"Home alone!" influence of nest parental attendance on offspring behavioral and hormonal stress responses in an Antarctic seabird, the snow petrel (Pagodroma nivea).

In altricial species, parents brood their chicks constantly before leaving them unattended sometimes for extended periods when they become thermally independent. During this second phase, there is sometimes important inter-individual differences in parental attendance and the fitness costs and benefits of parental strategies have previously been extensively investigated. However, the impact of parental presence on offspring behaviors and stress physiology has been overlooked. Here, we examined the influence of parental presence on offspring hormonal and behavioral stress sensitivities in snow petrel chicks. We demonstrated for the first time in a wild bird species that attended chicks had lower stress-induced corticosterone levels and a lower probability to show defensive behavior compared to the alone chicks. This reduced stress sensitivity is certainly explained by the well-known link between corticosterone and nutritional status, and by the recent delivery of meals to the attended chicks and the improvement of their nutritional status. It may also be explained by the parental protection against predators or inclement weather, or/and by the psychosocial comfort of parental presence for the offspring. Overall, these results suggest that the presence of a parent in the nest reduces offspring stress sensitivity in wild birds. Further studies would now be required to disentangle the impact of nutritional status and parental presence on stress sensitivity and to better understand the potential impact of parental presence and circulating corticosterone levels on growth and cognitive development in wild birds.

Does urbanization cause stress in wild birds during development? Insights from feather corticosterone levels in juvenile house sparrows (Passer domesticus).

Urban landscapes are associated with abiotic and biotic environmental changes that may result in potential stressors for wild vertebrates. Urban exploiters have physiological, morphological, and behavioral adaptations to live in cities. However, there is increasing evidence that urban exploiters themselves can suffer from urban conditions, especially during specific life-history stages. We looked for a link between the degree of urbanization and the level of developmental stress in an urban exploiter (the house sparrow, Passer domesticus), which has recently been declining in multiple European cities (e.g., London, UK). Specifically, we conducted a large-scale study and sampled juvenile sparrows in 11 urban and rural sites to evaluate their feather corticosterone (CORT) levels. We found that juvenile feather CORT levels were positively correlated with the degree of urbanization, supporting the idea that developing house sparrows may suffer from urban environmental conditions. However, we did not find any correlation between juvenile feather CORT levels and body size, mass, or body condition. This suggests either that the growth and condition of urban sparrows are not impacted by elevated developmental CORT levels, or that urban sparrows may compensate for developmental constraints once they have left the nest. Although feather CORT levels were not correlated with baseline CORT levels, we found that feather CORT levels were slightly and positively correlated with the CORT stress response in juveniles. This suggests that urban developmental conditions may potentially have long-lasting effects on stress physiology and stress sensitivity in this urban exploiter.

The behavioural and physiological stress responses are linked to plumage coloration in the rock pigeon (Columbia livia).

In wild vertebrates, conspicuousness often signals a high phenotypic quality and is therefore associated with fitness benefits ("the handicap principle hypothesis"). However, conspicuous individuals usually face a higher risk of predation because they are easier to detect. Therefore, conspicuous individuals are expected to modify their behavioural and their physiological stress responses to limit the negative effect of their conspicuousness on survival. We examined the link between plumage coloration and the stress response in a bird species (the rock pigeon, Columbia livia) by comparing two groups of pigeons with different plumage characteristics: 'blue-bar' birds (lighter birds) and 'checker' birds (darker birds). Specifically, we measured several behavioural and physiological components of the stress response (breath rate, corticosterone, and tonic immobility). Although breath rate did not differ between 'checker' and 'blue bar' birds, the corticosterone stress response of 'blue-bar' birds was greater relative to that of 'checker' birds. Moreover, 'checker' birds were more likely to initiate tonic immobility relative to 'blue bar' birds. Therefore, this study suggests that the behavioural and physiological stress responses of wild birds may differ according to their plumage coloration. To conclude, our results suggest that plumage colorations may be functionally linked to specific behavioural and physiological sensitivities to stressors.

Water deprivation increases maternal corticosterone levels and enhances offspring growth in the snake Vipera aspis.

Circulating glucocorticoid (GC) levels may increase as a result of reproductive effort or in response to unpredictable events. However, GC secretion can vary with the availability of vital trophic resources such as energy. While water represents another critical resource, the impact of water deprivation on GC secretion during reproduction has not yet been thoroughly investigated. Here, we examined the effects of water deprivation on plasma corticosterone (CORT) concentrations of female aspic vipers (Vipera aspis), and determined the impacts of water deprivation on offspring traits. We exposed both pregnant and non-reproductive females to a 20-day water deprivation and compared their pre- and post-deprivation CORT levels with those of control females. At the end of the treatment, only water-deprived pregnant females showed a significant increase in CORT levels. In pregnant females, changes in baseline CORT level were correlated with changes in female hydration state. Changes in baseline CORT levels were also negatively influenced by maternal reproductive effort in pregnant control females, while such a relationship was not apparent in pregnant water-deprived females. Finally, we found that offspring from water-deprived females had higher growth rates than offspring from control females. Offspring growth was also positively correlated with changes in both maternal osmolality and baseline CORT levels. Together, our results suggest that dehydration increases maternal CORT levels, which may subsequently influence offspring development. Further long-term field studies are therefore required to assess whether there is an adaptive significance of this response.

No evidence for an effect of traffic noise on the development of the corticosterone stress response in an urban exploiter.

Anthropogenic noise can have important physiological and behavioral effects on wild animals. For example, urban noise could lead to a state of chronic stress and could alter the development of the hypothalamus-pituitary-adrenal (HPA) axis. Supporting this hypothesis, several studies have found that human disturbance is associated with increased circulating corticosterone (CORT) levels. However, it remains unclear whether increased CORT levels are the result of anthropogenic noise or other anthropogenic factors. Here, we experimentally tested the impact of urban noise on the CORT stress response in an urban exploiter (the house sparrow, Passer domesticus) by exposing chicks to a traffic noise ('disturbed chicks') or not ('control chicks'). If noise exposure has a negative impact on developing chicks, we predicted that (1) disturbed chicks will grow slower, will be in poorer condition, and will have a lower fledging probability than controls; (2) disturbed chicks will have higher baseline CORT levels than control; (3) the CORT stress response will be affected by this noise exposure. Contrary to these predictions, we found no effect of our experiment on growth, body condition, and fledging success, suggesting that house sparrow chicks were not negatively affected by this noise exposure. Moreover, we did not find any effect of noise exposure on either baseline CORT levels or the CORT stress response of chicks. This suggests not only that house sparrow chicks did not perceive this noise as stressful, but also that the development of the HPA axis was not affected by such noise exposure. Our study suggests that, contrary to urban avoiders, urban exploiters might be relatively insensitive to urban noise during their development. Further comparative studies are now needed to understand whether such insensitivity to anthropogenic noise is a consistent phenomenon in urban exploiters and whether this is a major requirement of an urban way of life.

Does the stress response predict the ability of wild birds to adjust to short-term captivity? A study of the rock pigeon (Columbia livia).

Although the transfer of wild animals to captivity is crucial for conservation purposes, this process is often challenging because some species or individuals do not adjust well to captive conditions. Chronic stress has been identified as a major concern for animals held on long-term captivity. Surprisingly, the first hours or days of captivity have been relatively overlooked. However, they are certainly very stressful, because individuals are being transferred to a totally novel and confined environment. To ensure the success of conservation programmes, it appears crucial to better understand the proximate causes of interspecific and interindividual variability in the sensitivity to these first hours of captivity. In that respect, the study of stress hormones is relevant, because the hormonal stress response may help to assess whether specific individuals or species adjust, or not, to such captive conditions ('the stress response-adjustment to captivity hypothesis'). We tested this hypothesis in rock pigeons by measuring their corticosterone stress response and their ability to adjust to short-term captivity (body mass loss and circulating corticosterone levels after a day of captivity). We showed that an increased corticosterone stress response is associated with a lower ability to adjust to short-term captivity (i.e. higher body mass loss and circulating corticosterone levels). Our study suggests, therefore, that a low physiological sensitivity to stress may be beneficial for adjusting to captivity. Future studies should now explore whether the stress response can be useful to predict the ability of individuals from different populations or species to not only adjust to short-term but also long-term captivity.

Housing conditions and sacrifice protocol affect neural activity and vocal behavior in a songbird species, the zebra finch (Taeniopygia guttata).

Individual cages represent a widely used housing condition in laboratories. This isolation represents an impoverished physical and social environment in gregarious animals. It prevents animals from socializing, even when auditory and visual contact is maintained. Zebra finches are colonial songbirds that are widely used as laboratory animals for the study of vocal communication from brain to behavior. In this study, we investigated the effect of single housing on the vocal behavior and the brain activity of male zebra finches (Taeniopygia guttata): male birds housed in individual cages were compared to freely interacting male birds housed as a social group in a communal cage. We focused on the activity of septo-hypothalamic regions of the "social behavior network" (SBN), a set of limbic regions involved in several social behaviors in vertebrates. The activity of four structures of the SBN (BSTm, medial bed nucleus of the stria terminalis; POM, medial preoptic area; lateral septum; ventromedial hypothalamus) and one associated region (paraventricular nucleus of the hypothalamus) was assessed using immunoreactive nuclei density of the immediate early gene Zenk (egr-1). We further assessed the identity of active cell populations by labeling vasotocin (VT). Brain activity was related to behavioral activities of birds like physical and vocal interactions. We showed that individual housing modifies vocal exchanges between birds compared to communal housing. This is of particular importance in the zebra finch, a model species for the study of vocal communication. In addition, a protocol that daily removes one or two birds from the group affects differently male zebra finches depending of their housing conditions: while communally-housed males changed their vocal output, brains of individually housed males show increased Zenk labeling in non-VT cells of the BSTm and enhanced correlation of Zenk-revealed activity between the studied structures. These results show that housing conditions must gain some attention in behavioral neuroscience protocols.

Physiological and fitness correlates of experimentally altered hatching asynchrony magnitude in chicks of a wild seabird.

Nest-bound chicks depend entirely on their parents for food, often leading to high sibling competition. Asynchronous hatching, resulting from the onset of incubation before clutch completion, facilitates the establishment of within-nest hierarchy, with younger chicks being subject to lower feeding and growth rates. Because social and nutritional stresses affect baseline stress hormone levels in birds, younger chicks are expected to have higher levels of corticosterone than their siblings. As previous studies showed that hatching asynchrony magnitude influences the course of sibling competition, it should also affect baseline corticosterone. We measured baseline corticosterone at age 5 days in nestling black-legged kittiwakes (Rissa tridactyla) in 3 types of experimental broods: synchronous, asynchronous, and highly asynchronous. Sexual dimorphism takes place during chick-rearing and might also influence baseline corticosterone, we thus included chick sex in our analyses and also monitored chick growth and survival. Baseline corticosterone did not differ among A-chicks, but was higher in B-chicks from highly asynchronous broods compared with the other brood types, in line with the presumed increase in nutritional stress. In asynchronous broods, A-chicks had higher baseline corticosterone than their siblings, contrary to our expectations. We interpret that result as a cost of dominance among A-chicks. In line with previous studies, mass gain was negatively correlated with baseline corticosterone levels. We found that baseline corticosterone predicted survival in a sex-specific way. Regardless of hatching rank, males with higher baseline corticosterone suffered higher mortality, suggesting that males were more sensitive to high level of stress, independently of its cause.

Modulation of the prolactin and the corticosterone stress responses: do they tell the same story in a long-lived bird, the Cape petrel?

Over the last decades, the corticosterone stress response has been suggested as a major physiological tool to understand what strategy an individual might adopt in response to environmental perturbations. More recently, another hormone related to parental care--prolactin--has been suggested as a complementary tool to investigate this question. Indeed, both of these hormones are affected by stressors and are involved in parental decisions, such as deserting the nest. Because of these similarities, it remains unclear what the functional distinction between the prolactin and corticosterone stress responses is. Here, we investigated whether natural variations of the corticosterone and prolactin stress responses are functionally linked in free-living Cape petrel (Daption capense) parents. If prolactin and corticosterone mediate the same functional response to a stressor and are the proxies of the same response, we predict that corticosterone and prolactin stress responses (1) will be modulated according to the same factors; (2) will affect reproductive performances in the same way; and, (3) of course, will be correlated. Contrary to these predictions, we found that the corticosterone and prolactin stress responses were respectively modulated according to body condition and breeding status. Moreover, prolactin levels, but not corticosterone levels, were related to hatching success in this species. Finally, we did not find any significant correlation between these two stress responses under any circumstances (failed breeders, incubating or chick rearing birds) and this result was overall supported by a review of the existing literature. Therefore, these two stress responses do not seem to be tightly linked and we believe that they may provide complementary pieces of information on parental investment in birds.

Relationships between POPs and baseline corticosterone levels in black-legged kittiwakes (Rissa tridactyla) across their breeding cycle.

Chronic exposure to persistent organic pollutants (POPs) in wildlife might alter the response to environmental changes through interference with the regulation of stress hormones. Here, we examined the relationship between blood concentrations of several POPs and baseline plasma corticosterone levels in the black-legged kittiwake (Rissa tridactyla) during three distinct periods in the breeding season. The concentrations of POPs and corticosterone increased, whereas body mass decreased progressively from the pre-laying period to the incubation and the chick rearing period. ∑PCB (polychlorinated biphenyls) correlated positively with the baseline corticosterone levels during the pre-laying period, which might suggest that PCBs affect the regulation of corticosterone. However, this relationship was not found during the incubation or the chick rearing period. Possible explanations are discussed with emphasis on how total stress/allostatic load is handled during different periods and conditions.

Corticosterone levels in host and parasite nestlings: is brood parasitism a hormonal stressor?

Parasite chicks from non-evictor species usually try to monopolize host parental care, thereby increasing considerably the level of food competition in the nest. Here, we propose that brood parasitism is an important stressor for host and parasite nestlings and explore this hypothesis in the non-evictor great spotted cuckoo (Clamator glandarius) and its main hosts, the same-sized black-billed magpie (Pica pica) and the larger carrion crow (Corvus corone). We experimentally created 3-nestling broods of different brood compositions (only cuckoo chicks, only host chicks, or cuckoo and host chicks together) and measured baseline corticosterone levels of nestlings along their developmental period (early, middle and late). We found that brood parasitism increased corticosterone levels in magpie nestlings in the mid and late nestling period compared to those raised in unparasitized nests. Interestingly, carrion crow nestlings from parasitized nests only increased their corticosterone levels in the mid nestling period, when the competition for food with the cuckoo nestling was highest. Our results suggest that brood parasitism could be a potential physiological stressor for host nestlings, especially during the developmental stages where food requirements are highest. Conversely, cuckoo nestlings could be physiologically adapted to high competition levels since they did not show significant differences in corticosterone levels in relation to brood composition.

Measuring corticosterone in seabird egg yolk and the presence of high yolk gestagen concentrations.

Large inter-species differences have been found in yolk corticosterone amounts in avian eggs. While some studies have failed to detect significant amounts of corticosterone, in other species high amounts have been recorded, such as in a recent study of southern rockhopper penguins Eudyptes chrysocome chrysocome. However, attention has been drawn recently to the fact that many antibodies for corticosterone measurement cross-react with other steroids present in the yolk. In particular, progesterone and related substances can occur in yolk in high concentrations, such that also low cross-reactions of corticosterone assays may lead to measurement errors. We thus performed high-performance liquid chromatographic (HPLC) analyses of yolk extracts and determined the concentration of immunoreactive corticosterone, as well as cross-reacting progesterone and cortisol in egg yolks of southern rockhopper penguins and imperial shags Phalacrocorax atriceps albiventer. We found that high gestagen concentrations in the yolk result in large measurement errors for yolk corticosterone, even when the cross-reactivity seems small. This was observed for both species. We further found species-specific differences in the actual corticosterone amounts present in the egg yolks.

Stress and the timing of breeding: glucocorticoid-luteinizing hormones relationships in an arctic seabird.

In birds, stressful environmental conditions delay the timing of breeding but the underlying mechanisms are poorly understood. The stress hormone corticosterone appears to be a good candidate for mediating the decision to breed and when to start egg-laying, via a possible inhibition of luteinizing hormone (LH) and sex-steroids production. We used luteinizing hormone releasing hormone (LHRH) challenge in pre-laying male and female Black-legged kittiwakes (Rissa tridactyla) to test whether LH and testosterone secretion were depressed by elevated corticosterone levels. Females bearing high baseline corticosterone levels showed reduced baseline LH levels and a low ability to release LH, following LHRH challenge. Further, females bearing low baseline LH levels and elevated baseline corticosterone levels were more likely to skip breeding. However, non-breeding females were physiologically primed for breeding, since they mounted high LHRH-induced LH release. Egg-laying date was advanced in good body condition females but was unaffected by hormones secretion. In males, corticosterone levels had no effect on LH and/or testosterone secretion and did not affect their decision to breed. Interestingly, males with high LHRH-induced testosterone release bred early. Our study highlights clear sex-differences in the HPG sensitivity to stress hormones in pre-laying kittiwakes. Because females have to store body reserves and to build up the clutch, they would be more sensitive to stress than males. Moreover, intrasexual competition could force male kittiwakes to acquire reproductive readiness earlier in the season than females and to better resist environmental perturbations. We suggest that high testosterone releasing ability would mediate behavioural adjustments such as courtship feeding, which would stimulate early egg-laying in females.

Does testosterone influence activity budget in the male Greek tortoise (Testudo graeca graeca)?

The stimulatory effect of testosterone on male sexual activity is one of the clearest examples linking hormones and behaviors. However, this relationship is complex in Chelonians. We experimentally studied the influence of testosterone levels on the activity budget and space use in male Greek tortoises (Testudo graeca graeca) during the spring mating season. We first described the annual pattern of changes in plasma testosterone levels in free-ranging animals in Morocco. Two peaks, one in winter and one in summer, corresponded to periods of inactivity; whereas mating periods in spring and to a lesser extent in autumn were associated with low plasma testosterone levels. Second, we experimentally manipulated plasma testosterone levels in free-ranging males, and analyzed the behavioral consequences. The strong contrasts in plasma hormone levels induced by the experimental treatments did not result in changes in activity budget or space use, both in the short-term or more than one month after the beginning of the hormonal treatment. Our results suggest that testosterone levels did not influence directly behavioral activity in this species, either immediately or after a time delay of one month.

Patterns of aging in the long-lived wandering albatross.

How does an animal age in natural conditions? Given the multifaceted nature of senescence, identifying the effects of age on physiology and behavior remains challenging. We investigated the effects of age on a broad array of phenotypic traits in a wild, long-lived animal, the wandering albatross. We studied foraging behavior using satellite tracking and activity loggers in males and females (age 6-48+ years), and monitored reproductive performance and nine markers of baseline physiology known to reflect senescence in vertebrates (humoral immunity, oxidative stress, antioxidant defenses, and hormone levels). Age strongly affected foraging behavior and reproductive performance, but not baseline physiology. Consistent with results of mammal and human studies, age affected males and females differently. Overall, our findings demonstrate that age, sex, and foraging ability interact in shaping aging patterns in natural conditions. Specifically, we found an unexpected pattern of spatial segregation by age; old males foraged in remote Antarctica waters, whereas young and middle-aged males never foraged south of the Polar Front. Old males traveled a greater distance but were less active at the sea surface, and returned from sea with elevated levels of stress hormone (corticosterone), mirroring a low foraging efficiency. In contrast to findings in captive animals and short-lived birds, and consistent with disposable soma theory, we found no detectable age-related deterioration of baseline physiology in albatrosses. We propose that foraging efficiency (i.e., the ability of individuals to extract energy from their environment) might play a central role in shaping aging patterns in natural conditions.

Maternal deposition of yolk corticosterone in clutches of southern rockhopper penguins (Eudyptes chrysocome chrysocome).

High corticosterone levels can have deleterious effects in developing avian embryos and chicks. Therefore, it may be adaptive for avian mothers to reduce corticosterone transfer to their eggs. However, until now, data about the active or/and passive role of mothers in corticosterone transfer to eggs are inconclusive. Here, we study maternal investment into A- and B-eggs of southern rockhopper penguins (Eudyptes chrysocome chrysocome). This species exhibits reversed hatching asynchrony and provides a unique model to test whether there is a strategic investment in corticosterone favoring the B-chick, which is most likely to survive. We found that rockhopper penguins had the highest yolk concentrations of any wild bird species studied so far. Contrary to our expectations, B-eggs had more yolk corticosterone both in concentration and in quantity than A-eggs, independently of the laying period and the level of human disturbance. Additionally, females deposited more yolk corticosterone in their eggs when they were disturbed. However, this disturbance effect was particularly strong for A-eggs and for late-laid eggs. The present data support neither the predictions for an active regulation nor for a passive deposition, and hormone deposition mechanisms still need to be explored. The adaptive value, if any, of high yolk corticosterone is presently unknown.

Competition for resources modulates cell-mediated immunity and stress hormone level in nestling collared doves (Streptopelia decaocto).

Competitive stress imposed by hatching asynchrony may affect developmental trajectories of offsprings by regulating resource allocation between growth and other fitness-related traits. For instance, the down-regulation of immunity is a commonly observed phenomenon under stressful conditions. However, physiological mechanisms that regulate resources allocation to growth and immune functions in response to competition for resources, as well as inter-sexual differences in physiological strategies, are still poorly investigated. To partially fill this gap, we first conducted a descriptive study on chicks of the collared dove (Streptopelia decaocto), a species producing two chicks per brood. Our results show that first hatchlings (seniors) were bigger, showed lower baseline corticosterone levels (CORT) and showed a higher cell-mediated immunoresponsiveness (CMI) than late hatchlings (juniors). However, when controlling for body size, only CMI remained weaker in junior chicks suggesting differences in strategies of resource allocation between siblings. Interestingly, CORT in juniors increased with increasing within-brood hierarchy. But, while within-brood variation in CMI followed the opposite pattern of variation in CORT, we found no evidence that inter-individual variation in CMI was directly related to CORT. In addition hatching-rank related differences in body size, CMI and CORT was similar between sexes. To ensure that the lower phenotypic value expressed by juniors was not fully related to a lower quality of the late-laid egg, we experimentally suppressed the competitive stress experienced by juniors by removing the first-laid egg (i.e. the egg normally producing a senior chick). In the absence of their brood mates, juniors were bigger, had lower levels of CORT and showed a higher CMI than juniors raised in two-brood chicks, suggesting that body size, CMI and CORT in juniors were modulated by the competitive stress. Overall, this study suggests that juniors respond to within-brood competition by elevating CORT and down-regulating CMI. In this context, the role of CORT, as a mechanism regulating physiological strategies related to growth and immunocompetence is discussed.

Corticosterone and foraging behavior in a pelagic seabird.

Because endocrine mechanisms are thought to mediate behavioral responses to changes in the environment, examining these mechanisms is essential for understanding how long-lived seabirds adjust their foraging decisions to contrasting environmental conditions in order to maximize their fitness. In this context, the hormone corticosterone (CORT) deserves specific attention because of its major connections with locomotor activities. We examined for the first time the relationships between individual CORT levels and measurements of foraging success and behavior using satellite tracking and blood sampling from wandering albatrosses (Diomedea exulans) before (pretrip CORT levels) and after (posttrip CORT levels) foraging trips during the incubation period. Plasma CORT levels decreased after a foraging trip, and the level of posttrip CORT was negatively correlated with individual foraging success, calculated as total mass gain over a foraging trip. Pretrip CORT levels were not linked to time spent at sea but were positively correlated with daily distance traveled and maximum range at sea. In this study, we were able to highlight the sensitivity of CORT levels to variation in energy intake, and we showed for the first time that individual CORT levels can be explained by variation in foraging success. Relationships between pretrip CORT levels and daily distance traveled and maximum range were independent of pretrip body mass, suggesting that slight elevations in pretrip CORT levels might facilitate locomotor activity. However, because both foraging behavior and pretrip CORT levels could be affected by individual quality, future experimental studies including manipulation of CORT levels are needed to test whether CORT can mediate foraging decisions according to foraging conditions.

Plasma steroid and nutrient levels during the active season in wild Testudo horsfieldi.

Plasma concentrations of sex steroids (testosterone and progesterone), proteins (total protein and albumin), lipids (phospholipids, cholesterol, and triglycerides), and minerals (calcium and phosphorus) were measured in wild Testudo horsfieldi in Uzbekistan, during the short, 3-month activity period (March-May, 1998). Testosterone concentration in males was highest (52 ng/ml) when they had just emerged from brumation (hibernation) in mid-March, which was also the beginning of the mating period, and fell in April. In females, progesterone peaked in mid-April (at 10 ng/ml), just before ovulation of the first clutches at the end of April and beginning of May. Testosterone levels in females and progesterone levels in males were low (<3 ng/ml) throughout the activity period. In general, the plasma concentrations of proteins, lipids, and phosphorus increased slowly in males, but more rapidly in females, during the activity season. These increases were particularly strong in females in the second half of April, coinciding with the peaks in female hormone levels. The changes in plasma hormones and nutrients reflected the timing of the different behaviours. The four first weeks of above-ground activity (mid-March to mid-April) by males, when they had high testosterone levels, were primarily allocated to fighting other males, courtship, and mating, while females spent much of that time feeding. Thereafter, both sexes concentrated on feeding. Females were probably preparing to ovulate in late-April, when their progesterone levels were highest and when plasma nutrient levels increased considerably.