In 2001, 3269 unidentified individuals were found in a mass grave on the Northern part of Vilnius, Lithuania. Artifactual context indicates that these individuals were likely soldiers that were a part of Napoleon’s Grand Army. Stable oxygen isotope analysis was performed on bone apatite from 9 femoral bone samples to determine whether or not these individuals were Lithuanian locals and to test ratio variation. If individuals were foreigners, then geographical origins were approximated utilizing percentages of C4 plants from Holder (2013) and δ18O values that were extracted from bone apatite. The carbonate oxygen isotope compositions (δ18Ocarbonate) of bone apatite from the femoral samples (-4.4‰ to -6.2‰) indicate that these individuals were from central and western Europe (-4.0‰ to -6.9‰). It is significant that none of the individuals have values consistent with the area around Lithuania (-10.0‰ to -11.9‰), because it means that they all were non-local. It is also indicative that the Lithuanians were not burying their citizens in the grave and therefore strongly support that these individuals were Napoleonic soldiers. Additionally, although C4 percentages in the diet ranged from 17.8% to 31.7%, which overlaps with eastern European consumption patterns (approximately 15% to 25% of C4 plants) (Reitsema et al., 2010), the slight shift towards a higher C4 percentage is more representative of a central and western European diet. These results are significant because they provide stable isotopic evidence that these individuals were Napoleon’s soldiers whom participated in the Russian campaign of 1812.

Stable isotope analysis is often used to evaluate elements of the lives of past peoples, such as diet and health status, at a societal level. Analysis at an individual level is exceptionally rare, and has not been conducted using a variety of tissues representing both early life and life approximate to death. In this study, δ 13C and δ15N isotope signatures are used to create life histories for single individuals from Romano-Christian period Kellis 2 cemetery in the Dakhleh Oasis, Egypt. Samples are obtained from several different tissues, including tooth dentin, bone collagen, hair, nail, skin, and gut content, all of which have been previously researched, but have not been studied at such an individualistic level. By using data and previous research conducted by Drs. Tosha Dupras and Lana Williams, this research uses isotopic values from the aforementioned tissues, and the differing turnover rates of these tissues, to develop lifetime timetables for 15 individuals (female, male, and juvenile). Results show that individual analysis is possible, informative, and can enlighten researchers not just concerning the individual, but about the population as a whole. The methods presented can serve as a model for reconstructing individual life histories using isotope data from multiple tissues.

Although stable isotope analysis is routinely utilized in bioarchaeology and relies on principles that are well-understood, there are still fundamental issues that have not been thoroughly investigated. This thesis examines the relationship between inter-tissue spacing of carbon stable isotope values (δ13C) and age in juvenile human remains. Analyses of tissues within the same individual reveal disparate isotopic values for a variety of physiological and biological reasons discussed herein. This project examines the distance between the δ13C values in bone collagen, skin, hair, and nail, and examines how these distances vary between different age groups, utilizing data collected from 52 well-preserved human remains from the Dakhleh Oasis, Egypt: 28 individuals between the ages of 1 and 4 years, 14 between 5 and 10 years, and 10 between the ages of 11-15 years. The mean carbon isotope values for each tissue were compared across each age group, and used to calculate the differences between each tissue type. Although distances between tissues were found to vary across all age categories, the distances between collagen and hair, collagen and skin, and collagen and nail are all substantially greater in the 11-15 year old category than those in the 1-4 and 5-10 year categories. Possible physiological, developmental and social factors are discussed in an effort to explain this discrepancy

A juvenile’s dependency on their caregiver is significant to the overall development of nutritionally related pathological lesions. However, not all skeletal pathology is caused by nutritional stress; despite anemia being the usual inferred cause, the origin of Cribra Orbitalia (CO) – lesions on the orbital roofs of the cranium– is undetermined. The purpose of this research is to compare the reconstructed diets of juveniles with and without CO and explore connections to dietary patterns (inferred from stable isotope ratios of carbon and nitrogen). Rib bone samples of 79 juveniles with and without CO were sampled from the Kuelap archaeological site in Chachapoyas, Peru (AD 800–1532) – known for its archaeological diversity. Stable isotope analysis was conducted (δ13C and δ15N values) to statistically analyze each group’s values. Samples were further subdivided into age cohorts of infants (0–3 years), juveniles (4–11 years), and adolescents (12–18 years). The diets of juveniles with and without CO were determined to have no statistically significant difference between each other. However, a significant statistical difference did exist between the diets of the different juvenile age cohorts regardless of CO status, indicating that weaning and early dietary transitions through childhood affected the juvenile’s nutritional regime in the region. The research presented is the first study of the relationship between nutrition and CO from Kuelap; significantly, it further explores the lifestyle of past individuals in Chachapoyas through the understanding of childhood diets.

Before reaching adulthood, every individual experiences a period of dependency, the juvenile period, during which they rely on the older, more experienced members of their society for their security, subsistence and care. This juvenile period is an important stage of life for human physical and physiological development. In bioarchaeology, there has been limited research conducted on juveniles, particularly, the development of their own social identity and influences. The research method of stable carbon (C) and nitrogen (N) isotope analysis is used to reconstruct the paleodiet of juveniles to determine their dietary composition. Specifically, this research is focused on Kuelap, located in the highlands of Peru, a large settlement inhabited from 900-1535 AD, including pre-Inca (900-1469 AD) and Inca periods (1470-1535 AD). The primary aim of this research is to determine if juveniles consumed different foods through different time periods. Another aim of this research is to determine if juveniles were treated differently than their adult counterparts. Bone collagen samples, primarily ribs, from 32 juveniles were analyzed. The average δ13C value for the pre-Inca juveniles was –13.1‰, and –13.4‰ for the Inca period juveniles. There was no statistically significant difference in δ13C values between juvenile groups or between adult and juvenile subsamples. The average δ15N for the pre-Inca juveniles was +8.1‰ and +7.8‰ for the Inca period juveniles. The Mann-Whitney U test determined there was not a statistically significant difference in δ15N values between the juvenile burial groups; however, there was a statistically significant difference between the juvenile and adult subgroups. The findings suggest that there may have been preferential treatment toward or metabolic stressors on the juvenile. The results of this study offer insights to availability of dietary components, societal roles based on developmental age stages, and the potential role of parenting in Kuelap.

<p>Investigating the presence of microbial communities in the Earth's deep terrestrial subsurface and the metabolic processes taking place in these environments provides insight into the some of the ultimate limits for life on Earth, as well as the potential for microbial life to exist within the subsurface of other planetary bodies. This Master's thesis project utilized phospholipid fatty acid (PLFA) analysis, in combination with carbon isotope analyses (δ¹³C and Δ¹⁴C), to explore the presence and activity of microbial communities living within deep terrestrial subsurface fracture water systems and low permeability, deep sedimentary rocks. Deep fracture water systems, ranging from 0.9 to 3.2 km below land surface, were sampled for microbial communities via deep mine boreholes in the Witwatersrand Basin of South Africa. PLFA concentrations revealed low biomass microbial communities, ranging from 2x10¹ to 5x10⁴ cells per mL and the PLFA profiles contained indicators for environmental stressors, including high temperatures and nutrient deprivation. δ¹³C and Δ¹⁴C analyses of PLFAs and potential carbon sources (dissolved inorganic carbon (DIC), dissolved organic carbon (DOC) and methane) identified microbial utilization of methane in some systems and utilization of DIC in others. Evidence for microbial oxidation of methane and chemoautotrophy in these systems is consistent with a self-sustaining deep terrestrial subsurface biosphere that is capable of surviving independent of the photosphere. Viable microbial communities were also identified within deep (334 to 694 m depth) sedimentary rock cores sampled from the Michigan Basin, Canada. PLFA analyses revealed microbial cell densities ranging from 1-3 x 10⁵ cells/mL and identified PLFA indicators for environmental stressors. These results demonstrate the ubiquity of microbial life in the deep terrestrial subsurface and provide insight into microbial carbon sources and cycling in deep microbial systems which may persist in isolation over geologic timescales.</p> / Master of Science (MSc)

Assessing population diversity in southern Italy has traditionally relied on archaeological and historic evidence. Although informative, these lines of evidence do not establish specific instances of within lifetime mobility, nor track population diversity over time. In order to investigate the population structure of ancient South Italy I sequenced the mitochondrial DNA (mtDNA) from 15 Iron Age (7th – 4th c. BCE) and 30 Roman period (1st – 4th c. BCE) individuals buried at Iron Age Botromagno and Roman period Vagnari, in southern Italy, and analyzed δ18O and 87Sr/86Sr values from a subset of the Vagnari skeletal assemblage. Phylogenetic analysis of 15 Iron Age mtDNAs together with 231 mtDNAs spanning European prehistory suggest that southern Italian Iapygians share close genetic affinities to Neolithic populations from eastern Europe and the Near East. Population pairwise analysis of Iron Age, Roman, and mtDNA datasets spanning the pan-Mediterranean region (n=357), indicate that Roman maternal genetic diversity is more similar to Neolithic and Bronze Age populations from central Europe and the eastern Mediterranean, respectively, than to Iron Age Italians. Genetic distance between population age categories imply moderate mtDNA turnover and constant population size during the Roman conquest of South Italy in the 3rd century BCE. In order to determine the local versus non-local demographic at Vagnari, I measured the 87Sr/86Sr and 18O/16O of composition of 43 molars, and the 87Sr/86Sr composition of an additional 13 molars, and constructed a preliminary 87Sr/86Sr variation map of the Italian peninsula using disparate 87Sr/86Sr datasets. The relationship between 87Sr/86Sr and previously published δ18O data suggest a relatively low proportion of migrants lived at Vagnari (7%). This research is the first to generate whole-mitochondrial DNA sequences from Iron Age and Roman period necropoleis, and demonstrates the ability to gain valuable information from the integration of aDNA, stable isotope, archaeological and historic evidence. / Thesis / Doctor of Philosophy (PhD) / With biochemical information obtained from teeth, this study examines the population structure and geographic origins in two archaeological communities located in southern Italy. Analysis of classical remains has traditionally been the subject of historical and archaeological inquiry. However, new applications evaluate these population changes with integrated stable isotope and ancient DNA techniques. Overall, the biochemical results suggest that the pre-Roman communities harbor deep maternal ancestry originating from eastern Europe and the eastern Mediterannean. These results, when compared to the genetic diversity of Roman and broader Mediterranean populations, indicate that the Romans share closer genetic similarity with ancient Stone and Bronze Age communites from Europe and the eastern Mediterranean, than with the pre-Roman community studied here. Furthermore, tooth chemistry results indicate a predominantly local population buried in the Roman period cemetery.

No / Incremental dentine analysis utilizes tissue that does not remodel and that permits comparison, at the same age, of those who survived infancy with those who did not at high temporal resolution. Here, we present a pilot study of teeth from a 19th-century cemetery in London, comparing the merits of two methods of obtaining dentine increments for subsequent isotope determination. Covariation in ¿13C and ¿15N values suggests that even small variations have a physiological basis. We show that high-resolution intra-dentine isotope profiles can pinpoint short-duration events such as dietary change or nutritional deprivation in the juvenile years of life.

Yes / Identifying people of exotic origins with isotopes depends upon finding isotopic attributes that are inconsistent with the indigenous population. This task is seldom straightforward and may vary with physical geography, through time, and with cultural practices. Isotopes and trace elements were measured in four Viking Age (8th to 10th centuries A.D.) skeletons from Dublin, Ireland, and three from Westness, Orkney. These were compared with other data from these locations and contemporaneous skeletons from Britain. We conclude that the male skeletons from Dublin have disparate origins, two originating beyond the shores of Ireland, and that the female and two male skeletons from Westness are not indigenous to Orkney. However, the homeland of the female, in contrast to the males, is unlikely to be in Scandinavia.

Yes / The major components of human diet both past and present may be estimated by measuring the carbon and nitrogen isotope ratios (δ13C and δ15N) of the collagenous proteins in bone and tooth dentine. However, the results from these two tissues differ substantially: bone collagen records a multi-year average whilst primary dentine records and retains timebound isotope ratios deriving from the period of tooth development. Recent studies harnessing a sub-annual temporal sampling resolution have shed new light on the individual dietary histories of our ancestors by identifying unexpected radical short-term dietary changes, the duration of breastfeeding and migration where dietary change occurs, and by raising questions regarding factors other than diet that may impact on δ13C and δ15N values. Here we show that the dentine δ13C and δ15N profiles of workhouse inmates dating from the Great Irish Famine of the 19th century not only record the expected dietary change from C3 potatoes to C4 maize, but when used together they also document prolonged nutritional and other physiological stress resulting from insufficient sustenance. In the adults, the influence of the maize-based diet is seen in the δ13C difference between dentine (formed in childhood) and rib (representing an average from the last few years of life). The demonstrated effects of stress on the δ13C and δ15N values will have an impact on the interpretations of diet in past populations even in slow-turnover tissues such as compact bone. This technique also has applicability in the investigation of modern children subject to nutritional distress where hair and nails are unavailable or do not record an adequate period of time. / This study was supported by an Arts and Humanities Research Council grant funding to JB under AHRC Studentship AH/I503307/1.