Yes / Abstract: The reconstruction of the diet of past populations using the stable isotope analysis of bone collagen has become a well-established tool for examining their lifeways. For example, variations in foods ingested can demonstrate differences in the foods available to individuals of different sex, age, status and in some cases identifying migrants. However, because of the remodelling of bone throughout life, this produces average values which have been incorporated in the tissues over a period of time and gives a blurred picture of the diet. The analysis of the stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) from tiny increments of dentine 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 study of teeth from a Great Famine period workhouse cemetery in Kilkenny, Ireland, and a contemporary 19th-century cemetery in London, England and compare these with published data from early Neolithic individuals from Sumburgh, Shetland, Scotland. Covariation in δ13C and δ15N values suggests that even small variations have a physiological basis. We show that high-resolution intra-dentine isotope profiles can pinpoint shortduration events such as dietary change, and in the historical populations these can be related to known periods of nutritional deprivation in the juvenile years of life. We further suggest that the data from the Famine cemetery individuals suggest a physiological marker within these isotope profiles for a period of nutritional deprivation which could be utilised in other periods and geographical areas, particularly where there is a catastrophic cemetery assemblage with no known aetiology. This technique could also have applications in a forensic setting.

Yes / Objective: This project sought to investigate whether an association may be observed between isotopic stress indicators and skeletal evidence of pathological conditions. Materials: Deciduous and permanent teeth of 15 non-adults from two contemporaneous mid-19th century London burial grounds (City Bunhill, Lukin Street). Methods: δ13C and δ15N was measured in the incrementally sectioned dentine collagen. Isotopic profiles for each individual included death during tooth development. Results: Individuals with skeletal evidence of chronic pathological conditions (e.g., rickets, tuberculosis) exhibited raised δ15N values of 0.5-1.7‰ in the months prior to death. Isotopic change consistent with chronic physiological stress prior to death was also recorded in two individuals with no skeletal evidence of disease. An offset was observed between co-forming bone and dentine δ15N values in both populations, indicating that bone and dentine are not recording the same isotopic changes. Conclusions: Isotopic change consistent with chronic physiological stress was observed in both those with and without skeletal evidence of disease, suggesting that adaptation to chronic stress in childhood was not uncommon within these 19th century London populations. Significance: Chronic physiological stress prior to death may be seen in the incrementally sampled dentine of non-adults who die during tooth formation. Limitations: The temporal resolution of current dentine micro-sampling methods may mask or minimise visibility of shorter-term periods of stress or dietary change. Suggestions for further research: Future research should further explore the relationship between specific skeletal pathologies and isotopic evidence for stress.

This thesis examines breastfeeding, weaning, and the post-weaning diets of 18 adults (18y+) and two subadults (aged 10y – 14y) from the rural Imperial Roman (1st – 4th c. CE) site of Vagnari, located in southern Italy. The investigation used a new method to sample dentine sections that accounts for the oblique nature of dentine development and allowed for the assignment of age categories to diagenetically altered teeth without visible dentine lines. The results indicate Vagnari children were weaned by ~3.5y, and that some males appear to have been breastfed longer than females. Despite the similar ages-at-weaning across the sample, the individuals in this study demonstrated a variety of weaning rates (i.e., speeds or paces), post-weaning dietary trends, and changes in diet across the life course. Some individuals (n = 6) appear to have been weaned rapidly, marked by significant removal of breastmilk prior to 2.5y, with small amounts of breast milk remaining in the diet until ~3.5y. Other children (n = 9) were weaned gradually, with slow, consistent removal of breastmilk until as late as 5.0y. Throughout and after the weaning period, children were fed a diet based on C3 plants and terrestrial proteins such as wheat, goat/sheep, and their by-products. A comparison of early life dentine and adult bone collagen signals for 14 individuals revealed changes in diet with increasing age, in which most people had increased access to higher terrestrial food sources such as pork and/or small amounts of marine food later in life. However, there was notable variation in dietary trends and practices across the sample, suggesting diverse dietary patterns among people from Vagnari. / Thesis / Master of Arts (MA) / In this thesis, I used samples from tooth dentine to analyze longitudinal stable isotope data for breastfeeding, weaning, and post-weaning dietary signals in a sample of 20 individuals from the Roman Imperial estate of Vagnari (1st – 4th c. CE) located in southern Italy. On average, children were weaned by ~3.5 years of age. Despite the similar age-at-weaning across the sample, individuals exhibited different weaning patterns and diversity in early life dietary practices at Vagnari. During and after weaning, the isotope data indicate that children were fed with C3 plants (e.g., wheat) and terrestrial proteins such as sheep/goat and pig. I analyzed childhood and adult diet by comparing dentine stable isotope data to bone collagen results from a sub-sample of 14 individuals. There was variability between the childhood dentine data and the adult bone collagen data, where individuals appeared to eat more pork and small amounts of fish later in life. This is the first study to explore breastfeeding and weaning practices of rural Roman children in southern Italy using stable isotope analysis of tooth dentine.

Yes / Objectives: This study presents isotopic information for incremental dentine collagen and bone bulk collagen from individuals from the Canary Islands (Tenerife and Gran Canaria) to explore dietary differences during childhood life. Materials and Methods: Eight individuals have been studied, which comprises 122 δ15N and δ13C incremental dentine measurements and eight bulk bone collagen analyses. A baseline of potentially consumed food sources has been developed for comparative purposes. A FRUITS model of probable contributions of each food source towards the diet of each individual has been developed. All samples but one belongs to the later period of indigenous occupation of the archipelago. Results: The dentine collagen data are presented in correlated δ13C and δ15N plots per individual, showing the isotopic changes throughout time. δ15N values for each individual tend to be variable whereas δ13C data are generally more stable with a range of +9.1 to +14‰ for δ15N and –17.4 to –20.8‰ for δ13C. Conclusion: The isotopic analysis allows for the reconstruction of 8 dietary profiles, which allow us to estimate the different dietary protein sources. The FRUITS model shows different percentages of the primary food sources for each individual. Where both δ13C and δ15N are elevated, this could be indicative of a higher marine contribution to the diet. There appear to be two main dietary profiles identifiable in the dataset and these may be related to changes in status or place of residence. Short-term variations in δ13C and δ15N and opposing co-variance of isotopic values can be indicative of nutritional stress, although metabolic changes during growth are also considered. / PhD grant from Universidad de La Laguna—Fundación La Caixa (“Contratos Predoctorales para la formación de doctores ULL-2015”), MINECO (Ministerio de Economía y Competitividad) project “Guanches y europeos en Las Cañadas del Teide, Ocupación, Producción y Comunicación” (HAR2015-68323-P), “Erasmus+ Scholarship” given by the University of La Laguna, ERC Starting Grant project IsoCAN (grant 851733, European Comission) and the projects RTI2018-101923-J-I00 and RYC2019-028346 (Ministerio de Ciencia e Innovación)

Yes / Background: Carbon (δ13C) and nitrogen (δ15N) isotope ratios of collagen from teeth and bone are used to study human nutrition and health. As bones are constantly remodelling throughout life, isotopic values of bone collagen represent an average of several years. In contrast, human teeth do not remodel and their primary dentine contains only the isotopic data from the time of formation. In contrast to all other bones, human auditory ossicles also appear not to remodel. As they develop in utero and finish formation in the first 2 years of life, their collagen should also represent isotopic values of these two relatively short periods. Aim: By comparing δ13C and δ15N data from ossicles and incremental dentine, this study aims to investigate how two developmental periods of the ossicles, in utero and the first 2 years of life, reflect in collagen obtained from the ossicles. Subject and methods: Ossicle and tooth samples of 12 individuals aged 0.5 ± 0.4 years to 13 ± 1 years from the nineteenth century St. Peter’s burial ground in Blackburn were collected and processed to obtain bulk bone and incremental dentine collagen which was measured for δ13C and δ15N. Results: Averaged δ13C and δ15N of ossicles are lower when compared to every age group except after 3 years of age. Average offset between ossicles and dentine of different groups ranges from 0.4–0.9‰ for δ13C and from 0.3–0.9‰ for δ15N, with highest counterbalance at birth and after the first 5 months after birth. Conclusions: There appears to be a systematic offset between the dentine and ossicle data. It seems that the second phase of development does not influence the isotopic values of collagen significantly and the data we are obtaining from ossicles represents the in utero period. / Research grant from The Society for the Study of Human Biology.

Yes / There are few reliable osteological indicators to detect parity or infer puberty in skeletal remains. Nitrogen (δ15N) and stable carbon (δ13C) isotope ratios in human tissues can be affected by metabolically unbalanced states engendered by pregnancy or rapid growth, offering potential biomarkers. This pilot study explores the potential of incremental dentine-collagen isotope ratio analysis to identify puberty and gestation. Methodology: Incremental dentine δ15N and δ13C profiles were produced by analysing third molars extracted as part of dental treatment of 10 individuals living in Sudan. Demographic and anthropometric data at the time of tooth extraction was available. Medical histories were unknown. Results: Isotopic signatures potentially related to pubertal growth, with an average δ15N reduction of 0.78±0.29‰, are indicated. Six isotopic signals suggestive of pregnancy, with an average δ15N decrease of 0.48±0.22‰, are also observed. The timing, speed and amplitude of post-partum δ15N patterns seemingly infer infant feeding practices and maternal nutritional status. Conclusion: This pilot study highlights the potential of incremental dentine isotope analysis for the reconstruction of early reproductive histories in skeletal remains. However, controlled studies with larger human cohort are needed to validate these findings, establish isotopic signals linked to puberty and lactation, and improve chronology accuracy.

Yes / In 2015, a previously unknown enclosed settlement and burial ground was found near the summit of a low hill in Ranelagh townland, just north of Roscommon town. The site—officially designated Ranelagh 1, and hereafter referred to variously as ‘the Ranelagh site’, ‘the site at Ranelagh’ or simply ‘Ranelagh’—was excavated over a 54-week period by Excavation Director Shane Delaney for Irish Archaeological Consultancy (IAC) Ltd between October 2015 and October 20161 . Excavations revealed that the site was established during the fourth century AD; for over 1,000 years, until the final phase of burial activity proper concluded there shortly after AD 1400, the site would have been a prominent feature in both the geographical and psychological landscape of the time. Cillín (children’s) burials continued at the site until about AD 1650, further asserting this prominence.

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.

Yes / This paper presents a case study of a young infant, from a larger isotopic and osteological investigation of Bronze/Iron Age (14th-4th century BC) skeletal assemblages from Croatia and Slovenia. The osteological analysis of this infant identified pathological lesions including abnormal porosity and new bone formation consistent with malnutrition and phases of recovery. The distribution and appearance of these pathological lesions (i.e. diffuse micro-porosities and plaques of subperiosteal new bone formation on the skull and long bones) led to the conclusion that this infant probably suffered from scurvy (vitamin C deficiency). The diet and nitrogen balance of this individual were investigated by incremental dentine sampling and stable carbon and nitrogen isotope analysis. This sampling method provided a high resolution record of dietary and metabolic changes from pre-birth to around the time of death. The resulting isotope data exhibited unusually high δ13C values for this region and time period (between -11.3‰ and -12.6‰), while δ15N values were observed to be c. 3‰ above that of rib collagen sampled from contemporary adults recovered from the same site. The isotope profiles generated from the incremental dentine analysis show that δ13C and especially δ15N continue to increase until death. The evidence from the skeletal remains and high resolution isotopic data support the hypothesis that this infant suffered from severe malnutrition and an increasingly negative nitrogen balance. The paper discusses some scenarios which could have resulted in these unusual isotope ratios, whilst considering the diagnosis of possible metabolic disease. The paper also addresses the need for context when interpreting isotopic results. The isotope data should not be viewed in isolation, but rather as part of a multidisciplinary approach, considering the multiple causes of isotopic variability.