Timing of fledging is influenced by glucocorticoid physiology in Laysan Albatross chicks

Fledging is a major life transition for birds, when juveniles move from the safety of a nest into an environment where they must find food and avoid predators. The timing of fledging within a season can have significant effects on future survival and breeding success. Proximate triggers of fledging are unknown: though wing development is likely a primary factor, other physiological changes, such as elevated plasma corticosterone (CORT), may affect fledging behavior. Laysan Albatross (Phoebastria immutabilis) chicks have an extended post−hatching period during which they reach 150% of adult mass. However, approaching fledging, chicks fast for days to weeks and lose mass while still putting energy into feather growth. We evaluated chick morphology and physiology to elucidate proximate triggers of fledging. As in some other species, CORT increased as chicks fasted and lost body mass. At the same time, corticosteroid binding globulin (CBG) declined, thus amplifying free CORT prior to fledging. Once chicks reached a morphological threshold, free CORT levels predicted how long they stayed at the colony: chicks with higher free CORT fledged sooner. To perturb the relationship between body condition, endocrine physiology, and fledging behavior, we supplementally fed chicks for the month before fledging. Fed birds had a slower decrease in body mass, slower decrease in CBG, slower increase in free CORT, and stayed at the colony longer after reaching a morphological threshold. Our study suggests that as chicks lose mass, free CORT acts as a signal of energetic or nutritional state to adjust the timing of fledging.     

Mediation of a corticosterone-induced reproductive conflict

Current research in birds suggests that a conflict should exist during reproduction for the role of the glucocorticoid corticosterone (CORT). While elevated levels have been correlated with the increased energetic demand of raising offspring, elevated CORT levels have traditionally been implicated in reproductive abandonment. We examined the relationship between CORT and nest desertion in breeding wild female European starlings (Sturnus vulgaris) incorporating analyses of both total circulating levels and 'free', unbound CORT through analysis of corticosteroid-binding globulin (CBG). Free baseline CORT levels of nest-abandoning birds were significantly higher than nonabandoning birds within each stage, with chick-rearing birds exhibiting the highest free baseline CORT levels, while concurrently remaining the most resistant stage to nest desertion. Elevated free baseline CORT levels in chick-rearing birds were not due to increased total CORT secretion, but rather to a decrease in CBG levels. Overall, our results suggest that CORT and CBG interact to play a role in mediating the increased energetic demand of offspring, while minimizing the chances of nest desertion, thereby alleviating any potential behavioral conflict for CORT during reproduction. Furthermore, these results demonstrate that the traditional view of the role of CORT during reproduction is much more complex than previously appreciated. Together with mounting evidence, we suggest that elevated corticosteroid levels are an inherent and necessary part of reproduction in nonmammalian tetrapods.     

Circadian influences on feeding-induced changes in ACTH and corticosterone secretion in rats.

Food ingestion has a variable influence on pituitary-adrenal hormone secretion depending on the species studied and/or the experimental design. In this study, changes in plasma concentrations of ACTH and corticosterone following 30 min of food ingestion were compared in both 24 h fasted and ad libitum fed rats tested during either the early light cycle or the early dark cycle. In the dark, food ingestion caused significant decreases in both ACTH (to 80% of control) and corticosterone (to 32% of control) in both fasted and ad libitum fed rats. In contrast, in the light, food ingestion by the fasted animal resulted in a doubling of corticosterone concentrations. Such a response was not seen in ad libitum fed animals; however, these animals ate very little during the test. There were no significant changes in ACTH during the light phase. These data indicate that time of day has a significant impact on the responses of the pituitary-adrenal system to food ingestion. This circadian effect may be due to the influence of the endogenous levels of ACTH/corticosterone existing at the time of food ingestion.     

Effects of corticosterone treatment on responses to fasting in Japanese quail

Plasma concentrations of corticosterone, a glucocorticoid hormone, have been reported to increase during fasting in some species of birds. Although Japanese quail are widely used in laboratory studies, corticosterone responses to fasting have not been described for this species. We therefore investigated the effects of 24 h of fasting on plasma corticosterone concentrations in quail. Previous work has shown that the corticosterone response to handling in quail may be affected by treatment with corticosterone, so we also measured corticosterone responses to 24 h of fasting in quail treated with corticosterone in their drinking water, and determined the effects of corticosterone on food intake immediately after a fast. Plasma corticosterone concentrations were unaffected by 24 h of fasting in controls and in three out of four groups of birds with varying corticosterone intakes. Fasting in birds can be divided into three phases, and our results suggest that quail remained in phase I during the 24 h fast. Food intake in the 3 h immediately after fasting was higher in four groups of birds with varying corticosterone intakes than in the control group, and was greatest in quail with the highest corticosterone intake. The current results suggest that corticosterone can stimulate food intake in quail following a period of food deprivation.     

Fecal corticosterone, body mass, and caching rates of Carolina chickadees (Poecile carolinensis) from disturbed and undisturbed sites

We tested for hormonal and behavioral differences between Carolina chickadees (Poecile carolinensis) taken from a disturbed (recently logged) forest, an undisturbed forest, or a residential site. We measured fecal corticosterone and body mass levels in the field, and fecal corticosterone, body mass, and caching behavior in an aviary experiment. In the field, birds from the disturbed forest exhibited significantly higher fecal corticosterone levels than birds from either the undisturbed forest or from the residential site. Birds from the disturbed forest also exhibited lower body mass than those from the undisturbed forest but higher body mass than those from the residential site. Our aviary results suggest that these physiological differences between field sites are the result of short-term responses to ecological factors: neither body mass nor fecal corticosterone levels varied between birds captured at different sites. Aviary sample sizes were sufficient to detect seasonal variation in fecal corticosterone (lowest in summer), body mass (highest in spring), and rate of gain in body mass (highest in winter). Under "closed-economy" aviary conditions (all food available from a feeder in the aviary), there were no site differences in the percent of seeds taken from the feeder that were cached. However, under "open-economy" conditions (food occasionally available ad libitum), significantly fewer seeds were cached by birds from the disturbed forest compared to the undisturbed or residential sites. On average, there was only a two-fold difference in population levels of fecal corticosterone. This difference is about the same as an increase in fecal corticosterone induced by a 2-h increase in food deprivation and cannot be considered to be an acute stress response to disturbance.     

The role of corticosterone in supporting migratory lipogenesis in the dark-eyed Junco, Junco hyemalis: a model for central and peripheral regulation

The functional role of corticosterone (CORT) in regulating migratory hyperphagia and lipogenesis was investigated in an annual migrant, the dark-eyed junco (Junco hyemalis). Intraperitoneal injections of either dexamethasone (9 microg DXM/500 microL of 5% EtOH in saline, n=10) to inhibit an increase in baseline CORT or saline (5% EtOH, n=9) were given every 48 h for 15 d after transfer from short (10.5L:13.5D) to long (15.5L:8.5D) days. Food intake, body mass, furcular fat deposition scores, and nocturnal migratory activity were recorded for 29 d after photostimulation. Both groups showed the same increase in daily food intake over the study period (DXM=52%, control=41%). Controls began to increase baseline CORT and mass about 2 wk after photostimulation. DXM-treated birds maintained low CORT and did not increase mass or CORT until injections ceased, at which time they gained mass at the same rate shown earlier by controls. DXM-treated birds did not show greater levels of migratory activity despite experiencing an increase in energy intake during the CORT-inhibited period. Collectively, the results support the migration modulation hypothesis, illustrating how an increase in baseline CORT is needed to support the development of migratory condition. We address the apparent conflict with earlier studies on CORT and migratory food intake and propose a model in which migratory hyperphagia is supported by changes in centrally regulated responses to CORT that can occur even if CORT remains low and lipogenesis is regulated predominantly by peripheral mechanisms that require an increase in baseline CORT.     

Food restricted Tufted Puffin (Fratercula cirrhata) nestlings increase vocal activity during handling without modulating total or free corticosterone

In some species, corticosterone (CORT) appears to play a role in the control of begging behavior. Because of the potentially high costs associated with chronic elevation of CORT, it has also been proposed as a mechanism to ensure begging is an honest signal. We determined the effects of moderate food restriction (50% of high calorie treatment) on vocal behavior during handling, and on baseline levels of both total and ‘free’ unbound CORT in Tufted Puffin (Fratercula cirrhata) nestlings. Chick vocalizations during handling were similar to begging calls, and we assumed they were representative of begging behavior. We also measured total and free CORT in free-living Tufted Puffin chicks to determine if hormone levels in our experiment were comparable to natural levels. We found no effect of caloric restriction on either total or free baseline CORT, yet food-restricted nestlings vocalized more intensely during handling than chicks in the high calorie group. Mean plasma concentrations of total and free CORT in experimentally manipulated birds did not differ from levels in free-living nestlings. These results suggest that CORT does not play a role in modulating begging behavior in this species.     

Elevated plasma corticosterone increases metabolic rate in a terrestrial salamander

Plasma glucocorticoid hormones (GCs) increase intermediary metabolism, which may be reflected in whole-animal metabolic rate. Studies in fish, birds, and reptiles have shown that GCs may alter whole-animal energy expenditure, but results are conflicting and often involve GC levels that are not physiologically relevant. A previous study in red-legged salamanders found that male courtship pheromone increased plasma corticosterone (CORT; the primary GC in amphibians) concentrations in males, which could elevate metabolic processes to sustain courtship behaviors. To understand the possible metabolic effect of elevated plasma CORT, we measured the effects of male courtship pheromone and exogenous application of CORT on oxygen consumption in male red-legged salamanders (Plethodon shermani). Exogenous application of CORT elevated plasma CORT to physiologically relevant levels. Compared to treatment with male courtship pheromone and vehicle, treatment with CORT increased oxygen consumption rates for several hours after treatment, resulting in 12% more oxygen consumed (equivalent to 0.33 J) during our first 2 h sampling period. Contrary to our previous work, treatment with pheromone did not increase plasma CORT, perhaps because subjects used in this study were not in breeding condition. Pheromone application did not affect respiration rates. Our study is one of the few to evaluate the influence of physiologically relevant elevations in CORT on whole-animal metabolism in vertebrates, and the first to show that elevated plasma CORT increases metabolism in an amphibian.     

Acute embryonic exposure to corticosterone alters physiology,behaviour and growth in nestlings of a wild passerine

Maternally-derived glucocorticoids can modify the normal development of young animals. To date, little is known about maternal effects that are mediated by acute embryonic exposure to glucocorticoids. In birds, elevated maternal transmission of corticosterone (CORT) to egg albumen is mainly dependent on acute stress. In this study, we increased CORT levels in the egg albumen of a wild passerine, the great tit (Parus major), breeding in favourable deciduous and less suitable coniferous habitat. Subsequently we measured the somatic growth, baseline and acute glucocorticoid responses, immunity and behaviour of prenatally manipulated offspring with respect to control siblings. We found that prenatally CORT-exposed nestlings had lower baseline CORT levels, a more rapid decline in CORT during recovery from a standardized stressor, and a reduced heterophil/lymphocyte ratio compared with controls. Although stress-induced total CORT levels remained unchanged, free CORT levels were significantly lower and the levels of corticosteroid binding globulins (CBG) significantly higher in experimental offspring. Prenatally CORT-exposed offspring begged longer after hatching than controls. Stress-induced behavioural activity of fledglings did not differ between treatments, while its association with baseline CORT levels was significant in the control group only. The body mass and tarsus length of fledglings was positively affected by manipulation in unfavourable coniferous habitat only. We conclude that maternal effects related to elevated levels of albumen CORT modify diverse aspects of offspring phenotype and potentially increase offspring performance in resource poor environments. Moreover, our results indicate that maternal glucocorticoids may suppress the effect of hormones on behavioural responses.     

Nutritional stress affects corticosterone deposition in feathers of Caspian tern chicks

Stressful environmental conditions affect the adrenocortical function of developing animals, which can have consequences for their fitness. Discovery of the avian stress hormone corticosterone (CORT) in feathers has the potential to broaden the application of endocrine research in ecological and evolutionary studies of wild birds by providing a long‐term measure of CORT secretion. Mechanisms of CORT deposition in feathers are not well known and few studies have related feather CORT to circulating plasma CORT during feather growth. Our objective was to experimentally test the validity of using feather CORT as a measure of CORT secretion in developing birds experiencing nutritional stress. Caspian tern Hydroprogne caspia chicks were fed ad libitum or restricted (35% less than ad libitum) diets for four weeks. We measured CORT in feathers from these chicks to examine the relationship between feather CORT concentrations and nutritional limitation, circulating plasma CORT, and feather development. We found that feather CORT was higher in controls fed ad libitum than in restricted individuals, despite higher levels of plasma CORT in restricted chicks compared to controls. Feather mass and growth rates were strongly and positively related to feather CORT concentrations in both treatments. This is the first experimental study to show that feather CORT concentrations can be lower in response to nutritional stress, even when plasma CORT concentrations are elevated. Our results indicate that CORT deposition in feathers may be confounded when feather mass and growth rates are compromised by nutritional stress. We conclude that feather CORT can be used for assessing nutritional stress in growing birds, but the direction of response depends on how strongly stress affects feather development.     

Energy metabolism, testosterone and corticosterone in white-crowned sparrows

The influence of the steroid hormones testosterone and corticosterone on energy metabolism and activity of birds is largely enigmatic. We measured resting metabolic rate during night and day in 12 long-term castrated and 12 intact male white-crowned sparrows (Zonotrichia leucophrys gambelii) under short-day (8:16 SD), long-day (20:4 LD), LD+testosterone implant and LD−testosterone implant conditions. Each male was sequentially measured under all four conditions. Photostimulation increased testosterone, resting metabolic rate, food intake, hopping activity and body mass in castrates and intact males. Surprisingly, testosterone levels and metabolic rates did not differ between intact and castrated males. Testosterone implantation increased activity and food intake, but decreased body mass and resting metabolic rate in both groups. Removing testosterone implants reversed the effects on resting metabolic rate, activity and food intake. Corticosterone levels, measured immediately at the end of metabolism measurements, showed birds were not stressed. Corticosterone had no apparent relationship with resting metabolic rate and there was no interaction between corticosterone and testosterone. Overall, positive changes in testosterone levels resulted in a decrease of resting metabolic rate. We speculate that testosterone increases activity, and birds compensate for increased activity metabolism by reducing resting metabolic rate. Accepted: 18 July 1999     

The effects of variable foraging conditions on common murre (Uria aalge) corticosterone concentrations and parental provisioning

This study investigated how total corticosterone concentrations, chick-feeding rates, and adult body mass changed with food availability from 1998 to 2000 in the same individually marked common murres (Uria aalge). Capelin, the main prey species, arrived inshore by the onset of murre chick hatching in 1998 and 1999 (prey match years); whereas in 2000, hatching began approximately 1 week before the capelin arrived inshore to spawn (prey mismatch year). Serum corticosterone concentrations were higher in the same individuals in the prey mismatch year than they were in either of the match years. Birds sampled before peak capelin spawning in the mismatch year had higher corticosterone levels than murres sampled after peak spawning. Murres with higher corticosterone levels had higher chick-feeding rates and less mass loss in the mismatch year (compared to the match year 1999) than birds with lower levels. Corticosterone levels did not differ between birds that had not foraged for at least 12 h (brooded chick overnight) and those that had, suggesting that short-term food deprivation did not affect corticosterone concentrations. Taken together, these findings suggest that the difference between years reflects a baseline shift in corticosterone levels, particularly in the high-quality birds that were able to increase both corticosterone concentrations and foraging effort.     

Interaction of testosterone, corticosterone and corticosterone binding globulin in the white-throated sparrow (Zonotrichia albicollis)

Plasma binding globulins bind steroid hormones and are thought to regulate hormone access to tissues. Mammals have both sex steroid binding globulin (SSBG) and corticosteroid binding globulin (CBG). Birds, however, have no detectable SSBG, leading to the early conclusion that birds have no plasma regulation of sex steroids. CBG, however, can bind androgens with relatively high affinity. In birds, therefore, the control of androgenic effects may be tightly regulated by glucocorticoid physiology because glucocorticoids compete with androgens for CBG binding sites. We report levels of total testosterone (T), total corticosterone, CBG, and estimated free T in the males, the more aggressive morph had higher levels of total T; female morphs did not differ. Approximately 96% of T was bound to CBG, but a lack of morph or sex-specific differences in corticosterone titers or CBG capacity caused patterns of free T to mirror those of total T. While CBG has the potential to greatly influence T availability to tissues, in this species interactions between T, CBG and corticosterone do not appear to alter general patterns of T availability to tissues.     

Effects of aggressive encounters on plasma corticosteroid-binding globulin and its ligands in white-crowned sparrows

In birds, corticosteroid-binding globulin (CBG) binds corticosterone, progesterone and testosterone. The concentration of each ligand can alter the binding of the other ligands through competitive interactions. Thus, an increase in corticosterone or progesterone may displace testosterone bound to CBG, leading to an increase in bioactive free testosterone levels without affecting total testosterone levels in the circulation. Aggressive interactions increase plasma total testosterone levels in some birds but not in others. Here, we tested the hypothesis that aggressive encounters in the late breeding season would not increase total testosterone levels in plasma, but would alter CBG, total corticosterone or total progesterone levels in such a way as to modify the number of available binding sites and therefore occupancy by testosterone. A marked decrease in CBG occupancy by testosterone would indirectly suggest an increase in free testosterone levels in plasma. Wild male white-crowned sparrows were exposed to a simulated territorial intrusion (STI) or control for 30 min. Subjects were then caught and bled. We measured CBG using a ligand-binding assay and corticosterone, progesterone and testosterone using highly sensitive radioimmunoassays. STI significantly increased aggressive behaviors but did not affect plasma total testosterone levels. STI significantly increased plasma CBG and total corticosterone levels and decreased plasma total progesterone levels. We predict that CBG occupancy by corticosterone will increase slightly following an aggressive encounter. However, this small change is unlikely to increase free testosterone levels, because of the large number of seemingly unoccupied CBG binding sites in these subjects.     

Hormonal responses to male-male social challenge in the blue tit Cyanistes caeruleus: single-broodedness as an explanatory variable

The "challenge hypothesis" posits that when established social orders are challenged, plasma testosterone (T) in socially monogamous breeding male birds will temporarily increase to facilitate aggressive responses. However, not all birds conform to predictions. To expand upon past findings, we examined effects of direct territorial challenge on T levels in the blue tit (Cyanistes caeruleus). We found that simulated territorial intrusions caused a decline in plasma T during both territory establishment and laying/incubation. Conversely, corticosterone (CORT) levels dramatically increased. We also examined challenged blue tit males for levels of corticosterone-binding globulin (CBG), a carrier molecule that displays affinity for both CORT and T in birds. Although the CBG showed increased occupation by CORT during challenge, effects were not accompanied by a significant increase in the unbound T fraction. Thus, competitive hormone interactions on the CBG do not seem sufficient to explain changes in circulating T levels. To place our results within the context of past findings, we compared all socially monogamous birds tested to date for plasma levels of T during situational territorial intrusion experiments. We found that birds raising only one brood per season (e.g., the blue tit) consistently show no increase in plasma T but instead show elevations in circulating CORT. Thus, we suggest that single-broodedness plays an important role in determining patterns of hormone change and should be considered in future discussions of hormone-behavior interactions.     

A role for corticosterone and food restriction in the fledging of nestling White storks

Fledging is a critical period in the life of a bird, notably because at this stage under-development and lack of experience in searching for food may impair survival. The behavioral changes that accompany nest departure are therefore expected to be finely tuned to body condition and growth by endocrine processes. This study examines the possible involvement of corticosterone (CORT) in the stimulation of fledging in White storks through measurement of the changes in its plasma levels in relation to growth, nutritional status and the hatching rank of nestlings. For the first time in nest-bound chicks, we show that fledging is preceded by a marked and progressive 4 fold increase in baseline plasma CORT levels concomitant with an increase in locomotor activity (wing flapping) at the nest. Data on changes in body size, body mass, plasma metabolites and feeding frequency support the view that the increase in plasma CORT was induced by food restriction rather than being endogenously programmed. The timing and intensity of plasma CORT increase was dependent on the hatching rank within a brood, this increase being blunted in the less developed chicks possibly to avoid the impairment of final wing growth. These results show that an increase in plasma CORT as a result of food restriction and through the stimulation of locomotor activity is involved in the control of fledging in White storks. Moreover the CORT increase is adjusted to the hatching rank-related growth status of nestlings.     

Corticosterone administration does not affect timing of breeding in Florida scrub-jays (Aphelocoma coerulescens)

Providing supplemental food to Florida scrub-jays (Aphelocoma coerulescens) causes a reliable advance in clutch initiation of 1 to 2 weeks. In some years, supplemental food appeared to not only advance laying date but also decrease baseline concentrations of corticosterone (CORT) relative to controls. The coincidence of low CORT levels and early breeding led us to hypothesize that CORT serves to communicate information about environmental conditions to the hypothalamic-pituitary-gonadal (HPG) axis, which ultimately influences the timing of breeding. To test this hypothesis, we administered small oral doses of CORT three times each day to female breeders that were provisioned with supplemental food. We compared clutch initiation dates of the CORT-dosed females to females with supplementation but no exogenous CORT and to females with neither CORT nor supplemental food. CORT administration had a strong temporary effect on circulating CORT concentrations but clutch initiation did not differ between the two groups of supplemented birds, both of which laid eggs approximately 10 days earlier than nonsupplemented birds. Furthermore, during the year of our study we found no reduction in baseline CORT concentrations in our undosed supplemental groups, as had been observed in past studies.     

Plasma corticosterone increases during migratory restlessness in the captive white-crowned sparrow Zonotrichia leucophrys gambelli

Plasma corticosterone increases during the period of spring migration in a variety of bird species. Long-distance migrants show elevations in corticosterone specifically in association with the stage of flight, suggesting that corticosterone may support flight-related processes, for example, locomotor activity and/or energy mobilization. The pattern of corticosterone secretion as it relates to migratory flight has hitherto not been clearly described in migrants that frequently interrupt flight to refuel, for example, the Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii). The Gambel's white-crowned sparrow fuels by day and expresses peak migratory activity during the first few hours of night. To determine if plasma corticosterone increases in association with the stage of migratory flight also in this short-bout migrant, we induced captive white-crowned sparrows to enter into the migratory condition by placing photosensitive birds on long days (16L:8D) and then evaluated birds for plasma corticosterone and locomotor activity during four time points of the day. Patterns found in long-day birds were compared to those observed in short-day controls (8L:16D). Differences in energy metabolism as determined from plasma metabolites were also evaluated. We found that locomotor activity and corticosterone were significantly elevated at the onset of the dark period, but only in long-day birds. Plasma beta-hydroxybutyrate (a ketone body) was also elevated. Thus, findings suggest that plasma corticosterone and ketogenesis increase in association with migratory restlessness in a short-bout migrant. In fact, corticosterone may play a regulatory role, because it shows a trend to increase already before night-time activity.     

Effects of captivity and body condition on plasma corticosterone, locomotor behavior, and plasma metabolites in curve-billed thrashers

The acute stress response involves the secretion of catabolic glucocorticoids, such as corticosterone (CORT) in birds, that mobilize intrinsic energy stores primarily through a gluconeogenic pathway involving fat breakdown, thus linking body condition and stress. We measured changes in CORT and gluconeogenic metabolites (triglycerides, free glycerols, glucose) during handling stress in curve-billed thrashers Toxostoma curvirostre from two habitats (urban vs. desert) that may differ in food abundance in the wild, in captivity, and in response to both food restriction and subsequent recovery. Urban thrashers were heavier and secreted more CORT than desert birds in the field, but differences did not persist in captivity. Decreased access to food resulted in decreased body mass and a diminished ability to elevate plasma CORT in response to handling stress. However, the opposite effect was observed as these birds recovered from food restriction. Plasma levels of glucose and triglycerides did not change with stress. Food restriction also increased locomotor activity, which likely further exacerbated energy loss. These observations suggest that body condition and stress differences between urban and desert birds may be related to differences in their relative energetic states, possibly due to food availability. Body condition may affect the extent to which an individual can elevate CORT and use free glycerol as energy during acute stress.     

Does sex matter? Differential responses to corticosterone administration in the zebra finch

Because of potentially deleterious effects of chronic stress, physiological measurements of stress hormones (in birds, corticosterone (CORT)) are often used to determine the consequences of natural or human-induced change. Often, it is assumed that CORT levels will be similar between the sexes and the results are pooled. However, recent studies have reported sex differences in CORT concentrations in avian species. As zebra finches (Taeniopygia guttata) are one of the most widely used bird species in laboratory studies worldwide, potential sex-specific differences in hormone metabolism, as well as the clearance rate of oral doses of exogenous CORT, are highly relevant. The results of this study show that female zebra finches have a significantly higher baseline CORT than males, which could partially be a product of differential responses to semi-isolation. In addition, a single dose of exogenous CORT resulted in different blood profiles between the sexes over time, though exogenous CORT was cleared from blood within 90 min following treatment in both sexes. Interestingly, exposure to multiple doses of exogenous CORT resulted in elevated CORT levels 24 h after treatment in both sexes. These results highlight the need for further investigations into potential sex differences in hormone metabolism, as well as possible cumulative effects of repeated stress.