Population numbers of bumblebees has been on a steady decline leading to the endangerment of several species that are critical to the growth and pollination of crops and plants. Varying factors have contributed to this situation, one key factor being pathogens wiping out colonies. Studying pathogens that diminish bumblebee populations is vital to understanding how to combat the decline of these important species effectively. The aim of this study was to analyse five of the most common pathogens that exist in the abdomen of five different species of bumblebees. This was achieved by extracting DNA, from 176 previously acquired frozen samples, through homogenisation and varying washing steps before an analysis of the resulting nucleic acids. The analysis was done with quantitative polymerase chain reactions to quantify the amounts of each pathogen, as well as a passive reference nucleic acid, to then be able to calculate the absolute total -amount of pathogen carried by each individual bumblebee. The results showed that almost half of the bumblebees were infected by Nosema, one sixth were infected by Crithidia and Apicystis while only a small number of individuals had been infected by Bombus Densovirus and the Apis Mellifera Filamentous Virus. Bombus Lapponicus was the most infected species. The conclusion based on the results is that future research could put a focus on the more commonly found parasites Nosema, Crithidia and Apicystis. Additional research should also be undertaken into the factors contributing to the less common pathogens Bombus Densovirus and the Apis Mellifera Filamentous Virus.

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Lipids from the fat body of queens of Bombus terrestris species in different life stages were studied using chromatographic methods. High-performance liquid chromatography - mass spectrometry with electrospray ionization was used to analyze all lipids from the tissue. Semi- preparative thin-layer chromatography was used to isolate triacylglycerols (TGs) from the fat body tissue. The TGs were subsequently analyzed by high-performance liquid chromatography - atmospheric pressure chemical ionization mass spectrometry. Quantitative differences between different life stages have been found. Qualitative composition has not been changing significantly. Only minor differences have been found in the substances, which were present in amount less than 1%.

Bumble bees and other wild pollinators are crucial to the support of both natural and agricultural ecosystems. However, unprecedented declines of pollinator populations have been observed all over the world, raising concerns of a looming threat to both the human food supply, as well as sustainability of the biodiversity in local ecological niches. Though declines are well described, the cause behind these still evades scientists. Proposed contributors include anthropogenic-mediated environmental stress, including application of xenobiotics for pest control, and increase of pathogen diversity and abundance due to the shipment of infection human-managed colonies. This research examined these theories and attempted to quantify the threats they may pose. Through development of a chronic, oral toxicity experiment, susceptibility of all Bombus impatiens castes to clothianidin exposure was examined. This exposed a substantial increase in vulnerability of male bumble bees to realistic concentrations of neonicotinoid pesticides, highlighting the crucial need to examine all members of wild bumble bee life cycles before determining pesticide regulations. Additionally, sublethal effects on fitness-related foraging behaviors in Bombus impatiens were examined through development of a voluntary task switching assay. The results of this experiment suggest humoral immune stimulation, through pathogenic infection, leads to significant impairment of cognitive flexibility. Taken together, this data suggests that pesticides and pathogens are capable of causing severe detrimental effects, both lethally and sublethally, in wild bumble bees. I hope this data will eventually contribute to reassessment of environmental regulations and establishment of effective conservation strategies in order to sustain the critical populations of wild bumble bees.

Plants are under selection to simultaneously attract pollinators while deterring herbivores. This dilemma can lead to tradeoffs in floral traits, which are traditionally thought to be optimized for pollinators. My dissertation addresses the ecological costs and putative functional significance of nectar secondary metabolites, a paradoxical but widespread phenomenon in the angiosperms. I investigate this issue from the pollinator’s perspective using a series of controlled laboratory investigations focused primarily on the bumble bee Bombus impatiens and the nectar alkaloid gelsemine, from Gelsemium sempervirens. I begin by demonstrating that nectar enriched with the alkaloid gelsemine significantly deters visits from bumble bees at a range of natural alkaloid concentrations. However, this aversion can be mitigated by increasing the sucrose concentration such that the alkaloid-rich nectar is more rewarding than its alkaloid-free counterpart. I then demonstrate that the consumption of gelsemine-rich nectar can inhibit oocyte development and protein utilization in bees, but that this effect is limited to bees of suboptimal condition. Continuous consumption of the nectar alkaloid gelsemine also leads to a reduction in the pathogen load of bumble bees infected with Crithidia bombi, but direct interactions between the pathogen and the alkaloid have no impact on infection intensity. Gelsemine also fails to inhibit floral yeast growth, suggesting that nectar alkaloids may not be universally antimicrobial. Finally, I demonstrate that gross nectar cardenolides from the genus Asclepias are strongly correlated with gross leaf cardenolides and that the majority of individual cardenolides found in nectar are a subset of those identified in leaves. This pattern suggests that nectar cardenolides are a consequence of defense for Asclepias; however, they may not be a costly corollary because bumble bees show an overall preference for nectar cardenolides at mean concentrations. Altogether, my dissertation provides a new perspective on the role of chemical defenses against herbivores in plant-pollinator interactions.

Plants are under selection to simultaneously attract pollinators while deterring herbivores. This dilemma can lead to tradeoffs in floral traits, which are traditionally thought to be optimized for pollinators. My dissertation addresses the ecological costs and putative functional significance of nectar secondary metabolites, a paradoxical but widespread phenomenon in the angiosperms. I investigate this issue from the pollinator’s perspective using a series of controlled laboratory investigations focused primarily on the bumble bee Bombus impatiens and the nectar alkaloid gelsemine, from Gelsemium sempervirens. I begin by demonstrating that nectar enriched with the alkaloid gelsemine significantly deters visits from bumble bees at a range of natural alkaloid concentrations. However, this aversion can be mitigated by increasing the sucrose concentration such that the alkaloid-rich nectar is more rewarding than its alkaloid-free counterpart. I then demonstrate that the consumption of gelsemine-rich nectar can inhibit oocyte development and protein utilization in bees, but that this effect is limited to bees of suboptimal condition. Continuous consumption of the nectar alkaloid gelsemine also leads to a reduction in the pathogen load of bumble bees infected with Crithidia bombi, but direct interactions between the pathogen and the alkaloid have no impact on infection intensity. Gelsemine also fails to inhibit floral yeast growth, suggesting that nectar alkaloids may not be universally antimicrobial. Finally, I demonstrate that gross nectar cardenolides from the genus Asclepias are strongly correlated with gross leaf cardenolides and that the majority of individual cardenolides found in nectar are a subset of those identified in leaves. This pattern suggests that nectar cardenolides are a consequence of defense for Asclepias; however, they may not be a costly corollary because bumble bees show an overall preference for nectar cardenolides at mean concentrations. Altogether, my dissertation provides a new perspective on the role of chemical defenses against herbivores in plant-pollinator interactions.

At certain stages of a bumblebee colony life cycle workers lay eggs. Not all workers reproduce, however, since many continue to forage and care for the nest. This leads to questions regarding what differentiates a reproductive worker from a non-reproductive one. It is hypothesized that a form of reproductive competition takes place, where the most behaviourally dominant worker becomes reproductively dominant. The behaviour of orphaned Bombus impatiens pairs was recorded and aggression, social interactions, egg-laying, and ovarian development were identified. Experiment 1 examined the association between aggression and egg-laying. Contrary to the hypothesis, the most aggressive worker did not lay more eggs. When the ovarian development of workers was manipulated and two workers with developed ovaries were paired (Experiment 3), they were more aggressive than pairs with discouraged ovarian development. This provides support for the supposition that aggression and reproduction are related, however, it is only partial support as worker pairs with encouraged ovarian development did not lay more eggs. Since aggression is believed to be only one part of behavioural dominance, Experiment 2 studied the association between social interactions and aggression and reproduction. Results showed that when two socially active bees were paired they were more aggressive than pairs including one or two socially inactive bumblebees. No significant difference in ovarian development between socially active pairs and socially inactive pairs was found. Brood presence was also predicted to affect reproductive control. Experiment 1 found egg-laying and aggression were more likely to co-occur in the absence of brood. Results from Experiment 2 supplemented the first experiment since the absence of brood increased rates of aggression and ovarian development in pairs. Whereas the results confirm aggression has a role in worker reproduction the findings also reveal that behavioural dominance does not equate to reproductive dominance under all conditions. The primary contributions of this thesis were the development of a method to distinguish behavioural dominance from reproductive dominance and determining their relationship under different environments (brood presence) and experimental manipulations (ovarian development). These contributions further define dominance in Bombus impatiens.

This dissertation explored the way in which bumblebees' visual system helps them discover their first flower. Previous studies found bees have unlearned preferences for parts of a flower, such as its colour and shape. The first study pitted two variables against each other: pattern type: sunburst or bull's eye, versus the location of the pattern: shapes appeared peripherally or centrally. We observed free-flying bees in a flight cage using Radio-Frequency Identification (RFID) tracking. The results show two distinct behavioural preferences: Pattern type predicts landing: bees prefer radial over concentric patterns, regardless of whether the radial pattern is on the perimeter or near the centre of the flower. Pattern location predicts exploration: bees were more likely to explore the inside of artificial flowers if the shapes were displayed near the centre of the flower, regardless of whether the pattern was radial or concentric. As part of the second component, we implemented a mathematical model aimed at explaining how bees come to prefer radial patterns, leafy backgrounds and symmetry. The model was based on unsupervised neural networks used to describe cognitive mechanisms. The results captured with the results of multiple behavioural experiments. The model suggests that bees choose computationally "cheaper" stimuli, those that contain less information. The third study tested the computational load hypothesis generated by the artificial neural networks. Visual properties of symmetry, and spatial frequency were tested. Studying free-flying bees in a flight cage using motion-sensitive video recordings, we found that bees preferred 4-axis symmetrical patterns in both low and high frequency displays.

Cattleya é um gênero Neotropical com 114 espécies, sendo mais da metade destas exclusivas do Brasil. Embora as espécies de Cattleya sejam de grande importância horticultural, o número de trabalhos relativos à polinização, sistema reprodutivo e comportamento dos polinizadores é bastante reduzido. No presente trabalho foram analisadas onze populações para quatro espécies, das quais três são ameaçadas (de acordo com os critérios da IUCN) e nativas de Cattleya do Rio Grande do Sul, Brasil. Apresentamse resultados de caracterização morfológica, fenologia floral, sistema reprodutivo, remoção e deposição de polinários, sucesso na formação de frutos e interações com os polinizadores para C. coccinea, C. intermedia, C. tigrina e C. purpurata. Foi documentada pela primeira vez a polinização de C. intermedia, C. purpurata e C. tigrina por quatro espécies de abelhas dos gêneros Xylocopa, Bombus e Epicharis. A visitação exclusiva por beija-flores foi corroborada para C. coccinea. De um modo geral, as espécies mostraram-se autocompatíveis, mas polinizadores dependentes. Verificou-se nas quatro espécies ausência de néctar e um sistema de polinização baseado no mecanismo por engodo. Encontrou-se que em todas as espécies estudadas a taxa de visitas é baixa e a frutificação em condições naturais é infrequente. Verificou-se nas quatro espécies um maior sucesso na formação de frutos através de polinização cruzada. / Cattleya Lindl. is a Neotropical genus that comprises 114 species with more than a half restricted to Brazil. In spite of the horticultural importance of the Cattleya species their pollination, reproductive system, and behaviour of its pollinators is poorly known. In this work were analized eleven populations of four species, of which three are endangered (according to the IUCN criteria) native species of Cattleya from Rio Grande do Sul, Brazil. Here are shown data and results of morphological caracterization, floral phenology, reproductive system, pollinary removal and deposition, fruit set success, and interactions with the pollinator for C. coccinea, C. intermedia, C. tigrina and C. purpurata. It was documented for first time the pollination of C. intermedia, C. purpurata and C. tigrina by four bee species from the genus Xylocopa, Bombus and Epicharis. The exclusive visitation from the hummingbird was confirmed for C. coccinea. All studied species are selfcompatible, but pollinator dependent. There were also verified a lack of reward, and a pollination system based on deceit for the four species. The findings show that in all the studied species the visitation ratio is low, and the natural fruiting has a low frequency. All studies species showed higher fruit production after cross-pollination.

Cattleya é um gênero Neotropical com 114 espécies, sendo mais da metade destas exclusivas do Brasil. Embora as espécies de Cattleya sejam de grande importância horticultural, o número de trabalhos relativos à polinização, sistema reprodutivo e comportamento dos polinizadores é bastante reduzido. No presente trabalho foram analisadas onze populações para quatro espécies, das quais três são ameaçadas (de acordo com os critérios da IUCN) e nativas de Cattleya do Rio Grande do Sul, Brasil. Apresentamse resultados de caracterização morfológica, fenologia floral, sistema reprodutivo, remoção e deposição de polinários, sucesso na formação de frutos e interações com os polinizadores para C. coccinea, C. intermedia, C. tigrina e C. purpurata. Foi documentada pela primeira vez a polinização de C. intermedia, C. purpurata e C. tigrina por quatro espécies de abelhas dos gêneros Xylocopa, Bombus e Epicharis. A visitação exclusiva por beija-flores foi corroborada para C. coccinea. De um modo geral, as espécies mostraram-se autocompatíveis, mas polinizadores dependentes. Verificou-se nas quatro espécies ausência de néctar e um sistema de polinização baseado no mecanismo por engodo. Encontrou-se que em todas as espécies estudadas a taxa de visitas é baixa e a frutificação em condições naturais é infrequente. Verificou-se nas quatro espécies um maior sucesso na formação de frutos através de polinização cruzada. / Cattleya Lindl. is a Neotropical genus that comprises 114 species with more than a half restricted to Brazil. In spite of the horticultural importance of the Cattleya species their pollination, reproductive system, and behaviour of its pollinators is poorly known. In this work were analized eleven populations of four species, of which three are endangered (according to the IUCN criteria) native species of Cattleya from Rio Grande do Sul, Brazil. Here are shown data and results of morphological caracterization, floral phenology, reproductive system, pollinary removal and deposition, fruit set success, and interactions with the pollinator for C. coccinea, C. intermedia, C. tigrina and C. purpurata. It was documented for first time the pollination of C. intermedia, C. purpurata and C. tigrina by four bee species from the genus Xylocopa, Bombus and Epicharis. The exclusive visitation from the hummingbird was confirmed for C. coccinea. All studied species are selfcompatible, but pollinator dependent. There were also verified a lack of reward, and a pollination system based on deceit for the four species. The findings show that in all the studied species the visitation ratio is low, and the natural fruiting has a low frequency. All studies species showed higher fruit production after cross-pollination.