Thesis (Ph. D.)--University of Michigan, 1928. / Thesis note in foot-notes on p. [5]. Descriptive letterpress on versos facing the plates. Bibliography: p. 41-43.

Thesis (Ph. D.)--University of Michigan, 1928. / Thesis note in foot-notes on p. [5]. Descriptive letterpress on versos facing the plates. Bibliography: p. 41-43.

Pouco se conhece acerca dos aspectos fisiológicos da reprodução de ranfastídeos, sendo estas informações importantes para sua conservação e criação em cativeiro. Métodos não-invasivos permitem análises a longo prazo e em espécies pequenas, além de facilitar estudos em vida livre e minimizar o estresse causado aos animais. Este trabalho teve por intuito a dosagem de metabólitos de testosterona (machos e fêmeas), progesterona e estradiol (somente fêmeas) nas excretas de 3 casais de tucano-toco (Ramphastos toco) e 3 casais de tucano-de-bico-verde (Ramphastos dicolorus), dentro e fora da estação reprodutiva. Dados de observações comportamentais mostraram que os animais foram mais ativos durante a estação reprodutiva, porém comportamentos sociais e proximidade entre o casal se mostraram raros nas duas fases durante as seções de observação. Concentrações de metabólitos de progesterona foram obtidos para as duas espécies na Fase Reprodutiva, mas não foram detectados nos ensaios para a Fase Não-reprodutiva. Metabólitos de testosterona puderam ser dosados em ambas as espécies, nas duas fases, tanto em machos quanto em fêmeas; sendo que os resultados mostraram para a maioria dos indivíduos concentrações maiores na Fase Reprodutiva. Aumentos acima de variações basais de metabólitos de testosterona foram associados à ocorrência de posturas, que ocorreram durante a Fase Reprodutiva numa das fêmeas de R. toco. Não foram obtidos resultados de metabólitos de estradiol em nenhuma das duas espécies, possivelmente pela falha do anticorpo utilizado em reconhecer os metabólitos. No entanto, este trabalho mostrou que a técnica utilizada foi eficaz para dosagem de metabólitos de testosterona e progesterona nas excretas destas espécies, evidenciando variações sazonais nas concentrações hormonais e indicando uma relação entre a concentração de metabólitos de testosterona e as posturas. / Little is known about reproductive physiology in Ranfastidae and this knowledge is needed for the development of conservation programs and for successful captive breeding. Non-invasive methods allow long-term analysis in small animals, facilitate studies with free-living animals and minimize the handling stress. The purpose of this work was to measure the testosterone (males and females), progesterone and oestrogen (females) metabolites concentrations in the droppings of Toco-toucan (Ramphastos toco) and Red-billed-toucan (Ramphastos dicolorus), inside and outside the reproductive season. Three captive couples of each species were studied. Collected behavioural data showed that the animals were more active during reproductive season, but social behaviours and the couple proximity were rare in the two phases during the observation sessions. Progesterone metabolites concentrations could be obtained in the Reproductive Phase for both species, but weren`t detected in the Nonreproductive Phase. Testosterone metabolites could be measured for both species in the two phases, in males and females; and the results showed higher concentrations in most of individuals in the Reproductive Phase. Concentrations of testosterone metabolites above basal levels were associated to egg laying, which occurred during Reproductive Phase in one of the R. toco females. Results of oestrogen metabolites concentrations could not be obtained in both species, probably because the fail of the antibody in detecting these metabolites. However, this work showed that the utilized technique was efficient to measure testosterone and progesterone metabolites in those species droppings, evidencing seasonal variations in hormonal concentrations and indicating a relationship between testosterone concentrations and oviposition.

Pouco se conhece acerca dos aspectos fisiológicos da reprodução de ranfastídeos, sendo estas informações importantes para sua conservação e criação em cativeiro. Métodos não-invasivos permitem análises a longo prazo e em espécies pequenas, além de facilitar estudos em vida livre e minimizar o estresse causado aos animais. Este trabalho teve por intuito a dosagem de metabólitos de testosterona (machos e fêmeas), progesterona e estradiol (somente fêmeas) nas excretas de 3 casais de tucano-toco (Ramphastos toco) e 3 casais de tucano-de-bico-verde (Ramphastos dicolorus), dentro e fora da estação reprodutiva. Dados de observações comportamentais mostraram que os animais foram mais ativos durante a estação reprodutiva, porém comportamentos sociais e proximidade entre o casal se mostraram raros nas duas fases durante as seções de observação. Concentrações de metabólitos de progesterona foram obtidos para as duas espécies na Fase Reprodutiva, mas não foram detectados nos ensaios para a Fase Não-reprodutiva. Metabólitos de testosterona puderam ser dosados em ambas as espécies, nas duas fases, tanto em machos quanto em fêmeas; sendo que os resultados mostraram para a maioria dos indivíduos concentrações maiores na Fase Reprodutiva. Aumentos acima de variações basais de metabólitos de testosterona foram associados à ocorrência de posturas, que ocorreram durante a Fase Reprodutiva numa das fêmeas de R. toco. Não foram obtidos resultados de metabólitos de estradiol em nenhuma das duas espécies, possivelmente pela falha do anticorpo utilizado em reconhecer os metabólitos. No entanto, este trabalho mostrou que a técnica utilizada foi eficaz para dosagem de metabólitos de testosterona e progesterona nas excretas destas espécies, evidenciando variações sazonais nas concentrações hormonais e indicando uma relação entre a concentração de metabólitos de testosterona e as posturas. / Little is known about reproductive physiology in Ranfastidae and this knowledge is needed for the development of conservation programs and for successful captive breeding. Non-invasive methods allow long-term analysis in small animals, facilitate studies with free-living animals and minimize the handling stress. The purpose of this work was to measure the testosterone (males and females), progesterone and oestrogen (females) metabolites concentrations in the droppings of Toco-toucan (Ramphastos toco) and Red-billed-toucan (Ramphastos dicolorus), inside and outside the reproductive season. Three captive couples of each species were studied. Collected behavioural data showed that the animals were more active during reproductive season, but social behaviours and the couple proximity were rare in the two phases during the observation sessions. Progesterone metabolites concentrations could be obtained in the Reproductive Phase for both species, but weren`t detected in the Nonreproductive Phase. Testosterone metabolites could be measured for both species in the two phases, in males and females; and the results showed higher concentrations in most of individuals in the Reproductive Phase. Concentrations of testosterone metabolites above basal levels were associated to egg laying, which occurred during Reproductive Phase in one of the R. toco females. Results of oestrogen metabolites concentrations could not be obtained in both species, probably because the fail of the antibody in detecting these metabolites. However, this work showed that the utilized technique was efficient to measure testosterone and progesterone metabolites in those species droppings, evidencing seasonal variations in hormonal concentrations and indicating a relationship between testosterone concentrations and oviposition.

Master of Science / Department of Diagnostic Medicine/ Pathobiology / Lisa M. Pohlman / Background: Iron storage disease has proven to be a serious health concern for captive toucans. Physiologic mechanisms to efficiently extract iron from naturally iron-deficient diets appear the likely cause of iron overload when fed iron-sufficient diets in captivity. Iron overload can result in diabetes, heart failure, and even death. Serum ferritin concentrations are considered the most reliable screening tool to predict total body iron stores in many species, but an assay has not been available to measure serum ferritin in toucans. Objective: The purpose of this study was to develop an enzyme-linked immunosorbent assay (ELISA) to measure serum ferritin in toucans using a polyclonal antibody in a sandwich arrangement. Methods: Ferritin was isolated from toucan liver and used as a standard. A rabbit polyclonal anti-toucan antibody was used as the capture antibody and as a detection antibody conjugated to horseradish peroxidase. Linearity of toucan ferritin standards, effect of serum dilution, recovery of added ferritin standards, and intra- and inter-assay variability were determined. Results: Ferritin standards were linear from 0 to 50 ng/ml. The relationship between serum dilution and serum ferritin concentration was also linear. When 10, 20, 30, 40, or 50 ng/ml of purified toucan ferritin were added to diluted serum, the recoveries varied from 69% to 104%. The intra-assay variability for four test serum samples averaged 11% and the inter-assay variability for the same four samples averaged 11%. Conclusions: Although the results from the linearity and recovery studies are promising for assay development when viewed independently, preliminary ferritin concentrations from all toucans studied are much higher than expected. Upon further evaluation including Dot blot assays, Western blot assays, SDS-PAGE, and protein determination of the ferritin stock solution, it was determined that the ferritin stock solution did not contain a pure protein and therefore likely renders the assay invalid. Further testing is needed to confirm these findings.