Nutrition

We studied the physiological and molecular responses of lab-grown Daphnia pulex to shifts in dietary (carbon, nitrogen, phosphorus, and cyanobacteria) and growth media (calcium) nutrient supply using a set of potential biomarkers via qPCR. Each examined nutrient had a strong effect on Daphnia mass-specific growth rate, and we found significant dose-dependent effects of treatment level (medium and low) on the gene expression of selected indicator genes. Furthermore, linear discriminant analysis models using different combinations of treatment levels could separate the animals between nutritional treatments with 86-100% prediction accuracy. This would suggest that Daphnia appear able to respond to nutrient limitation by adjusting their growth rate and associated molecular pathways to deal with an insufficient supply of nutrients. While this study provides valuable information regarding Daphnia's ability to adjust physiological and molecular processes under controlled laboratory conditions, more validation needs to be conducted before applying these potential biomarkers to wild populations to assess the type and intensity of nutritional stress.

Due to climate change and managed care, birds may face interacting effects of novel diets and environmental temperatures. I examined how captive zebra finches (Taeniopygia guttata) responded to a 6-month acclimation to either thermoneutral or cool temperatures when provided with saturated or unsaturated fatty acid enriched diets. Body mass was lower on the unsaturated fatty acid diet, but only in cool conditions. Individuals in cool temperatures and saturated fatty acid enriched diets had heavier organs, but pectoralis mass and metabolic rate did not display an effect of diet. While birds on each treatments illustrated similar initial increases in plasma fatty acid unsaturation, composition and concentration of constituent fatty acid varied based on their diet and temperature. Individuals on the unsaturated fatty acid enriched diet and cool treatments had lower survivorship. Collectively, my research will allow researchers to better predict how birds will be impacted by shifting or novel environments.

Author Keywords: chronic acclimation, fatty acids, metabolic rate, Taeniopygia guttata, temperature, unsaturation

Organismal nutrition lies at the interface between biotic and abiotic factors in an ecosystem, dictating the transfer of energy and nutrients across trophic levels. Our ability to detect nutritional limitation in consumers is reliant on a priori knowledge of dietary history due to our inability to differentiate nutrient stress based on body-wide responses. Molecular physiological responses are increasingly being used to measure physiological stress with high levels of specificity due to the specific modes of action ecological stressors have on organismal molecular physiology. Because animal consumers respond to varying nutrient supplies by up- and down-regulating nutrient-specific metabolic pathways, we can quantify nutritional status by quantifying the expression of those pathways. Here I present an investigation into the use of transcriptomics to detect nutritional stress in the keystone aquatic herbivore, Daphnia pulex, I use RNAseq and quantitative PCR (qPCR) identify nutritional indicator genes. I found that nutritional status could be determined with 100% accuracy with just ten genes. Additionally, the functional annotation of those genes uncovered previously unidentified responses to dietary stress. Further testing and validation of the selected indicator genes is required however these findings have the potential to revolutionize our ability to measure and monitor consumer nutritional stress.

Author Keywords: Biomarkers, Daphnia, Gene expression, Nutrigenomics, Nutritional ecology, RNAseq

Aquatic consumers frequently face nutritional limitation, caused in part, by imbalances between the nutrients supplied by primary producers and the metabolic demands of the consumers. These nutritional imbalances alter many ecological processes including consumer life-history traits, population dynamics, and food web properties. Given the important ecological role of organismal nutrition, there is a need to have precise and specific indicators of nutritional stress in animals. Despite this need, current methods used to study nutrition are unable to distinguish between different types of nutritional limitation. Here I studied nutritional metabolism in the freshwater zooplankter, Daphnia. A greater understanding of nutritional metabolism would allow for the development of dietary bio-indicators that could improve the study of the nutritional ecology of animal consumers. Specifically, I addressed the question: What affects the biochemical composition of a generalist aquatic consumer? My overall hypothesis was that the quantity and quality of the diet affects the biochemical composition in a nutrient specific manner. To test this hypothesis, I examined various response variables involved in nutrient metabolism such as alkaline phosphatase activity, whole metabolome, and free amino acid composition. For each response variable, I grew Daphnia under various nutritional stressors and determined if responses are nutrient specific or are a general stress response. I found the current method of measuring alkaline phosphatase was not a phosphorus specific indicator, as activity increased in all nutrient stressed treatments. Analyzing the whole metabolome resulted in nutritional stressors being separated in multivariate space, with many identified metabolites being significantly different from nutrient rich Daphnia. Upon further examination the daphnids free amino acids profiles are caused by differences between the supply of amino acids from the algae and the demand within the Daphnia. These differences in supply and demand resulted in the ability to classify the nutritional status of Daphnia with the use of discriminant analysis, a classification multivariate model. In addition to a deeper understanding and advanced knowledge of the physiological changes caused by nutrient limitation, this research has provided strong evidence for the application of nutritional biomarkers/profiles to identified the nutritional status of Daphnia.

Author Keywords: Bio-indictor, Ecological stoichiometry, Metabolism, Nutritional limitation, Nutritional status

Environmental variation can affect consumer trait expression and alter ecological and evolutionary dynamics in natural populations. However, although dietary nutrient content can vary by an order of magnitude in natural ecosystems, intra-specific differences in consumer responses to food quality have not been thoroughly investigated. Therefore, the purpose of my dissertation was to examine the influence of dietary nutrition and other environmental factors on consumer phenotypic variation using the freshwater cladoceran Daphnia. I conducted a series of complementary laboratory and field studies where I examined the effects of dietary phosphorus (P) content and additional biological/environmental variables (multi-elemental limitation, genetic variation, and temperature) on daphnid life-history, biochemistry, body elemental composition, and population growth. In general, phenotypic expression within a species varied significantly in response to all experimental variables, but the relative influence of each was highly context dependent. In my first chapter, I found that dietary P content and environmental calcium (Ca) concentrations both altered Daphnia body Ca:P ratios and growth rates of individuals and affected intrinsic rates of increase at the population level. However, food quality appeared to have a much larger effect on trait expression, and body Ca:P ratios were highly sensitive to other forms of dietary nutrient limitation. Next, I documented significant quantitative genetic variation and phenotypic plasticity in daphnid P content, growth, and P use efficiency of field collected animals grown across dietary P gradients. Trait expression was also influenced by genotype X diet interactions suggesting that consumer responses to dietary nutrient limitation can be heritable and may be adaptive in different nutrient environments. Finally, I found that temperature appeared to override food quality effects and decouple P metabolism in natural Daphnia populations, but total biomass production was affected by both dietary P content and temperature, depending on the nutrient content of the lake. Overall, my dissertation shows that consumer responses to nutrient limitation can vary significantly within a species and that changes in trait expression may be modified by other environmental variables. These results should be incorporated into existing stoichiometric models and used to investigate the eco-evolutionary consequences of consumer phenotypic variation in response to nutritional stress.

Author Keywords: ecological stoichiometry, evolution, life-history, nutrient limitation, nutrient metabolism, zooplankton

This project is a pamphlet (A Food Sourcing Ethnography for ... the Seasoned Spoon cafe), which describes the Seasoned Spoon, and discusses the products available at this cafe.

The Seasoned Spoon Co-Operative Cafe (the Spoon) is a student run organization (at Trent University) which provides vegetarian and vegan food produced using ingredients that are as local and organic.