Corticosterone Concentrations In Feathers Research Articles

Increasing concentrations of dietary threonine, tryptophan, and glycine improve growth performance and intestinal health with decreasing stress responses in broiler chickens raised under multiple stress conditions

The current study aimed to compare the effects of increasing concentrations of dietary threonine (Thr), tryptophan (Trp), and glycine (Gly) on productive performance, stress biomarkers, and intestinal function in broiler chickens under multiple stress conditions. Five hundred sixty 21-d-old broiler chickens were randomly allotted to 5 treatments with 8 replicates. Birds in a positive control (PC) treatment were raised under low stock density (16.9 birds/m2) with recommended environmental conditions, whereas birds in 4 treatments were subjected to multiple stress conditions: a cyclic heat stress of 30 ± 0.3 °C for 10 h and 23 ± 0.2 °C for 14 h per day with high stock density (25.3 birds/m2). A basal diet was assigned to both PC and negative control (NC) treatments. Three additional diets were individually formulated to contain double concentrations of digestible Thr, Trp, or Gly + Ser compared with their concentrations in the basal diet. The experiment lasted for 14 d. Results showed that NC treatment had less growth performance (P < 0.001), goblet cell counts (P = 0.018), and trans-epithelial electrical resistance (TEER; P < 0.001), but greater (P = 0.026) feather corticosterone (CORT) concentrations than PC treatment. Thr treatment showed the least (P < 0.001) feed conversion ratio (FCR) among treatments under multiple stress conditions. Thr, Trp, and Gly treatments had less (P = 0.026) feather CORT concentrations, but had greater (P < 0.001) TEER than NC treatment. In conclusion, increasing concentrations of dietary Thr, Trp, or Gly improve the growth performance and intestinal health of broiler chickens with decreasing stress response under multiple stress conditions.

Juvenile Northern Spotted Owls with higher mass and intermediate levels of corticosterone have greater long-term survival

Abstract Early life experiences have carry-over effects that manifest in later life stages. Challenging rearing environments result in more energy invested in immediate survival and less energy on growth and maturation, which can decrease survival in both the short- and long-term. One way to measure differences in energy allocation to growth between individuals is through physiological indices, such as concentrations of the metabolic hormone corticosterone, and body condition. Corticosterone increases in response to challenges to homeostasis and mobilizes stored fat and muscle to meet energetic demands. Maintaining elevated corticosterone can result in poor body condition and decreased survival. Juvenile or pre-breeding age classes are typically a substantial portion of the total population and serve key functions in population dynamics in many long-lived species. Thus, understanding how the rearing environment may influence demographics across life-history stages is crucial to understanding larger population dynamics. Yet, demographic models and conservation planning often lack vital rate estimates for early life stages because, prior to breeding, animals cannot be effectively sampled (i.e., they are unobservable). We used corticosterone concentrations in feathers and body mass of fledged juvenile Northern Spotted Owls (Strix occidentalis caurina) as indicators of potential energetic limitations early in life and used a multi-state modeling framework to quantify the effect of these indicators on apparent annual survival prior to claiming a territory (pre-territorial birds) and the probability of recruiting into the territorial population. Apparent annual survival for pre-territorial birds was higher for birds with greater mass, earlier banding dates, and intermediate levels of corticosterone. Birds with greater mass at banding were also more likely to recruit into the territorial population. Our results demonstrate the importance of early development and traditionally excluded life-history stages on long-term demographics. Although early life stages are difficult to observe, monitoring and conservation efforts may be improved by increasing studies on pre-territorial animals, control of Barred Owls, and conservation of forest structures important for Spotted Owls. This may contribute to increased juvenile survival and recruitment.

Provision of black soldier fly larvae (Hermetia illucens) in different ways benefits broiler welfare and performance, with largest effects of scattering live larvae

Including black soldier fly larvae (BSFL) in broiler diets has the potential to benefit broiler welfare and increase production performance, but the effects of dietary BSFL likely depend on the way BSFL are provided. In this study we aimed to discern the effects of different BSFL forms and provisioning methods by providing male broilers with no BSFL (CON), processed BSFL meal and oil incorporated in the feed pellets (INC-F), dried BSFL in the feeder on top of the feed (D-F), or dried or live BSFL scattered through the pen (D-S and L-S, respectively), and evaluating various indicators of broiler welfare and production performance. In all dietary BSFL treatments 8% of the total dietary dry matter content was replaced with BSFL. Dried and live larvae were provided in four equal daily portions at 08:00, 11:00, 14:00, and 17:00. Compared to a diet without BSFL, scattering dried or live larvae through the pen increased active behaviors, though only live larvae increased the time broilers spent standing. Broilers in the D-F, D-S and L-S treatments had higher average daily body weight gain during some periods, and they had higher final weights, despite L-S broilers having a lower total dry matter intake than CON broilers. Furthermore, the dry matter conversion ratio of INC-F, D-S and L-S broilers was reduced. At the end of the rearing period, pens in all dietary BSFL treatments had better litter quality than CON pens. Furthermore, food pad dermatitis was less severe for INC-F and D-S broilers than for CON broilers, and for L-S broilers than for broilers in all other treatments, and hock burn severity was less for L-S than for CON broilers. Broiler lameness, cleanliness, plasma natural antibody titers, and whole blood serotonin were not influenced by dietary BSFL treatment. Feather corticosterone concentrations were affected by treatment, though without any significant post-hoc differences. Our results indicate that BSFL meal and oil, and dried and live BSFL are all promising feed ingredients for broilers as they all benefit some aspects of broiler welfare and production performance. Scattering BSFL through the pen results in more welfare benefits than providing BSFL in the feeder, with live BSFL having the most beneficial effects on broiler welfare.

Comparison of stress biomarkers in laying hens raised under a long-term multiple stress condition

The objective of the current experiment was to compare various stress biomarkers including the heterophil to lymphocyte ratio (H:L ratio) in blood and the corticosterone (CORT) concentrations in feathers, claws, and egg yolk and to find the potential stress biomarkers in laying hens exposed to a long-term multiple stress condition. A total of 24 Hy-Line Brown laying hens at 47 wk of age were allotted to 2 distinct conditions including normal condition (NC) and multiple stress condition (MC) with 8 replicated cages. In NC treatment, 8 hens were raised individually in the cage (0.16 m2/hen) under the temperature of 21 ± 0.6°C. In MC treatment, 16 hens were raised with 2 hens per cage to decrease space allowance (0.08 m2/hen) and the temperature was maintained at 31 ± 1.6°C. The experiment lasted for 8 wk. The common diets and water were fed on an ad libitum basis during the experiment. Results indicated no interactions between stress conditions and duration of stress exposure for all measurements. Hens in MC treatment had a greater (P < 0.01) blood H:L ratio than those in NC treatment. The greater (P < 0.05) H:L ratio for MC treatment was observed at all weeks. Hens in MC treatment had greater (P < 0.05) feather CORT concentrations than those in NC treatment. Feather CORT concentrations were increased (P < 0.05) with duration of the experiment. However, stress conditions did not influence claw CORT concentrations. A tendency (P = 0.081) was observed for greater yolk CORT concentrations in MC treatment than in NC treatment. In conclusion, the H:L ratio in blood and CORT concentrations in feathers and egg yolks are considered potential stress biomarkers in laying hens exposed to stress conditions, although each measurement has its respective limitations. However, CORT concentrations in the claw appear to be insensitive to a long-term stress exposure.

Mercury toxicity risk and corticosterone levels across the breeding range of the Yellow-breasted Chat

Mercury (Hg) is an environmental contaminant that can negatively impact human and wildlife health. For songbirds, Hg risk may be elevated near riparian habitats due to the transfer of methylmercury (MeHg) from aquatic to terrestrial food webs. We measured Hg levels in tail feathers sampled across the breeding range of the Yellow-breasted Chat (Icteria virens), a riparian songbird species of conservation concern. We assessed the risk of Hg toxicity based on published benchmarks. Simultaneously, we measured corticosterone, a hormone implicated in the stress response system, released via the hypothalamus-pituitary-adrenal axis. To better understand range-wide trends in Hg and corticosterone, we examined whether age, sex, subspecies, or range position were important predictors. Lastly, we examined whether Hg and corticosterone were correlated. Hg levels in chats were relatively low: 0.30 ± 0.02 µg/g dry weight. 148 out of 150 (98.6%) had Hg levels considered background, and 2 (1.6%) had levels considered low toxicity risk. Hg levels were similar between sexes and subspecies. Younger chats (<1 year) had higher Hg levels than older chats (>1 year). Hg levels were lowest in the northern and central portion of the eastern subspecies’ range. Corticosterone concentrations in feathers averaged 3.68 ± 0.23 pg/mm. Corticosterone levels were similar between ages and sexes. Western chats had higher levels of corticosterone than eastern chats. Hg and corticosterone were not correlated, suggesting these low Hg burdens did not affect the activity of the hypothalamus-pituitary-adrenal axis. Altogether, the chat has low Hg toxicity risk across its breeding range, despite living in riparian habitats.

Feather Corticosterone Measurements and Behavioral Observations in the Great White Pelican (Pelecanus onocrotalus) Living under Different Flight Restraint Conditions in German Zoos.

Simple SummaryThe welfare of zoo birds kept under flight restraint is a frequently discussed topic. Therefore, this study was conducted with one of the most regularly kept types of deflighted birds in German zoos, the great white pelican, to find scientific data regarding welfare assessments of deflighted birds. The detection of corticosterone in feathers (CORTf) as a stress indicator for birds is an almost completely noninvasive form of measurement meant to evaluate the effects of deflighting birds in zoos. Three groups of animals were compared: irreversibly deflighted pelicans that were pinioned or extirpated, reversibly deflighted individuals that were feather-clipped, and airworthy pelicans that were able to fly. Combining two independent research methods, behavioral observation and the measurement of CORTf levels of great white pelicans, we aimed to obtain an objective overview of whether deflighted birds showed differences in CORTf or behavior compared to airworthy birds. As a result of the analysis, we found no significant differences in CORTf between flight-restricted and airworthy birds. However, reversibly deflighted pelicans had higher CORTf values than irreversibly deflighted and airworthy pelicans. In addition, pelicans living in groups consisting of more than 10 individuals showed lower CORTf values than pelicans in groups of less than 10 individuals. “Fluttering” behavior was significantly associated with higher CORTf values. In conclusion, the flight restriction of great white pelicans does not seem to impact the welfare indicators assessed in this study, adrenal activity, or behavior. The data show that the living conditions of pelicans (such as group size) may influence the welfare of these birds. To confirm this, further studies on other ground- and water-based birds are needed to provide more scientific data on animal welfare and living conditions in zoos.The pinioning of birds was previously one of the most-accepted forms of mutilation in zoos. Despite a lack of knowledge on the effects of deflighting procedures with regard to the well-being of deflighted birds, pelicans are often reversibly deflighted by feather-clipping to keep them in open enclosures, including those with ponds without netting. In the present study, we focused on the welfare implications of flight restraint on one of the most commonly kept types of birds in German zoos, the great white pelican. A combination of behavioral observations and feather corticosterone concentrations (CORTf) of pelicans with different deflighting statuses (i.e., irreversibly deflighted, reversibly deflighted, and airworthy) was used to evaluate the effects of deflighting status on pelican welfare. We observed 215 individuals in 21 different German zoos. The pelicans lived in differently designed exhibits. An ethogram for these species was developed and their behavior was evaluated by scan sampling. Feather samples from 182 individuals were collected to determine if different deflighting conditions influenced the CORTf and therefore stress levels. The hypothesis was that the CORTf values of airworthy pelicans differ from those of deflighted pelicans. Tendencies with regard to the flight status groups were found. Conversely, reversibly deflighted pelicans had higher CORTf levels than irreversible deflighted and airworthy pelicans. Tendencies with regard to CORTf values and the group size of the kept pelicans were observed. The CORTf values were lower in groups consisting of more than 10 animals. In addition, the frequency of fluttering behavior was positively associated with CORTf values. Pelicans that frequently showed fluttering had higher CORTf values. Therefore, fluttering behavior might be considered a sign of stress levels in pelicans. This study is one of the first important steps in assessing the impact of deflighting procedures on the welfare of great white pelicans kept in zoos.

Comparative effects of dietary functional nutrients on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions.

ObjectiveThe objective of the present study was to investigate the comparative effects of dietary functional nutrients including glutamine (Gln), chromium picolinate (Cr picolinate), vitamin C (Vit C), betaine (Bet), and taurine (Tau) on growth performance, meat quality, immune responses, and stress biomarkers in broiler chickens raised under heat stress conditions.MethodsA total of 420 21-d-old Ross 308 male broiler chickens (initial body weight = 866±61.9 g) were randomly allotted to 1 of 7 treatment groups with 6 replicates. One group was kept under thermoneutral conditions and was fed a basal diet (PC, positive control). Other 6 groups were exposed to a cyclic heat stress condition. One of the 6 groups was fed the basal diet (NC, negative control), whereas 5 other groups were fed the basal diet supplemented with 0.5% Gln, 500 ppb Cr picolinate, 250 mg/kg Vit C, 0.2% Bet, or 1.0% Tau. The diets and water were provided ad libitum for 21 d.ResultsBroiler chickens in NC group had decreased (p<0.05) growth performance and immune responses measured based on cutaneous basophil hypersensitivity (CBH), but increased (p<0.05) stress responses measured based on feather corticosterone concentrations and blood heterophil:lymphocyte than those in PC group. However, none of dietary functional nutrients had a positive effect on growth performance of broiler chickens. Dietary supplementation of 250 mg/kg Vit C improved (p<0.05) CBH responses of broiler chickens, but other functional nutrients had no such an improvement in CBH responses. All functional nutrients decreased (p<0.05) stress responses of broiler chickens.ConclusionFunctional nutrients including Gln, Cr picolinate, Vit C, Bet, and Tau at the supplemental levels used in this study decrease stress responses of broiler chickens to a relatively similar extent. However, this reduction in stress responses could not fully ameliorate decreased productive performance of broiler chickens raised under the current heat stress conditions.

Effects of stocking density and dietary vitamin C on performance, meat quality, intestinal permeability, and stress indicators in broiler chickens.

The objective of the current study was to investigate the effects of stocking density (SD) and dietary supplementation of vitamin C on growth performance, meat quality, intestinal permeability, and stress indicators in broiler chickens. The study was conducted using a completely randomized design with a 2 × 2 factorial arrangement consisting of 2 different SD and 2 supplemental levels of dietary vitamin C. A total of 1,368 Ross 308 broiler chickens of 21 days of age with similar body weights (BW) were randomly allotted to 1 of 4 treatments with 6 replicates each. Different numbers of birds per identical floor pen (2.0 m × 2.4 m) were used to create 2 different SD levels of low SD (9 birds/m2) and high SD (18 birds/m2). The basal diet was formulated with no supplemental vitamin C to meet or exceed nutrient recommendations of the Ross 308 manual. The other diet was prepared by supplementing 200 mg/kg vitamin C in the basal diet. The study lasted for 14 days. At the end of the study, 3 male birds per replicate were selected to analyze meat quality, intestinal permeability, and stress indicators such as blood heterophil:lymphocyte (H:L) and feather corticosterone (CORT) concentrations. Results indicated that there were no interactions between different SD and dietary supplementation of vitamin C for all measurements. For the main effects of SD, birds raised at high SD had less (p < 0.01) BW, BW gain, and feed intake with increasing stress responses including greater blood H:L and feather CORT concentrations (p < 0.01) than those raised at low SD. Transepithelial electrical resistance in the jejunal mucosa was decreased (p < 0.05) at high SD, indicating an increase in intestinal permeability. However, the main effects of dietary supplementation of 200 mg/kg vitamin C were insignificant for all measurements. In conclusion, high SD of broiler chickens impairs growth performance and intestinal barrier function with increasing stress responses. However, dietary supplementation of vitamin C may have little beneficial effects on broiler chickens raised at the high SD condition used in the present study.

The Effect of Egg Laying on Feather and Plasma Corticosterone Concentrations in Turkey (Meleagris gallopavo) Hens.

Simple SummaryIt is not known if an energetically demanding process, such as egg laying, can affect corticosterone measured in turkey feathers and blood plasma, or if there are differences between feather types and genetic lines. The objective of this study was to compare hormone levels from feathers and plasma taken before and during egg laying. We found that the corticosterone in the feathers were higher during egg laying, but only for one genetic line. This genetic line produces more eggs and, so, the elevated corticosterone may reflect the higher metabolic investment in producing eggs. Increases in corticosterone levels were found in different feather types; however, the overall hormone level differed between feather types. Unlike the feathers, we found a decrease in plasma hormone measures between the two periods which might reflect the influence of gonadal development or suppression of acute hormone production during lay. From this study, we conclude that feathers can be used to detect increases in corticosterone during periods of high energy demand, but the genetic line needs to be considered, and the results may not correspond with plasma measures. This study also reinforces the requirement for consistent feather sampling when using corticosterone levels in feathers as non-invasive biomarkers.Phenological differences in energy demand (i.e., reproductive status) might influence the measurement of corticosterone. The objective of this study was to compare corticosterone concentrations in feathers (FCORT) and plasma (PCORT) for turkey hens before and during egg laying. Secondary feathers 1 and 3, and a plasma sample were collected from 50 hens at 30 weeks (before egg laying) from two purebred lines. The hens were reexamined during lay (45 weeks) to collect regrown feathers and plasma samples. Corticosterone concentrations were measured using an ELISA. Linear mixed models were used to assess the effect of genetic line (A vs. B) and period (pre-lay vs. lay) on FCORT and PCORT levels. An increase in FCORT during lay was detected for line B (p < 0.0001), but not line A (p = 0.3076). An increase in FCORT during lay was detectable in both feather types, but there was a difference between secondary 1 and 3 in FCORT concentration within each line studied. Conversely, PCORT decreased between the pre-lay and lay periods for both lines, although the decrease was more substantial for line A (p < 0.0001). Differences in metabolic investment in egg production between the two genetic lines may explain the different FCORT response during lay. The results from this study provide insight into how periods of high energy demand may influence corticosterone which should be considered when interpreting results.

Early-Life Corticosterone Body Condition Influence Social Status and Survival in a Food-Caching Passerine.

Individuals undergo profound changes throughout their early life as they grow and transition between life-history stages. As a result, the conditions that individuals experience during development can have both immediate and lasting effects on their physiology, behavior, and, ultimately, fitness. In a population of Canada jays in Algonquin Provincial Park, Ontario, Canada, we characterized the diet composition and physiological profile of young jays at three key time points during development (nestling, pre-fledge, and pre-dispersal) by quantifying stable-carbon (δ13C) and -nitrogen (δ15N) isotopes and corticosterone concentrations in feathers. We then investigated the downstream effects of early-life diet composition, feather corticosterone, and environmental conditions on a juvenile's social status, body condition, and probability of being observed in the fall following hatch. Across the three time points, the diet of Canada jay young was composed primarily of vertebrate tissue and human food with the proportion of these food items increasing as the jays neared dispersal. Feather corticosterone concentrations also shifted across the three time points, decreasing from nestling to pre-dispersal. Dominant juveniles had elevated corticosterone concentrations in their feathers grown pre-dispersal compared with subordinates. High body condition as nestlings was associated with high body condition as juveniles and an increased probability of being observed in the fall. Together, our results demonstrate that nestling physiology and body condition influence the social status and body condition once individuals are independent, with potential long-term consequences on survival and fitness.

Psychological and Physiological Stress in Hens With Bone Damage.

Abnormalities in bone development in humans and non-humans can lead to impaired physical and psychological health; however, evidence is lacking regarding the role of individual psychosocial factors in the development of poor bone conditions. Addressing this lack of knowledge, we used low-productive laying hens (n = 93) and assessed behavioral responses to an open-field test [at 17, 18, 29, 33 weeks of age (wa)], an aerial predator test (at 39 wa), and a social reinstatement test (at 42 wa). Bone condition was assessed using a palpation technique on five occasions (at 16, 29, 33, 45, 58 wa), with half of the hens experiencing damage (deviations, fractures, or both) at 29 wa and all hens by 58 wa. Corticosterone (CORT) concentration in feathers (at 16, 33, 58 wa) and body weight (at 23, 47, 58 wa) were also investigated. We hypothesized that lighter birds (at 23 wa) with higher CORT (at 16 wa) and open field-induced fear collected before the onset of lay (at 17 and 18 wa) are associated with a worse bone condition when in lay. We also hypothesized that those birds with more damage at the peak of laying (at 29 wa) would be lighter at 47 and 58 wa and more fearful by showing higher open field-induced (at 29 and 33 wa) and predator-induced fear responses, however, acting less socially toward conspecifics. These hens were also expected to have higher CORT (at 33 and 58 wa). Our results show no association between open-field fear level and fear behavior, CORT concentration, or body weight on the one hand (all measured before starting to lay) and bone damage at 29 wa on the other. When in lay, bone damage was associated with more pecking and less crossing zones when faced with an open-field situation at 29 wa and improved sociality at 42 wa. This study provides the first evidence of a relationship of bone health with fear, sociality, and stress response. When in poor bone condition, our hens had enhanced psychological stress measured by fear behavior reactivity but not physiological stress measured as feather CORT concentration.

Corticosterone in feathers: Inter- and intraindividual variation in pullets and the importance of the feather type

Measuring corticosterone concentrations in feathers of poultry may be suitable to determine birds' exposure to stress. It is thinkable, that in laying hens such information could be helpful as an animal welfare indicator to evaluate adverse husbandry conditions and to predict the risk of developing behavioral disorders, such as feather pecking and cannibalism. Yet, there are some fundamental issues which remain unclear. Therefore, the objective of the current pilot study was to examine the inter- and intraindividual variation of pullets at the end of the rearing period, when most of the feathers are fully grown and animals are reaching sexual maturity. Flight feathers from both wings (n = 4), the tail (n = 2 – 3), and body feathers (n = 1 pool of 3 – 5 feathers) were taken from pullets (n = 10), genetics Lohmann Brown, at an age of 19 weeks who were reared in the same flock (N = 728). Corticosterone analysis was performed applying a validated protocol for laying hens. Results indicate not only high intraindividual, but also high interindividual variation. Mean over all samples was 75.2 pg/mg (± 38.58 pg/mg, n = 76), showing higher intraindividual variation (between feather types; SD: 23.75 pg/mg – 49.38 pg/mg; n = 10 pullets) than interindividual variation (within feather types; SD: 11.91 pg/mg – 49.55 pg/mg; n = 6 feather types). The variation between different feather types within one bird was higher than the variation within one feather type between different birds, indicating that birds a) may respond differently when exposed to stressors and b) corticosterone measurements should be done with the same feather type.

Effects of feather processing methods on quantity of extracted corticosterone in broiler chickens.

Corticosterone is known as a biological stress index in many species including birds. Feather corticosterone concentration (FCC) has increasingly been used as a measure for chronic stress status in broiler chickens. As sample preparation is the first step of analytical process, different techniques of feather matrix disruption need to be validated for obtaining better result in analysing corticosterone extraction. The current study was a validation of pulverizing the feather by bead beater (BB) and surgical scissors (SS) processing prior to corticosterone extraction in feather of broiler chickens. The type of feather processing prior to the hormone extraction may alter the final output. Thereby, finding a standard method according to laboratory facilities is pivotal. This study carried out to determine the effects of feather pulverization methods on the extraction amount of corticosterone in broiler chickens. Feathers were sampled from four weeks old Ross 308 broiler chickens (n = 12 birds). All broiler chickens were kept under the same environmental condition and had access to feed and water. Feather samples were assigned to one of the following processing methods 1) using a BB for pulverizing and 2) using a SS for chopping into tiny pieces. Each sample was duplicated into two wells during enzyme immunoassay (EIA) analysis to improve the accuracy of the obtained data. The results showed lower standard errors and constant output of FCC by using the BB method compared with the SS method. Overall comparison of FCC showed a significantly higher (p < 0.001) amount of the FCC in the BB compared with the SS. Overall, using the BB method is recommended over the SS method for feather processing due to the ability to homogenize a large number of samples simultaneously, ease of use and greater extraction of feather corticosterone.

Research Note: Quantifying corticosterone in turkey (Meleagris gallopavo) feathers using ELISA

The measurement of corticosterone in feathers is an appealing tool for assessing glucocorticoids in wild and domestic bird species. Feather corticosterone measurements can be performed noninvasively and can provide a means for comparing glucocorticoid secretion between individual birds; thus, such measurements can be used to assess the welfare of domestic poultry. The focus of this study was to assess the validity of detecting corticosterone in turkey (Meleagris gallopavo) feathers using an ELISA and investigate differences between genetic lines. Primary feather 9 was obtained at a processing plant from 123 individuals from 3 different purebred turkey lines (line A [N = 46], line B [N = 24], and line C [N = 53]). Assay validation tests were performed using a commercially available ELISA kit (Cayman Chemicals, Cedarlane Labs, Canada). Indicators of accuracy, recovery, precision, and sensitivity were sufficient. Significant differences in feather corticosterone concentration between the 3 lines were observed. Line C had significantly higher feather corticosterone than line A (P < 0.0001) and line B (P = 0.036). These results indicate that the quantification of feather corticosterone using an ELISA is a valid method for assessing glucocorticoid levels in turkeys. This is the first report of differences in feather corticosterone between different purebred turkey lines. Differences observed between purebred lines provide an intriguing basis for further investigation into the genetic parameters of glucocorticoid levels in turkeys.

Corticosterone in feathers of laying hens: an assay validation for evidence-based assessment of animal welfare

Studies indicate that the evaluation of animal welfare in birds may be carried out with the measurement of the stress-related hormone corticosterone in feathers. However a standardized procedure for corticosterone measurements in feathers is lacking, a validation needs to be carried out for each new species before implementation. The aim of the present study was to establish a valid method to measure corticosterone concentrations in feathers of laying hens in a precise and repeatable manner using an established and commercially available ELISA. Validation was performed with feather pools of tail and interscapular feathers of commercial Lohmann Brown laying hens. Assessment groups, consisting of 5 replicates, were created. All replicates of an assessment group were processed at the same time. Each replicate was run in 4 repetitions by ELISA. Intra-assay and interassay CV was 7.5 and 6.4%, respectively. The serial dilution showed linearity and parallelism. Examining the hormone extraction efficiency by using different methanol volumes resulted in no statistical differences (P > 0.05). Pulverized feathers showed higher corticosterone values than minced feathers (P > 0.05). Differences were shown between 2 feather types (tail vs. interscapular feathers; P < 0.05), as well as between vane and rachis (P < 0.05). Performance of a freeze–thaw cycle led to a decrease of corticosterone concentrations in the samples. A possible effect of UV-A radiation on the stability of corticosterone in the feathers was not found (P > 0.05). With the present study, a valid protocol, feasible for analyzing feather pools of laying hens, was developed. It may provide fundamentals for further investigations on corticosterone in feathers as a noninvasive indicator to evaluate aspects of animal welfare.

Phenotypic signatures of urbanization are scale-dependent: A multi-trait study on a classic urban exploiter

Understanding at which spatial scales anthropogenic selection pressures operate most strongly is a prerequisite for efficient conservation and management of urban biodiversity. Heterogeneity in findings on the strength and direction of urbanization effects may result from a lack of consensus on which spatial scales are most adequate when studying biotic effects of urbanization. Therefore, here, using the house sparrow (Passer domesticus) as model, we test the hypothesis that more than one spatial scale will explain variation among phenotypic stress markers. By applying a unique hierarchical sampling design enabling us to differentiate between local and regional effects of urbanization, we here show that the strength and direction of relationships with the percentage of built-up area – a simple structural measure of urbanization - vary among phenotypic stress markers and across the spatial range over which urbanization is measured. While inverse relationships with scaled body mass and bill height of adult house sparrows (Passer domesticus) were strongest when the degree of urbanization was quantified at city-level, similar relationships with corticosterone concentrations in feathers were only detected at the scale of individual home ranges. In contrast, tarsus length, wing length, and two measures of feather development were not significantly related to urbanization at any spatial scale. As the suite of phenotypic stress markers applied in this study revealed signatures of urbanization over a broad spatial range, we conclude that measures aimed at mitigating impacts of urbanization on free-ranging populations should best be implemented at multiple spatial scales too.

Feather corticosterone in Northern Bald Ibis Geronticus eremita: a stable matrix over time able to predict reproductive success

Measuring corticosterone concentrations in feathers (FCC) has been increasingly used as an integrated index of bird’s response to stressors, offering valuable insights into subsequent carry-over effects. However, it is still unclear to what extent corticosterone levels deposited in feathers during the moulting period relate to individual physiology in high-energetic demanding situations. In addition, the stability and repeatability of FCC over the same feather generation and across successive generations are still unresolved questions. In this study, we assessed FCC in 24 captive Northern Bald Ibis, Geronticus eremita. We sampled body feathers on three occasions during two consecutive years (2015–2016) to explore within-individual stability of FCC in feathers grown during the same generation and test whether FCC was repeatable or varied between two consecutive generations. Then, we explored whether FCC (a) was associated with individual health status, (b) correlated with previous reproductive success and (c) was related to future reproductive success. Results showed high stability of FCC among feathers of the same generation, while FCC differed between successive generations. We also observed that FCC, age and health status were negatively associated with subsequent reproductive success the following season, while no association was found with previous reproductive success. Overall, our study supports the use of feathers as a stable measure of corticosterone over the same feather replacement generation and emphasizes the usefulness of FCC as a biomarker of subsequent reproductive success.

Corticosterone mediates telomere length in raptor chicks exposed to chemical mixture

Stressors experience early in life by animals may have carry over impacts on life-traits over the life cycle. Accelerated telomere attrition induced by stress during development and growth could play a role in such delayed effects. Among stressors, exposure to chemicals may modify telomere dynamic but, to date, the trends evidenced between exposure and telomere shortening remains inconsistent. Moreover, the role of corticosterone as a possible mediator of chemical impact on telomere is not yet clearly established. Here, we investigated in wild populations of Red kite whether nestling exposure to metals and pesticides was related to corticosterone concentrations in feathers and telomere length measured in 47 individuals. Lead and mercury concentrations in blood ranged from 2.3 to 59.0 µg L−1 and to 1.4 to 115.7 µg L−1, respectively, and were below the toxicity thresholds proposed for wildlife. Rodenticides were detected in 30% of the chicks. Corticosterone increased with mercury and lead in interaction, showing a synergistic effect of these 2 non-essential metals on this stress hormone. Telomere length was not linked to metals and/or rodenticide exposure while it was related negatively to corticosterone. The relationship between telomere and corticosterone was modulated by nestling’s age, which suggests that the rate of telomere shortening is higher when corticosterone increases. Our findings propose an effect of low exposure of Red Kite nestlings to mercury and lead mixture to raise baseline corticosterone in feathers. The relationships established suggest the hypothesis that telomere attrition could be an indirect consequence of metal exposure mediated by corticosterone.

The Effect of Extensive Human Presence at an Early Age on Stress Responses and Reactivity of Juvenile Ostriches towards Humans.

Simple SummaryHusbandry practices for rearing ostriches in commercial farming environments are currently not optimized. Ostrich chicks may experience stressful episodes and fear of humans during routine farm management practices such as handling, which ultimately may impact on their welfare and explain the poor production performance observed in this species. However, extensive human presence and regular gentle handling has been demonstrated to alleviate stress sensitivity during handling by lowering the fear of humans in other species; be they kept as livestock, in a laboratory, or as pet animals. In this study, ostrich chicks exposed to extensive human presence and gentle handling showed lower stress sensitivity when handled for feather harvesting and clipping and were more inclined to associate with familiar humans at a later stage of their life compared with chicks that had limited human presence and care. This suggests that providing ostrich chicks with extensive human presence and gentle handling at a young age can assist in improving ostrich welfare.The effect of extensive human presence and regular gentle handling performed at an early age (0–3 months old) on stress responses and reactivity of juvenile ostriches towards humans was investigated. A total of 416 ostrich chicks over two years were exposed to one of three treatments for three months after hatching; namely, Human Presence 1 (HP1, N = 144): extensive/prolonged human presence with physical contact (touch, stroking), gentle human voice, and visual stimuli; Human Presence 2 (HP2, N = 136): extensive/prolonged human presence without physical contact, but with gentle human voice and visual stimuli; and the Standard treatment (S, N = 136): human presence limited to routine feed and water supply as a control. At 7.5 months of age, the plasma heterophil/lymphocyte (H/L) ratio was measured before and 72 h after feather harvesting and feather clipping to determine acute stress responses, while chronic stress was measured by quantification of corticosterone (CORT) concentrations in the floss feathers of the birds. Birds’ behavioural response towards a familiar or an unfamiliar handler was evaluated at 12 months using docility and fear tests, and through behavioural observations conducted on random days between the ages of 8–13 months. Willingness to approach, and to allow touch interactions, aggressiveness, and exhibition of sexual display towards the handler, was recorded. No difference in the H/L ratios before and after feather harvesting and clipping was observed in HP1 birds, whereas H/L ratios showed a significant increase 72 h post feather harvesting and clipping in HP2 and S birds (p < 0.05). Birds from the S treatment exhibited a significantly (p < 0.05) higher feather CORT concentration compared with HP1 birds, while HP2 birds had intermediate responses. Birds’ reactivity towards humans and temperament as evaluated using behavioural observations, docility, and fear tests was not affected by treatment (p > 0.05). However, HP1 and HP2 birds were more inclined (p < 0.05) to approach a familiar rather than an unfamiliar handler during the behavioural observations, indicating an ability to distinguish between a familiar and an unfamiliar handler. Overall, the results indicate that early gentle human interactions with ostrich chicks can be beneficial in reducing physiological stress sensitivity later in life and facilitate the ability of ostriches to distinguish between familiar and unfamiliar handlers.

Can variability in corticosterone levels be related to POPs and OPEs in feathers from nestling cinereous vultures (Aegypius monachus)?

Persistent organic pollutants (POPs) are still globally distributed and some have been shown to interact with the endocrine system of birds. However, the relationship between POPs and the stress response mediated by the hypothalamic-pituitary-adrenal (HPA) axis is still poorly understood. Raising concerns are now focused on the toxic properties of emergent organophosphate ester flame retardants (OPEs), but whether OPEs interact with the HPA axis response has not yet been investigated. We measured corticosterone concentrations in feathers (CORTf) as a long-term biomarker of the bird HPA axis response and we investigated their relationship with POP and OPE concentrations in down feathers of nestling cinereous vultures (Aegypius monachus). We also examined whether high contaminant burden and high CORTf concentrations impacted the duration of chick development. The most predominant compounds were the following: p,p′-DDE (3.28 ± 0.26 ng g−1 dw) > γ-HCH (0.78 ± 0.09 ng g−1 dw) > BDE-99 (0.73 ± 0.09 ng g−1 dw) > CB-153 (0.67 ± 0.04 ng g−1 dw). The most persistent POP compounds (CB-170, -177, -180, -183, -187, -194 and p,p′-DDE) were associated (P = 0.02) with high concentrations of CORTf (range: 0.55–6.09 pg mm−1), while no relationship was found when OPEs were tested (P > 0.05). Later egg-laying was positively associated to high levels of CORTf (P = 0.02) and reduced duration of chick development (P < 0.001), suggesting a beneficial effect of the HPA axis response on the growth of the chicks. In addition, males with high concentrations of the most persistent POP compounds tended to show a reduced duration of the nestling period (P = 0.05) and an equal fledging success than chicks with lower levels. These findings suggest that POPs, but not OPEs, may interact with the HPA axis response of chicks, although levels were not high enough to cause detrimental consequences.