Thesis (Ph. D.)--University of Wisconsin--Madison, 1960. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1951. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 67-70).

Sun-compass orientation tests were performed on adults and larvae of the spring peeper (Hyla crucifer) and the western chorus frog (Pseudacris triseriata triseriata). All animals were tested outdoors in a circular test pool from which all visible landmarks were obscured. Response on the Y-axis varied with the life history stage, and significant interspecific differences in sun-compass orientation behavior were noted. Statistical analysis was performed with Batschelet's V-test.Adult frogs were captured as they migrated to the breeding pond and tested under sunny skies. Chorus frogs oriented on the deep-water Y-axis of the nearest shoreline (n=30, p <0.005) but spring peepers did not orient (n=42, p >0.10).Frogs were also captured in the breeding pond at weekly intervals throughout the breeding season and tested under sunny skies. Chorus frogs oriented toward shore on the Y-axis (n=45, p <0.005); spring peepers did not orient (n=96, p >0.10). Adult chorus frogs kept in outdoor enclosures for a month after the breeding season were still oriented toward shore on the Y-axis (n=6, p <0.05). Chorus frogs tested under sunny skies after ten days of constant temperature, dim-light conditions in the laboratory did not orient (n=10, p >0.10).Adult chorus frogs captured as they left the breeding pond and tested under sunny skies did not orient on the Y-axis of the nearest shoreline (n=33, p >0.10), and neither did spring peepers (n=38, p >0.10).Larval western chorus frogs raised in outdoor enclosures oriented toward deep water on the Y-axis within five days after hatching (n=29, p< 0.005) and during late premetamorphosis (n=33, p <0.001). Chorus frog larvae did not orient during mid-premetamorphosis (n=32, p >0.10). Newly metamorphosed juveniles oriented on the Y-axis toward shore within a week after emergence (n=25, p< 0.01).Larval spring peepers oriented on the Y-axis toward shore within 18 days after hatching (n=37, p <0.0001) and also late in metamorphosis (n=29, p< 0.0001). Newly metamorphosed juveniles were not tested.

Magister Scientiae (Biodiversity and Conservation Biology) - MSc (Biodiv and Cons Biol) / Studies on snake competitive interactions have relatively been well documented globally, however, those examples tend to be dominated by non-African examples. Africa has a large and spectacular reptile diversity and yet robust and empirical studies on snake population ecology remain poorly understood or documented. Given the close phylogenetic relationship between the two species, as well as the remarkable similarities in overall appearance, morphology, reproductive biology, and most importantly geographic distribution, Psammophis crucifer and Psammophylax rhombeatus offered an ideal study system in which to ask questions related to interspecific competition and niche partitioning.

This thesis investigates two aspects of phytoalexin metabolism by the phytopathogenic fungus <i>Sclerotinia sclerotiorum</i> (Lib) de Bary: (i) determination of detoxification pathways of structurally different molecules; (ii) design and synthesis of potential inhibitors of enzyme(s) involved in detoxification steps.<p>First, the transformations of important cruciferous phytoalexins by the economically important stem rot fungus, <i>S. sclerotiorum</i>, were investigated. During these studies a number of new metabolic products were isolated, their chemical structures were determined using spectroscopic techniques, and further confirmed by synthesis. The metabolic products did not show detectable antifungal activity against <i>S. sclerotiorum </i> which indicated that these metabolic transformations were detoxification processes. Overall, the results of these transformations suggested that <i>S. sclerotiorum</i> produces various enzymes that can detoxify cruciferous phytoalexins via different pathways. While the detoxifications of strongly and moderately antifungal phytoalexins such as brassilexin, sinalexin, and 1-methoxybrassinin were fast and led to glucosylated products, the transformations of the weakly antifungal phytoalexins brassicanal A, spirobrassinin and 1-methoxyspirobrassinin were very slow and yielded non-glucosylated compounds.<p>Next, the design of potentially selective inhibitors of the brassinin detoxification enzyme, BGT, was sought. Two sets of potential inhibitors of BGT were designed: (i) a group was based on the structure of brassinin, where the indole ring of brassinin was replaced with benzofuran, thianaphthene, 7-azaindole and pyrazolo[1,5-a]pyridine and/or the position of side chain was changed from C-3 to C-2; and (ii) another group based on the structure of camalexin where the thiazole ring of camalexin was replaced with a phenyl group. The syntheses and chemical characterization of these potential detoxification inhibitors, along with their antifungal activity, as well as screening using fungal cultures and cell-free extracts of <i>S. sclerotiorum</i>, were examined. The results of these screening indicated that 3-phenylindoles, 3-phenylbenzofuran, 5-fluorocamalexin, methyl (indol-2-yl)methyl-dithiocarbamate, methyl (benzofuran-3-yl)methyldithiocarbamate and methyl (benzo-furan-2-yl)methyldithiocarbamate could slow down the rate of detoxification of brassinin in fungal cultures and also in cell-free extracts of <i>S. sclerotiorum</i>. Among the designed compounds, 3-phenylindole appeared to be the best inhibitor both in fungal cultures and in cell-free extracts. Metabolism studies of all the designed compounds using fungal cultures of <i>S. sclerotiorum</i> indicated that they were metabolized by <i>S. sclerotiorum</i> to glucosyl derivatives, although at much slower rates.<p>It is concluded that some inhibitors that can slow down the rate of metabolism of brassinin could be good leading structures to design more active inhibitors of BGT.

This thesis investigates two aspects of phytoalexin metabolism by the phytopathogenic fungus <i>Sclerotinia sclerotiorum</i> (Lib) de Bary: (i) determination of detoxification pathways of structurally different molecules; (ii) design and synthesis of potential inhibitors of enzyme(s) involved in detoxification steps.<p>First, the transformations of important cruciferous phytoalexins by the economically important stem rot fungus, <i>S. sclerotiorum</i>, were investigated. During these studies a number of new metabolic products were isolated, their chemical structures were determined using spectroscopic techniques, and further confirmed by synthesis. The metabolic products did not show detectable antifungal activity against <i>S. sclerotiorum </i> which indicated that these metabolic transformations were detoxification processes. Overall, the results of these transformations suggested that <i>S. sclerotiorum</i> produces various enzymes that can detoxify cruciferous phytoalexins via different pathways. While the detoxifications of strongly and moderately antifungal phytoalexins such as brassilexin, sinalexin, and 1-methoxybrassinin were fast and led to glucosylated products, the transformations of the weakly antifungal phytoalexins brassicanal A, spirobrassinin and 1-methoxyspirobrassinin were very slow and yielded non-glucosylated compounds.<p>Next, the design of potentially selective inhibitors of the brassinin detoxification enzyme, BGT, was sought. Two sets of potential inhibitors of BGT were designed: (i) a group was based on the structure of brassinin, where the indole ring of brassinin was replaced with benzofuran, thianaphthene, 7-azaindole and pyrazolo[1,5-a]pyridine and/or the position of side chain was changed from C-3 to C-2; and (ii) another group based on the structure of camalexin where the thiazole ring of camalexin was replaced with a phenyl group. The syntheses and chemical characterization of these potential detoxification inhibitors, along with their antifungal activity, as well as screening using fungal cultures and cell-free extracts of <i>S. sclerotiorum</i>, were examined. The results of these screening indicated that 3-phenylindoles, 3-phenylbenzofuran, 5-fluorocamalexin, methyl (indol-2-yl)methyl-dithiocarbamate, methyl (benzofuran-3-yl)methyldithiocarbamate and methyl (benzo-furan-2-yl)methyldithiocarbamate could slow down the rate of detoxification of brassinin in fungal cultures and also in cell-free extracts of <i>S. sclerotiorum</i>. Among the designed compounds, 3-phenylindole appeared to be the best inhibitor both in fungal cultures and in cell-free extracts. Metabolism studies of all the designed compounds using fungal cultures of <i>S. sclerotiorum</i> indicated that they were metabolized by <i>S. sclerotiorum</i> to glucosyl derivatives, although at much slower rates.<p>It is concluded that some inhibitors that can slow down the rate of metabolism of brassinin could be good leading structures to design more active inhibitors of BGT.

This research investigated the phenology of the striped flea beetle (Phyllotreta striolata (Fabricius) (SFB) and crucifer flea beetle (Phyllotreta cruciferae (Goeze)) (CFB), and examined the efficacy and the persistence of biological activity of the reduced-risk insecticides cyantraniliprole, clothianidin, and thiamethoxam applied as seed treatments to spring canola. Yellow sticky cards were placed around field perimeters in order to assess if CFB and SFB have synchronous phenology. It was found that CFB and SFB have largely asynchronous populations in southern Ontario. In laboratory bioassays, while flea beetle control with cyantraniliprole was comparable to that provided by thiamethoxam or clothianidin at the cotyledon stage, control did not persist through the 1st and 2nd leaf stages. Thiamethoxam and clothianidin both provided moderate control of flea beetles in both 1st and 2nd leaves. Field studies suggested that cyantraniliprole may offer control of flea beetles comparable to that provided by thiamethoxam and clothianidin. / E.I. DuPont Canada Co.

Masters of Science / Studies on snake competitive interactions have relatively been well documented globally, however, those examples tend to be dominated by non-African examples. Africa has a large and spectacular reptile diversity and yet robust and empirical studies on snake population ecology remain poorly understood or documented. Given the close phylogenetic relationship between the two species, as well as the remarkable similarities in overall appearance, morphology, reproductive biology, and most importantly geographic distribution, Psammophis crucifer and Psammophylax rhombeatus offered an ideal study system in which to ask questions related to interspecific competition and niche partitioning. Specifically I asked (1) whether broad scale geographic sympatry is facilitated by fine-scale allopatry through separation of space-use, and (2) whether the diets of the two species provided evidence of partitioning along the dietary niche. To answer these questions, I first quantified relative abundance of the two species at a fine scale, and secondly used existing dietary data to quantify dietary niche overlap. Even though P. rhombeatus was always more abundant in my sample, I found no evidence of space-use partitioning in this study, instead it showed a positive correlation in their abundance, and therefore suggesting space was not a limiting resource. Pianka niche overlap analysis showed significant differences in their feeding habits whereby P. rhombeatus had a broader diet which included mammals and birds, whereas P. crucifer predominantly fed on lizards and other snakes. In conclusion, my study suggests that across multiple geographic scales these two snakes use the same spatial resources and are able to co-exist by partitioning food resources. Lastly, my study serves to provoke more African studies of this nature with suitable candidate snake species.

Communities are a critical link that impact how species-level population dynamics translate into ecosystem functions, and thus, understanding community assembly is an important goal of ecology. Variation in the relative importance of the four processes of drift, selection, speciation, and dispersal likely govern much of the variation that is observed in community structure across landscapes. Microbial communities provide critical functions across an array of environments, but only recently have technological advances in DNA sequencing allowed us to study these communities with higher resolution. My dissertation research has investigated community assembly in host-associated microbial communities, with a focus on understanding how stochasticity in dispersal that leads to priority affects can impact bacterial community assembly in amphibian embryos. In chapter 1, I experimentally show that priority effects resulting from stochastic dispersal can be observed in the microbiome of newly-hatched hourglass treefrog (Dendropsophus ebraccatus) tadpoles. Changes in microbiome composition due to priority effects could be observed in a simple two bacteria system and when the inoculation by the initial bacteria is followed by a more diverse community inoculum. Outcomes of my two taxa system in co-culture do not strictly mirror those observed in treefrog embryos, highlighting that priority effect outcomes are context dependent. Additionally, these results provide support that priority effects do not benefit all bacterial species equally and the magnitude of these effects will be dependent on the traits of individual colonists. In chapter 2 I demonstrate that priority effects are not unique to the hourglass treefrog system but can be observed in spring peeper (Pseudacris crucifer) tadpoles as well. This study demonstrates the applicability of priority effects in increasing the abundance of target probiotic taxa; a benefit to amphibian populations facing threats by a lethal fungal pathogen. By treating embryos with a priority inoculation of Janthinobacterium lividum, a bacterial species known to inhibit fungal pathogen growth, I increased the relative abundance of J. lividum on newly hatched tadpoles. I also provide evidence that closely-related species of bacteria can effectively co-exist regardless of priority inoculation. An understanding of variation in the amphibian microbiome across life stages in the wild is required to better understand the long-term impacts of priority effects in embryos. My final chapter, therefore, examined compositional changes in the microbiomes of locally occurring amphibians in Virginia across the egg, tadpole, and juvenile developmental stages. In this study, I show characterize the initial egg microbiome across amphibian species and demonstrate that egg microbiomes, are distinct between species but are more similar across species than tadpole or juvenile microbiomes. Additionally, I show that minor differences in host environment can lead to differences in the microbiome structure of conspecific tadpoles. Overall, my dissertation empirically demonstrates the role of dispersal, and more specifically priority effects, in the assembly of the vertebrate microbiome. / Doctor of Philosophy / An ecological community is a set of species that occur at a given site. Communities have been a fundamental focus of ecological research, as communities serve to link the population dynamics of individual species to ecosystem level processes provided by species. Microbial communities, in particular, are of interest because of the wide range of important functions they provide across a variety of systems, yet relatively little is known about how these communities initially come together and are maintained. This is particularly true for the microbial communities that live in and on plants and animals, which are called "host-associated" communities. Host-associated microbial communities contribute many important functions to their hosts, including guiding host development, assisting with nutrient assimilation, and providing disease resistance. Four processes are thought to govern how ecological communities assemble across landscapes at local sites or habitat patches: selection, dispersal, speciation, and drift. Variation in the relative importance of these processes is thought to drive the variation in community composition across sites, or in the case of host-associated microbial communities, across hosts. Selection occurs at a local level when environmental variables or the presence of other species impact where a species occurs. Dispersal of individuals among habitat patches can also impact what species occur at a local site, and speciation gives rise to new species in communities over time. Drift is the stochastic, or random, element of species abundance that is driven by variation in the birth and death rates of a population at a site. I have investigated the assembly of host-associated microbial communities using amphibians as a study system. In chapter 1, I experimentally demonstrate that stochasticity in dispersal that impacts which species arrive first to a site (priority effects) can be observed in the host-associated bacterial communities of newly-hatched treefrog (Dendropsophus ebraccatus) tadpoles. This can be observed in a simplified system where only two bacterial species are used, and also when a single bacterial species arrives and is followed by a more diverse community of bacteria. However, not every bacterial species is able to take advantage of priority, and these results seem to be context dependent, as the outcomes in treefrog embryos do not exactly mirror the outcomes when the bacteria are grown in a nutrient broth together. In chapter 2, I show that priority effects are not unique to the hourglass treefrog system; priority effects can also be observed in spring peeper (Pseudacris crucifer) tadpoles. In this study, I also demonstrated that we may be able to apply our knowledge of priority effects to benefit amphibian populations threatened by a potentially lethal fungal pathogen by manipulating the abundances of bacteria on the skin during development. Priority treatment of embryos with Janthinobacterium lividum, a bacterial species known for its ability to inhibit growth of this fungal pathogen, resulted in increased relative abundance of J. lividum in the tadpoles following hatching. Additionally, I found that even closely-related bacterial species can have differing abilities to take advantage of priority effects and can co-exist on tadpoles. To determine long-term impacts of priority effects in embryos requires an understanding of the variation associated with amphibians in the wild across different life stages. My final chapter, therefore, focused on examining changes in the bacterial communities associated with locally occurring amphibians in Virginia across the egg, tadpole, and juvenile stages of development. Specifically, I characterize the initial communities associated with eggs across different species, including predicted associations with algal symbionts, and examine patterns of host-associated communities among species and across development. Overall, my dissertation showcases the role that dispersal, but more specifically priority effects, can play in the development of the vertebrate microbiome.