Made available in DSpace on 2014-06-11T19:26:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-10-29Bitstream added on 2014-06-13T19:33:17Z : No. of bitstreams: 1 hata_me_me_jabo.pdf: 405459 bytes, checksum: 3bd879e643e736a926ba7adff70cd7b0 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O medo é uma característica comportamental importante em espécies domesticadas e pode ser incluído no programa de seleção, pois responde à seleção artificial e tem conseqüências importantes ao bem estar e desempenho das aves domésticas. A reação de medo pode ser avaliada pelo tempo de permanência em imobilidade tônica (IT), que é o período em que o animal fica em estado catatônico induzido manualmente pelo homem. Quanto menos tempo permanecer neste estado, menor é o medo do animal e mais adaptado este se mostra a viver em cativeiro. A característica IT é bem representativa do nível de medo do indivíduo e também pode estar relacionada com a relação heterófilo/linfócito (H/L). Assim, o objetivo deste trabalho foi determinar efeitos de ambiente e estimar parâmetros genéticos da característica tempo de permanência em IT de perdiz (Rhynchotus rufescens) criada no ambiente de cativeiro. A análise realizada pelo método dos quadrados mínimos revelou a influência da época de nascimento dentro de geração e o peso corporal, sendo que animais mais pesados permaneceram maior tempo em IT (b = 0,32 ± 0,14g, p<0,05). O método de máxima verossimilhança restrita possibilitou a estimação do coeficiente de herdabilidade da característica IT, apresentando valor igual a 0,29 evidenciando influência do ambiente sobre o tempo de permanência em IT. Contudo, a seleção pode ser eficiente para alterar as médias desta característica / Fear is an important behavior trait in domesticated species and can be included in the selection program, and have important consequences to the welfare and performance of poultry. The fear reaction can be measured by the time spend in tonic immobility (TI), which is the period where the animal stays in catatonic state induced by human hand. The less time remaining in this state, smaller is the fear of the animal and it shows more adapted to living in captivity. The trait IT is well representative of the level of fear of the individual and may also be related to the heterophil to lymphocyte ratio (H/L). The objective of this study was to determine the effects of environment and estimate the genetic characteristic of tonic immobility time in Red-winged Tinamou (Rhynchotus rufescens) hosed in captivity environment. The method of least squares analysis resulted the influence of season within generation and body weight, whose heavier animals showed longer period in IT (b = 0.32 ± 0.14 g, p <0.05). The restricted maximum likelihood method was applied to estimate heritability of TI with value of 0.29 indicating environmental influence However, the selection can be effective to change the means of this trit

Orientador: Sandra Aidar de Queiroz / Banca: Valter Udler Cromberg / Banca: Danísio Prado Munari / Resumo: O medo é uma característica comportamental importante em espécies domesticadas e pode ser incluído no programa de seleção, pois responde à seleção artificial e tem conseqüências importantes ao bem estar e desempenho das aves domésticas. A reação de medo pode ser avaliada pelo tempo de permanência em imobilidade tônica (IT), que é o período em que o animal fica em estado catatônico induzido manualmente pelo homem. Quanto menos tempo permanecer neste estado, menor é o medo do animal e mais adaptado este se mostra a viver em cativeiro. A característica IT é bem representativa do nível de medo do indivíduo e também pode estar relacionada com a relação heterófilo/linfócito (H/L). Assim, o objetivo deste trabalho foi determinar efeitos de ambiente e estimar parâmetros genéticos da característica tempo de permanência em IT de perdiz (Rhynchotus rufescens) criada no ambiente de cativeiro. A análise realizada pelo método dos quadrados mínimos revelou a influência da época de nascimento dentro de geração e o peso corporal, sendo que animais mais pesados permaneceram maior tempo em IT (b = 0,32 ± 0,14g, p<0,05). O método de máxima verossimilhança restrita possibilitou a estimação do coeficiente de herdabilidade da característica IT, apresentando valor igual a 0,29 evidenciando influência do ambiente sobre o tempo de permanência em IT. Contudo, a seleção pode ser eficiente para alterar as médias desta característica / Abstract: Fear is an important behavior trait in domesticated species and can be included in the selection program, and have important consequences to the welfare and performance of poultry. The fear reaction can be measured by the time spend in tonic immobility (TI), which is the period where the animal stays in catatonic state induced by human hand. The less time remaining in this state, smaller is the fear of the animal and it shows more adapted to living in captivity. The trait IT is well representative of the level of fear of the individual and may also be related to the heterophil to lymphocyte ratio (H/L). The objective of this study was to determine the effects of environment and estimate the genetic characteristic of tonic immobility time in Red-winged Tinamou (Rhynchotus rufescens) hosed in captivity environment. The method of least squares analysis resulted the influence of season within generation and body weight, whose heavier animals showed longer period in IT (b = 0.32 ± 0.14 g, p <0.05). The restricted maximum likelihood method was applied to estimate heritability of TI with value of 0.29 indicating environmental influence However, the selection can be effective to change the means of this trit / Mestre

Chronic stress can result in elevated circulating levels of glucocorticoid hormones in vertebrates, which can affect their stress response, their immune function, and eventually their fitness. I tested the effect of chronic corticosterone (CORT) elevation on the acute stress responsiveness, immune function, and body condition of free-living painted turtles (Chrysemys picta) in Gatineau Park, using slow-release exogenous CORT administration. While Silastic implants did not predictably elevate circulating CORT concentrations in painted turtles, injections of CORT-laden cocoa butter kept circulating levels elevated for up to 3 weeks, to concentrations likely physiologically and ecologically relevant for the species. I measured the acute CORT stress response, parasitaemia, heterophil-to-lymphocyte ratios, and total leukocyte counts after 1 week and 3 weeks, and determined body condition after 1 week, 3 weeks, and 1 year. Compared to sham and control turtles, I observed no effect of treatment on these hormonal, immune, and body condition metrics of stress, possibly because CORT mediates resource allocation only in the presence of additional immune or energy challenges, because of the masking effect of extrinsic factors, or because free, not total, CORT appears to be biologically active.

<p>  </p> <p>Glucocorticoids (GC) play a critical role in regulating the physiological response to stress. Disruptions to baseline levels due to stress can have negative implications on a variety of factors including growth and development, physical body conditions, metabolism, immune functions, and expression of normal behaviors, although this list is not exhaustive. When birds are unable to adapt to the stressor and return to homeostasis, the energy expenditure associated with the failed attempt at coping can lead to significant declines in the overall health, welfare, production, and performance of the bird. This can go on to impact producers and consumers as well, indicating the extensive repercussions of stress. Recently, scientists have been investigating thorough and efficient methods of quantifying stress in birds, such as measuring heterophil-to-lymphocyte ratio (HLR) or detecting glucocorticoid levels through enzyme-linked immunoassays (ELISA). However, the precise mechanism behind HLR increase during stress is unknown and ELISAs may not provide accurate results depending on when the blood is being measured. </p> <p><br></p> <p>GC are differentially released and exert their effects in a manner that is dependent on sex, age, and time. However, before investigating this, it was critical to validate the GC kits to ensure that they were measuring cortisol and corticosterone separately along with zero cross reactions with other precursors. Chapter 2 had 4 experiments carried out. The objective of experiment 1 was to validate ELISAs to ensure that they were measuring the GC accurately and separately since both cortisol and corticosterone were being measured. To do this, duck serum was pooled and charcoal-stripped to remove the presence of steroids. 3 standard curves were run to confirm that there was no cross reactivity. The objective of experiment 2 was to further validate the ELISA kits with mass spectrometry by checking for both glucocorticoids in the pooled samples. Once the validation process was complete, experiment 3 was carried out to look at the effect of ACTH stimulation on GC release. 16-week-old drakes and hens were given either intramuscular (IM) injections of cosyntropin (0.06 mg/kg) or saline as control. The cosyntropin dose was chosen according to previous studies reporting relatively high physiological responses, therefore, we wanted to replicate this. N was 10/sex/treatment. Blood was then collected at 0, 1, and 2 hours after injections and serum was analyzed by ELISAs. Lastly for experiment 4, 14-week-old developer drakes and hens at Maple Leaf Farms were assessed for a transportation stress experiment. Blood from 10 ducks/sex/time/barn were collected at 24 hours before transport to the breeder barn, immediately after a 1-hour transport, 24 hours after, and 1 week after transport. The results from experiment 1 yielded that both cortisol and corticosterone can be measured without the presence of unwanted contaminants or other products. Experiment 2 identified the greater sensitivity of mass spectrometry when reading GC levels, although the differences were linear. Experiment 3 showed that serum corticosterone levels were significantly increased at 1 hour after ACTH injections in both drakes and hens, with levels continuing to increase for the drakes. Serum cortisol levels were significantly increased at 1 hour after ACTH injections in both sexes, however, the hens had greater levels compared to the drakes. Serum cortisol levels returned to levels similar to that of saline-injected ducks at the 2-hour mark. Lastly, the transportation stress portion showed that cortisol was released at about 1/3 of corticosterone levels in both sexes. Hens showed increased levels of serum corticosterone compared to drakes at all time points except for 1 week after transport, and also had significantly increased serum cortisol levels at all time points. In conclusion, the ELISA kits were verified for future use when measuring GC as well as mass spectrometry. GC were detected in the ACTH and transportation stress experiments with hens displaying a greater sensitivity to GC release due to increased circulating levels compared to drakes. Although it was nonsignificant, there was a trend for GC to increase in response to transport. </p> <p><br></p> <p>There are sex differences in GC release and HLR for Pekin ducks and various challenges from the studies support this. With hens showing increased sensitivity to stress and drakes with more transient and gradual levels, we have consistently seen that both GC have differential roles in the stress response and not only is it critical to study both hormones, the timing of when measurements are taken are important as well to get a clear understanding of when the stress response is initiated. </p> <p><br></p> <p>Chapter 3 went further to understand the response of GC and HLR. The objective was to  investigate the release of cortisol and corticosterone in response to an ACTH dose response challenge. In Chapter 2, only one dose of cosyntropin was used and sample collection times only went to 2 hours after injections. In this study, 2 additional doses and an extra hour of sample collection were added to obtain more information. Pekin ducks were either given IM cosyntropin injections or saline for control, with an N of 10/sex/treatment. There were 3 treatment doses: High (0.06 mg/kg), medium (0.03 mg/kg), and low (0.015 mg/kg). All injections were given promptly at 0730 hours. Blood was collected at 0, 1, 2 and 3 hours after injections from the tibia veins to obtain serum for ELISAs. Blood smears were done to analyze HLR and sent to an independent lab to obtain values. The results indicated that both GC had significant sex x dose x time interactions. The low dose injection had no effect on corticosterone in hens with a slight increase for drakes at the first hour. The high dose for hens led to a spike in corticosterone levels at the first hour with a gradual decrease, and drakes had an increase that lasted for 2 hours until they returned to baseline at the last hour. The high dose in drakes stimulated cortisol release during the first 2 hours after injection with a similar effect in hens. However, hens had elevated levels compared to drakes. Finally, there was no dose response effect for HLR, although interestingly, the low dose injection elevated HLR even though there was no effect in GC. There were sex differences in the HLR response where the drakes given the high dose had levels that plateaued by the third hour, while the hens still had elevated levels. In conclusion, the ACTH dose-response test identified that ACTH has a dose-dependent effect in both GC and sex differences in their release. HLR also showed sex differences that did not depend on the dose given.</p> <p><br></p> <p>Chapter 4 observed acute exposure of GC in ducks. Pekin ducks were assigned 10/sex/treatment to receive either IM control, cortisol, or corticosterone injections. In addition, a low-dose cortisol treatment was given to represent the endogenous levels of cortisol compared to corticosterone. The control injections contained safflower oil, which was chosen as vehicle due to the low levels of genistein present. This is important as genistein is a plant estrogen and this could interact with the GC and alter the results. Blood was collected at 0, 1, 2, and 3 hours after injections for serum analysis with ELISAs, and blood smears were collected for complete blood count (CBC) differentials. Significant sex x treatment x time interactions were notable in both GC. Hens had significant increases at the first hour after injections in all treatments compared to controls, and drakes had increases at 2 hours after injections in all treatments except the low-dose cortisol. </p> <p><br></p> <p>After observing the effect of acute stress in ducks, the next step was to investigate the effects of chronic stress in chapter 5. Adult breeder Pekin ducks were randomly distributed into 3 groups: corticosterone, cortisol, or control treatments. The GC were in crystalline steroid form distributed through 2 capsules that were subcutaneously implanted on the backs of the neck. The ducks in the control group were given empty capsules. Blood smears, blood draws for serum, egg collection, body weights, and organ samples were collected over a period of 2 weeks. For the results, the corticosterone implants elevated corticosterone levels in both sexes. Interestingly, cortisol levels were elevated in both GC treatments in both sexes. Cortisol elevated HLR in drakes 1 day after implants with no effect from corticosterone. Hens had elevated HLR from both GC at all timepoints throughout the experiment. There were no significant differences in morphometrics in either sex. Corticosterone was not present in eggs, but cortisol was elevated in the albumen on day 7 and 14 of the experiment. Overall, there were sex differences in HLR where hens had greater levels in both GC treatments.</p>