Vários estressores, incluindo poluição e patógenos, têm contribuído para o declínio/extinção da população de anfíbios levantando a questão da necessidade de ampliar o conhecimento dos aspectos da imunidade inata destes animais. Este trabalho teve por objetivo investigar os efeitos da inflamação sistêmica induzida por LPS na produção de mediadores que participam da montagem e resolução de respostas inflamatórias em anfíbios anuros. Adultos machos da espécie Rhinella icterica coletados em Janeiro de 2015 (São Luiz do Paraitinga, SP, Brazil (Licença do IBAMA: 29896) foram mantidos individualmente em caixas plásticas com circulação de ar, 22±2°C, LD 12:12h, água ad libitum e alimentados uma vez por semana com grilos e baratas. As variações diárias de CORT e MEL foram determinadas no plasma coletado durante 24 h por punção cardíaca. Para a indução de uma inflamação sistêmica, os animais foram injetados no saco linfático dorsal com solução salina ou LPS (de Escherichia coli, serotype 0127:B8; 2.0mg/Kg; ZT15) e eutanaziados após duas horas. O padrão temporal de liberação de CORT foi evidenciado pelos níveis aumentados no período de atividade dos animais, com pico de liberação na transição claro-escuro. O padrão temporal noturno de produção de MEL foi evidenciado atingindo um pico de liberação 6 h após a entrada do escuro. A inflamação sistêmica induzida por LPS levou a um aumento nos níveis da citocina IL-1?, ativação do eixo hipotálamo-hipófise-interrenal com aumento de 10x nos níveis de CORT e uma diminuição no conteúdo de MEL plasmática e ocular. Desta forma, os dados apresentados evidenciam uma variação circadiana na produção dos hormônios CORT e MEL, ainda não demonstrados em Rhinella icterica, bem como a existência de um mecanismo regulatório bidirecional entre os sistemas imune e endócrino em anfíbios, apontando para a presença de um Eixo Imune-Pineal ativo no controle de processos patológicos em anfíbios tal como já demonstrado em mamíferos / Several stressors, including pollution and pathogens, have contributed to the decline/extinction of the amphibian population raising the question of the need to increase knowledge of the aspects of the innate immunity of these animals. This work aimed to investigate the effects of systemic inflammation induced by LPS on the production of mediators that participate in the assembly and resolution of inflammatory responses in amphibians. Male adults of the specie Rhinella icterica collected in January 2015 (São Luiz do Paraitinga, SP, Brazil (IBAMA License: 29896) were individually kept in plastic boxes with air circulation, 22 ± 2 ° C, LD 12: 12h, water ad libitum and fed once a week with crickets and cockroaches.The daily variations of CORT and MEL were determined in plasma collected for 24 h by cardiac puncture.In order to induce a systemic inflammation, the animals were injected into the dorsal lymph sac with LPS (Escherichia coli, serotype 0127: B8, 2.0 mg/kg, ZT15) and euthanized after two hours. The temporal pattern of CORT release was evidenced by the increased levels in the period of activity of the animals, with a peak release at the transition light/dark cycle. The nocturnal temporal pattern of MEL production was evidenced reaching a peak of release 6 h after the dark entrance. LPS-induced systemic inflammation led to an increase in the cytokine IL-1? levels, activation of the hypothalamic-pituitary-interrenal axis with a 10x increase in CORT levels and a decrease in plasma and ocular MEL content. Thus, the data presented evidence a circadian variation in the production of CORT and MEL hormones, not yet demonstrated in Rhinella ictérica, as well as the existence of a bidirectional regulatory mechanism between the immune and endocrine systems in amphibians, pointing to the presence of an Immune-Pineal Axis active in the control of pathological processes in amphibians as already demonstrated in mammals

Em resposta a diversos estressores, o hipotálamo de anfíbios secreta o hormônio liberador de corticotropina (CRH), o qual estimula a hipófise a liberar o hormônio adrenocorticoctrópico (ACTH) na corrente sanguínea. O ACTH, por sua vez, atua na glândula inter-renal estimulando a liberação de glicocorticoides, sendo a corticosterona o principal glicocorticoide em anfíbios. Embora a modulação exercida pelos glicocorticoides de diversas respostas fisiológicas de estresse seja fundamental para a manutenção da homeostase em curto prazo, a ativação muito frequente e/ou prolongada do eixo hipotálamo-hipófise-interrenais (HHI) pode inibir processos importantes em longo prazo, tais como a reprodução e a imunocompetência e afetar a síntese de melatonina atuando diretamente sobre a pineal. O objetivo do presente estudo foi avaliar possíveis alterações nos níveis plasmáticos de corticosterona, testosterona, níveis oculares de melatonina, hematócrito e aspectos imunitários (contagem total de leucócitos e capacidade antimicrobiana plasmática - contra Escherichia coli) de machos da perereca Hypsiboas faber em resposta à injeção de ACTH, bem como o curso temporal destas respostas. Amostras de sangue e plasma foram obtidas a partir de 30 machos capturados em campo, dentro de 3 minutos da captura. Os animais foram trazidos para o laboratório e, após dois meses, divididos em seis grupos para serem amostrados em três horários distintos 1 h, 3h e 6h após o tratamento. Para cada horário um grupo controle e um grupo experimental foram adicionados, recebendo uma injeção de 105 μL de solução salina ou de solução de ACTH (0,446 μg de ACTH/g de massa corpórea (grupo experimental)). O tratamento com ACTH aumentou significativamente os níveis plasmáticos de corticosterona e diminuiu os níveis plasmáticos de testosterona bem como os níveis oculares de melatonina após 1 hora. Após 6 h da injeção de ACTH, houve um aumento da porcentagem do hematócrito e dos níveis de melatonina ocular. O tratamento com ACTH não afetou os aspectos imunitários estudados dentro do intervalo amostrado. Os resultados confirmam o papel da ativação do eixo HHI na inibição da secreção de testosterona e na modulação da secreção de melatonina, embora os possíveis efeitos imunomodulatórios possam se dar em intervalos temporais mais tardios / In response for several stressors, the amphibian hypothalamus release the corticotrophin release hormone (CRH), which stimulates pituitary to release the adenocorticotropic hormone (ACTH) in the bloodstream. The ACTH, in your turn, acts in inter renal glands stimulating the glicocorticoids releasing, being the corticosterone the main glicocorticoid in anphibians. While the modulation exercised by the glicocorticoids of diverses physiological responses of the stress is fundamental for de maintainance of homeostasis in short period, the frequent and/or prolongued activation of the hypothamlamus pituitary interenal axis (HHI) could inhibit important process in long periods, as the reproduction and immunocompetence and affect melatonin synthesis acting directly in pineal gland. The objective of the present study was evaluate possible alterations in plasmatic corticosterone levels, testosterone, eyes melatonin levels, hematocrit and immunity aspects (Total count leucocytes and antimicrobian plasmatic capacity - against Escherichia coli) from males of the treefrog Hypsiboas faber in response to ACTH injection, as well, the temporal course of these responses. Blood and plasma samples were obtained from the 30 males captured in the field, whitin 3 minutes from the capture. The animals were transferred to the laboratory and, after two months, divided in six groups to be sampled in three different times 1 h, 3 h and 6 h after treatment. For each time one control group and one experimental group were added, receiving one injection of 105 μL of saline or ACTH solution (0,446 μg de ACTH/g of body mass (experimental group)). The ACTH treatment increase significantly the plasmatic corticosterone levels and decrease the plasmatic testosterone levels, as well, the eye melatonin levels after one hour. After six hours of the ACTH injection, occurred the increase of the hematocrit percentage and the eye melatonin levels. The ACTH treatment did not affect the immunity aspects studied in the sampling interval. The results confirm the role of the activation on the axis HHI in the inhibition of the testosterone secretion and in the modulation of melatonin secretion, althougth the possible immunomodutatory effects could happen in temporal intervals later

O objetivo do presente trabalho foi investigar as repercussões de uma relação social estável sobre diferentes parâmetros de comportamento, neuroquímica e atividade imune de camundongos dominantes e submissos. Machos adultos (com aproximadamente 90 dias de idade) mantidos em duplas desde o desmame, foram determinados como dominantes ou submissos, após três avaliações consecutivas do comportamento, onde foram observadas a presença ou ausência de ataques ou fugas e posturas de submissão. Em alguns experimentos, grupos de cinco animais mantidos em uma mesma caixa foram utilizados para comparação com resultados obtidos de animais que conviveram em duplas. Foram utilizadas apenas as duplas de camundongos onde a hierarquia social foi claramente observada. Os resultados mostraram que os animais submissos apresentaram em relação aos dominantes: 1) diminuição no tempo gasto na zona central do campo aberto; 2) diminuição no número de entradas nos braços abertos e diminuição no tempo gasto na exploração dos braços abertos do labirinto em cruz elevado; 3) aumento no tempo gasto na exploração dos braços fechados do labirinto em cruz elevado; 4) diminuição no número de entradas e no tempo gasto na exploração do terço final dos braços fechados do labirinto em cruz elevado; 5) aumento na taxa de renovação de dopamina no hipotálamo; 6) diminuição da taxa de renovação de dopamina no corpo estriado; 7) maior número de metástases induzidas pelo melanoma murino experimental B16F10; 8) aumento do percentual de células T CD8+ no timo após 14 dias de inoculação do mesmo melanoma; 9) diminuição no burst oxidativo basal de neutrófilos e monócitos sangüíneos, mas não naquele induzido por bactérias; 10) menor atividade de células NK presentes no baço e no sangue. Em relação aos animais mantidos em número de cinco, os animais submissos apresentaram: menor percentual de células NK no sangue. Já os animais dominantes, apresentaram em relação aos animais mantidos em grupos: 1) aumento da taxa de renovação de noradrenalina no hipotálamo; 2) aumento na taxa de renovação de dopamina no corpo estriado; 3) menor percentual de células NK no sangue. O status social, no entanto, não provocou diferenças: 1) nos níveis absolutos de dopamina, noradrenalina e serotonina; 2); nos metabólitos de serotonina; 3) nos níveis séricos basais de corticosterona; 4) no peso e número de células do baço e timo; 5) no percentual de células T CD4+ e CD8+ no baço e 6) no percentual de linfócitos, neutrófilos e monócitos sangüíneos. Em conjunto, os presentes resultados mostraram que animais dominantes e submissos mantidos por 90 dias em uma hierarquia social estável, apresentaram diferenças comportamentais e neuroquímicas, e responderam de forma diferente a um mesmo estímulo imune, no caso, o desenvolvimento de metástases induzida nos pulmões pela administração do melanoma experimental murino B16F10. Estes resultados sugerem que diferentes mecanismos, que não a ativação do eixo HPA, estejam envolvidos com o aumento de susceptibilidade ao desenvolvimento do tumor observado nos indivíduos submissos / The objective of the present work was to investigate the repercussions of a stable social relationship on different parameters of the behavior, neurochemical and immune activity of dominant and submissives mice. Adult males (with approximately 90 days of age) kept in pairs since wean it, had been determined as dominant or submissives, after three consecutive evaluations of the behavior, where presences or absences of attacks or escapes and positions of submission had been observed. In some experiments, groups of five animals kept in one same box had been used to compare the results gotten between these and the animals coexisting in pairs. The pairs had been used only where the social hierarchy clearly was observed. The results had shown that the submissives animals in relation to the dominant ones had presented: 1) reduction in the time spent in the central zone of the open field; 2) reduction in the number of entrances in the open arms and reduction in the time spent in the exploration of the open arms of the plus maze; 3) increase in the time spent in the exploration of the closed arms of the plus maze; 4) reduction in the number of entrances and time spent in the exploration of the final third of the closed arms of the plus maze; 5) increase in the turnover of dopamine in the hypothalamus; 6) reduction in the turnover of dopamine in the corpus striatum; 7) increased number of metastasis in the lungs induced by murino melanoma experimental B16F10; 8) increase of the percentage of cells T CD8+ in the thymus after 14 days of inoculation of the same melanoma; 9) reduction in the basal oxidative burst of neutrophil and monocytes sanguine, but not in the induced by bacteria; 10) decreased NK cells activity measured in the blood and spleen. In relation to the animals kept in number of five, the submissives animals had presented: 11) reduction in the percentile of NK cells in the blood. While the dominant animals had presented in relation to the animals kept in groups: 1) increase in the turnover of norepinephrine in hypothalamus; 2) increase in the turnover of dopamine in the fluted body; 3) reduction in the percentile of NK cells in the blood. The social status, however, did not provoke differences: 1) in the absolute levels of dopamine, norepinephrine and serotonin; 2) in the metabolites of serotonin; 3) in the serum levels of corticosterone; 4) in the weight and number of cells of the spleen and thymus; 5) in the percentage of cells T CD4+ and CD8+ in the spleen and 6) in the percentage of lymphocytes, neutrophil and monocytes in the blood. Together, the results obtained had shown that dominants and submissives animals kept 90 days living in a stable social hierarchy had presented behavior and neurochemical differences, and had answered of different form to one same immune stimulation, in this case, the induced development of metastasis in the lungs for experimental melanoma murino B16F10, where the submissives had been more susceptible than the dominant ones. This results suggest that other mechanisms, different of HPA activation, may be involved with the decreased resistance of submissive mice to B16F10 tumor dissemination

Recently, the European Union published regulations regarding the welfare of horses during transport requiring that horses be transported in individual stalls separated by partitions. The objective of this study was to determine if concentrations of cortisol, corticosterone, or dehydroepiandrosterone (DHEA) differed among horses with no prior transport experience when transported in individual stalls versus loose groups. Twenty na�ve yearlings were assigned to either individual stalls or a loose group, then transported for 6 hours. Ten horses were transported per day (5 in stalls and 5 in a loose group) over a two day trial. The experiment was replicated with a second trial 35 days later, and utilized a switchback design where the horses exchanged treatments between trials. Blood samples were collected and analyzed for cortisol, corticosterone, and DHEA concentrations at pre-transport, 2, 4, and 6 h of transport, and at 2 and 4 h after unloading. After Trial 2, the horses? changed housing from group paddocks to stalls and a follow-up experiment was conducted. The data were analyzed using a mixed model repeated measures ANOVA with the animal as the subject, with trial, treatment, sample time, and treatment-sample time interaction in the model with unstructured covariance (SAS 9.1). Differences between sample times within each trial, and pre-transport concentrations between trials, were analyzed using paired t-tests (SPSS 12.0.1). No significant differences were found in hormone concentrations for horses transported in individual stalls versus in loose groups. Horses exhibited a significant elevation in cortisol and corticosterone during transport which returned to pre-transport concentrations by 2 hr after transport (P < 0.01). Mean pre-transport cortisol concentrations rose significantly in Trial 3 (7.87 ng/ml) from Trials 1 (2.71 ng/ml) and 2 (2.84 ng/ml) (P < 0.001). Pre-transport concentrations of DHEA in Trials 1 (482 pg/ml) and 2 (392 pg/ml) also rose significantly in Trial 3 (1607 pg/ml) (P < 0.01). Changes in cortisol and DHEA indicated that transportation was a significant stressor for horses, however, being transported in a loose group versus individual stalls was not different. Also, housing changes from paddocks to stalls resulted in significant increases in pre-transport concentrations of stress-related compounds.

It was hypothesized that alterations in plasma concentrations of corticosterone (CS) and prolactin (PRL) may be at least partially responsible for polychlorinated biphenyl (PCB)-induced immunosuppression. A 2 by 2 factorial experiment examined the interactions of PCB and P815, an allogeneic tumor, on plasma concentrations of CS and PRL, and on body, spleen, and thymus weights. The PCB dosage used (10 mg/kg) was previously shown to suppress immune response to the tumor. The four study groups were: Group A (vehicle control), Group B (tumor only), Group C (PCB only), and Group D (tumor plus PCB). Mice received one dose of PCB (Groups C and D) or carrier (Groups A and B) on day -1; tumor (Groups B and D) or carrier (Groups A and C) was injected intraperitoneally on day 0. In Experiment 1, animals were killed on days -1, -0.6, 0 through 10, 21, 42, and 84. Body, spleen, and thymus weights were measured. Plasma samples were obtained for CS and PRL measurements. In Experiment 2, the study was repeated with samples obtained only on days 3 and 10. Group A body weights increased steadily throughout Experiment 1. Relative to Group A, the weight gain in Group B was significantly (p < 0.05) higher. Group C lost weight on days 0 through 6, and gained significantly (p < 0.05) less weight than Group A. Group D gained significantly (p < 0.05) less weight than Groups A, B, and C. As a percent of body weight, spleen weight remained constant over 21 days in Experiment 1 in both Groups A and D. Compared to Group A, Group B showed significantly (p < 0.05) increased spleen percent body weight while Group C showed significantly (p < 0.05) decreased spleen percent body weight. As a percent of body weight, thymus weight remained constant for 21 days in Experiment 1 in Group A. Groups Band C were similar (p > 0.05) and showed a decreased thymus percent body weight compared to Group A. Group D showed significantly (p < 0.05) decreased thymus percent body weight relative to the other three groups. Mean CS concentrations in Experiment 1 in Groups A and B were similar (p < 0.05). Relative to Groups A and B, Group C CS concentrations were elevated, with a peak of 126.1 ng/ml on day 4. Group D CS concentrations were higher than the other three groups, peaking at 294.1 ng/ml on day 10. There was no significant difference in PRL concentrations in Groups A, B, and C in Experiment 1 (p > 0.05). Mean PRL concentration in Group D was significantly (p < 0.05) lower than in the other three groups. The results of Experiment 2 validated those of Experiment 1. Although absolute values differed, the pattern of changes seen in body and organ weights and in CS and PRL concentrations was similar. An acute exposure to PCB and tumor resulted in an increase in circulating CS concentration and a decrease in circulating PRL concentrations. These changes may contribute to PCB-induced immunosuppressioin. / Graduation date:1992

Stress has long lasting effects on physiology, development, behavior, reproductive success and survival. These effects are mediated by glucocorticoids, such as corticosterone (Cort), via glucocorticoid receptors (GR), though the exact mechanisms underlying these effects are unknown. Early developmental stress affects the size of the avian song control nuclei (particularly HVC; proper name) and song quality in many songbirds, suggesting a direct link between brain and behavior. HVC is required for song learning and production. The complexity of the male zebra finch (Taeniopygia guttata) courtship song is important in female mate choice. Although the mechanisms behind the effects of developmental stress on song nuclei size and song quality are unknown, it is likely that elevated levels of Cort via GR within brain song nuclei play a significant role. We investigated the distribution, quantity, and subcellular-localization of GR- immunoreactive (GR-ir) neurons in the brains of male zebra finches 10 days post-hatch and in adulthood using immunohistochemistry. There was wide distribution of GR-ir neurons including two song nuclei HVC and robust nucleus of the arcopallium (RA). Distribution did not vary between the two ages but there were significant differences in the overall number of GR-ir neurons and their subcellular localization. We hypothesized that early Cort treatment would reduce song quality and HVC size in adult males. We inserted Cort implants in males at four days post-hatch and quantified the effects of early Cort treatment on adult song quality. Early Cort treatment decreased song similarity between the tutor and tutee’s songs and resulted in poorer copies of tutor song, but did not alter mean amplitude or song duration. Early Cort treatment reduced the HVC size in both juvenile and adult birds. This result suggests that the effect of developmental stress on the HVC size may be mediated through Cort via activation of GR within HVC as a mechanism by which HVC size and song quality are altered in developmentally stressed birds. These results suggest a potential role for Cort in mediating adverse effects of developmental stress in adult male zebra finches and highlight the developmental plasticity of the zebra finch brain.

Recently, the European Union published regulations regarding the welfare of horses during transport requiring that horses be transported in individual stalls separated by partitions. The objective of this study was to determine if concentrations of cortisol, corticosterone, or dehydroepiandrosterone (DHEA) differed among horses with no prior transport experience when transported in individual stalls versus loose groups. Twenty na�ve yearlings were assigned to either individual stalls or a loose group, then transported for 6 hours. Ten horses were transported per day (5 in stalls and 5 in a loose group) over a two day trial. The experiment was replicated with a second trial 35 days later, and utilized a switchback design where the horses exchanged treatments between trials. Blood samples were collected and analyzed for cortisol, corticosterone, and DHEA concentrations at pre-transport, 2, 4, and 6 h of transport, and at 2 and 4 h after unloading. After Trial 2, the horses? changed housing from group paddocks to stalls and a follow-up experiment was conducted. The data were analyzed using a mixed model repeated measures ANOVA with the animal as the subject, with trial, treatment, sample time, and treatment-sample time interaction in the model with unstructured covariance (SAS 9.1). Differences between sample times within each trial, and pre-transport concentrations between trials, were analyzed using paired t-tests (SPSS 12.0.1). No significant differences were found in hormone concentrations for horses transported in individual stalls versus in loose groups. Horses exhibited a significant elevation in cortisol and corticosterone during transport which returned to pre-transport concentrations by 2 hr after transport (P < 0.01). Mean pre-transport cortisol concentrations rose significantly in Trial 3 (7.87 ng/ml) from Trials 1 (2.71 ng/ml) and 2 (2.84 ng/ml) (P < 0.001). Pre-transport concentrations of DHEA in Trials 1 (482 pg/ml) and 2 (392 pg/ml) also rose significantly in Trial 3 (1607 pg/ml) (P < 0.01). Changes in cortisol and DHEA indicated that transportation was a significant stressor for horses, however, being transported in a loose group versus individual stalls was not different. Also, housing changes from paddocks to stalls resulted in significant increases in pre-transport concentrations of stress-related compounds.

It has been known that light input can affect the emotions of a person. The depressive syndrome Seasonal Affective Disorder (SAD) is an effective example of the power of light in changing the mood of a person. Patients with SAD have recurring depressive episodes that follow seasonal changes, which is due to the changing daylight hours. This phenomenon suggests that there would be receptors in the retina that would not simply be responsible for vision, but also for the regulation of non-visual signals such as emotion. In many animals, projections have been found from the retina to the dorsal raphe nucleus (DRN). This brain region is a serotonergic area and has been found to be involved in the occurrence of depression. As such, the cells in the retina which were found to have projections to the DRN have a high possibility to be involved in emotion regulation. Retinal Ganglion Cells (RGCs) are classified into many types. A specific type known as an alpha cell is suspected to be the DRN-projecting subtype. This study uses Lycium Barbarum Polysaccharide (LBP) as a treatment in protecting the large RGCs from corticosterone (CORT) -induced damage. The aim is to observe if LBP will provide neuroprotection to large sized RGCs damaged by 40mg/kg or 50mg/kg CORT, and hence if LBP can be further investigated as a possible anti-depressant drug. This study observed that although LBP did not reduce large cell deaths, it reduced cell atrophy of the RGCs under high dosage of CORT (50mg/kg). For the same number of cells counted, treatment groups with a high dose CORT injection found more cells over 300μm2 in area than cells under 300μm2. Also, it was found that the temporal quadrants were more sensitive to cell size change than the nasal quadrants, paving way for more in-depth research of the spatial sensitivity to CORT or to LBP. The findings in this study indicate that LBP does indeed have a neuroprotective effect on large RGCs, although this effect is limited and as of yet seems conditional, as this study ignores the effect of CORT and LBP on other large cell properties such as the dendritic field size and the amount of synapses. Further studies are needed to determine the mechanism of the neuroprotective effect of LBP and to determine the exact site of action LBP works on. / published_or_final_version / Anatomy / Master / Master of Medical Sciences

Major depression and Alzheimer’s disease (AD) are highly prevalent psychiatric disorders. Further investigation demonstrated that depression itself is a risk factor for AD, and several associated genetic mutations have been found Moreover, significant proportion of AD patients suffer also suffer from depression. These findings generated interests in finding the neurobiological linkages between depression and AD. The elucidation of pathophysiological mechanisms common in both disorders would be important, as the knowledge could provide additional insights regarding the pathogeneses of the disorders and possible interventions. The present study proposes that synaptic degeneration plays a central role in the pathogenesis of depression and AD. Using in vitro disease models, this study demonstrated abnormalities in pre-synaptic and cytoskeletal proteins, which leads to impaired synaptic function. Further investigation into the upstream events demonstrated the involvement of ubiquitin-mediated protein degradation mechanism and the preferential activation of the autophagic-lysosomal pathway. This study also investigated the neuroprotective properties of the antidepressants imipramine and escitalopram. Antidepressants have originally been thought to exert their therapeutic effects through monoaminergic system modulation. Interestingly, results in this study showed that these two agents were able to ameliorate the observed synaptic protein changes, thereby implicating other possible mechanism of action for antidepressants. In conclusion, this study provides evidence that similar synaptic pathologies exist between depression and AD, which could be responsible for the development of these two disorders. Furthermore, antidepressants may be exerting its effects through alleviating synaptic degeneration. / published_or_final_version / Psychiatry / Doctoral / Doctor of Philosophy

This thesis investigates the use of an enzyme immunoassay to measure keratin glucocorticoid concentrations in mammalian hair, bird feathers and reptilian shed skins. Keratin glucocorticoid concentrations were compared to fecal glucocorticoid concentrations produced during the period of keratin growth in the African Elephant (Loxodonta africana), the Western Lowland Gorilla (Gorilla gorilla gorilla), the Sumatran Orangutan (Pongo abelii), the Domestic Chicken (Gallus gallus domesticus), the Eastern Loggerhead Shrike (Lanius ludovicianus migrans), the African House Snake (Lamprophis fuliginosus) and the Eastern Massasauga Rattlesnake (Sistrurus catenatus catenatus). Complete biochemical validation was performed for the feather and shed skin corticosterone and fecal corticosterone enzyme immunoassays in the Domestic Chicken and the African House Snake. Biological validation was performed in the Domestic Chicken. Biological and physiological validation were attempted in the African House Snake. African Elephant, Western Lowland Gorilla and Sumatran Orangutan hair cortisol concentrations ranged from 2.10 - 312.70 ng/g. African Elephant hair corticosterone concentrations ranged from 2.68 - 20.70 ng/g. Domestic Chicken and Eastern Loggerhead Shrike feather corticosterone concentrations ranged from 1.31 - 8.09 pg/mm and from 1.09 - 6.59 pg/mm, respectively. African House Snake and Massasauga Rattlesnake shed skin corticosterone concentrations ranged from 4.42 - 124.35 ng/g and 3.82 - 22.85 ng/g, respectively. In the majority of cases, a statistically significant association was not found between summary statistical measures of fecal and keratin glucocorticoid concentrations. A statistically significant positive association was detected between hair cortisol and the coefficient of variation of fecal corticosterone in the African Elephants. A statistically significant negative association was detected between feather corticosterone and the 75th percentile and coefficient of variation of fecal corticosterone in the Eastern Loggerhead Shrikes. A statistically significant positive association was also detected between shed skin corticosterone and the mean fecal corticosterone from 3 weeks before to 1 week after the previous ecdysis in the African House Snake. Feather corticosterone concentrations were significantly higher in feathers from Domestic Chickens that were socially housed with roosters than in feathers from chickens housed individually in laying cages. A statistically significant difference was not detected between the shed skin corticosterone concentrations of the minimally handled control and the weekly handled experimental African House Snakes. Adrenocorticotropic hormone stimulation did not result in the physiological validation anticipated for shed skin corticosterone concentrations in the African House Snake. / Toronto Zoological Foundation and Ontario Ministry of Agriculture, Food and Rural Affairs