Integrating cortisol and isotopic analyses of archeological hair: reconstructing individual experiences of health and stress.

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.

Intraskeletal isotopic compositions (δ(13) C, δ(15) N) of bone collagen: nonpathological and pathological variation.

Paleodiet research traditionally interprets differences in collagen isotopic compositions (δ(13) C, δ(15) N) as indicators of dietary distinction even though physiological processes likely play some role in creating variation. This research investigates the degree to which bone collagen δ(13) C and δ(15) N values normally vary within the skeleton and examines the influence of several diseases common to ancient populations on these isotopic compositions. The samples derive from two medieval German cemeteries and one Swiss reference collection and include examples of metabolic disease (rickets/osteomalacia), degenerative joint disease (osteoarthritis), trauma (fracture), infection (osteomyelitis), and inflammation (periostitis). A separate subset of visibly nonpathological skeletal elements from the German collections established normal intraindividual variation. For each disease type, tests compared bone lesion samples to those near and distant to the lesions sites. Results show that normal (nonpathological) skeletons exhibit limited intraskeletal variation in carbon- and nitrogen-isotope ratios, suggesting that sampling of distinct elements is appropriate for paleodiet studies. In contrast, individuals with osteomyelitis, healed fractures, and osteoarthritis exhibit significant intraskeletal differences in isotope values, depending on whether one is comparing lesions to near or to distant sites. Skeletons with periostitis result in significant intraskeletal differences in nitrogen isotope values only, while those with rickets/osteomalacia do not exhibit significant intraskeletal differences. Based on these results, we suggest that paleodiet researchers avoid sampling collagen at or close to lesion sites because the isotope values may be reflecting both altered metabolic processes and differences in diet relative to others in the population.

Integrating cortisol and isotopic analyses of archaeological hair: Elucidating juvenile ante-mortem stress and behaviour.

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±402ng/g) compared to Peruvian adults (281±35ng/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.

Carbon and nitrogen isotopic survey of northern peruvian plants: baselines for paleodietary and paleoecological studies.

The development of isotopic baselines for comparison with paleodietary data is crucial, but often overlooked. We review the factors affecting the carbon (δ(13)C) and nitrogen (δ(15)N) isotopic compositions of plants, with a special focus on the carbon and nitrogen isotopic compositions of twelve different species of cultivated plants (n = 91) and 139 wild plant species collected in northern Peru. The cultivated plants were collected from nineteen local markets. The mean δ(13)C value for maize (grain) was -11.8±0.4 ‰ (n = 27). Leguminous cultigens (beans, Andean lupin) were characterized by significantly lower δ(15)N values and significantly higher %N than non-leguminous cultigens. Wild plants from thirteen sites were collected in the Moche River Valley area between sea level and ∼4,000 meters above sea level (masl). These sites were associated with mean annual precipitation ranging from 0 to 710 mm. Plants growing at low altitude sites receiving low amounts of precipitation were characterized by higher δ(15)N values than plants growing at higher altitudes and receiving higher amounts of precipitation, although this trend dissipated when altitude was >2,000 masl and MAP was >400 mm. For C(3) plants, foliar δ(13)C was positively correlated with altitude and precipitation. This suggests that the influence of altitude may overshadow the influence of water availability on foliar δ(13)C values at this scale.

Stable isotope biogeochemistry of seabird guano fertilization: results from growth chamber studies with maize (Zea mays).

BACKGROUND: Stable isotope analysis is being utilized with increasing regularity to examine a wide range of issues (diet, habitat use, migration) in ecology, geology, archaeology, and related disciplines. A crucial component to these studies is a thorough understanding of the range and causes of baseline isotopic variation, which is relatively poorly understood for nitrogen (δ(15)N). Animal excrement is known to impact plant δ(15)N values, but the effects of seabird guano have not been systematically studied from an agricultural or horticultural standpoint. METHODOLOGY/PRINCIPAL FINDINGS: This paper presents isotopic (δ(13)C and δ(15)N) and vital data for maize (Zea mays) fertilized with Peruvian seabird guano under controlled conditions. The level of (15)N enrichment in fertilized plants is very large, with δ(15)N values ranging between 25.5 and 44.7‰ depending on the tissue and amount of fertilizer applied; comparatively, control plant δ(15)N values ranged between -0.3 and 5.7‰. Intraplant and temporal variability in δ(15)N values were large, particularly for the guano-fertilized plants, which can be attributed to changes in the availability of guano-derived N over time, and the reliance of stored vs. absorbed N. Plant δ(13)C values were not significantly impacted by guano fertilization. High concentrations of seabird guano inhibited maize germination and maize growth. Moreover, high levels of seabird guano greatly impacted the N metabolism of the plants, resulting in significantly higher tissue N content, particularly in the stalk. CONCLUSIONS/SIGNIFICANCE: The results presented in this study demonstrate the very large impact of seabird guano on maize δ(15)N values. The use of seabird guano as a fertilizer can thus be traced using stable isotope analysis in food chemistry applications (certification of organic inputs). Furthermore, the fertilization of maize with seabird guano creates an isotopic signature very similar to a high-trophic level marine resource, which must be considered when interpreting isotopic data from archaeological material.

Trophic level and macronutrient shift effects associated with the weaning process in the Postclassic Maya.

The weaning process was investigated at two Maya sites dominated by Postclassic remains: Marco Gonzalez (100 BC-AD 1350) and San Pedro (1400-AD 1650), Belize. Bone collagen and bioapatite were analyzed from 67 individuals (n < or = 6 years = 15, n > 6 years = 52). Five isotopic measures were used to reconstruct diet and weaning: stable nitrogen- and carbon-isotope ratios in collagen, stable carbon- and oxygen-isotope ratios in bioapatite, and the difference in stable carbon-isotope values of coexisting collagen and bioapatite. Nitrogen-isotope ratios in infant collagen from both sites are distinct from adult females, indicating a trophic level effect. Collagen-to-bioapatite differences in infant bone from both sites are distinct from adult females, indicating a shift in macronutrients. Oxygen-isotope ratios in infant bioapatite from both sites are also distinct from adult females, indicating the consumption of breast milk. Among infants, carbon- and nitrogen-isotope ratios vary, indicating death during different stages in the weaning process. The ethnohistoric and paleopathological literature on the Maya indicate cessation of breast-feeding between ages 3-4 years. Isotopic data from Marco Gonzalez and San Pedro also indicate an average weaning age of 3-4 years. Based on various isotopic indicators, weaning likely began around age 12 months. This data set is not only important for understanding the weaning process during the Postclassic, but also demonstrates the use of collagen-to-bioapatite spacing as an indicator of macronutrient shifts associated with weaning.