Thesis (M.S.)--University of Missouri-Columbia, 2005. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file viewed on (January 10, 2007) Includes bibliographical references.

Thesis (Ph. D.)--Oregon State University, 2011. / Printout. Includes bibliographical references (leaves 85-90). Also available on the World Wide Web.

Thesis (Ph. D.)--University of Missouri-Columbia, 2007. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on September 25, 2007) Vita. Includes bibliographical references.

Anurans are declining worldwide because of the spread of Batachochytrium dendrobatidis (Bd), the fungus that causes chytridiomycosis. However, some frogs are resistant to this disease, and understanding why may be critical to saving those that are susceptible. In Peru, Gastrotheca excubitor is resistant to chytridiomycosis while Gastrotheca nebulanastes is susceptible. Two anuran skin defenses, symbiotic bacteria and antimicrobial peptides (AMPs), have demonstrated the ability to inhibit Bd in vitro when isolated from certain frogs. We tested if these defenses can explain the difference in susceptibility between the two Gastrotheca species. The cutaneous bacteria and AMPs of both species were collected, tested for their abilities to inhibit the growth of Bd, and analyzed for their compositions. Results indicate that 34%of the strains of skin bacteria from G. excubitor were able to inhibit the growth of Bd whereas only 10% isolated from G. nebulanastes were effective. Gastrotheca excubitor also has stronger anti-Bd skin bacteria. Neither frog species has peptide mixtures capable of completely inhibiting Bd, and overall species did not differ in the anti-Bd abilities of their peptides. These results suggest that the chytridiomycosis resistance experienced by G. excubitor may be attributed to its skin bacteria.

Made available in DSpace on 2014-06-11T19:22:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-08-09Bitstream added on 2014-06-13T20:28:40Z : No. of bitstreams: 1 teixeira_pc_me_jabo.pdf: 886510 bytes, checksum: 7e701e9c09d806beb2b0ac2590cbdfaa (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Estudar o sangue significa monitorar a saúde dos animais reduzindo, desta forma, as perdas por doenças ou injúrias, seja frente a um determinado tipo de estressor, por sistemas de criação não apropriados, por manejos inadequados ou ainda por alterações ambientais. O objetivo deste trabalho foi avaliar a resposta fisiológica de girinos de rã-touro (Rana catesbeiana) em diferentes densidades e após exposição aérea. O experimento foi realizado em duas etapas utilizando-se de animais entre os estágios 31 a 39, na fase de pró-metamorfose; na primeira etapa, realizada no CAUNESP - UNESP/SP, avaliou-se somente a exposição aérea nos tempos de 0, 15, 30,45,60 e 90 minutos. Na segunda etapa, realizada no Instituto de Pesca – APTA/ SP foi avaliada a densidade de estocagem sendo testados 1 girino/L (Tratamento I), 5 girinos/L (Tratamento 2) e 10 girinos/L (Tratamento 3), conduzidos em 3 réplicas simultâneas, e a exposição aérea nos dias 0,14(6 horas), 4, 8 e 12 dias. O sangue foi retirado por rompimento do vaso caudal na condição de Normóxia - N (tempo zero) e Hipóxia - H (tempo de 15 minutos de exposição ao ar). Avaliou¬se, na primeira etapa, somente o cortisol plasmático e na segunda etapa, cortisol plasmático, glicemia, hematócrito, hemoglobina, número de eritrócitos, os índices hematimétricos absolutos ¬VCM, HCM e CHCM, contagem diferencial de leucócitos e contagem total de leucócitos e trombócitos. Foi observado um aumento nos valores de cortisol, aos 4 e 8 dias de exposição aérea retomando aos valores basais ao final do experimento, apesar de não haver diferenças significativas. A glicemia foi influenciada no tratamento com 10 girinos/L mostrando forte correlação com a hipóxia. Os parâmetros hematológicos, principalmente, o número de linfócitos, neutrófilos e eosinófilos mostraram diferença significativa aos 12 dias de experimentação... / Studying blood means monitoring animals health reducing, losses by disease or injury caused by certain kinds of stress related to improper rearing systems, handling or alterations in the environment. The aim of this study was to evaluate bullfrog's tadpole physiological answer (Rana catesbeiana) to different densities and air exposition. The experiment was carried out in two parts using animals, at the pro-metamorphosis phase, on the first part carried out at CAUNESP - Aquaculture Center, São Paulo State University was evaluated only air exposition, which animals were sampled on zero, fifteen, thirty, forty five, sixty and ninety minutes and on second part, carried out at Fisheries Institute – APTA/SP was evaluated different densities and air exposition, which stocking densities was 1 tadpole/L (treatment one), 5 tadpoles/L (treatment two) and 10 tadpoleslL (treatment three) with three simultaneous replicates and animals were sampled on zero, 114 (six hours), four, eight and twelve experimental days. Blood was collected by breaking tail blood vessel at norrnoxic (zero time) and hipoxic (fifteen minutes by air exposition) conditions. Only cortisol was evaluate on the first part and on second, cortisol, glicemy, hematocryts, haemoglobin tax, couting of erytrocyte's number, absolute hematimetrycs indices -MCV, MCH and MCHC, differenctial couting of leucocytes, total counting of leucocytes and trombocytes were evaluated. An increase in plasmatic cortisol was observed at four and eight days of the experiment, retuming to initial values after 12 days. Glicemy values increased at treatrnent. three, showing strong correlation with hypoxia. Hematological parameters as linphocytes, neutrophils and eosinophils numbers at the 12th day of the experiment showed differences comparing to zero day; concluding that stocking densities until 10 tadpoles/L and air exposition ...(Complete abstract, click electronic access below)

Orientador: Marta Verardino de Stéfani / Banca: Claudia Maris Ferreira Mostério / Banca: Luciano Hauschild / Resumo: Melhorias no manejo alimentar devem ser implementadas na ranicultura, visando diminuir os efeitos indiretos da alimentação inadequada, resultando em melhores taxas de crescimento e deposição de nutrientes, consequentemente obtendo animais de melhor qualidade. O objetivo do presente estudo foi estabelecer curvas de crescimento morfométrico de girinos de rã-touro e sua deposição de nutrientes na carcaça. Foram utilizados 6.480 girinos de rã-touro no estágio 25 de Gosner, alimentados com dieta experimental (26,23% PD - 32,68 PB) e comercial (37,92% PB), oferecida "ad libitum. O modelo utilizado para descrever a curva de crescimento foi de Gompertz. Os girinos alimentados com a dieta experimental, além de apresentarem uma deposição protéica final maior, o modelo sigmoidal apresentou-se muito mais homogêneo, mostrando uma taxa de deposição protéica diária mais constante. O modelo de Gompertz apresentou um ótimo ajuste para descrição da curva de crescimento morfométrico e deposição de nutrientes na carcaça para girinos de rã-touro, mostrando que os girinos alimentados com a dieta experimental, apresentaram melhor taxa de crescimento e deposição de nutrientes na carcaça / Abstract: Improvements in food handling must be implemented in frog culture in order to reduce the indirect effects of inadequate nutrition, resulting in better rates of growth and deposition of nutrients, thus obtaining better quality animals. The objective of this study was to establish morphometric growth curves of bullfrog tadpoles and deposition of nutrients in the substrate. We used 6480 of bullfrog tadpoles in stage 25 of Gosner, fed experimental diets (DP 26,23% - 32,68 CP) and commercial (37,92% CP) offered "ad libitum. The model used to describe the curve of growth was the Gompertz. Tadpoles fed the experimental diet, in addition to having a higher final protein deposition, the sigmoidal model presented is much more homogeneous, showing a daily protein deposition rate constant. The Gompertz model showed a good fit for describing the growth curve morphometric and deposition of nutrients in the substrate for bullfrog tadpoles, showing that the tadpoles fed the experimental diet showed better growth rate and deposition of nutrients in the substrate / Mestre

In my thesis I was concerned with pectoral girdle muscular system in selected frog species, especially with the muscles of ventral side, which probably play a crucial role in frog jumping abilities (mainly in landing phase). I find out a lot of differences between particular species and I mean that some of those differences are straightly connected with jumping movements.

One of the biggest attractions of the tropics is its stunning diversity; the origin and determinants of this diversity are amongst the most important questions in evolution and biogeography. There are two ways of looking at diversity: diversity as observed in clades on the tree of life and diversity as distributed in space. Various processes drive patterns in clades leading to a gradient of evolutionary radiations. These radiations are amongst the major contributors to Earth's biological diversity and act as good model systems to test the relative significance of processes governing macroevolutionary patterns of diversification. My research presented here revolves around one such evolutionary radiation of frogs that we uncovered from the Western Ghats, a tropical mountain system in Peninsular India. The broad aim of my thesis is to understand the causes and consequences of diversification, the primary process that drives evolutionary radiations and diversity differences in clades. The ultimate goal is to use a model system in a regional setting to explore diversification process¬es in clades against the backdrop of existing theory and ultimately paint a broader picture to further our understanding of diversity patterns in space. I have organized this thesis into six chapters. The first chapter serves as an introduction to the concepts, models and problems addressed, while chapters two to five form the crux of this thesis, and the final chapter pro¬vides a synthesis of results and directions to carry this research forward. In the first chapter, I have provided a conceptual link between diversity in Glades, evolutionary radiations and diversification and its drivers, all of which form the backbone for the following chapters. I review the potential historical processes that acted independently or in combination to give rise to one of the global biodiversity hotspots, the Western Ghats of the Indian sub-continent. I also introduce the model system, bush frogs, and provide a brief overview of their current taxonomic and systematic status. Finally, I have described the goal and questions addressed in this thesis. In chapter 2, I address the problem of delimiting lineages and provide a base for the remaining chapters 3, 4 SE 5, which form the core of this research. In this chapter, I address two major issues in biogeography. namely the 'Linnean shortfall' (the problem of undescribed lineages) and the 'Wallacean shortfall' (the lack of distribution data), and use an extensive sampling regime and a phylogenetic framework to delimit lineages. When I began my sampling, 21 lineages had been described from the Western Ghats. Three years of sampling across 13 major massifs has led to an increase in the number of the lineages to 70. These lineages were delimited using a hierarchical multi-criteria approach using a haplotype phylogenetic tree, genetic distance, geography and morphology. Lineages recovered in this study range from shallow to deep divergences and are spatially distributed from lowlands to Massif summits. Further, inclusion of geography as a variable in the hierarchical approach increased efficacy in delimiting lineages. This approach was used to establish a working hypothesis of lineages for later chapters, where I address questions related to the drivers behind the diversification of these Lineages. The goal in chapter 3 was to examine the evolutionary and biogeographic processes underlying in-situ Glade diversification in the bush frog clade at broad temporal and spatial scales. First, I examined the Glade for characteristics of an evolutionary radiation. One of the fundamental challenges in characterizing evolutionary radiations is to discern independent colonization from in-situ diversification. Phylogenetic reconstructions shows that bush frog lineages in the Western Ghats belong to two clades: a larger Glade composed of 67 lineages, with their ancestral origin centered in the Western Ghats Escarpment, and a set of three lineages with their distribution in Western Ghats, but phylogenetically nested within the Sri La.nkan bush frog Glade. The larger Western Ghats clade recovered here lends strong support for in-situ radiation of bush frogs, the origin of which can be dated to Eocene - Oligocene boundary. The bush frogs are a relatively young radiation compared to recent reports of ancient lineages and potential radiations whose origins date to the Cretaceous. Within the Western Ghats, bush frogs make up the largest vertebrate radiation, and when considered with the sister clade in Sri Lanka, constitute the largest vertebrate radiation in South Asia. These clades attain greater significance because of the limited geographical space in which the whole radiation has occurred. Temporal patterns of line-age diversification in the Western Ghats bush frog Glade show strong signatures o f an early burst, considered as one of the characteristic features of evolutionary radiations, especially adaptive radiations. Considering that a similar pattern could arise from the initial formation of geographical isolates, I tested the Glade for the role of regional biogeographic barriers in the initial diversification of the Glade. Ancestral range reconstruction showed that the initial diversification of the bush frog Glade is coincident with the Palghat Gap, a prominent geographical break in the Escarpment. Two sister clades, North and South, could be discerned, with their respective centres of origin north and south of the Palghat Gap. There was limited dispersal across the gap and a number of these dispersal events resulted in diversification in the adjoining regions. These results, for the first time, lend strong evidence for the biogeographic significance of the Palghat gap for amphibian diversification. The discovery of the North and South sister-clades, apart from elucidating the significance of Palghat gap as a barrier in the initial diversification of the Glade, provided a unique setting to gain further insights into diversification processes. using the sister clades. The South Glade shows support for the models of early burst and subsequent diversity-dependent decline, while the North Glade did not differ from a constant pure-birth model of diversification. The sister clades also showed strong differences in the number of lineages and in the ecological and morphological diversity of component lineages. In terms of the ecological diversity of the clades, the lineages in the South Glade are predominantly closed canopy dwellers and occupy all the vertical strata of the forest from understorey to canopy. In contrast, the North Glade is characterized by the absence of canopy lineages and also in the relative increase in the number of open canopy lineages. I excluded area as a causal factor, considering the relatively smaller area of the region of the South Glade, south of Palghat Gap, compared to the North Glade. Taken together, an early burst in lineage diversification, high species richness and Glade-level endemism, narrow geographical range of the Glade, restriction of canopy lineages to this Glade and persistence of a historical rainforest refugium in this area suggested a potential role for ecological opportunity in the adaptive radiation of the South Glade. The results from Chapter 3 brought out the strong contrast in the ecological diversity of extant lineages among the bush frog sister clades occurring in this region. One major ecological pattern is the predominance of closed canopy lineages in the South Blade, while the North Glade was characterized by relatively more open canopy lineages. However, what caused the origin of open canopy lineages and the predominance of such lineages in the North Glade was not known. In chapter 4, I explore whether signatures of the global climate mediated emergence of open canopy biomes can be discerned in the habitat shift and in the observed contrast in ecological disparity between the sister clades. The results show multiple habitat shifts among bush frog lineages in the Western Ghats, with a strong contrast between the sister clades; the North Glade exhibited a greater number of habitat shifts compared to the South Blade. A unique shift to the bamboo reed Ochlandra could be discerned in the South Glade. The study provided the first evidence for independent shifts to grass associated habitats among bush frogs and frogs in general. Examination of ecomorphology brought out further interesting insights. For the same body size, lineages occurring in grass-associated habitats, Oehiandra reeds and grasslands have independently evolved smaller hind limbs compared to lineages that occur in the forest and shrub-land habitats. This convergence in limb-size and a. pattern of reduced limb-size provides support for the adaptive nature of this trait in relation to grass associated habitat shifts among bush frog lineages. Examination of this morphological trait in time, using disparity through time plots, showed high disparity within the North Glade as expected from the patterns of greater habitat shift. These results lend support for orbitally forced habitat dynamics as a driver for the intra-cladal patterns of ecological disparity in bush frog sister clades. Researchers in the past have largely tested evolutionary radiations for adapt signatures under the assumption of ecological opportunity. One of the drawbacks of temporal models of diversification is the exclusion of geography. Mountain systems slid-the -Western Ghats provide an ideal setting to explore diversification in space. In chapter took a biogeographic approach and tested the relative importance of Quaternary glaciatic ecological gradients and Earth related processes on the spatial patterns of lineage and s dada! diversification in a mountain setting. This was done using a set of a-priori predict', on the phylogenetic, spatial and temporal patterns of lineage diversification based on geological and climatic history of the Western ghats. These were tested using sister-lines and latitudinal and elevational range data obtained from field sampling across 13 mass Independent methods were used to test for the alternative processes. A majority of recer diverged bush frog sister - lineages were characterized by non-overlapping geographi ranges, and were isolated on adjacent massifs, indicating allopatric speciation eves Independent transitions along elevational gradients among sub-clades during the Mioc' lend support for diversification mediated by the uplift of the escarpment. In-situ diversificat in the montane zone of the two highest massifs shows the effect of past - climate media forest-grassland dynamics, but divergence times lend less support for the role of Quatern glaciations. These results highlight the important role of geological processes in the line; diversification of bush frogs. By examining patterns in a large radiation, I was also able bring out the temporal and spatial significance of different processes. Diversification in s Glades shows that recent non-adaptive processes have masked the effect of initial adapt radiation. Based on these findings, we recognize the massifs in this Escarpment as cent of lineage diversification and generators of diversity and the lowland and medium elevat habitats in the southern Western Ghats, with deeply divergent lineages, as museums. Through this research, I have provided a number of insights — many for the first time — into the macroevolutionary patterns and historical processes behind this large vertebrate radiation in a global biodiversity hotspot. The thesis also sets a foundation to explore the processes driving ecological and biogeographic patterns — species richness, endemism and community assembly — in space.

Environmental stressors are ubiquitous. Animals will face a variety of natural and anthropogenic stressors throughout their life cycle. The physiological mechanisms that mediate stressful stimuli can have pleiotropic effects on life history traits, such as reproduction and development. Furthermore, these phenotypic changes can affect larger-scale ecosystem dynamics, like nutrient cycling and disease epizootics. Animals are not equally susceptible to stressors across all stages of their life cycles. Critical windows of development, common in young and developing animals, are time periods when stressors have an outsized effect and can permanently alter phenotype. Larval amphibians use a critical window in late larval development wherein activation of the hypothalamic-pituitary-adrenal/interrenal (HPA/I) axis speeds their development, allowing them to metamorphose and escape suboptimal aquatic habitat. Accelerated development comes at a cost, the glucocorticoid (GC) hormones secreted by the HPA/I axis affect other systems such as growth, immune function, metabolism, and nutrient use. In chapter two, I investigated the effects of GC hormones on metabolism, energetics, and nutrient oxidation in larval wood frogs. The results show that GC hormones increase metabolism, reduce lipid stores, and increase protein oxidation during metamorphic climax. Chapter three focuses on the effects of GC hormones on immune function and r susceptibility. I found that chronic exposure to GC hormones increased ranavirus replication in infected wood frog larvae, but did not affect survival time. This could contribute to ranavirus epizootics. Acute GC hormone exposure increased survival, possibly by activing the inflammatory response. Finally, chapter four investigates the effects of GC hormones on nutrient stoichiometry. I found that larvae treated with GC hormones had lower nitrogen to phosphorus ratios, possibly because disrupted skeletal ossification. They also had reduced phosphorus excretion, which could affect ecosystem-level processes like nutrient cycling and decomposition.

The retinal pigment epithelium (RPE) is a transporting epithelium with polarized membrane domains. A unique characteristic of these cells is that their apical surface does not face a lumenal space, but is directly apposed to a layer of neurons (photoreceptors) and their associated extracellular matrix. Because the interaction occurring at this site is important for retinal attachment and particle phagocytosis, an attempt was made to identify epithelial molecules which potentially could mediate cell-cell or cell-matrix adhesion. In the present report, the subcellular localization of β1-integrins, the main receptors for extracellular matrix ligands, has been examined within Xenopus RPE. Several previously characterized antibodies were used in this analysis including: two rabbit polyclonal antibodies directed against purified chick muscle fibronectin receptor (pAbs No. 3818 and No. 2999), and a monoclonal antibody specific for Xenopus β1-integrin subunit (mAb 8C8). In Western blots of whole epithelial cell extracts, each of the antibodies intensely labeled a 115 kDa band, consistent with β1-integrin reactivity. One of the reagents (pAb No. 3818) also weakly stained unidentified bands of 50 and 100 kDa. Pre-clearing experiments demonstrated that pAb No. 3818 and mAb 8C8 both recognize the same detergent-soluble integrin: when cell extracts were depleted of β1-integrin by immunoprecipitation with mAb 8C8, the 115 kDa antigen recognized by pAb No. 3818 was not observed. Consistent with their similar immunochemical reactivities, each of the antibodies produced equivalent immunocytochemical staining of many eyecup tissues, including extraocular skeletal muscle cells, scleral and choroidal fibroblasts and vascular endothelium of the choroid and neural retina. In the native RPE, and isolated sheets of epithelium, however, qualitative differences in labeling between these antibodies were evident. Analysis by confocal microscopy showed that, while all three antibodies stained the basal surface of the epithelium, pAb No. 3818 also strongly labeled the apical microvillar surface. As the adjacent photoreceptors did not cross-react with this antibody in control experiments, the apical RPE staining could not be accounted for as contamination with retinal tissues during isolation. Furthermore, when the apical cell surface was selectively biotinylated in situ, and biotinylated proteins precipitated by streptavidin-agarose, β1-integrin was detected by immunoblotting with both mAb 8C8 and pAb No. 3818. This domain-specific material, however, represented only a fraction of the whole cell surface integrin: substantially greater amounts of tagged molecules could be detected when isolated epithelial sheets were biotinylated, most likely representing the basal protein. Based on these results, it can be concluded that β1-integrin is present in both basal and apical RPE plasma membranes. Molecules present in the apical, membrane may represent components of adhesion receptors responsible for retina-epithelium interactions.