John F. Cockrem

Stress, corticosterone responses and avian personalities

Birds are constantly responding to stimuli from their environment. When these stimuli are perceived as threatening, stress responses are initiated, with activation of the hypothalamo-pituitary-adrenal axis and the release of corticosterone from the adrenal gland. The basic emotion of fear is also experienced during a stress response. Corticosterone stress responses and behavioural responses to stimuli vary markedly between individual birds, raising questions about the significance of these individual differences and about the relationship between corticosterone responses and fearfulness in birds. Although fearfulness can be challenging to measure, data from several species indicate that corticosterone responses and fear behaviour responses are linked in individual birds. Consistent profiles of behavioural responses of birds to a wide range of stimuli can be identified and are called personalities. Personalities vary along a continuum, but are usually classified as proactive or reactive. Individual corticosterone and behaviour responses depend on each bird’s personality. Birds with proactive personalities have relatively active behavioural responses and relatively low corticosterone stress responses, whilst birds with reactive personalities have relatively passive behavioural responses and large corticosterone responses. Relationships between the physiological and behavioural characteristics of avian personalities can now be explored in detail to determine the significance of individual differences in stress responses and personalities in birds.

Individual variation in glucocorticoid stress responses in animals

When stimuli from the environment are perceived to be a threat or potential threat then animals initiate stress responses, with activation of the hypothalamo-pituitary-adrenal axis and secretion of glucocorticoid hormones (cortisol and corticosterone). Whilst standard deviation or standard error values are always reported, it is only when graphs of individual responses are shown that the extensive variation between animals is apparent. Some animals have little or no response to a stressor that evokes a relatively large response in others. Glucocorticoid responses of fish, amphibian, reptiles, birds, and mammals are considered in this review. Comparisons of responses between animals and groups of animals focused on responses to restraint or confinement as relatively standard stressors. Individual graphs could not be found in the literature for glucocorticoid responses to capture or restraint in fish or reptiles, with just one graph in mammals with the first sample was collected when animals were initially restrained. Coefficients of variation (CVs) calculated for parameters of glucocorticoid stress responses showed that the relative magnitudes of variation were similar in different vertebrate groups. The overall mean CV for glucocorticoid concentrations in initial (0 min) samples was 74.5%, and CVs for samples collected over various times up to 4 h were consistently between 50% and 60%. The factors that lead to the observed individual variation and the extent to which this variation is adaptive or non-adaptive are little known in most animals, and future studies of glucocorticoid responses in animals can focus on individual responses and their origins and significance.

Corticosterone responses in birds: Individual variation and repeatability in Adelie penguins (Pygoscelis adeliae) and other species, and the use of power analysis to determine sample sizes

Plasma corticosterone concentrations increase when birds experience a stressor, and in this study we quantified variation in corticosterone responses for the first time in a species of free-living bird. Adelie penguins (Pygoscelisadeliae) nesting at Cape Bird on Ross Island in Antarctica were sampled on three occasions. Penguins with relatively low or high corticosterone responses on the first occasion had consistently low or high responses, as previously found for great tits and chickens. A model for birds is proposed in which birds with low corticosterone responses and proactive personalities are likely to be more successful (have greater fitness) in constant or predictable conditions, whilst birds with reactive personalities and high corticosterone responses will be more successful in changing or unpredictable conditions. There is thus no linear relationship between the size of a corticosterone response and fitness. Whilst the absolute magnitude of corticosterone responses varies markedly across species of birds, coefficients of variation are similar. Individual corticosterone responses are generally repeatable, with significant statistical repeatabilities for 30 min corticosterone concentrations and integrated corticosterone concentrations in the Adelie penguin, great tit and chicken. Coefficients of variation in corticosterone responses between birds and power analyses were used to provide a rule of thumb for determining differences between groups of birds in mean corticosterone concentrations to enable statistical analyses to have acceptable levels of statistical power for given sample sizes. It is suggested that power analyses and this rule of thumb be adopted in future investigations of corticosterone responses in birds.

Competition for male reproductive investment elevates testosterone levels in female dunnocks, Prunella modularis

In many songbirds, females occasionally sing in contexts of high female–female competition. Testosterone may be involved in the activation of song, because testosterone implants elicit female song in many species with rare female song. A possible mechanism for the hormonal control of female song is provided by the challenge hypothesis, which predicts a rise in testosterone in response to aggressive interactions during socially unstable situations. We tested this by comparing faecal testosterone levels in polygynandrous and monogamous female dunnocks. In groups with two to three females (polygynandry and polygyny) males provide less help at each nest than in groups with a single female (monogamy and polyandry). Polygynandrous and polygynous females are aggressive towards one another and attempt to expel rivals. Polygynandrous females had significantly higher testosterone levels than monogamous females. Competition between females that was induced by removal of males caused testosterone levels to rise. Further, female testosterone levels were correlated with the rate of ‘tseep’ calls, which are produced during aggressive encounters between females. Finally, polygynandrous and polygynous females sang significantly more than monogamous females. To the best of our knowledge, these results provide the first experimental support for the challenge hypothesis in female birds, and suggest that testosterone can regulate facultative female song in songbirds.

Conservation and behavioral neuroendocrinology

The total number of threatened species of vertebrates is likely to be more than 10,000, with approximately one quarter of the worlds mammal species, one eighth of the birds and one third of the amphibians threatened with extinction. The rate of loss of animal species and hence of biodiversity is increasing and may become even greater as ecosystems become affected by climate change due to global warming. Behavioral neuroendocrinology, which considers interactions between behavior and neuroendocrine function in animals from all vertebrate taxa, can contribute to animal conservation. Research with laboratory animals can address questions in basic biology relevant to conservation and develop methods for use with threatened animals. Field work with free-living animals considers the basic biology of new species and the use of endocrine tools to assess the susceptibility of species to threats. Non-invasive measurements of hormone concentrations, especially fecal steroids, are extensively used to assess reproductive function and the stress status of animals in captive breeding programs and in the wild. Biodiversity and natural selection both depend on individual variation, and conservation programs often work with animals on an individual basis. The consideration of data from individuals is essential in conservation endocrinology. Direct contributions to conservation programs are challenging as study situations are determined by practical conservation concerns. Indirect contributions such as the provision of scientific input to conservation plans and participation in public education programs offer significant benefits for conservation programs. Directly and indirectly, there are many opportunities for behavioral neuroendocrinologists to contribute to conservation.

Corticosterone and fear behaviour in white and brown caged laying hens

1. Physiological and behavioural measures of stress in caged hens on a commercial farm were compared between White Leghorn and brown Hyline strains, and between three tiers of cages. 2. Blood and faecal samples were collected from undisturbed birds for corticosterone measurements. Plasma corticosterone responses to a stressor were measured by the collection of blood samples after 15, 30, and 60 min of a handling stressor. Tonic immobility and novel object tests were used to measure fear behaviour. 3. Plasma corticosterone in undisturbed hens and faecal corticosterone did not differ between White Leghorn and brown Hyline hens, whereas the plasma corticosterone response to a handling stressor was greater in White Leghorns. The duration of tonic immobility, latency to first head movement and number of head movements in tonic immobility tests were greater in white than brown birds, whereas the number of inductions was less for tonic immobility tests. There were no differences between the strains in their responses to a novel object. 4. There were no differences between tiers in plasma corticosterone or corticosterone responses or tonic immobility responses, and no consistent differences in responses of birds to a novel object. 5. This is the first study in which plasma and faecal corticosterone concentrations and fear behaviour have been measured together in laying hens, and the first description of plasma corticosterone responses to handling over 60 min for caged laying hens on a commercial farm. The study has shown the value of measuring endocrine and behavioural variables together to provide objective data on characteristics of different strains of hens.

Sulphatoxymelatonin excretion in older people: Relationship to plasma melatonin and renal function

Baskett JJ, Cockrem JF, Antunovich TA. Sulphatoxymelatonin excretion in older people: Relationship to plasma melatonin and renal function. J. Pineal Res. 1998; 24:58–61.

Urinary corticosterone metabolite responses to capture, and annual patterns of urinary corticosterone in wild and captive endangered Fijian ground frogs (Platymantis vitiana)

This study was based on the development of a non-invasive glucocorticoid enzyme-immunoassay for the assessment of stress in wild and captive endangered Fijian ground frogs (Platymantis vitiana). Enzyme-immunoassays were developed and validated for the first time to non-invasively measure both cortisol and corticosterone metabolites in frog urine. Frog urine showed parallel displacement with corticosterone but not cortisol standards, therefore corticosterone enzyme immunoassays were used to examine stress in wild and captive frogs. Urinary corticosterone metabolite concentrations increased in frog urine (n = 4) at 6 h, 1 day and 2 days after injection with adrenocorticotropic hormone (0.44 μg g–1 bodyweight), indicating that the corticosterone enzyme-immunoassay could detect changes in circulating corticosterone in frogs. Urinary concentrations of corticosterone were measured in wild frogs (n = 18) after capture in the field. The first measurement beyond the initial sample was at 2–3 h. Mean urinary corticosterone concentrations rose after the initial sample and were significantly elevated in samples collected 3–4 h after capture. This is the first demonstration of a urinary corticosterone response to capture in amphibians. Urinary corticosterone metabolite concentrations for all months combined were lower in captive males than in wild males, and differed between vitellogenic, non-vitellogenic and captive females. Concentrations did not differ between captive and wild females. In conclusion, urinary corticosterone enzyme immunoassays can be used in frogs for assessing stress responses to capture and natural stress profiles of both captive and wild populations.

Faecal measurements of oestradiol and testosterone allow the non‐invasive estimation of plasma steroid concentrations in the domestic fowl

1. Radioimmunoassays were validated for oestradiol and testosterone in faecal and plasma samples from domestic fowls. Faecal samples were prepared in phosphate buffer. A delay of up to 24 h between defaecation and the freezing of a dropping did not affect faecal steroid concentrations. 2. Differences in plasma concentrations of testosterone and oestradiol between cockerels and hens were clearly reflected in faecal hormone concentrations. Faecal concentrations of oestradiol were low in males and did not overlap with much higher concentrations in females, whereas there was some overlap between the sexes in faecal concentrations of testosterone. 3. The correlation coefficients (r2) between plasma and faecal steroid concentrations were 0.464 for testosterone and 0.852 for oestradiol (log-transformed data). The coefficients did not increase when the mean hormone concentrations for several droppings produced by each bird during a collection period were used. 4. Faecal steroid concentrations can be used as a measure of plasma steroid concentrations and hence of gonadal activity in chickens. The resolution of the faecal steroid method is less than if plasma measurements were used, but it offers a practical alternative to blood sampling that is non-invasive and does not require birds to be handled.

Urinary corticosterone metabolite responses to capture and captivity in the cane toad (Rhinella marina)

Urinary corticosterone metabolite responses to capture have recently been shown for the first time in amphibians, and in the present study urinary corticosterone metabolite responses to capture and to confinement in captivity were measured in adult cane toads (Rhinella marina) in Queensland, Australia. An adrenocorticotropic hormone (ACTH) challenge was used to provide a biological validation for urinary corticosterone metabolite concentrations measured by radioimmunoassay (RIA). Urinary corticosterone metabolite increased 1-2 days after ACTH but not saline injection and then returned to initial values, indicating that the RIA could detect changes in corticosterone secretion in toads. Urinary corticosterone metabolite responses to short-term capture and restraint in plastic bags were first apparent 2h after capture of wild toads. Toads held communally in captivity for 5 days had elevated urinary corticosterone metabolite concentrations. Mean corticosterone concentrations declined significantly after a further 7 days in individual housing chambers. There was no sex difference in urinary corticosterone metabolite responses of toads to ACTH challenge, short-term capture or captivity. The relative amount of variation in the mean corticosterone responses was quantified by calculating coefficients of variation (CV) for each mean corticosterone response. Mean corticosterone at 0 min was more variable for captive toads than wild toads. Furthermore, initial corticosterone concentrations (0 min) were more variable than concentrations during the ACTH challenge, short-term capture and captivity. There was little change in the amount of variation of mean corticosterone levels between male and female toads with increasing time in captivity (12-29 days). This study has shown individual corticosterone responses of amphibians for the first-time, and has provided a novel method for quantifying the relative amount of variation in amphibian corticosterone responses.