POTENTIAL EFFECTS OF PARENTAL HEAT STRESS EXPOSURE ON HYPOTHALAMIC-PITUITARY-ADRENAL AXIS SENSITIVITY THROUGH EPIGENETIC PROCESSES.
Creators: name: Oluwagbenga, Esther Mary
Publisher: name: Purdue University Graduate School
Year: 2023
Subjects: Animal welfare
Environmental studies in animal production
Animal management

description: Heat stress affects breeder ducks raised in North America and other parts of the world, but the effects of such stress on the progenies is not known. Therefore, the objectives of this study were to investigate: 1) The objectives of this thesis were to first investigate the effect of heat stress or exposure to exogenous glucocorticoid (GC) on fertility, production performance, egg biochemistry, egg quality, and welfare of breeder Pekin ducks. 2) the effects of maternal GC on phenotypic plasticity and behavior of the F1 generation. Three studies were carried out to investigate these objectives. The first experiment was conducted to test the hypothesis that chronic treatment with low levels of either corticosterone or cortisol would alter heterophil to lymphocyte ratio (HLR) and immune organ morphometrics. Further, we wanted to determine if chronic treatment with either GC would elicit an increase in cortisol levels in egg albumen. To test our hypotheses, we implanted silastic capsules subcutaneously under the skin of the neck of adult ducks (n = 5/sex/dose) using propofol anesthesia. Capsules contained corticosterone, cortisol, or empty capsules as controls. Over the course of 2 weeks, blood serum, blood smears, body weights, and egg quality data were collected. After 2 weeks, ducks were euthanized using pentobarbital (FatalPlus, 396 mg/ml/kg) and body weight, weights of spleens, livers, and the number of active follicles were recorded. Blood smears were analyzed for HLR by a lab unaware of the treatment groups. Albumen GC levels were assessed using mass spectrometry. Data were analyzed using a 2- or 3-way ANOVA as appropriate and <em>post hoc </em>with Fishers protected least squares difference (PLSD). There were no treatment effects on egg quality measures or body weight. Corticosterone treatment did elicit an increase in serum corticosterone (p &lt; 0.05), but not cortisol levels, compared to controls in both sexes. Both cortisol and corticosterone treatments increased (p &lt; 0.05) serum levels of cortisol compared to controls. Relative spleen weights were higher (p &lt; 0.05) in hens following corticosterone but not cortisol treatment. No other organs showed any differences among the treatment groups. Both GCs elicited an increase (p &lt; 0.001) in HLR in hens at all time-points over the 2-week treatment period compared to controls. Cortisol, not corticosterone, elicited an increase in HLR for drakes (p &lt; 0.05) compared to controls at day 1 after implants. Chronic treatment with cortisol, but not corticosterone, elicited an increase (p &lt; 0.01) in egg albumen cortisol levels compared to other groups. Corticosterone was not detected in any albumen samples. The goal of our second experiment was to test the hypothesis that heat stress (HS) would alter welfare, egg quality, and morphometrics of breeder ducks. Furthermore, we wanted to test if HS would increase cortisol levels in egg albumen due to recent exciting findings that cortisol, not corticosterone, is isolated in egg albumen. To test our hypothesis, adult Pekin ducks were randomly assigned to two different rooms at 85% lay with 60 hens and 20 drakes per room. Baseline data including body weight, body condition scores (BCS) (such as footpad quality, eyes, nostrils, feather cleanliness, and feather quality scores), and egg production/quality were collected the week preceding heat treatment. Ducks were subjected to cyclic HS of 350C for 10h/day and to 29.50C for the remaining 14h/day for 3 weeks while the control room was maintained at 220C. Eggs were collected daily, and body weights were taken on days 0 and 21 relative to the onset of heat treatment. BCS were collected weekly. Eggs were collected weekly for quality assessment and albumen glucocorticoid (GCs) levels assessment using mass spectrometry. One week before the exposure to HS, 10 hens and 5 drakes were euthanized and the same number again after 3 weeks of HS or control exposures using pentobarbital and birds necropsied. Body weight, weights of the liver, spleen, and the number of maturing follicles were recorded. Data analyses were done by 2- or 3-way ANOVA as appropriate with a Tukey-Kramer post hoc test. BCS were analyzed using a chi-squared test. A p value ≤ 0.05 was considered significant. Circulating levels of corticosterone were significantly (p &lt; 0.01) elevated at week 1 only in the HS hens while there was no significant difference in the circulating levels of corticosterone in drakes compared to the controls. The circulating levels of cortisol increased significantly at week 1 (p &lt; 0.05), week 2 (p &lt; 0.05), and week 3 (p &lt; 0.01) in the hens and at week 2 and 3 only (p &lt; 0.05) in the drakes compared to the controls. Feather quality scores (p &lt; 0.01), feather cleanliness scores (p &lt; 0.001) and footpad quality scores (p &lt; 0.05) increased significantly in the HS group compared to controls, higher BCS indicate a decline in welfare. HS elicited a significant (p &lt; 0.001) decrease in egg production at weeks 1 and 3 and a descriptive decrease in the number of fertile eggs upon candling at 10 days of incubation, numeric decrease hatchability and increase in the number of dead embryos in the HS group after the incubation period. Hens in the HS group showed a significantly decreased BW (p &lt; 0.001), and number of ovarian follicles (p &lt; 0.05) compared to controls. Shell weight decreased significantly at week 1 (p &lt; 0.05) compared to controls. Yolk weight decreased significantly at week 3 (p &lt; 0.01) compared to controls. HS elicited a significant increase in albumen cortisol levels at week 1 (p &lt; 0.05) and week 3 (p &lt; 0.05). The third experiment was conducted to determine if parental exposure to heat stress would impair performance, hypothalamic pituitary adrenal (HPA) axis response, welfare, or behavior of their offspring. To achieve these goals, we treated adult drakes and hens at peak lay to heat stress or control temperature for 3 weeks and incubated eggs collected from the last 3 days of the experiment. A total of 76 ducklings were placed into pens from each parental treatment group: control (CON-F1) and heat stress (HS-F1) and raised as grow-out ducks. Weekly data for body weights, body condition scores (BCS), and novel object test (NOT) were collected weekly. At 3 weeks of age, ducks (n = 6 per treatment group) were subjected to adrenocorticotropic hormone (ACTH) (ACTH/cosyntropin, 0.0625 mg/kg) challenge or vehicle as control. Blood samples were collected from the metatarsal vein into serum-separator tubes at 0, 1, 2, 3, and 4 hours relative to treatment for the determination of serum glucocorticoids. Blood smears were also produced from these same samples to determine heterophil to lymphocyte ratios (HLR). All injected birds were euthanized with pentobarbital on the second day relative to ACTH administration, spleen and bursa were removed and weighed immediately. Duck level analyses were completed using 1-, or 2 -way ANOVA as appropriate. BCS were analyzed using a chi-squared test. We observed that HS-F1 had a lower hatch weight (p &lt; 0.05) compared to CON-F1. However, growth rates during the 5-week grow-out period were not significantly different between the two flocks. NOT (N = 4) analyses showed that the HS-F1 had a greater fear response (P&lt; 0.001) compared to CON-F1. Similarly, an ACTH stimulation test showed that the HS-F1 ducks had significantly heightened corticosterone and HLR responses compared to CON-F1 ducks (p &lt; 0.05). The HS-F1 showed altered baseline and ACTH-stimulated levels of cortisol compared to controls. In conclusion, GC elicit differential effects and although corticosterone has been stated to be the predominant GC in avian species, cortisol may provide critical information to further understand and improve welfare. HS decreased performance, fertility, and productivity of breeder ducks. In addition, HS and exogenous GC elicited a selective deposition of cortisol, not corticosterone, in the egg albumen. The maternal cortisol deposited in eggs alter the hypothalamic-pituitary adrenal (HPA) axis and behavioral responses of the F1 generation. This suggests that maternal hormones can alter the phenotypic plasticity of the offspring and can be used to produce offspring that have better adaptation to the rising temperatures as a result of climate change. Finally, the measure of cortisol in egg albumen can be used as a non-invasive marker of stress.