A genetic selection study to determine the effects on egg hatchability and subsequent chick performance of Japanese quail (Coturnix japonica) eggs incubated at 100 F dry bulb temperature (Control, Line C) when compared to other eggs incubated at 102 F (Selected, Line S) was conducted over 10 consecutive generations. Eggs from a randomly mated population (designated as Generation 0) of Japanese quail maintained at the Oregon Agricultural Experiment Station were randomly allocated to two treatment groups (Lines C and S) and incubated at the different temperatures in separate but identical Jamesway 252 machines. On day 14 of incubation all eggs were transferred to a common hatcher (98.5 F). Using family-based selection, the chicks that hatched from the two lines were subsequently used as breeders (25 paired matings per line) and the resulting eggs from each line incubated at their respective temperatures for 10 consecutive generations. Following the 10th generation percent egg fertility and percent hatch of fertile eggs were greater in Line C vs. Line S (p<O.O3 and p<O.0001, respectively). Embryo development time was shortened in Line S by 24 hours and mean 4- or 5- week body weights were greater (p<0.001) in Line S. Ten-day post-hatch mortality increased greatly in Line S vs. Line C after generation 6 (p<0.001) and hen-day egg production decreased after generation 4 in Line S vs. Line C (p<0.0001). The results indicate that embryo development time can be reduced by high temperature incubation, but at the expense of reproductive traits such as egg production, fertility, and hatchability of fertile eggs. / Graduation date: 2005

Hypoxic responses in quail development differ depending upon stage, duration and level of oxygen partial pressure of embryo. Incubation was switched to/from 110mmHg partial pressure (hypoxia), to/from 150mmHg (normoxia) during different stages in development, and control was incubated in normoxia throughout. Hatchability and embryo survival resulted in no hatchlings in continuous hypoxia. Responses to various hypoxic exposures throughout development resulted in recovery/repair of hypoxic damage by hatch. Heart and body mass, beak and toe length, hemoglobin, and hematocrit were measured to determine embryo responses to hypoxia during development at days 10, 15, and hatch. Hypoxia seemed to have the most deleterious effects on eggs in continuous hypoxia. Collectively, data indicate critical developmental windows for hypoxia susceptibility, especially during mid-embryonic development.

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Appetite is a complex behavior which can be influenced by factors within the animal's body as well as the environment around it. Internal factors include hormonal and nutrient concentrations found in the blood stream and subsequent neuropeptide and neurotransmitter signaling in the hypothalamus. External factors, such as high ambient temperature (HAT), can indirectly affect appetite regulation through other neuroendocrine systems such as the hypothalamo-pituitary-adrenal (HPA) axis. Understanding the physiological responses to endogenous factors and HAT exposure in birds will have implications in both the agricultural and biomedical fields. Thus, the purpose of this dissertation research was to explore the hypothalamic molecular mechanisms associated with food intake in broiler type chickens and Japanese quail, and the effect of HAT exposure on food intake in broiler type chickens. Broiler type chickens have undergone intense artificial selection for traits that promote rapid growth, consequently driving them to consume feed incessantly. It is hypothesized that broiler type chickens lack a mechanism that signals satiety, causing them to eat significantly more than layer type chickens. Selection for rapid growth of meat (muscle tissue) has made the broiler more susceptible to the deficits associated with HAT exposure, as animals composed primarily of muscle dissipate less heat while also producing more heat than those composed of fat. The Japanese quail have undergone relatively minor artificial selection compared to the chicken, suggesting that use of this model may provide insight into the mechanisms of appetite regulation in wild-type bird species. This research involved administrating appetite associated factors into the avian brain via an intracerebroventricular (ICV) injection including gastrin releasing peptide, vasoactive intestinal polypeptide, neuropeptide AF, and prostaglandin D2. Additionally, I explored the effects of HAT on food intake, and on the efficacy of several ICV administered appetite associated factors including neuropeptide Y, corticotropin releasing factor and α-melanocyte stimulating hormone. After treatment administration, I measured changes in food intake and behavior, activation of hypothalamic nuclei including the arcuate nucleus, dorsomedial nucleus, lateral hypothalamus, paraventricular nucleus, and the ventromedial nucleus, and the nucleus of the hippocampal commissure. I then measured changes in gene expression in both whole hypothalamic samples and specific hypothalamic nuclei. The data from non-HAT associated studies provided information on the hypothalamic nuclei which respond to the various appetite associated factors and the molecular mechanisms mediating changes in appetite. The data from the HAT study provided information on the hypothalamic nuclei involved in the avian response to HAT exposure, and the molecular mechanisms involved in the effect on food intake. Overall, these data provide insight on the mechanisms associated with short-term regulation of appetite, and pathways associated with stress and food intake. / Doctor of Philosophy / Appetite regulation can be affected by factors both in the body and out in the environment. Understanding how both internal and external factors affect appetite regulation can have positive implications in both the agriculture industry as well as the biomedical field. In agriculture, animals exposed to high ambient temperatures often exhibit several deficits including immunosuppression, decreased body weight, and ultimately an increased risk of mortality. It is hypothesized that the factor linking negative wellbeing to heat exposure is a reduction in food intake. Animals aside, the prevalence of eating disorders has doubled worldwide every 6 years since the year 2000. These numbers have increased even more during the recent COVID-19 pandemic. In order to improve the wellbeing of both humans and animals exposed to stressing stimuli, it is imperative we understand how individual appetite associated factors affect food intake, and how external stressors can impact the normal physiology of the hypothalamus. Thus, the purpose of this dissertation was to elucidate the hypothalamic mechanisms mediating appetite regulation using broiler type chickens and Japanese quail as models. Related pathways and molecular mechanisms were explored for several appetite associated factors including gastrin releasing peptide, vasoactive intestinal polypeptide, neuropeptide AF, and prostaglandin D2. Additionally, the effect of high ambient temperature on food intake, on the efficacy of several appetite associated factors including neuropeptide Y, corticotropin releasing factor and α-melanocyte stimulating hormone, and the hypothalamic pathways and molecular mechanisms mediating heat-induced anorexia were assessed.

Adrenal corticosterone and aldosterone content, body weight changes and serum osmolality were examined during water restriction and water deprivation. Progressive water deprivation resulted in increased serum osmolality and decreased body weight; adrenal aldosterone content did not change. Adrenal corticosterone content tended to be elevated during early water deprivation indicating a stress response, but tended to decrease after seven days of water deprivation suggesting adrenal fatigue. During water restriction, after the period of weight loss, adrenal elevated corticosterone content and serum osmolality were elevated. As the birds began to gain weight aldosterone did not change, but adrenal corticosterone content and serum osmolality approached control values, suggesting the birds were beginning to adapt to the water restriction. The lack of an aldosterone response suggests that high sodium in the diet was more important than the water regimes in regulating aldosterone. The adrenal was sensitive to ACTH as indicated by the elevated adrenal aldosterone and corticosterone content after ACTH injection, however sodium status probably affects the aldosterone response to ACTH. / Master of Science

Feeding behavior is a complex trait that is regulated by various hypothalamic neuropeptides and neuronal populations (nuclei). Understanding the physiological regulation of food intake is important for improving nutrient utilization efficiency in agricultural species and for understanding and treating eating disorders. Knowledge about appetite in birds has agricultural and biomedical relevance and provides evolutionary perspective. I thus investigated hypothalamic molecular mechanisms associated with appetite in broilers, layers, chicken lines selected for low (LWS) or high (HWS) body weight, and Japanese quail, which provide a unique perspective to understanding appetite. Broiler-type chicks have been genetically selected for rapid growth and consume much more feed than do layer-type chicks which have been selected for egg production. Long-term selection has caused the LWS chicks to have different severities of anorexia while the HWS chicks become obese, thus making these lines a valuable model for metabolic disorders. Lastly, the Japanese quail have not undergone as extensive artificial selection as the chicken, thus this model may provide insights on how human intervention has changed the mechanisms that regulate feeding behavior in birds. This research involved applying a variety of different treatments including fasting and refeeding, diets differing in macronutrient composition, and/or central administration of neuropeptide Y, xenopsin, neuropeptide K, oxytocin, mesotocin, gonadotropin-inhibitory hormone, and prolactin-releasing peptide, after which I measured feeding behavior and various aspects of hypothalamic physiology. I measured nuclei activation in hypothalamic appetite-associated regions including the lateral hypothalamus, paraventricular nucleus, ventromedial hypothalamus, dorsomedial nucleus, and arcuate nucleus and I measured gene expression of various appetite-associated factors in the whole hypothalamus and individual nuclei. These data provided information about the regions of the brain involved in mediating effects on appetite and the molecular pathways involved in the effect on appetite. There were differences in dose threshold sensitivity to various injected factors in the different stocks, differential responses to fasting and refeeding, and differences in nuclei and genes that were activated in response to the various treatments. These data provide valuable insights on the molecular mechanisms that are associated with the short-term regulation of feeding behavior and pathways that may be genetically stock-dependent. / PHD

Many aspects of biological function are affected by hormones, from physiology to behavior, and the synthesis and release of hormones in vertebrates are regulated by the endocrine axes of control. A growing body of research shows that the mechanisms underlying the endocrine axes of control are complex and interconnected, with many hormones having multiple effects, and with many interactions between axes. In this study, I examined the effects of decreased thyroid function on the glucocorticoid stress response in Japanese quail, a potential interaction between the hypothalamic-pituitary thyroid (HPT) and hypothalamic-pituitary adrenal (HPA) axes of control. I used the thyroid inhibitor ammonium perchlorate (AP) for 2 weeks and 5 weeks to induce two states of decreased thyroid function: a thyroid challenged state, in which birds have depleted thyroidal T4 content, but still maintain euthyroid (normal) concentrations of plasma T4, and a hypothyroid state, in which birds have depleted thyroidal T4 content and decreased concentrations of plasma T4. Thyroid function was assessed by measuring plasma T4 concentrations, thyroidal T4 content, and thyroid gland mass. I took blood samples from birds both immediately prior to and immediately following a 30 minute confinement and agitation stressor to evaluate the effects of decreases in thyroid function on basal and stress-induced plasma corticosterone and plasma T4 concentrations. I found two key results: First, although baseline levels of plasma corticosterone were unchanged, the corticosterone stress response was significantly blunted in both the thyroid challenged and hypothyroid birds as compared to controls. This finding suggests that the HPT and HPA axes are functionally connected in birds, and other evidence suggests this connection is likely at the pituitary or hypothalamic level. Second, in hypothyroid birds, plasma T4 concentrations were elevated (into the euthyroid range) in response to the experimental stressor, although no change in plasma T4 was observed in thyroid challenged or control birds. This finding suggests that plasma T4 may have a permissive role in mounting a stress response. / Master of Science in Life Sciences