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 / The Early Bronze Age barrows at Irthlingborough and Gayhurst in central England are notable for the large number of cattle (Bos taurus) remains associated with their human Beaker burials. Previous work using strontium isotope analysis has indicated that most of the cattle analysed, and one aurochs (Bos primigenius), were of local origin (Towers et al. 2010). In this study, stable isotope analysis of enamel and bone was carried out to investigate whether the mature cattle had experienced similar husbandry practices, climate and environment. Bulk carbon, nitrogen and sulphur isotope analysis of collagen suggested most were consuming similar sources of plant protein from environments probably local to the sites and this was supported by high resolution intra-enamel carbon isotope profiles. Oxygen isotope profiles indicated the aurochs and most of the cattle experienced similar climatic regimes: the only exception being an animal with a non-local strontium isotope ratio. However, a comparison of seasonality profiles of the local animals using estimated tooth formation times showed that there was no consistency in season of birth: the animals appeared to have been born throughout the year. Cattle can breed throughout the year but it requires considerable human effort and intervention to successfully overwinter young stock; it is therefore unlikely to have been carried out without good reason and benefit if winters were harsh. One reason is to ensure a continuous supply of milk. Measuring oxygen isotope profiles to identify year-round calving may thus be a potential indicator of dairying economies.

Yes / The results of carbon and nitrogen isotope analysis from two medieval populations are presented here, in a study investigating dietary habits within a medieval hospital population in England. We used δ13C and δ15N measurements of bone collagen in order to attempt to identify a distinct group diet within the medieval hospital of St. Giles, Brough, Yorkshire, and examine the reasons why the dietary habits within the institution may have been noticeably different from that of a comparative lay population. Following the results and tentative conclusions of a study conducted by Müldner and Richards (2005), it was hypothesised that religious fasting rules would result in there being evidence of greater consumption marine fish at St. Giles than at the rural township of Box Lane, Pontefract, Yorkshire. While more dietary variation was found at the hospital, it can be seen that the differences in δ13C and δ15N isotope values vary in relation to the animal baselines. Thus, differences between the human populations can be attributed to geological and environmental factors as opposed to dietary differences.

Yes / Establishing the age at which prehistoric individuals move away from their childhood residential location holds crucial information about the socio dynamics and mobility patterns in ancient societies. We present a novel combination of strontium isotope analyses performed on the over 3000 year old “Skrydstrup Woman” from Denmark, for whom we compiled a highly detailed month-scale model of her migration timeline. When combined with physical anthropological analyses this timeline can be related to the chronological age at which the residential location changed. We conducted a series of high-resolution strontium isotope analyses of hard and soft human tissues and combined these with anthropological investigations including CT-scanning and 3D visualizations. The Skrydstrup Woman lived during a pan-European period characterized by technical innovation and great social transformations stimulated by long-distance connections; consequently she represents an important part of both Danish and European prehistory. Our multidisciplinary study involves complementary biochemical, biomolecular and microscopy analyses of her scalp hair. Our results reveal that the Skrydstrup Woman was between 17–18 years old when she died, and that she moved from her place of origin -outside present day Denmark- to the Skrydstrup area in Denmark 47 to 42 months before she died. Hence, she was between 13 to 14 years old when she migrated to and resided in the area around Skrydstrup for the rest of her life. From an archaeological standpoint, this one-time and one-way movement of an elite female during the possible “age of marriageability” might suggest that she migrated with the aim of establishing an alliance between chiefdoms. Consequently, this detailed multidisciplinary investigation provides a novel tool to reconstruct high resolution chronology of individual mobility with the perspective of studying complex patterns of social and economic interaction in prehistory. / Carlsberg Foundation through the project entitled "Tales of Bronze Age Women" CF-15 0878 to KMF (http://www. carlsbergfondet.dk/en).

Doctor of Philosophy / Department of Biology / David A. Haukos / Mottled ducks (Anas fulvigula) are dabbling waterfowl species native to coastal wetlands of the Gulf of Mexico of the United States and Mexico. Although closely related to common waterfowl species such as the mallard (A. platyrhynchos) and American black duck (A. rubripes), the mottled duck exhibits unique behavior, mainly in its life history as a non-migratory species. As such, because of population declines caused by predation, habitat destruction, and environmental contaminants, this species requires specialized conservation concerns and species-specific management to protect population numbers. The goal of this study was to assess ongoing effect of observed lead (Pb) contamination and exposure issues in mottled ducks and their habitats, which I achieved by conducting assessments that will provide managers habitat and organism level metrics to detect and mitigate lead in mottled ducks and their environments. My field study was conducted at the Texas Chenier Plain National Wildlife Refuge Complex (TCPC), which was the area of greatest mottled duck density on the Texas Coast. I first created a body condition index to provide managers a tool to monitor population health, and a proxy for lead exposure and avian health without destructively sampling individuals. I then used presence-only maximum entropy (MaxENT) and multivariate statistical modeling procedures in conjunction with mottled duck movement data to elucidate sets of habitat conditions that were conducive to predicting the occurrence of mottled ducks and environmental lead “hot spots”. MaxENT analyses suggested that lead in the top portion of the soil column is similarly related to all environmental variables considered, may be increasingly available after large-scale environmental disturbances. Lack of variation in coarse-scale habitat use between breeding and non-breeding seasons may further point to a food-based exposure pathway for lead as mottled ducks switch from an invertebrate to plant diet, either as a result of changing age classes or normal adult phenology, during the period of increased lead exposure. Using stable isotope ratio analysis, I then tested environmental samples of soil and vegetation as well as mottled duck blood to determine isotopic signatures that were consistent with particular sources of lead deposition (e.g., lead shot pellets, leaded fossil fuel combustion, industrial effluents). Comparisons suggested a great deal of similarity to lead shot reference values in vegetation and blood samples, especially in blood samples with higher concentrations of lead present. Last, I conducted a formal Ecological Risk Assessment (ERA) procedure to quantify the risk to mottled ducks from lead exposure in their current habitat and direct managers towards effective mitigation and habitat management strategies to reduce exposure in the future. One scenario suggested that mottled ducks were at greatest risk from eating an invertebrate-based diet, but lead content values at the TCPC suggest that a plant-based diet may provide a higher lead exposure risk for mottled ducks, depending on true levels of bioavailability in environmental media. Overall, I determined that mottled ducks experience greatest lead exposure risk from lead shot pellets on the TCPC or in nearby habitat, while potentially also experiencing low levels of exposure from several other sources. Additionally, management efforts that focus on plants that do not provide food resources for mottled ducks as a potential environmental sink for lead contamination, such as phytoremediation, may prove effective in reducing the overall lead load from historical activities that likely deposited much of the lead in this ecosystem.

Amphibians are regarded as sensitive indicators of environmental change and are therefore excellent subjects for use in ecotoxicology. The Phongolo River floodplain is South Africa’s most diverse natural floodplain system and hosts more than 40 frog species. It is also a malaria endemic region and is subjected to active spraying with Dichlorodiphenyltrichloroethane (DDT) through means of indoor residual spraying over the summer months. The upper Phongolo River runs through agricultural landscape and is subjected to runoff from forest plantations, orchards and sugar cane plantations. In this study residue levels of 22 different organochlorine pesticides (OCPs) were analysed in selected amphibian species from in and around the Ndumo Nature Reserve coupled with 12 different biomarker response assays to determine environmental exposure levels and possible sublethal effects in amphibians from the lower Phongolo River floodplain. Seasonal change, direct influence of anthropogenic activity and the influence of species’ aquatic preference in habitat selection were all factors considered during this assessment. Stable Isotope analyses were performed on 11 different food web components In order to determine the food web structure pertaining to Xenopus muelleri (Müller's platanna). Samples were collected during both high and low flow seasons from inside and outside Ndumo Nature Reserve. Organochlorine pesticide bioaccumulation was analysed in whole frog samples using a GC-μECD. Results indicated significant seasonal variation in OCP levels and exposure composition. Significant differences between inside and outside sites were also noted. Dichlorodiphenyltrichloroethane in its different isomer forms and their metabolites along with the hexachlorocyclohexane (HCH) isomers was the two main contributing OCP groups detected. Total OCP levels from all sample sets ranged between 8.71 ng/g lipid and 21,399.03 ng/g lipid. An increase in OCP accumulation was observed for X. muelleri over a period of one year. Organochlorine pesticides are known to have neurotoxic effects causing imbalances in Na+, K+, and Ca+ ion exchange. Hyperactivity has been reported in Rana temporaria (European Common frog) tadpoles exposed to p,p-DDT concentrations above 110,000 ng/g lipid. Despite OCP levels measured in frogs from this study being lower than reported toxic levels, the biomarker response assays indicated definite oxidative stress responses correlating to OCP bioaccumulation, with other minor responses shown. Cellular energy allocation showed a shift in the main energy source type from proteins to lipids correlating to increased OCP bioaccumulation. A slight inhibition response was noted in the hepato-somatic index correlating to γ-HCH bioaccumulation. Stable isotope analyses indicated food web structure differences between inside and outside the reserve, with outside showing less clear distinction between trophic groups and nitrogen enrichment of primary producers. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

Amphibians are regarded as sensitive indicators of environmental change and are therefore excellent subjects for use in ecotoxicology. The Phongolo River floodplain is South Africa’s most diverse natural floodplain system and hosts more than 40 frog species. It is also a malaria endemic region and is subjected to active spraying with Dichlorodiphenyltrichloroethane (DDT) through means of indoor residual spraying over the summer months. The upper Phongolo River runs through agricultural landscape and is subjected to runoff from forest plantations, orchards and sugar cane plantations. In this study residue levels of 22 different organochlorine pesticides (OCPs) were analysed in selected amphibian species from in and around the Ndumo Nature Reserve coupled with 12 different biomarker response assays to determine environmental exposure levels and possible sublethal effects in amphibians from the lower Phongolo River floodplain. Seasonal change, direct influence of anthropogenic activity and the influence of species’ aquatic preference in habitat selection were all factors considered during this assessment. Stable Isotope analyses were performed on 11 different food web components In order to determine the food web structure pertaining to Xenopus muelleri (Müller's platanna). Samples were collected during both high and low flow seasons from inside and outside Ndumo Nature Reserve. Organochlorine pesticide bioaccumulation was analysed in whole frog samples using a GC-μECD. Results indicated significant seasonal variation in OCP levels and exposure composition. Significant differences between inside and outside sites were also noted. Dichlorodiphenyltrichloroethane in its different isomer forms and their metabolites along with the hexachlorocyclohexane (HCH) isomers was the two main contributing OCP groups detected. Total OCP levels from all sample sets ranged between 8.71 ng/g lipid and 21,399.03 ng/g lipid. An increase in OCP accumulation was observed for X. muelleri over a period of one year. Organochlorine pesticides are known to have neurotoxic effects causing imbalances in Na+, K+, and Ca+ ion exchange. Hyperactivity has been reported in Rana temporaria (European Common frog) tadpoles exposed to p,p-DDT concentrations above 110,000 ng/g lipid. Despite OCP levels measured in frogs from this study being lower than reported toxic levels, the biomarker response assays indicated definite oxidative stress responses correlating to OCP bioaccumulation, with other minor responses shown. Cellular energy allocation showed a shift in the main energy source type from proteins to lipids correlating to increased OCP bioaccumulation. A slight inhibition response was noted in the hepato-somatic index correlating to γ-HCH bioaccumulation. Stable isotope analyses indicated food web structure differences between inside and outside the reserve, with outside showing less clear distinction between trophic groups and nitrogen enrichment of primary producers. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

This paper deals with isotope analyses of bones and teeth of two individuals from a medieval cemetery in Västerhus, Jämtland with the purpose to ascertain the diet and how much fish the people ate. The stable isotopes of carbon and nitrogen have been analysed with isotope-ratio mass sepectrometry and the diet of the people in Västerhus has been studied. The results show that most of the people had a terrestrial diet and by comparing with Sigtuna, Uppland and Varnhem, Västergötland it was possible to conclude that they ate meat and vegetable feed.

Food web structure is of underlying importance to ecological functions and processes. Whilst it is understood that a range of biotic and abiotic factors affect structure, relatively little is known of the role of biodiversity per se in structuring food webs. In this thesis I utilise novel multi-dimensional estimates of food web structure based on stable isotope ratios of nitrogen (δ15N) and carbon (δ13C) to quantify structural responses to changing community diversity. I additionally investigate methodological aspects of sample preparation and stable isotope quantifications of food chains. Using an arthropod prey-predator system, in chapter 2 I demonstrate that tissue selection and lipid extraction are important methodological procedures for deriving accurate δ15N and δ13C signatures. In chapter 3 I test the utility of δ15N to quantify food chain length, and δ13C to trace primary energy sources through to end consumers. Bayesian resampling of variance in sample means for plant and arthropod food chains produces robust isotopic estimates that match known food chain length well despite some error variance, and estimates of δ13C-range that trace trophic transfers. Chapter 4 represents a change in system from lab to field as I determine δ15N and δ13C signatures for plant and invertebrate species within three grassland communities representing a gradient of biodiversity. Quantifications of community bivariate isotopic space using isotopic metrics revealed that greater taxonomic richness increased both diversity of resource space exploited and overlap in resource space. These results therefore suggest that loss of diversity affected structure through altering relative patterns of niche partitioning in resource exploitation amongst community members. In chapter 5, I additionally find evidence that grassland management mediated changes in food web compartmental structure that were associated with differences in generalist invertebrate predator feeding habits. Taken together, these findings develop and demonstrate the utility of isotopic approaches to quantifying food web structure, and provide evidence of important mechanisms by which biodiversity affects food web structure. I conclude that the preservation of natural food web structure and trophic dynamics are further reasons for halting loss of biodiversity.

Nitrogen (N) is an essential element to plants for growth, maintenance and reproduction, however most N does not exist in a form that is biologically available to plants. In order to maximise the acquisition and retention of N, plants have evolved a variety of morphological and physiological adaptations and life history strategies, as well as the ability to respond plastically to changes in resource availability in ecological time. Determining the ecophysiological responses of plants to changes in root N availability is crucial to further understanding of the mechanisms underlying competitive interactions between plants, and between plants and other organisms, that ultimately contribute to community structure and ecosystem functioning. Carnivorous plants are ideal systems for investigating ecophysiological responses to N availability as:- (i) they share a unique adaptation for obtaining supplemental N from captured prey, therefore ecological stoichiometry and energetic cost/benefit models may be explored; (ii) the trait of botanical carnivory is widely considered to have independently co-evolved as a response to N-deficient, sunny and wet environments, therefore resource allocation trade-offs between plant investment in N and carbon (C) acquisition may be observed, and (iii) they are extremely sensitive to changes in root N availability in ecological time. In this research, the carnivorous plant Drosera rotundifolia (round-leaved sundew) was used to address several unanswered ecophysiological and evolutionary questions relating to patterns and processes of prey capture and the N nutrition of carnivorous plants. Furthermore, the potential for reducing uncertainty in the calculation of plant reliance on carnivory using a δ15N natural abundance multi-level linear mixing model was explored. A combined approach of in-situ and ex-situ studies was employed, using co-occurring non-carnivorous plants or carnivorous plant species with differing evolutionary lineages or prey capture mechanisms respectively to provide context. Results show that the adaptations of carnivory, high reproductive investment and a relatively short life span enable Drosera rotundifolia to survive and thrive in an extreme, N deficient environment. Phenotypically plastic responses by the plant to light and root N availability provide evidence of resource allocation trade-offs between investment in carnivory for N acquisition and in photosynthesis for C acquisition. Plants invested less heavily in prey capture (measured as the stickiness of leaf mucilage) as N availability increased or light availability decreased. These results show that the energetic costs associated with carnivory are avoided by the plant when less costly sources of N are available for uptake and that the production of carbon-rich mucilage is only made under nutrient-limited and well-lit conditions. Results obtained from the comparison of captured insect prey with background invertebrates of potential prey indicate that Drosera rotundifolia is a dietary generalist, where the quantity of prey captured per plant is positively correlated with leaf stickiness and total leaf area. Plant reliance on prey-derived N decreased with increasing root N availability, providing evidence that carnivory is only of net benefit to the plant in N-deficient and well-lit environments, as the photosynthetic costs of investment in the trait are not exceeded by the energetic gain from prey N uptake in shady or dry habitats. A more accurate and precise method for calculating plant reliance on botanical carnivory is presented which incorporates the insect diet of the plant. This method has wider significance for reducing uncertainty in the calculation of relative source contributions to a mixture for most natural abundance applications using a multi-level linear mixing model. To conclude, results from this research further understanding of the ecophysiological mechanisms underlying plant responses to changes in resource availability and the selective pressures driving the evolution of plant adaptations. These results therefore assist with predicting how plants and plant communities may respond to sustained N deposition inputs and future environmental scenarios.