Haemoproteus infected birds have increased lifetime reproductive success.

The impact of haematozoan infection on host fitness has received substantial attention since Hamilton and Zuk posited that parasites are important drivers of sexual selection. However, short-term studies testing the assumption that these parasites consistently reduce host fitness in the wild have produced contradictory results. To address this complex issue, we conducted a long-term study examining the relationship between naturally occurring infection with Haemoproteus and Plasmodium, and lifetime reproductive success and survival of Mountain White-crowned Sparrows. Specifically, we tested the hypothesis that birds infected with haematozoan parasites have reduced survival (as determined by overwinter return rates) and reproductive success. Contrary to expectation, there was no relationship between Haemoproteus and Plasmodium infection and reproduction or survival in males, nor was there a relationship between Plasmodium infection and reproduction in females. Interestingly, Haemoproteus-infected females had significantly higher overwinter return rates and these females fledged more than twice as many chicks during their lifetimes as did uninfected females. We discuss the impact of parasitic infections on host fitness in light of these findings and suggest that, in the case of less virulent pathogens, investment in excessive immune defence may decrease lifetime reproduction.

Assessing the role of reproduction and stress in the spring emergence of haematozoan parasites in birds.

A spring emergence of avian haemosporidian infections is nearly universal among temperate zone birds and is often described as a cost of reproductive effort. We take advantage of the opportunistic (i.e. aseasonal) breeding schedule of the red crossbill (Loxia curvirostra) to determine the relative contributions of season versus host physiology to the timing and intensity of Haemoproteus infections in the temperate zone. Despite breeding activity in both the winter and summer, Haemoproteus infections were highly seasonal--occurring largely from May through September--and measures of host physiology (i.e. reproductive condition and stress parameters) did not explain parasite prevalence. However, within the spring-summer peak, infection intensity (i.e. parasite density) was positively correlated with plasma levels of testosterone and free corticosterone and negatively correlated with corticosterone binding globulin capacity. These data are discussed in terms of the behavioral ecology of host and vector, and suggest that both seasonal increases in vector activity and relapse of latent (i.e. dormant) infections contribute to the spring emergence in birds. Relapse of latent infections does not appear to be induced by reproductive activity or increased allostatic (i.e. energy) load, but rather by a season-specific change in host or parasite physiology (e.g. melatonin or endogenous rhythms).

Natural selection and glucocorticoid physiology.

Glucocorticoid hormones are considered potent modulators of trade-offs between reproduction and survival. As such, selection should affect glucocorticoid physiology, although relatively little is known about how selection may act on glucocorticoid profiles. In general, the evolution of physiology is less studied and less well understood than morphological or life history traits. Here, we used a long-term data set from a population of mountain white-crowned sparrows to estimate natural selection on glucocorticoid profiles. Our study suggests that survival selection favours higher hormone concentrations for multiple components of glucocorticoid physiology (both baseline and stress-induced glucocorticoid levels). Fecundity selection varies depending on the component of hypothalamic-pituitary-adrenal physiology; greater reproductive output was associated with higher baseline glucocorticoid levels, but lower stress-induced glucocorticoid levels. Additionally, the selection gradient was greater for glucocorticoids than for a morphological trait (wing length). These results support the hypothesis that stress-induced glucocorticoids increase survival over reproduction within a wild population (the CORT-trade-off hypothesis). Taken together, these results add to our knowledge of how selection operates on physiological traits and also provide an evolutionary and ecological perspective on several key open issues in the field of glucocorticoid physiology.

Corticosterone exposure during development has sustained but not lifelong effects on body size and total and free corticosterone responses in the zebra finch.

Animals exposed to stress during development experience sustained morphological, physiological, neurological, and behavioral consequences. For example, elevated glucocorticoids (GCs) during development can increase GC secretion in adults. Studies have examined the sustained effects of elevated developmental GCs on total GC responses, but no study to date has examined the effect of developmental stress on corticosteroid binding globulin (CBG). CBG is a protein which binds to GCs and facilitates their transportation in blood. When bound to CBG, GCs are unavailable to interact with target tissues. Exposure to stress can decrease CBG capacity and, thus, increase free GCs (the portion of unbound GCs). We examined the long-term effects of elevated corticosterone (CORT) during development (12-28days post-hatch) on acute stress responses, negative feedback, and CBG capacity at 30, 60, and 90days post-hatch in zebra finches. Additionally, we evaluated the effect of CORT treatment on body size and condition at 28, 60, and 90days post-hatch. CORT exposed birds had higher acute stress responses at 30days post-hatch compared to control birds. However, there was no treatment effect at 60 or 90days post-hatch. CBG levels were not affected by treatment, and so free CORT estimations reflected patterns in total CORT. CORT treatment decreased growth and condition in zebra finches at 28days post-hatch, but these differences were not present at later life history stages. However, brood size had a sustained effect on body size such that birds reared in medium sized broods were larger at 28, 60, and 90days post-hatch. These results demonstrate the complexity of early environmental effects on adult phenotype and suggest that some conditions may have stronger programmatic effects than others.

Contributions of endocrinology to the migration life history of birds.

Migration is a key life cycle stage in nearly 2000 species of birds and is a greatly appreciated phenomenon in both cultural and academic arenas. Despite a long research tradition concerning many aspects of migration, investigations of hormonal contributions to migratory physiology and behavior are more limited and represent a comparatively young research field. We review advances in our understanding of the hormonal mechanisms of migration with particular emphasis on the sub-stages of the migration life history: development, departure, flight and arrival. These sub-stages vary widely in their behavioral, ecological and physiological contexts and, as such, should be given appropriate individual consideration.

Proximity to a high traffic road: glucocorticoid and life history consequences for nestling white-crowned sparrows.

Roads have been associated with decreased reproductive success and biodiversity in avian communities and increased physiological stress in adult birds. Alternatively, roads may also increase food availability and reduce predator pressure. Previous studies have focused on adult birds, but nestlings may also be susceptible to the detrimental impacts of roads. We examined the effects of proximity to a road on nestling glucocorticoid activity and growth in the mountain white-crowned sparrow (Zonotrichia leucophrys oriantha). Additionally, we examined several possible indirect factors that may influence nestling corticosterone (CORT) activity secretion in relation to roads. These indirect effects include parental CORT activity, nest-site characteristics, and parental provisioning. And finally, we assessed possible fitness consequences of roads through measures of fledging success. Nestlings near roads had increased CORT activity, elevated at both baseline and stress-induced levels. Surprisingly, these nestlings were also bigger. Generally, greater corticosterone activity is associated with reduced growth. However, the hypothalamic-pituitary-adrenal axis matures through the nestling period (as nestlings get larger, HPA-activation is greater). Although much of the variance in CORT responses was explained by body size, nestling CORT responses were higher close to roads after controlling for developmental differences. Indirect effects of roads may be mediated through paternal care. Nestling CORT responses were correlated with paternal CORT responses and paternal provisioning increased near roads. Hence, nestlings near roads may be larger due to increased paternal attentiveness. And finally, nest predation was higher for nests close to the road. Roads have apparent costs for white-crowned sparrow nestlings--increased predation, and apparent benefits--increased size. The elevation in CORT activity seems to reflect both increased size (benefit) and elevation due to road proximity (cost). Whether or not roads are good or bad for nestlings remains equivocal. However, it is clear that roads affect nestlings; how or if these effects influence adult survival or reproduction remains to be elucidated.

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.

Plasma testosterone correlates with morph type across breeding substages in male white-throated Sparrows.

White-throated sparrows (Zonotrichia albicollis) exhibit a genetic polymorphism that affects plumage and behavior in both sexes. White-striped morphs are more territorially aggressive, whereas tan-striped morphs provision nestlings at a higher rate. We investigated testosterone physiology in this species in an effort to understand hormonal mechanisms for the observed differences in aggression and parental care between the morphs. We found a small but significant difference in plasma testosterone between free-living white-striped and tan-striped males over the course of the breeding season. This difference correlates with previously observed differences in aggressive behavior and suggests that testosterone may mediate these differences. Testosterone remained higher in white-striped males relative to tan-striped males when males were provisioning nestlings and fledglings. Thus, testosterone may also contribute to the relatively reduced levels of parental care exhibited by white-striped males. In contrast to males, plasma testosterone did not differ between free-living white-striped and tan-striped females, which suggests that testosterone does not mediate differences in aggression between female morphs. Injection with gonadotropin-releasing hormone led to greater testosterone secretion in both captive and free-living males and captive females but did not differ by morph. Therefore, we conclude that differences in plasma testosterone between the morphs are due to differences in testosterone regulation upstream of the pituitary.

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.

Plasma-binding globulins and acute stress response.

Within studies of acute stress physiology an increase in glucocorticoid secretion is thought to be the primary mediator of tissue response to stress. Corticosteroid-binding globulin may regulate tissue availability of steroids, but has not been considered a dynamic component of the acute stress response. Here, we examined CBG level over the common 60-minute time frame in an acute capture and handling protocol to investigate whether CBG capacity is dynamic or static over short stressors. Using a comparative approach, we measured CBG response to capture and handling stress in nine species of birds, representing five orders and nine families. CBG capacity significantly declined within 30 - 60 minutes of capture in five of the nine species examined. This decline may serve to significantly increase the level of corticosterone reaching tissues during acute stress.

Differential mechanisms for regulation of the stress response across latitudinal gradients.

We examined plasticity of the stress response among three populations of the white-crowned sparrow (Zonotrichia leucophrys). These populations breed at different elevations and latitudes and thus have breeding seasons that differ markedly in length. We hypothesize that in populations where birds raise only one or rarely two broods in a season, the fitness costs of abandoning a nest are substantially larger than in closely related populations that raise up to three broods per season. Thus individuals with short breeding seasons should be less responsive to stressors and therefore less likely to abandon their young. In our study, baseline and handling-induced corticosterone levels were similar among populations, but corticosteroid-binding globulins differed, leading to a direct relationship between stress-induced free corticosteroid levels and length of breeding season. There were also population-specific differences in intracellular low-affinity (glucocorticoid-like) receptors in both liver and brain tissue. Although investigations of population-based differences in glucocorticoid secretion are common, this is the first study to demonstrate population-level differences in binding globulins. These differences could lead to dramatically different physiological and behavioral responses to stress.

Plasma binding proteins as mediators of corticosteroid action in vertebrates.

Stressors elicit a complex but variable suite of endocrine events. Comparative studies of the stress response have focused primarily on the adrenocortical response to stress, in particular the measurement of plasma levels of glucocorticoids. However, a number of other factors contribute to and modify cellular and organismal responses to glucocorticoids. Notably, plasma corticosteroid binding globulins (CBGs) can regulate the general availability of steroid to tissues, and/or direct the delivery of hormones to specific sites. In this paper, we discuss possible functions of CBG and mechanisms of CBG action, review CBG characteristics among vertebrates, and discuss our recent studies indicating that CBG may indeed modulate responses to stressors. For example, in house sparrows, we found that basal and stress-induced concentrations of total corticosteroid (cortisol or corticosterone) (CORT) vary seasonally, but CBG concentrations change proportionally, so that free CORT concentrations appear static year-round. In contrast, in white-crowned sparrows and tree lizards, CBG concentrations change under conditions when total CORT levels do not, resulting in significant changes in circulating free CORT. These differences in free CORT are masked if CBG is not accounted for. We have also found that the binding properties of CBG vary considerably between species and need to be determined empirically. Such studies led to the observation that CBG in several species may also serve as a functional androgen binding protein; this is especially important for birds, because previous studies had concluded that birds lack androgen binding globulins. We propose that consideration of CBG is paramount to understanding the role of glucocorticoids in mediating behavioral and physiological responses to stress.

Seasonal regulation of membrane and intracellular corticosteroid receptors in the house sparrow brain.

A number of studies have demonstrated seasonal regulation of the adrenocortical response to stress, or of corticosteroid binding globulins, but very few studies have examined seasonal regulation of corticosteroid receptor levels. As a result, there have been few attempts to produce an integrated picture of seasonal plasticity of the stress response. We measured baseline and stress-induced corticosterone (CORT), corticosteroid binding globulin and neuronal cytosolic and membrane corticosteroid receptor levels in male and female, wild-caught house sparrows (Passer domesticus) during three different seasons over the annual cycle (nesting, molting and winter). We identified three neuronal corticosteroid receptors in the house sparrow brain: two intracellular receptors and one membrane-associated receptor. Little is known about corticosteroid receptors in neuronal membranes of avian and mammalian species, but we found that the levels of membrane corticosteroid receptors varied seasonally, being lowest during the nesting season. Cytosolic corticosteroid receptor numbers (both low and high affinity receptors) also varied seasonally. In contrast to the membrane bound receptors, however, the numbers of low and high affinity cytosolic receptors were lowest during winter. In addition, mean levels of total basal and stress-induced CORT in the plasma varied seasonally. Both basal and stress-induced levels of total CORT were significantly higher during nesting than during winter or molt. Finally, corticosteroid binding globulin levels in plasma were also seasonally regulated, in a pattern similar to total CORT, so that estimated free CORT levels did not vary between seasons. These data indicate that multiple components of the stress response are seasonally regulated in birds obtained from wild populations. Interactions between these regulated components provide a basis for seasonal differences in behavioural and physiological responses to stress.

Pharmacological adrenalectomy with mitotane.

The potential of mitotane (ortho, para'-DDD, commonly used to treat adrenal carcinomas in humans and dogs) was investigated as an alternative to surgical adrenalectomy in birds, salamanders, and lizards. House sparrows (Passer domesticus) were injected twice daily with vehicle or one of two doses of mitotane (225 or 450 mg/kg), and basal and stress-induced levels of corticosterone (CORT) were measured 3 and 5 days after injections. Mitotane reduced basal CORT levels to nondetectable and abolished stress-induced CORT increases by the 3rd day of treatment. In another study, a single injection of mitotane was effective in lowering endogenous CORT levels 36 h later, but levels had apparently recovered by 10 days after the injection. Mitotane did not effect testicular weights and had no detectable effect on testosterone levels. In contrast to its effects on house sparrows, mitotane did not lower endogenous CORT levels in either tiger salamanders (Ambystoma tigrinum) or tree lizards (Urosaurus ornatus), even at doses much higher than those used in house sparrows.

Rapid behavioral response to corticosterone varies with photoperiod and dose.

The magnitude of the adrenocortical response to stress can be modulated by a variety of environmental and physiological factors, such as daylength and body condition. To determine the consequences of this modulation for the organism, one also needs to investigate behavioral sensitivity to glucocorticoids. We examined the behavioral response of Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii) to elevated glucocorticoids. Using a behavioral assay in which a rapid and transient dose of corticosterone (CORT: the avian glucocorticoid) rapidly increases perch hopping, we first investigated the photoperiodic regulation of this behavioral response. Intermediate levels of CORT ( approximately 24 ng/ml), which increased activity in sparrows exposed to a long-day (breeding) photoperiod, had no behavioral effect in sparrows exposed to a short-day (winter) photoperiod. Hence, the neural mechanisms which regulate the behavioral response to CORT appear to be less sensitive under a winter photoperiod. Using the same behavioral assay, we also measured a dose-response curve for CORT's effects on activity in sparrows exposed to the long-day photoperiod. Intermediate levels (24 and 40 ng/ml) increased activity to threefold background levels, whereas high physiological levels (65 and 97 ng/ml) had no effect. Given that the behavioral response does not increase linearly with CORT, we can no longer assume that modulation of the adrenocortical response to stress will result in linear changes in behavior.

Diel rhythms of basal and stress-induced corticosterone in a wild, seasonal vertebrate, Gambel's white-crowned sparrow.

Glucocorticoids have a wide array of actions in vertebrates. Daily fluctuations in basal levels of glucocorticoids are thought to regulate homeostatic mechanisms. In contrast, elevated levels secreted in response to stress stimulate changes in physiology and behavior. These changes are thought to aid an animal in avoiding chronic stress or death. Twenty-four-hour rhythms in basal and stress-induced glucocorticoids have been detected in laboratory mammals, but studies in wild, seasonal vertebrates are rare. Identification of plasticity in hormone secretion in wild vertebrates is critical to understanding the effects of hormones on physiology and behavior, and therefore the success of an animal in its natural environment. In the present study, we characterized diel patterns of basal and stress-induced corticosterone (the avian glucocorticoid) under two photoperiods in Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii). In contrast to previous findings in the white-crowned sparrow, we demonstrated a robust rhythm in basal corticosterone secretion, in which corticosterone reaches peak levels at the end of the inactive period, and has returned to trough levels just after the active period has begun. We also demonstrated a diel rhythm in secretion of corticosterone in response to a stressor, showing the greatest response at the beginning of the active period. Patterns of CORT secretion were similar under both photoperiods. These patterns show interesting similarities and differences to classical mammalian rhythms.

Noninvasive corticosterone treatment rapidly increases activity in Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii).

Recent evidence supports the hypothesis that corticosteroids influence behavioral changes associated with stressful events. Most investigations into this relationship focus on the long-term behavioral effects of corticosterone. Because many behavioral responses to environmental perturbations occur within minutes, we determined what rapid effects corticosterone may have on behavior. With this goal in mind, we devised and evaluated a method of corticosterone delivery which allowed us to examine immediate effects of corticosterone on behavior in a noninvasive manner. White-crowned sparrows (Zonotrichia leucophrys gambelii) were allowed access to mealworms (Tenebrio molitor) injected with corticosterone. Once ingested, the corticosterone moves across the digestive epithelium into the circulation. This method was evaluated using two vehicles: dimethyl sulfoxide and peanut oil. We tested the efficiency and consistency of corticosterone transfer into the circulation for both vehicles. Dimethyl sulfoxide gave a more efficient transfer of corticosterone. Injecting mealworms with corticosterone (carried in dimethyl sulfoxide) and feeding those worms to white-crowned sparrows increased circulating corticosterone to a discrete, repeatable level which peaked within 7 min and was cleared within 60 min. Using this method, we demonstrated that intermediate levels of corticosterone caused an increase in perch hopping in white-crowned sparrows within 15 min of hormone administration. An increase in perch hopping indicated elevated locomotor activity that is consistent with behavioral responses to natural perturbations. High levels of corticosterone did not induce this behavioral change. In light of the rapid effect of corticosterone on behavior, we propose that corticosterone was acting through a nongenomic mechanism.

Seasonal plasticity and sexual dimorphism in the avian song control system: stereological measurement of neuron density and number.

Differences in neuron density and number are associated with seasonal plasticity and sexual dimorphism in the avian song control system. In previous studies, neuron density and number in this system have been quantified primarily through nonstereological approaches in thick tissue sections by using the nucleolus as the unit of count. The reported differences between seasons and sexes may be inaccurate due to biases introduced by neuron splitting during sectioning. We used the unbiased optical disector technique on tissue from three previous studies (two investigations of seasonal plasticity and one investigation of sexual dimorphism in avian song nuclei) to assess seasonal and sex differences in neuron density and number. In two song nuclei, HVc and the robust nucleus of the archistriatum (RA), the optical disector yielded intergroup differences in neuron density and number that coincided well with the three previous reports. We also estimated neuron number and density with a random, systematic, nonstereological counting protocol that used the neuronal nucleolus as the unit of count. We compared this method directly to the optical disector. In all cases, the two neuron-counting methods produced similar estimates of neuron number and density; the differences between treatment groups were equally discernible regardless of the counting method used. This study confirms previously reported seasonal and sex differences in the HVc and the RA by use of stereology and indicates that a random, systematic, nonstereological neuron-counting protocol is accurate and is well suited to the study of these phenomena in the avian song control system.