Glucocorticoid metabolites in an ex situ nocturnal bird, the tropical screech owl Megascops choliba: effects of sex, activity period and inter-individual variation (original) (raw)

Measuring Fecal Glucocorticoid Metabolites in Mammals and Birds: The Importance of Validation

Annals of the New York Academy of Sciences, 2005

In recent years, the noninvasive monitoring of steroid hormone metabolites in feces of mammals and droppings of birds has become an increasingly popular technique. It offers several advantages and has been applied to a variety of species under various settings. However, using this technique to reliably assess an animal's adrenocortical activity is not that simple and straightforward to apply. Because clear differences regarding the metabolism and excretion of glucocorticoid metabolites (GCMs) exist, a careful validation for each species and sex investigated is obligatory. In this review, general analytical issues regarding sample storage, extraction procedures, and immunoassays are briefly discussed, but the main focus lies on experiments and recommendations addressing the validation of fecal GCM measurements in mammals and birds. The crucial importance of scrutinizing the physiological and biological validity of fecal GCM analyses in a given species is stressed. In particular, the relevance of the technique to detect biologically meaningful alterations in adrenocortical activity must be shown. Furthermore, significant effects of the animals' sex, the time of day, season, and different life history stages are discussed, bringing about the necessity to seriously consider possible sex differences as well as diurnal and seasonal variations. Thus, comprehensive information on the animals' biology and stress physiology should be carefully taken into account. Together with an extensive physiological and biological validation, this will ensure that the measurement of fecal GCMs can be used as a powerful tool to assess adrenocortical activity in diverse investigations on laboratory, companion, farm, zoo, and wild animals.

A Generalized Fecal Glucocorticoid Assay for Use in a Diverse Array of Nondomestic Mammalian and Avian Species

General and Comparative Endocrinology, 2000

Noninvasive fecal glucocorticoid analysis has tremendous potential as a means of assessing stress associated with environmental disturbance in wildlife. However, interspecific variation in excreted glucocorticoid metabolites requires careful selection of the antibody used in their quantification. We compared four antibodies for detecting the major fecal cortisol metabolites in yellow baboons following 3 H cortisol administration, ACTH challenge, and HPLC separation of fecal glucocorticoid metabolites. The most effective antibody (ICN corticosterone RIA; Cat. No. 07-120102) demonstrated relatively high cross-reactivities to the major cortisol metabolites present in feces during peak excretion, following both radiolabel infusion and ACTH challenge. This same anti-body also detected increased fecal glucocorticoid metabolites after ACTH administration in the African elephant, black rhinoceros, Roosevelt elk, gerenuk, scimitarhorned oryx, Alaskan sea otter, Malayan sun bear, cheetah, clouded leopard, longtailed macaque, and northern spotted owl. Results suggest that (1) fecal glucocorticoid assays reliably detect endogenous changes in adrenal activity of a diverse array of species and (2) where comparisons were made, the ICN corticosterone antibody generally was superior to other antibodies for measuring glucocorticoid metabolites in feces.

Measurement of free glucocorticoids: quantifying corticosteroid binding capacity and its variation within and among mammal and bird species

Conservation Physiology, 2020

Plasma glucocorticoid (CORT) levels are one measure of stress in wildlife and give us insight into natural processes relevant to conservation issues. Many studies use total CORT concentrations to draw conclusions about animals'stress state and response to their environment. However, the blood of tetrapods contains corticosteroid-binding globulin (CBG), which strongly binds most circulating CORT. Only free CORT (CORT not bound by CBG) leaves the circulation and exerts biological effects on CORT-sensitive tissues. Measuring free CORT concentrations provides insight to an animal's stress response that cannot be revealed by simply measuring total CORT. To calculate free CORT concentrations in plasma or serum samples, one needs three measurements: the binding affinity of CBG for CORT (which varies by species), the total CORT concentration in the sample and the maximum corticosteroid binding capacity (MCBC) of CBG in the sample. Here, we detail the measurement of CBG binding capacity. We compare and contrast the three main methods to measure MCBC: charcoal, cell harvester and dialysis. Each is defined by the means by which free and bound CORT are separated. We weigh the relative merits and challenges of each. We conclude that sample volume, species and taxon binding specificity, and availability of equipment are the primary considerations in selecting the appropriate separation method. For most mammals, the charcoal method is recommended. For birds, the harvester method has critical advantages over the charcoal method. The dialysis method is widely regarded as the gold standard and has lower equipment costs but is more time-intensive and costly in terms of radioactive isotope needed and is less suited to processing large numbers of samples. The binding capacity of CBG varies tremendously within and among the bird and marine mammal species studied, and we discuss the implication of this variation for understanding the role of stress in wildlife.

Diurnal variation in corticosterone release among wild tropical forest birds

Frontiers in Zoology, 2016

Background: Glucocorticoids are adrenal steroid hormones essential to homeostatic maintenance. Their daily variation at low concentrations regulates physiology and behavior to sustain proper immunological and metabolic function. Glucocorticoids rise well above these baseline levels during stress to elicit emergency-state responses that increase short-term survival. Despite this essence in managing life processes under both regular and adverse conditions, relationships of glucocorticoid release to environmental and intrinsic factors that vary at daily and seasonal scales are rarely studied in the wild. Methods: This study on 41 passerine species of the Ecuadorian Chocó applied a standardized capture-and-restraint protocol to examine diurnal variation in baseline and stress-related release of corticosterone, the primary avian glucocorticoid. Tests for relationships to relative body mass, hemoglobin concentration, molt status and date complemented this evaluation of the time of day effect on corticosterone secretion in free-living tropical rainforest birds. Analyses were also partitioned by sex as well as performed separately on two common species, the wedgebilled woodcreeper and olive-striped flycatcher. Results: Interspecific analyses indicated maximum baseline corticosterone levels at the onset of the active phase and reductions thereafter. Stress-related levels did not correspond to time of day but accompanied baseline reductions during molt and elevations in birds sampled later during the September-November study period. Baseline corticosterone related negatively to hemoglobin in the wedge-billed woodcreeper and stress-related levels increased with body mass in the olive-striped flycatcher. There were no substantial sex-related differences. Conclusions: The results of this study suggest a diurnal rhythmicity in baseline corticosterone release so robust as to emerge in pooled analyses across a highly variable dataset. While this detection in nature is singular, correspondent patterns have been demonstrated outside of the tropics in captive model species. Congruity in daily rhythms and links to physiological and life-history state across disparate taxa and environments may promote the yet unresolved utility of corticosterone release as a global metric for population health. However, certain results of this study also deviate from laboratory and field research at higher latitudes, cautioning generalization. Environmental distinctions such as high productivity and tempered seasonality may precipitate unique life-history strategies and underlying hormonal mechanisms in tropical rainforest birds.

A Validation of Extraction Methods for Noninvasive Sampling of Glucocorticoids in Free-Living Ground Squirrels

Physiological and Biochemical Zoology, 2005

Fecal hormone assays provide a powerful tool for noninvasive monitoring of endocrine status in wild animals. In this study we validated a protocol for extracting and measuring glucocorticoids in free-living and captive Belding's ground squirrels (Spermophilus beldingi). We first compared two commonly used extraction protocols to determine which performed better with commercially available antibodies. We next verified the preferred extraction method by correlating circulating and fecal glucocorticoid measures from a group of individuals over time. For this comparison, we used both a cortisol and a corticosterone antibody to determine which had greater affinity to the fecal metabolites. Cortisol was the primary circulating glucocorticoid, but both hormones were present in well above detectable concentrations in the blood, which does not occur in other sciurids. In addition, the cortisol antibody showed greater binding with the fecal extracts than did the corticosterone antibody. Finally, we used adrenocorticotropic hormone and dexamethasone challenges to demonstrate that changes in adrenal functioning are reflected in changing fecal corticoid levels. These results suggest that our extraction protocol provides a fast, reliable assay of stress hormones in free-living ground squirrels without the confounding influence of short-term rises in glucocorticoid concentrations caused by handling and restraint stress and that it can facilitate ecological and evolutionary studies of stress in wild species.

Do seasonal glucocorticoid changes depend on reproductive investment? A comparative approach in birds

Integrative and comparative biology, 2018

Animals go through different life history stages such as reproduction, moult, or migration, of which some are more energy-demanding than others. Baseline concentrations of glucocorticoid hormones increase during moderate, predictable challenges and thus are expected to be higher when seasonal energy demands increase, such as during reproduction. By contrast, stress-induced glucocorticoids prioritize a survival mode that includes reproductive inhibition. Thus, many species down-regulate stress-induced glucocorticoid concentrations during the breeding season. Interspecific variation in glucocorticoid levels during reproduction has been successfully mapped onto reproductive investment, with species investing strongly in current reproduction (fast pace of life) showing higher baseline and lower stress-induced glucocorticoid concentrations than species that prioritize future reproduction over current attempts (slow pace of life). Here we test the "glucocorticoid seasonal plasticity ...

Glucocorticoid response to food availability in breeding barn swallows (Hirundo rustica)

General and Comparative Endocrinology, 2008

Parents feeding altricial nestlings have to trade-off the competing demands of self-maintenance and reproductive investment over their lifetime. Corticosterone, a glucocorticoid hormone released by birds in response to stressors, might play a key role in regulating parental investment when conditions unexpectedly deteriorate. However, birds breeding in unpredictable environmental conditions have been shown not to increase circulating levels of corticosterone as a response to bad weather to avoid nest abandonment when investment in offspring is high or when the probability of re-nesting is low. We investigated whether parent barn swallows Hirundo rustica, a passerine bird whose aerial insect food varies greatly in abundance depending on weather, also belongs to those species or whether it responds with an activation of the hypothalamo-pituitary-adrenal axis to natural variation in insect availability. We correlated plasma corticosterone levels of parents with weather conditions, the availability of aerial insects and parental body condition. Plasma corticosterone concentrations increased when mean daytime temperature declined, and consequently insect availability decreased and body condition of the parents deteriorated. Low temperatures also had a negative effect on body mass of the nestlings and there was a negative relationship between circulating corticosterone of parents and body mass of nestlings. We conclude that corticosterone is probably involved in the regulation of parental investment.

Heritable variation in circulating glucocorticoids and endocrine flexibility in a free-living songbird

Journal of evolutionary biology, 2017

Phenotypic flexibility is a central way that organisms cope with challenging and changing environments. As endocrine signals mediate many phenotypic traits, heritable variation in hormone levels, or their context-dependent flexibility, could present an important target for selection. Several studies have estimated the heritability of circulating glucocorticoid levels under acute stress conditions, but little is known about the potential for either baseline hormone levels or rapid endocrine flexibility to evolve. Here we assessed the potential for selection to operate on the elevation (circulating hormone levels) and flexibility of glucocorticoid reaction norms to acute restraint stress. Multivariate animal models revealed low but significant heritability in baseline (h(2) =0.13-0.14) and stress-induced glucocorticoids (h(2) =0.18), and moderate heritability in glucocorticoid flexibility in response to acute stress (h(2) =0.38) in free-living juvenile tree swallows (Tachycineta bicol...

Bird Hormones and Bird Migrations: Analyzing Hormones in Droppings and Egg Yolks and Assessing Adaptations in Long-Distance Migration

In recent years, the noninvasive monitoring of steroid hormone metabolites in feces of mammals and droppings of birds has become an increasingly popular technique. It offers several advantages and has been applied to a variety of species under various settings. However, using this technique to reliably assess an animal's adrenocortical activity is not that simple and straightforward to apply. Because clear differences regarding the metabolism and excretion of glucocorticoid metabolites (GCMs) exist, a careful validation for each species and sex investigated is obligatory. In this review, general analytical issues regarding sample storage, extraction procedures, and immunoassays are briefly discussed, but the main focus lies on experiments and recommendations addressing the validation of fecal GCM measurements in mammals and birds. The crucial importance of scrutinizing the physiological and biological validity of fecal GCM analyses in a given species is stressed. In particular, the relevance of the technique to detect biologically meaningful alterations in adrenocortical activity must be shown. Furthermore, significant effects of the animals' sex, the time of day, season, and different life history stages are discussed, bringing about the necessity to seriously consider possible sex differences as well as diurnal and seasonal variations. Thus, comprehensive information on the animals' biology and stress physiology should be carefully taken into account. Together with an extensive physiological and biological validation, this will ensure that the measurement of fecal GCMs can be used as a powerful tool to assess adrenocortical activity in diverse investigations on laboratory, companion, farm, zoo, and wild animals.