Sex differences in serum glucocorticoid levels and heterophil:lymphocyte ratios in adult pekin ducks (Anas platyrhynchos domesticus).

It is becoming more common for poultry scientists to utilize direct and indirect measures of stress hormones to monitor bird welfare. However, it has been clear that our understanding of the avian hypothalamic-pituitaryadrenal axis (HPA) is insufficient as evidenced by the many conflicting reports regarding stress responses, such as transportation stress, in poultry. It has long been assumed that the poultry HPA functions similarly to that of mammals, but now we know that there are considerable differences in the avian HPA compared to mammals. Synthesis and release of glucocorticoids (GC) are stimulated by adrenocorticotropin hormone (ACTH); GC are synthesized from a common pathway that begins with cholesterol and pregnenolone. The synthesis of one of the glucocorticoids does not depend upon the synthesis of the other. The purpose of our study was to test the hypothesis that ACTH will stimulate both corticosterone and cortisol release in ducks. To test this hypothesis, we injected artificial ACTH (cosyntropin; 0.0625 mg/kg, 0.031 mg/kg, or 0.016 mg/kg or saline as control) intramuscularly into adult drakes and hens (N = 10/sex/dose). Both glucocorticoids (GC) were assayed in serum using previously verified ELISAs. Blood smears were also assessed for heterophil to lymphocyte ratios (HLR). Data were analyzed by repeated measures 3-way ANOVA with Fishers PLSD as an ad hoc test. We observed that both GC were secreted in significantly (p = 0.0002) different patterns in a dose-dependent manner compared to controls, and that there was a significant (p = 0.0001) sex difference in both GC compared to saline controls. Further, we observed that all doses of ACTH elicited a significant (p = 0.004) sex difference in the HLR response compared to controls, but no dose-dependent effects were noted. Our data suggest that ducks, at least, may utilize more than just corticosterone to maintain physiological homeostasis in response to stress. Further, the time course of the stressor to release GC and subsequent HLR response may be dependent upon sex. More detailed analyses of the HPA are necessary in all avian species to better understand stress responses as we utilize biological bases for welfare assessments and stress responses.

Immunolesion of melanopsin neurons causes gonadal regression in Pekin drakes (Anas platyrhynchos domesticus).

Several light sensitive receptors have been described in the avian brain that are thought to regulate the reproductive axis independently from the eyes and pineal gland. Recently, our lab has described the presence of three of these photoneuroendocrine systems in the Pekin duck: opsin, opsin 5, & melanopsin. We set out to test the hypothesis that melanopsin receptive neurons are necessary to maintain seasonal reproductive status along with growth and development in the Pekin drake. To accomplish these goals we first investigated 50-week-old Pekin drakes that were housed in the aviary at Hope College under long day length (18h lights on) conditions in floor pens. To specifically lesion melanopsin-receptive neurons, 3µl of an anti-melanopsin-saporin conjugate (MSAP, 100ng/ul) was injected into the lateral ventricle (n=10). Control drakes were injected with 3µl of equimolar unconjugated anti-melanopsin and saporin (SAP, n=10). Reproductive behaviors were analyzed weekly in a test pen with adult hens and MSAP drakes showed a significant (p<0.01) reduction in reproductive behaviors after week 2. After 5weeks, drakes were euthanized and body weights were measured, and brains, pituitaries, and testes collected and stored for analyses. Mature MSAP-treated drakes had significantly (p<0.001) reduced relative teste weights compared to SAP controls. qRT-PCR analyses of hypothalamus showed a significant reduction (p<0.001) in GnRH and melanopsin mRNA levels, but not opsin 5, vertebrate ancient opsin, or opsin 2 (rhodopsin). Immunocytochemical analyses showed a significant reduction (p<0.01) in tyrosine hydroxylase-immunoreactivity in the PMM. These data suggest that although blue light alone is not able to maintain testicular function, the blue-light sensitive melanopsin activity is critical to maintain gonadal function.

Increased water contamination and grow-out Pekin duck mortality when raised with water troughs compared to pin-metered water lines using a United States management system.

Controversy has developed as to whether or not pin-metered water lines or water troughs are more appropriate for Pekin ducks. We hypothesized that water troughs would show improved duck body conditions and environmental quality compared to pin-metered water lines. To test this hypothesis, we housed ducks in 2 barns, one with water lines and one with water troughs. Water troughs were constructed to meet RSPCA guidelines for number and density of ducks and with recently described verandas. Ducks were divided into 4 pens per barn (n=1,000 ducks/pen). The study was then repeated (n=8 pens per water source) in a cross-over design so the barns each contained the opposite water source to the first experiment. We scored the ducks' body condition using an established scoring rubric and analyzed using SAS Proc GLM-Mix as binomial data. Ducks housed with water troughs showed higher (thus worse condition; P<0.001) scores for eyes, nostrils, feather quality, feather cleanliness, and foot pads. We also compared water condition, water quality, and duck mortality using a Student t test for both water sources each week. We found that the water troughs showed higher iron (P<0.001), nitrites (P<0.001), pH (P<0.01), and bacterial growth (P<0.001). The bacterial growth was shown to have higher (P<0.001)E. coli, coliforms, and Staphylococcusin the water troughs. Water lines typically showed no bacterial growth in culture-based assays. Ducks housed with water troughs used greater (P=0.001) volumes of water compared to ducks housed with water lines. Ducks with water troughs also showed a greater percent (P=0.008) mortality at all ages compared to ducks with water lines. These data suggest that water troughs may not be beneficial for duck welfare and could adversely impact both environment and duck or human health.

Effects of different wavelengths of light on the biology, behavior, and production of grow-out Pekin ducks.

Previous research has shown that red light conditions may improve growth and decrease aggressive behaviors in chickens and turkeys; however, more recent studies suggest that blue-green light may improve production of broilers over red light. To date, no research has been conducted to examine whether different wavelengths of light have an impact on production in the Pekin duck. To determine this, we raised Pekin ducks under aviary conditions that were similar to standard commercial barns. The ducks were kept in 3 different pens: red light (approximately 625 nm), blue light (approximately 425 nm), and white light. Light sources in each pen were standardized to produce a peak energy at 1.6 × 10³ µM photons/m²/s at the level of the ducks' heads. Ducks were given ad libitum access to water and commercial duck diet, and were housed on pine shavings at a density of 0.43 m²/duck. Ducks were evaluated weekly for BW and condition and a subjective measure of the duck's anxiety levels was determined. We found that ducks housed under blue light had significantly (P < 0.01) reduced BW at every age until the end of the study (processing age; 35 d). Unlike ducks housed under red or white light, ducks housed in the blue pen showed a higher level of anxiety; while evaluators were in the pen a majority of them began panting, they were much less inquisitive than other ducks, they took longer to exhibit normal social behavior once evaluation was completed, and they frequently "swarmed" when no people were present. There were no differences in any measurements between the red and white-lighted pens. These data suggest that unlike the chicken, blue lights may be inappropriate for raising Pekin ducks in a commercial setting.

The effects of environmental enrichment devices on feather picking in commercially housed Pekin ducks.

Like other poultry species, Pekin ducks occasionally show an auto-mutilation behavior referred to as feather picking. Self-picking can lead to further pecking by conspecifics and ultimately to reduced feather quality and poor overall health of the bird. Although the reasons underlying feather picking are not clear, it appears to occur when the ducks are transitioning between downy feathers and adult plumage, between 17 and 22 d of age. We hypothesized that giving Pekin ducks a substitute outlet for this behavior in the form of environmental enrichment devices (EED) would decrease feather picking and improve feather quality and duck health and welfare. The EED were plastic Wiffle-style balls, each threaded with 4 zip-ties. In the first experiment, we set out to determine that placement of EED would not induce fear or harmful behaviors. Five barns were each divided into 2 pens, holding an average combined total of approximately 4,500 ducks with one pen per barn used as control. Upon placement of the EED in one pen per barn, both pens were videotaped for a total of 2 h per day. The physical characteristics of 100 ducks per pen were scored at age 7, 21, 28, and 35. Results showed a decrease (P = 0.034) in both self-picking and conspecific-pecking in pens with EED compared with pens without EED. Although no differences in body condition scores were observed at 7 d of age, by d 21 ducks with EED showed better (P = 0.021) feather quality and cleanliness scores compared with ducks without EED. In a second experiment, we set out to determine if ducks had a color preference for blue/green, red, or white EED. Again, ducks with EED showed reduced (P = 0.038) feather picking compared with ducks without EED. Ducks interacted considerably more (P = 0.00089) frequently with blue/green EED then either red or white EED. These results suggest that providing environmental enrichment may minimize feather picking and improve feather quality and duck welfare.

Sex differences in intestinal morphology and increase in diencephalic neuropeptide Y gene expression in female but not male Pekin ducks exposed to chronic heat stress.

The impact of heat stress (HS) on production is intricately linked with feed intake. We investigated the effects of HS on intestines and diencephalic genes in Pekin ducks. One hundred and sixty adult ducks were allocated to two treatment rooms. The control room was maintained at 22°C and the HS room at 35°C for the first 10 h of the day then reduced to 29.5°C. After 3 weeks, 10 hens and 5 drakes were euthanized from each room and jejunum and ileum collected for histology. Brains were collected for gene expression analysis using qRT-PCR. Intestinal morphology data were analyzed with two-way ANOVA and diencephalic gene data were analyzed with Kruskal-Wallis test. There was an increase in villi width in the ileum (p = .0136) and jejunum (p = .0019) of HS hens compared to controls. HS drakes showed a higher crypt depth (CD) in the jejunum (p = .0198) compared to controls. There was an increase in crypt goblet cells (GC) count in the ileum (p = .0169) of HS drakes compared to HS hens. There was higher villi GC count (p = .07) in the jejunum of HS drakes compared to controls. There was an increase in the crypt GC density (p = .0054) in the ileum, not jejunum, of HS drakes compared to HS hens. Further, there were no differences in the proopiomelanocortin gene expression in either sex but there was an increase in the expression of neuropeptide Y (NPY) gene in HS hens (p = .031) only and a decrease in the corticotropin releasing hormone gene in the HS drakes (p = .037) compared to controls. These data show that there are sex differences in the effect of HS on gut morphology while the upregulation in NPY gene may suggest a role in mediating response to chronic HS.

Vocalizations of the Pekin duck (Anas platyrhynchos domesticus): how stimuli, sex, and social groups affect their vocal repertoire.

Pekin ducks are exposed to stressors such as heat stress, enteric pathogens, mycotoxins, and other environmental stressors. We know from wild bird literature that birds communicate through vocalizations. We hypothesized that Pekin ducks would have a diverse repertoire that is affected by the sex, social group, and specific stimuli. We utilized adult Pekin ducks to develop a vocal repertoire. We placed 1 to 4 ducks of varying sexes into a sound chamber with various stimuli used to encourage new vocalizations. Birds were recorded for 20 min with several variations of number and sexes of ducks. Once the ducks were recorded each vocalization that was clipped was named based on a predetermined naming system. We characterized the vocal system of the ducks under each stimulus and social treatment in 4 ways: overall call rates, call diversity, call repertoire, and call spectral properties. In all cases, normality of residuals and homogeneity of variances for GLM and ANOVA models were confirmed using Proc Univariate (SAS v9.4) where a p ≤ 0.05 was considered significant. We found that Pekin ducks produce up to 16 different vocalizations. The treatments had a significant effect on the overall rate of calls given by the ducks (ANOVA: F6,31 = 8.55, p < 0.0001). Ducks produced the most calls by far when someone was sitting in the chamber with them (30.04 ± 4.45 calls/min). For call diversity, we found that there was a significant main effect of hen number (F218 = 12.21, p = 0.0004) but no main effect of drake number (F3,18 = 3.04, p = 0.0555). Cluster analyses indicated that certain types of calls were given under specific conditions. There were generally 6 major clusters of vocal repertoires (R-square = 0.899, Cubic Clustering Criterion = 9.30). Our results suggest that Pekin ducks are affected by the types of stimuli and social environment in how much they vocalize and in the properties of the calls they use. In addition, males and females differ somewhat in the repertoire of the calls they use, and in the spectral properties of their calls.

Preening cups in duck housing are associated with an increase in central dopamine activity that suggests a negative affective state.

Preening cups are a form of environmental enrichment that provides Pekin ducks a semi-open water source to express their natural behaviors. We recently observed that preening cups may increase feather pecking behaviors in ducks. Thus, we set out to determine if this form of enrichment can impact the affective state of Pekin ducks. To accomplish this goal, we evaluated the effect of preening cups on serotonin (5-HT) and dopamine (DA) turnover via mass spectrometry and their respective synthetic enzyme gene expression via qRT-PCR. Our study investigated the link between aggressive pecking with levels and activity of brain 5-HT and DA. Brain 5-HT and DA levels and activity have been established for decades to be associated with affective states. Grow-out Pekin ducks (n = 260) were housed at Purdue and raised per industry standards. On day 18, brains were collected from ducks in pens before preening cups were placed (PRE, n = 6) and, again on day 43, in pens with (PC, n = 6) and without (CON, n = 6) preening cups. Brains were dissected into right and left halves, then further microdissected into 4 brain areas: caudal mesencephalon (CM), rostral mesencephalon (RM), diencephalon (DI), and forebrain (FB). The right hemisphere was used for mass spectrometry to determine the neurotransmitter concentration (ng/mg of tissue) and those concentrations were applied to neurotransmitter turnover equations. There were no differences across treatments for 5-HT turnover in any brain area. There were differences in DA turnover across age (P = 0.0067) in the CM and across treatments (P = 0.003) in the RM. The left hemisphere of the brain was used to perform qRT-PCR on the genes of 5-HT and DA production enzymes. Within the CM, day 43 duck brains had increased (P = 0.022) tryptophan hydroxylase and tyrosine hydroxylase relative mRNA levels. All other brain areas showed no differences. Our data suggest that ducks housed with preening cups and that showed increased feather pecking are associated with increased brain DA activity. The increased DA in the brain may lead to a predisposition for increased aggression in the form of feather pecking.

Effects of different dietary vitamin D combinations during the grower phase and the feed restriction phase on growth performance and sternal morphology, mineralization, and related genes expression of bone metabolism in Pekin ducks.

Our study aimed to investigate the effects of different dietary vitamin D (VD) combinations during the grower (1-32 d of age) and feed restriction (33-52 d of age) phases on growth performance. We also evaluated sternal morphology, mineralization, and related genes expression of bone metabolism as well as absorption of calcium and phosphorous in duodenal mucosa and kidney in Pekin ducks. During the grower phase, we used 2 VD regimes (Group A: 3,160 IU/kg VD3; Group B: 400 IU/kg VD3 + 69 µg/kg 25-OH-D3). Each dietary treatment had 50 replicate pens of 10 ducks per pen. During the feed restriction phase, 30 replicate pens selected from Group A and Group B, repetitively, were redivided into 5 different dietary VD regimes to form a 2 × 5 experimental design. Each group consisted of 6 replicates, each with 10 ducks. During the feed restriction phase, we evaluated 5 different dietary VD combinations were as follows: T1: 2,000 IU/kg VD3 ; T2: 5,000 IU/kg VD3; T3: 3,620 IU/kg VD3 + 34.5 µg/kg 25-OH-D3; T4: 2,240 IU/kg VD3 + 69 µg/kg 25-OH-D3; T5: 1,800 IU/kg VD3 + 80 µg/kg 25-OH-D3). Results showed that Group B combinations with T5 had a better growth performance and breast meat deposition (P < 0.1). Regardless of 5 dietary VD regimes during the feed restriction phase, Group B significantly increased (P < 0.05) 52 d sternal depth and tended to increase (P < 0.1) 52 d sternal defatted weight, ash content, and phosphate (P) content of ducks. A significant interactive effect (P < 0.05) was observed on the mRNA abundance of DMP1 and Sost1 as well as RANKL/OPG in sternum and of VDR in duodenal mucosa of ducks at 52 d of age between dietary VD combinations during 2 phases. These results indicated that dietary VD regimes during the grower phase could affect the effectiveness of dietary VD regimes during the feed restriction phases; Dietary VD combinations of both phases could affect the genes expression of bone formation and the absorption as well as reabsorption of calcium and phosphorus in duodenum and kidney.

Distribution and sequence of gonadotropin-inhibitory hormone and its potential role as a molecular link between feeding and reproductive systems in the Pekin duck (Anas platyrhynchos domestica).

The reproductive status of adult Pekin drakes is very sensitive to nutritional status. Thus, the purpose of this study was to increase our understanding of the neurobiology underlying the depressive effect of fasting on the secretion of reproductive hormones. It was hypothesized that this effect was mediated by gonadotropin-inhibitory hormone (GnIH). Networks of GnIH fibers were present throughout the diencephalon, and cell bodies were present primarily, in the hypothalamic paraventricular nucleus (PVN). The duck GnIH gene was cloned and sequenced and found to encode GnIH and two GnIH-related peptides (GnIH-RP1, GnIH-RP2) which have a similar identity to those found in other avian species. Intracerebroventricular injection of GnIH, but not of GnIH-RP1, depressed plasma LH and stimulated feeding. Fasting for 48h depressed plasma LH and induced fos expression in about half the population of GnIH-ir neurons. These data suggest that GnIH neurons are mediators between feeding and reproductive systems in Pekin drakes.

Influence of raised plastic floors compared with pine shaving litter on environment and Pekin duck condition.

Commercial poultry production management practices have been under increased public scrutiny driven by concerns for food safety and animal welfare. Within the United States, wood shavings and raised plastic floors are common flooring systems used in duck production. It is intuitive that each flooring type would present different management challenges influencing physical characteristics of growing ducks. This study evaluated the relationship between flooring type and duck condition during the winter. Random samples of 20 ducks from 5 predetermined areas (n = 100) were examined in commercial duck houses (n = 9, litter; n = 11, raised plastic slats). Ducks were assessed at 7, 21, and 32 d of age for eye, nostril, and feather cleanliness, feather and foot pad quality, and gait. The data were analyzed to determine the proportion of ducks with a given score. In both housing types, the proportion of 0 scores for foot pad quality improved during the production cycle (P < 0.0001). Feather hygiene declined with age in ducks reared on litter flooring, whereas ducks reared on slatted flooring had cleaner feathers at d 32 (P < 0.011). With the exception of foot pad scores, the majority of ducks had no detectable problems for any single trait. The only main effect due to flooring pertained to feather quality with the proportion of ducks having a 0 or 1 score greater in litter flooring systems than slats (P < 0.05). Overall, the condition of ducks reared, regardless of flooring system, was considered to be good.

Influence of plastic slatted floors compared with pine shaving litter on Pekin Duck condition during the summer months.

The management and well-being of commercial Pekin ducks has been studied in the European Union where straw is the predominant litter source. In the United States, however, the most prevalent litter is wood shavings, with a recent trend toward using plastic slatted flooring. A previous study in the United States evaluated the relationship between flooring type (litter, slats) and duck condition during winter months and found very few differences between the 2 in terms of overall duck condition. The purpose of the current study was to reevaluate the 2 flooring systems during the summer months to determine if seasonal differences would interact with flooring type to have an impact on duck condition. Eighteen commercial barns that produce Pekin ducks for Maple Leaf Farms Inc. (Leesburg, IN), located in northern Indiana and southern Wisconsin (n = 9 litter; n = 9 raised slatted floor), were used for this study. Twenty ducks were randomly selected from 5 predetermined areas within each house (n = 100 total) and scored for eye condition, nostril and feather cleanliness, and feather and foot pad quality at 7, 21, and 32 d of age. Environmental data, including carbon monoxide, ammonia, RH, and temperature, were also obtained at each collection day. The only statistical differences in body condition occurred at 7 d; there were more ducks with clear eyes and eye rings on the litter flooring, whereas average nostril scores were better on the plastic slatted floors. Live weight, weight gain per day, flock mortality, and condemnations at the plant were collected, and the only statistical difference was a higher gain per day for ducks reared on slatted floors compared with litter (P < 0.05). There were no differences between flooring systems in the environmental parameters measured within the barns. In summary, there were very few differences between the litter and slatted flooring systems, indicating that there may not be clear advantages for one particular flooring system over the other from the point of view of duck well-being and production.

Research Note: Retinal cryptochrome gene expression is not altered by presence of light in incubators.

Cryptochromes are blue-, or ultraviolet-, light-absorbing proteins involved in the circadian clock, blue/ultraviolet light perception and potentially magnetoreception. At least 4 separate cryptochrome genes have been identified in avian species. The purpose of this study was to first determine if cryptochrome genes are expressed in the developing duck retina, and second to determine if the presence of lights in incubators affects the expression of cryptochrome genes. To accomplish these goals, duck eggs were placed in one of 2 commercial incubators (Buckeye, Single Stage Incubator, Model, SS-112) at Maple Leaf Farms, Inc., one with "poultry" LEDs obtained from a commercial source (Once Innovation, Agrishift) and the other in the absence of light (dark). Eggs in the incubators were placed on a reciprocating tray, tilting to 45° to simulate the rotation of eggs; thus all eggs spent 50% facing the light source and the other 50% of time facing 45° away from light source. Temperature gradients and humidity were maintained at industry standards. Retinal tissue samples from light and dark incubators were collected on days 3, 7, 11, 16, and 21 of incubation (extraction day, ED) known to be anatomical hallmarks of visual system development (n = 9-18 treatment group/ED timepoint). Samples were prepped and assayed for Cry2 and GAPDH gene transcription using qRT-PCR. Data were analyzed by using 2-ddCt method and a 2-way ANOVA was performed. No significant differences in Cry2 gene expression were observed between the lighted or dark incubator (P > 0.10). When combining light and dark treatment groups there is a significant 9 P < 0.05) increase in retinal Cry2 at ED 21, compared to ED 3 and 7. The presence of cryptochrome does not necessitate a migratory drive as evidenced by the fact that the Cry2 expression has been shown in non-migratory birds. However, since blue/ultraviolet wavelengths also activate the Cry2 photoreceptor, its presence could explain reports that suggest duck welfare can be improved if housed under lights that include ultraviolet wavelengths.

Heat stress and poultry production: a comprehensive review.

The impact of global warming on poultry production has gained significant attention over the years. However, our current knowledge and understanding of the mechanisms through which heat stress (HS) resulting from global warming affects the welfare, behavior, immune response, production performance, and even transgenerational effects in poultry are still incomplete. Further research is needed to delve deeper into these mechanisms to gain a comprehensive understanding. Numerous studies have investigated various biomarkers of stress in poultry, aiming to identify reliable markers that can accurately assess the physiological status and well-being of birds. However, there is a significant amount of variation and inconsistency in the results reported across different studies. This inconsistency highlights the need for more standardized methods and assays and a clearer understanding of the factors that influence these biomarkers in poultry. This review article specifically focuses on 3 main aspects: 1) the neuroendocrine and behavioral responses of poultry to HS, 2) the biomarkers of HS and 3) the impact of HS on poultry production that have been studied in poultry. By examining the neuroendocrine and behavioral changes exhibited by poultry under HS, we aim to gain insights into the physiological impact of elevated temperatures in poultry.

Chronic treatment with glucocorticoids does not affect egg quality but increases cortisol deposition into egg albumen and elicits changes to the heterophil to lymphocyte ratio in a sex-dependent manner.

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 < 0.05), but not cortisol, levels compared to controls in both sexes. Both cortisol and corticosterone treatments increased (p < 0.05) serum levels of cortisol compared to controls. Relative spleen weights were higher (p < 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 < 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 < 0.05) compared to controls but only at day 1 after implants. Chronic treatment with cortisol, but not corticosterone, elicited an increase (p < 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.

Preening cups increase apparent wet preening behaviors, but have no impact on other behaviors, body condition, growth, or body morphometrics of grow-out Pekin ducks.

Preening cups may be a form of open water that would allow ducks to express preening behaviors. We set out to test the hypothesis that preening cups would not have detrimental effects on ducks or their environment. Control pens (N = 6, 65 ducks/pen) had nipple lines while experimental pens (N = 6, 65 ducks/pen) had the same nipple line plus one preening cup (PC). Body weights of 30 ducks per pen, and body condition scores on 50 ducks per pen were recorded weekly. On d 18 and 43, 5 ducks per pen were euthanized and their spleens, Bursas, liver, and uropygial glands were weighed. Behavior data were collected using scan sampling with video being recorded for 72 continuous hours at 4 different ages: 25 d, 30 d, 36 d, and 40 d. Body morphometrics were analyzed by 2-way ANOVA with repeated measures. Body condition scoring was analyzed by Pearson's chi-square. The GLIMMIX procedure (SAS 9.4) was used for behavioral analyses to examine treatment differences in the proportion of ducks performing dry preening, wet preening, eating, drinking, standing, and laying down. Feather pecking, feather picking, preening conspecifics (also known as allopreening), dunking head, and drinking from preening cup were analyzed using PROC LOGISTIC with the Firth bias correction for quasi-complete separation and odds ratios were calculated. More PC ducks housed with PC performed wet preening compared to control ducks (25 d: F1,26 = 6.90, P = 0.0143; 30, 36, and 40 d; F1,78 = 24.53, P < 0.0001). Ducks in the PC group were also more likely to lay down compared to controls (25 d: F1,33 = 4.95, P = 0.0330). No differences were observed for any other behavior, body condition score, body weight or morphometrics at any age. Although ducks in the preening cup group showed an increase in wet preening, our data suggest that open water is not necessary to maintain feather condition or uropygial gland size.

Sex differences in glucocorticoid responses to shipping stress in Pekin ducks.

Some concerns have been raised recently about the assay of corticosterone vs. cortisol in poultry species. Thus, we tested the hypothesis that ducks secrete both glucocorticoids. First, we validated two commercially ELISA kits for the two glucocorticoids by first charcoal stripping duck serum in order to remove all steroid hormones. We ran serial dilutions of spiked, charcoal-stripped serum on kits of opposite glucocorticoid as well as a serial dilution using the respective ELISA buffer of the opposite assay kit. We found that the glucocorticoid standard curve in duck serum matched the respective curve in that kit's own buffer. However, when the opposite hormone was run in each kit in both duck serum or ELISA buffer, a near zero slope was obtained. Second, we further validated the presence of both glucocorticoids using mass spectrometry. Third, we tested the hypothesis that exogenous ACTH would stimulate the release of both corticosterone and cortisol. And, fourth, we tested the hypothesis that each glucocorticoid would have different serum levels in response to shipping stress. To test this hypothesis, we collected serum from 10 drakes and 10 hens from 2 flocks (N = 20 per time point per sex): 24 h prior to shipping, at shipping as ducks were walked off the truck, 24 h after shipping, and 1 wk after shipping. Data were analyzed by 2-way repeated measures ANOVA. Surprisingly, we also observed a sex difference in both glucocorticoid levels in that hens showed higher (P < 0.01) serum levels than did drakes at all-time points in response to either ACTH or transportation. Finally, no differences were observed in either glucocorticoid levels associated with shipping in either sex. The fact that both glucocorticoids are released in measurable amounts lends to the possibility that they may be differentially regulated, or at least there is a sex difference in the neural pathways associated with glucocorticoid release in ducks. Although corticosterone is the likely predominate glucocorticoid in ducks, serious attention should be given to the role of cortisol in poultry. Further consideration of sex, age, and timing of blood collection to stressor needs to be considered when assessing glucocorticoid levels in any avian species.

Research Note: Sex difference in changes in heterophil to lymphocyte ratios in response to acute exposure of both corticosterone and cortisol in the Pekin duck.

Poultry scientists have utilized both direct and indirect measures of stress hormones for monitoring the state of avian welfare. For decades, it has been assumed that the mammalian and avian hypothalamic pituitary adrenal (HPA) function similarly to one another. However, there are considerable differences between the 2. Further, it has been assumed that the predominate glucocorticoid (GC) in birds was corticosterone, but recent studies have suggested that both corticosterone and cortisol are secreted. GC release is associated with an increase in blood heterophils due to increased migration from the lymph nodes and a decrease in lymphocytes due to marginalization. Both actions account for an increase in heterophil to lymphocyte ratios (HLR). The goal of this project was to determine the effect of each GC on HLR over time. To achieve this, we intramuscularly injected 2.0 mg/kg of corticosterone or cortisol, a lower dose cortisol treatment (0.5 mg/kg), or safflower oil as vehicle control. Blood was collected prior to intramuscular (IM) injections and blood collected 3 more times at every hour. Blood smears were also collected to assess HLR at the same four time points. HLR assays were completed by avian pathologists from an independent lab who were unaware of the treatments. Data were analyzed by 3-way repeated measures ANOVA with a P < 0.05 considered significant. We found significant sex (P < 0.001) x treatment (P < 0.001) x time (P < 0.001) effects with significant interactions (P = 0.0055). In hens, both GC resulted in significant increase in HLR at 1 h after injection compared to controls. In drakes, however, both GC showed a significant increase in HLR but not until 2 h after injection. The low dose cortisol had no significant effect on HLR in either sex. These data suggest that sex differences need to be considered when assessing duck welfare, and that cortisol may play a role in the HPA axis in ducks.

Chronic heat stress part 1: Decrease in egg quality, increase in cortisol levels in egg albumen, and reduction in fertility of breeder pekin ducks.

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.

Centennial Review: History and husbandry recommendations for raising Pekin ducks in research or commercial production.

By some accounts, ducks were domesticated between 400 and 10,000 yr ago and have been a growing portion of the poultry industry for decades. Ducks specifically, and waterfowl in general, have unique health, housing, nutrition and welfare concerns compared to their galliform counterparts. Although there have been many research publications in regards to health, nutrition, behavior, and welfare of ducks there have been very few reviews to provide an overview of these numerous studies, and only one text has attempted to review all aspects of the duck industry, from breeders to meat ducks. This review covers incubation, hatching, housing, welfare, nutrition, and euthanasia and highlights the needs for additional research at all levels of duck production. The purpose of this review is to provide guidelines to raise and house ducks for research as specifically related to industry practices.