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In recent years, it has become evident that limitations exist in our ability to meaningfully assess palaeodiet using stable isotope compositions. These limitations in part arise because many of the fundamental assumptions about tissue-diet relationships are poorly understood. In order to redress this deficiency, a controlled feeding experiment was undertaken to define the impact of terrestrial- vs. marine-derived dietary protein consumption on consumer tissue carbon isotopic compositions (δ13C). Two generations of pigs were raised on one of five feeds with varying proportions of terrestrial (soy) and marine (fish meal) protein. A comprehensive range of tissues and fluids from 49 pigs was submitted for δ13C analysis. The observed tissue–whole diet and tissue–dietary protein carbon isotopic offsets were found to be highly dependent on the percentage of marine protein in diet. We suggest that the trend in δ13C offsets most likely derives from the increased routing of non-essential amino acids, especially glycine, with the increasing proportion of marine protein in the diet. These findings demonstrate that solely using bulk δ13C compositions not only masks considerable information about diet, but may also lead to erroneous representations of marine and terrestrial resource consumption in the past.

Sulphur-isotope determinations are becoming increasingly useful for palaeodietary reconstruction, but knowledge of isotopic discrimination between diet and various tissues remains inadequate. In this study, we explore the sensitivity of δ34Stissue values to changes in δ34Sdiet values, sulphur isotopic discrimination between diet and consumer, and the potential impact of terrestrial vs. marine protein consumption on these discrimination offsets. We present new δ34S values of bone collagen, muscle, liver, hair, milk and faeces from ten mature sows, ten piglets and fifteen adolescent pigs from a controlled feeding study. The δ34Stissue values were found to co-vary with the δ34Sdiet values, the δ34Stissue – δ34Sdiet isotopic offsets (Δ34Stissue-diet) are small but consistent, and dietary protein source does not systematically alter the Δ34Stissue-diet isotopic discrimination. The outcomes of this study are of particular relevance to questions that are difficult to resolve using carbon and nitrogen stable isotopes alone, and will also be useful in regions where terrestrial, freshwater, and marine resources could have all potentially contributed to human diet.

Compound-specific amino acid carbon-isotope compositions have shown particular promise for elucidating dietary behaviors in complex environmental contexts, and may also be able to mitigate the effect of many of the limitations inherent to palaeodietary reconstructions. Here, we investigate the efficacy of compound-specific amino acid isotopic proxies in characterizing the consumption of different dietary protein sources using amino acid carbon-isotope compositions for humans and fauna from Rössberga (Early to Middle Neolithic), Köpingsvik (Mesolithic and Middle Neolithic), and Visby (Medieval Period), Sweden. We also assess the explanatory capabilities of an isotopic mixing model when used with essential amino acid carbon-isotope compositions of humans and local fauna. All three isotopic proxies distinguished among humans from the three sites consistently and informatively, and were able to enhance the broad interpretations made using bulk isotopic compositions. The mixing model palaeodietary reconstruction revealed considerable diversity in relative protein source contributions among individuals at both Köpingsvik and Visby. Comparing the mixing model for bulk carbon- and nitrogen-isotope compositions to the model for essential amino acid isotopic compositions further demonstrated the likelihood of underestimation and overestimation of marine protein consumption for both aquatic-dominant and mixed marine-terrestrial diets when using bulk isotopic compositions.

In this study, femoral collagen, rib collagen, femoral muscle, loin muscle and liver samples from sows, piglets and pigs raised in a controlled feeding study are analysed for their nitrogen-isotope compositions. The objectives of this research are to investigate the relationship between tissue and dietary δ15N values across age categories under controlled feeding and housing conditions, and to assess tissue 15N-enrichment relative to diet when pigs of different ages are consuming terrestrial, marine, or mixed terrestrial-marine dietary protein. There is a strong linear relationship between all tissue δ15N values and the amount of marine protein consumed, but the δ15N values do not become consistently elevated for all individuals consuming the same diet until at least 25% of the dietary protein source is marine-derived. Adolescent pigs also had consistently lower δ15N values than either piglets or sows consuming the same diet for collagen and muscle, which is most likely caused by the differences in growth rate among the age categories. Further, for some tissues and animals, a linear relationship between the amount of marine protein consumed and the Δ15NTissue – Whole Diet offset was also observed. We suggest that this variability results from both age-associated growth rates and differential incorporation of amino acids from terrestrial and marine dietary protein into rapidly growing tissue.

Of central importance to palaeodietary reconstruction is a clear understanding of relative contributions of different terrestrial (i.e., C3 vs. C4 plants) and aquatic (i.e., freshwater vs. marine) resources to human diet. There are, however, significant limitations associated with the ability to reconstruct palaeodiet using bulk collagen stable isotope compositions in regions where diverse dietary resources are available. Recent research has determined that carbon-isotope analysis of individual amino acids has considerable potential to elucidate dietary protein source where bulk isotopic compositions cannot. Using δ13CAA values for human and faunal remains from Zvejnieki, Latvia (8th – 3rd millennia BCE), we test several isotopic proxies focused on distinguishing freshwater protein consumption from both plant-derived and marine protein consumption. We determined that the Δ13CGly-Phe and Δ13CVal-Phe proxies can effectively discriminate between terrestrial and aquatic resource consumption, and the relationship between essential δ13CAA values and the Δ13CGly-Phe and Δ13CVal-Phe proxies can differentiate among the four protein consumption groups tested here. Compound-specific amino acid carbon-isotope dietary proxies thus enable an enhanced understanding of diet and resource exploitation in the past, and can elucidate complex dietary behaviour.

Archeological hair from 14 adults from the Nasca Region, Peru (c. AD1-1000) was analyzed for carbon and nitrogen isotopic compositions and cortisol levels. We investigated the relationship between isotopic compositions, which reflect diet, and cortisol, which reflects biogenic cortisol production and chronic stress. Using a case study approach, we determined that there are consistent changes in cortisol production associated with the rapid dietary change characteristic of local mobility. Moreover, changes in nitrogen- and carbon-isotope compositions, when integrated with cortisol levels, enabled inferences to be made about nitrogen metabolism and carbon routing, and elucidated the nature of potential stressors in the months before death. The isotopic and cortisol data suggested a relatively high rate of exposure to stress that is consistent with what is known about the Nasca Region social and physical environments. Of the 14 adults included in this study, six likely suffered from illness/trauma before death, and a further three experienced stress without an observable associated change in isotopic composition. Five individuals also experienced increased stress related to local mobility, inferred from co-occurring changes in cortisol production and dietary shifting. The integration of cortisol and isotopic data revealed individual characteristics of hidden frailty and risk that would not be apparent using more traditional methods of evaluating health status. This approach will provide a powerful enhancement to the understanding of stress, morbidity, and well-being developed through skeletal analysis.

Understanding childhood experiences of health and well-being is essential to reconstructing ancient life-ways. Here, archaeological hair samples from five juveniles from Cahuachi and near Huaca del Loro in the Nasca Region, Peru, are analyzed for their carbon- and nitrogen-isotope compositions and cortisol levels. Stable isotopic data are used to investigate dietary change and nitrogen metabolism, and cortisol levels are used to infer exposure to stress. Using a case study approach, we determined that juveniles have distinct, individualized experiences of diet and morbidity, which are, in some cases, similar to adult experiences in the same region. Overall, Nasca Region juveniles have high systemic cortisol levels (1444 ± 402 ng/g) compared to Peruvian adults (281 ± 35 ng/g; Webb et al., 2010 ). Younger juveniles have comparatively high δ15N values that decrease over several months, suggesting transition from breast milk to a weaning diet. Older juveniles exhibit patterns of dietary shifting similar to those determined for adults in the region, or suggestive of particular socioeconomic roles. This study demonstrates the value of applying biomolecular methods to juvenile mummified remains to better understand the life histories of children in archaeological contexts.