Global warming poses detrimental effects on poultry production leading to substantial economic losses. The goal of our experiment was to test the hypothesis that heat stress (HS) would alter welfare and egg quality (EQ) of breeder ducks. Furthermore, we wanted to test if HS would increase cortisol levels in egg albumen. 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), and egg production/quality were collected the week preceding heat treatment. Ducks were subjected to cyclic HS of 35°C for 10h/day and 29.5°C for the remaining 14h/day for 3 weeks while the control room was maintained at 22°C. Eggs were collected daily and analyzed weekly for quality assessment, and for albumen glucocorticoid (GCs) levels 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 and birds necropsied. Data analyses were done by 1- or 2-way ANOVA as appropriate with a Tukey-Kramer post hoc test. BCS were analyzed using a chi-squared test. A p ≤ 0.05 was considered significant. Circulating levels of corticosterone were significantly (p < 0.01) elevated at week 1 only in the HS hens. The circulating levels of cortisol increased significantly at week 1 and 2 (p < 0.05), and week 3 (p < 0.01) in the hens and at weeks 2 and 3 only (p < 0.05) in the drakes. Feather quality scores (p < 0.01), feather cleanliness scores (p < 0.001) and footpad quality scores (p < 0.05) increased significantly in the HS group. HS elicited a significant (p < 0.001) decrease in egg production at weeks 1 and 3. Hens in the HS group showed significantly decreased BW (p < 0.001) and number of follicles (p < 0.05). Shell weight decreased significantly at week 1 only (p < 0.05) compared to controls. Yolk weight decreased significantly at week 3 (p < 0.01) compared to controls. HS elicited a significant increase in albumen cortisol levels at week 1 (p < 0.05) and week 3 (p < 0.05). Thus, cortisol may provide critical information to further understand and to improve welfare.

The effects of HS on the welfare of poultry have been reported to have a transgenerational effect on phenotype plasticity. The goal of our experiment was to determine whether parental exposure to HS would impair the performance, HPA axis response, or behavior of their offspring. We treated adult drakes and hens (n = 80 ducks/treatment) at peak lay with HS or the control temperature for 3 weeks and incubated eggs collected from the last 3 days of the experiment. We utilized 76 ducklings/parental treatment group: control (CON-F1) and HS (HS-F1). Weekly data for body weights, body condition scores (BCSs), and novel object test (NOT) were collected. At 3 weeks of age, the ducks (n = 6/treatment) were subjected to adrenocorticotropic hormone (ACTH/cosyntropin, 0.0625 mg/kg) challenge or vehicle as the control. Blood samples were collected at 0, 1, 2, 3, and 4 h relative to treatment for serum glucocorticoid and heterophil-to-lymphocyte ratio (HLR) analyses. All injected birds were euthanized with pentobarbital on the second day relative to ACTH administration, and the spleen and bursa were removed and weighed immediately. Duck level analyses were completed using one- or two-way ANOVA as appropriate. BCSs were analyzed using a chi-squared test. The HS-F1 ducks had a lower hatch weight (p < 0.05) compared with the CON-F1 ducks but no significant difference in growth rates during the 5-week period. NOT (n = 4) analyses showed that the HS-F1 ducks had a greater fear response (p < 0.001) compared with the CON-F1 ducks. Similarly, an ACTH stimulation test showed that the HS-F1 ducks had significantly (p < 0.05) heightened corticosterone and HLR responses compared with the CON-F1 ducks. The HS-F1 ducks showed altered baseline and ACTH-stimulated levels of cortisol compared with the controls. Our data suggest that parental exposure to HS impacts the HPA response and fearfulness of the F1 generation in Pekin ducks.

During chronic stress, there is an initial increase in glucocorticoid (GC) levels, but they then return to low, albeit not baseline, levels. Recent studies have renewed interest in cortisol in that it may also have important roles in the stress response. The purpose of our study was to test the hypothesis that chronic treatment with low levels of either corticosterone or cortisol would alter 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 that contained corticosterone, cortisol, or empty capsules as controls (N = 5/sex/treatment). Blood serum, smears, body weights, and egg quality data were collected. Ducks were then euthanized and body weight, weights of spleens, livers, and the number of active follicles were recorded. Albumen GC levels were assessed using mass spectrometry. Data were analyzed using a 2- or 3-way ANOVA as appropriate and post-hoc with Fishers PLSD. No treatment elicited differences in egg quality measures or body weight compared to controls. 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, only elicited an increase in HLR for drakes (p &lt; 0.05) compared to controls but only 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. Our results suggest that glucocorticoids 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 bird welfare.

Rapid “fight-or-flight” responses to stress are largely orchestrated by the catecholamines. Moreover, catecholamines and catecholamine precursors are widely recognized to act as interkingdom signaling molecules among host and microbiota, as well as to serve as chemotactic signals for bacterial foodborne pathogens. While albumen and yolk concentrations of glucocorticoids have received extensive attention as non-invasive indicators of hen response to stress, little is known regarding the impact of the hen’s stress response on in ovo catecholamine and catecholamine precursor concentrations. The aim of the present study was to determine norepinephrine and L-dopa concentrations in albumen and yolk of eggs laid by hens maintained under normal or heat stress conditions. Norepinephrine and L-dopa concentrations were also measured in oviductal tissue. Breeder ducks (∼35 weeks/age) were kept under normal (22°C) conditions or subjected to cyclical heat stress (35°C day/29.5°C night) for 3 weeks. Eggs (n = 12 per timepoint/group) were collected on a weekly basis. Hens were sacrificed at baseline or after 3 weeks of heat stress for oviductal tissue collection. Albumen, yolk, and oviduct concentrations of norepinephrine and L-dopa were determined using ultra high-performance liquid chromatography with electrochemical detection. Norepinephrine and L-dopa were detected in oviductal tissue as well as egg albumen and yolk. Norepinephrine concentrations were elevated (p &lt; 0.05) in the yolk of eggs laid by the heat stress group compared to those of the control group. Norepinephrine concentrations in albumen were elevated (p &lt; 0.05) in the heat stress group compared to control group at week 2. L-dopa concentrations were not significantly affected (p &gt; 0.05) by heat stress in albumen, yolk, or oviductal tissue. Together, the present study provides the first evidence of the stress neurohormone, norepinephrine, in duck eggs and identifies that hen exposure to heat stress can affect in ovo norepinephrine concentrations. These data highlight the potential utility of in ovo catecholamine concentrations as non-invasive measures of the hen’s response to stress, as well as warrants future research into whether hen deposition of stress-related neurochemicals into the egg could serve as a chemotactic signal in the vertical transmission of foodborne pathogens.