geographical variation in the stable carbon isotope ratios of total dissolved inorganic carbon (¹³C_POC) and suspended matter (¹³C_POC) in the freshwater part of the River Scheldt. Two major sources of particulate organic matter (POM) occur in this riverine system: riverine phytoplankton and terrestrial detritus. In winter the lowest ¹³C_DIC values are observed due to enhanced input of CO₂ from decomposition of ¹³C-depleted terrestrial plant detritus (average ¹³C_DIC = –/14.3). During summer, when litter input from terrestrial flora is the lowest, water column respiration on POM of terrestrial origin is also the lowest as evidenced by less negative ¹³C_DIC values (average ¹³C_DIC = –9.9). In winter the phytoplankton biomass is low, as indicated by low chlorophyll a concentrations (Chl a < 4.5 gl^–1), compared to summer when chlorophyll a concentrations can rise to a maximum of 54 gl^–1. Furthermore, in winter the very narrow range of ¹³C_POC (from –26.5 to –27.6) is associated with relatively high C/N ratios (C/N > 9) suggesting that in winter a major fraction of POC is derived from allochthonous matter. In summer ¹³C_POC exhibits a very wide range of values, with the most negative values coinciding with high Chl a concentrations and low C/N ratios (C/N < 8). This suggests predominance of phytoplankton carbon in the total particulate carbon pool, utilising a dissolved inorganic carbon reservoir, which is already significantly depleted in ¹³C. Using a simple two source mixing approach a reconstruction of the relative importance of phytoplankton to the total POC pool and of ¹³C/¹²C fractionation by phytoplankton is attempted.     

Technical Note: Calcium and carbon stable isotope ratios as paleodietary indicators

Calcium stable isotope ratios are hypothesized to vary as a function of trophic level. This premise raises the possibility of using calcium stable isotope ratios to study the dietary behaviors of fossil taxa and to test competing hypotheses on the adaptive origins of euprimates. To explore this concept, we measured the stable isotope composition of contemporary mammals in northern Borneo and northwestern Costa Rica, two communities with functional or phylogenetic relevance to primate origins. We found that bone collagen δ¹³C and δ¹⁵N values could differentiate trophic levels in each assemblage, a result that justifies the use of these systems to test the predicted inverse relationship between bioapatite δ¹³C and δ⁴⁴Ca values. As expected, taxonomic carnivores (felids) showed a combination of high δ¹³C and low δ⁴⁴Ca values; however, the δ⁴⁴Ca values of other faunivores were indistinguishable from those of primary consumers. We suggest that the trophic insensitivity of most bioapatite δ⁴⁴Ca values is attributable to the negligible calcium content of arthropod prey. Although the present results are inconclusive, the tandem analysis of δ⁴⁴Ca and δ¹³C values in fossils continues to hold promise for informing paleodietary studies and we highlight this potential by drawing attention to the stable isotope composition of the Early Eocene primate Cantius. Am J Phys Anthropol 154:633–643, 2014. © 2014 Wiley Periodicals, Inc.     

Sources of organic carbon in the littoral of Lake Gooimeer as indicated by stable carbon isotope and carbohydrate compositions

The relative importance of potential carbon sources in the littoral of Lake Gooimeer, a lake in the centre of the Netherlands, was studied using a combination of ¹³C/¹²C-ratio analysis and carbohydrate composition analysis. The littoral is covered on the land side by a 80 m wide Phragmites australis bed. Potential carbon sources were macrophyte litter, seston and benthic algae. Samples of potential carbon sources, sediments and benthic macrofauna from inside and outside the bed were analyzed for their¹³C/¹²C-ratio and some for their carbohydrate composition. Results indicate that inside the bed, macrophyte litter was the main source of carbon for both the sediment organic matter and the benthic macrofauna, and that algal material was of minor importance. Outside the bed, production by benthic algae was the main carbon source, with seston as a second source. No macrophyte derived material could be detected outside the reed bed.Abbreviations DOC (Dissolved Organic Carbon) - SOM (Sediment Organic Matter)     

A stable carbon and nitrogen isotope study of lake food webs in Canada's Boreal Plain

1.  Stable carbon and nitrogen isotope and fish stomach content analyses were used to investigate food webs in five relatively undisturbed lakes on the Boreal Plain of Canada. Stable isotope analysis was also used to determine the importance of external and internal carbon sources.
2.  Overlap in the carbon and nitrogen signatures of primary producers made it difficult to determine unambiguously the feeding habits of many invertebrates. However, isotope analysis suggested that external carbon inputs were detectable in the aquatic food chains of the one lake with a short water residence time («1 year). In the other four lakes, with water residence times ≥1 year, autochthonous carbon was the only detectable carbon source in the food webs.
3.  Food webs in these lakes spanned a range of four to five trophic levels. Both invertebrates and fish appeared to eat a variety of food, often feeding at more than one trophic level.
4.  With the exception of one lake (SPH20), top predators in these lakes, northern pike ( Esox lucius ) and fathead minnows ( Pimephales promelas ), occupied similar trophic positions despite large differences in body size and trophic morphology. In SPH20, where there were two additional fish species, pike occupied a higher trophic position. However, all the top predators in each lake appeared to be omnivores and generalists.
5.  The prevalence of omnivory and the apparent generalist feeding habits of fish in these lakes suggest that organisms are flexible in their feeding habits and that these food webs will be resilient to disturbance.     

Determination of isotope ratios in human tissues enriched with zinc stable isotope tracers using inductively coupled plasma-mass spectrometry (ICP-MS)

Enriched fecal and urine samples were prepared using ion-exchange column chromatography for analysis by Inductively Coupled and Fast Atom Bombardment Mass spectrometry (ICP-MS, FAB-MS) to compare precision between methods. Unenriched samples of human milk, feces, and whole blood were prepared similarly to monitor instrumental precision and analytical error. A least squares fit of the ICP-MS results vs the FAB-MS for⁷⁰Zn/⁶⁴Zn gave a slope of 0.98, with a relative standard deviation (RSD) of only 0.7%. The results for⁶⁸Zn/⁶⁴Zn gave a slope of 0.82, with a RSD of 14%. For unenriched tissues, all potential interferences were removed by the preparation procedure with no significant differences between preparation for isotope ratios of⁷⁰Zn/⁶⁴Zn,⁶⁸Zn/⁶⁴Zn,⁶⁷Zn/⁶⁴Zn, and⁶⁶Zn/⁶⁴Zn. Poisson counting statistics are a major contribution to the total analytical error indicating the usefulness of this procedure for enrichment studies.     

Using Bayesian stable isotope mixing models to estimate wolf diet in a multi-prey ecosystem

Stable isotope analysis (SIA) of wolf (Canis lupus) tissues can be used to estimate diet and intra-population diet variability when potential prey have distinct δ¹³C and δ¹⁵N values. We tested this technique using guard hairs collected from 44 wolves in 12 northwestern Montana packs, summer 2009. We used hierarchical Bayesian stable isotope mixing models to determine diet and scales of diet variation from δ¹³C and δ¹⁵N of wolves and potential prey, white-tailed deer (Odocoileus virginianus), mule deer (Odocoileus hemionus), elk (Cervus canadensis), moose (Alces alces), snowshoe hare (Lepus americanus), and other prey. As a check on SIA results, we conducted a separate diet analysis with temporally matched scats (i.e., collected in summer 2008) from 4 of the same packs. Wolves were centered on the ungulate prey in the isotope mixing space. Both methods revealed differences among pack diets and that wolves may consume moose in greater proportions than predicted by available biomass. Stable isotope analysis, and scat results were not entirely concordant; assumptions related to tissues of use in SIA, hair growth period in wolves, and scat sampling may have contributed to a mismatch between methods. Incorrect fractionation values, insufficient separation of prey in the isotope mixing space, choice of prior information in the Bayesian mixing models, and unexplained factors may have distorted diet estimates. However, the consistently high proportion of moose in pack diets suggests that increased population monitoring would benefit management of moose and wolves. Our results also support suggestions of other researchers that species-specific fractionation values should be used whenever possible, and that SIA may sometimes only provide indices of use for general groups of prey (e.g., large ungulates). © 2012 The Wildlife Society.     

Stable carbon and oxygen isotopes in human tooth enamel: Identifying breastfeeding and weaning in prehistory

This paper investigates the utility of stable carbon and oxygen isotopes in human dental enamel to reveal patterns of breastfeeding and weaning in prehistory. Enamel preserves a record of childhood diet that can be studied in adult skeletons. Comparing different teeth, we used δ¹³C to document the introduction of solid foods to infant diets and δ¹⁸O to monitor the decline of breastfeeding. We report enamel carbonate δ¹³C and δ¹⁸O of 33 first molars, 35 premolars, and 25 third molars from 35 burials from Kaminaljuyú, an early state in the valley of Guatemala. The skeletons span from Middle Preclassic through Late Postclassic occupations, ca. 700 B.C. to 1500 A.D. Sections of enamel were removed from each tooth spanning from the cusp to the cemento-enamel junction. Stable isotope ratios were measured on CO₂ liberated by reaction of enamel with H₃PO₄ in an automated carbonate system attached to a VG Optima mass spectrometer. Within a skeleton, teeth developing at older ages are more enriched in ¹³C and more depleted in ¹⁸O than teeth developing at younger ages. Premolars average 0.5% higher in δ¹³C than first molars from the same skeleton (P = 0.0001), but third molars are not significantly enriched over premolars. The shift from first molars to premolars may be due to the shift to solid foods from lipid-rich milk. After 2 years, when premolars begin to mineralize, the δ¹³C in childhood diets did not change systematically. First molars and premolars are similar in δ¹⁸O, but third molars average 0.7% lower than first molars (P = 0.0001) and 0.5% lower than premolars (P = 0.0003). First molar and premolar δ¹⁸O is heavier, because breast milk is more enriched in ¹⁸O than is drinking water. Hence, many children continued to nurse during the period of premolar formation. Together, these results indicate that Kaminaljuyú children had begun to eat solid maize foods before the age of 2 years but continued to drink breast milk until much later. Am J Phys Anthropol 106:1–18, 1998. © 1998 Wiley-Liss, Inc.     

Comparing pellet and stable isotope analyses of nestling Bonelli's Eagle Aquila fasciata diet

Diet analyses are central to the study of avian trophic ecology, and stable isotope analyses have made an increasing contribution in the last two decades. Few isotopic studies have assessed the diet of raptor species, which are more frequently analysed by conventional diet methods such as pellet analysis. In this study, we compare prey consumption estimates of nestling Bonelli's Eagles Aquila fasciata from conventional pellet analysis (in terms of items and biomass) and stable isotopic mixing models (SIAR) using δ¹³C, δ¹⁵N and δ³⁴S of feathers. The pellet analysis showed that European Rabbits Oryctolagus cuniculus, pigeons (mainly Common Wood Pigeons Columba palumbus and Domestic Pigeons Columba livia dom.), Red‐legged Partridges Alectoris rufa, passerines, Yellow‐legged Gulls Larus michahellis and Eurasian Red Squirrels Sciurus vulgaris were the main prey, so they were selected for diet reconstructions in SIAR. At the population level, mean prey consumption estimates were similar for pellets (both items and biomass) and SIAR. At the territory level, the weighted kappa statistic showed good ordinal scale agreement in main prey consumption between items or biomass and SIAR. Although the intraclass correlation coefficient showed poor method agreement when considering all prey in the same analysis, the intraclass correlation coefficients for each prey category showed significant agreement between pellets and SIAR when estimating the consumption of Rabbits, pigeons and Gulls, with lower agreement for passerines and Squirrels. Lastly, there was poor method agreement for estimates of Partridges. Our results suggest an overall agreement between the pellet analysis and SIAR when estimating nestling Bonelli's Eagle diet at both the population and, to a lesser extent, the territory level, supporting the usefulness of isotopic mixing models when identifying the terrestrial and marine components of raptor diets.     

Morphology,diet, and stable carbon isotopes: On the diet of Theropithecus and some limits of uniformitarianism in paleoecology

Geladas were long supposed to be the only living primates feeding almost entirely on graminoids and accordingly display dramatic dental and manual adaptive traits. A recent study of Theropithecus gelada, the first in a relatively undisturbed habitat, revealed a more diverse diet, also incorporating large quantities of forbs. The peculiar adaptive traits of T. gelada are also observed in extinct Theropithecus as early as 3.7 Ma. Stable carbon isotopic data of extinct Theropithecus from eastern Africa indicate that specimens older than 3 Ma consumed a significant proportion of C₃ plants (on average ca. 40% of total food intake) whereas specimens younger than 2 Ma consumed more C₄ plants (on average ca. 80%). Recent paleobotanical evidence suggests that C₃ herbaceous plants were still present in non‐negligible proportions in Plio‐Pleistocene lowland tropical ecosystems. Together, the shared morphological adaptive traits of extant and extinct Theropithecus and the varied diets of extant T. gelada suggest that the paleodiets of Theropithecus may have been dominated by herbaceous plants, comprising both C₃ forbs and graminoids and C₄ graminoids. The changes in stable carbon isotopes could correspond to a replacement of C₃ plants by C₄ plants within the herbaceous strata rather than a shift from C₃ woody vegetation to C₄ graminoids. This synthesis highlights the need for a more exhaustive knowledge of the ecology of extant species to achieve meaningful paleodietary and paleoenvironmental reconstructions. A strong selectivity for food resources that are rare in the landscapes (as in T. gelada) should also be considered when interpreting stable carbon isotopes of extinct African mammals (and notably hominids).     

Stable isotope variability of meso-zooplankton along a gradient of dissolved organic carbon

1.  The δ¹³C and δ¹⁵N signatures of zooplankton vary with dissolved organic carbon (DOC), but inconsistent and limited taxonomic resolution of previous studies have masked differences that may exist among orders, genera or species and are attributable to dietary and/or habitat differences. Here we investigate differences among the isotopic signatures of five zooplankton taxa ( Daphnia , Holopedium , large Calanoida, small Calanoida and Cyclopoida) in Precambrian shield lakes with a sixfold range of DOC concentration.
2.  δ¹³C signatures of Daphnia , small calanoids and large calanoids became more depleted with increasing lake DOC, whereas Holopedium and cyclopoid δ¹³C became enriched with increasing DOC concentration.
3.  The variability of δ¹³C and δ¹⁵N isotopic signatures among zooplankton groups was reduced in high-DOC, compared to low-DOC lakes, especially for δ¹³C. Differences in δ¹³C and POM-corrected δ¹⁵N accounted for up to 33.7% and 19.5% of the variance, respectively, among lakes of varying DOC concentration.
4.  The narrow range of signatures found in higher DOC lakes suggests that different taxa have similar food sources and/or habitats. In contrast, the wide range of signatures in low-DOC lakes suggests that different taxa are exploiting different food sources and/or habitats. Together with the variable trends in zooplankton isotopic signatures along our DOC gradient, these results suggest that food web dynamics within the zooplankton community of temperate lakes will change as climate and lake DOC concentrations change.     

长江宜宾江段铜鱼属鱼类种间食物关系

采用碳、氮稳定同位素技术对2012年4-5月长江宜宾江段铜鱼和圆口铜鱼的食物组成进行了分析,并利用多元统计分析方法探讨了铜鱼和圆口铜鱼种间食物关系,包括饵料相似性指数、重叠系数及2种鱼的摄食器官形态差异对食物组成的影响.结果表明:1)铜鱼的δ13C和δ15N变化范围分别为-21.15‰～-20.31‰和9.67‰～ 10.21‰,为偏动物性的杂食性鱼类;圆口铜鱼的δ13C和δ15N分别为-23.30‰～-21.18‰和7.40‰～ 9.21‰,相对铜鱼为偏植物性的杂食性鱼类,铜鱼和圆口铜鱼均与传统肠含物分析的结果存在一定的差异;2)两种鱼的食物相似性指数和食物重叠系数分别为78.7％和55.6％;3)主成分分析(PCA)表明,铜鱼和圆口铜鱼摄食器官形态特征存在一定的差异.长江宜宾江段铜鱼和圆口铜鱼种间竞争不激烈,作为向家坝蓄水前最后一次对长江宜宾江段铜鱼和圆口铜鱼食性及种间食物关系的调查,研究结果可为分析向家坝蓄水对宜宾江段鱼类营养结构的影响提供数据参考.