Perspectives on environmental heterogeneity and seasonal modulation of stress response in neotropical birds.

Corticosterone (CORT), the main glucocorticoid in birds, regulates physiological and behavioral traits linked to predictable and unpredictable environmental fluctuations (i.e., stressors). Baseline and stress-induced CORT concentrations are known to fluctuate seasonally, linked to life history stages (LHS) such as breeding, molt, and wintering stage. These variations have been relatively well described in North American birds, but poorly addressed in neotropical species. To fill this gap, we explored how baseline and stress-induced CORT variation by LHS was affected by seasonality and environmental heterogeneity (i.e., frequency of unpredictable events such as droughts, flashfloods, etc) within the Neotropics using two approaches. First, we reviewed all currently available data about CORT concentrations for neotropical bird species. Second, we performed an in-depth analysis comparing the CORT responses of the two most common species of the Zonotrichia genus from North and South America (Z. leucophrys and Z. capensis, respectively) and their subspecies to seasonality and environmental heterogeneity. These species have been analyzed with the same methodology, allowing for an in-depth comparison of CORT variations. Despite scant data on neotropical bird species, we observed overlap between molt and breeding, and lower fluctuations of CORT among LHS. These patterns would be considered atypical compared to those described for North temperate species. Further, we found no significant associations between environmental heterogeneity and the stress-responses. In Zonotrichia we observed a positive association between baseline and stress-induced concentrations of CORT and latitude. We also observed differences by LHS. Both baseline and stress-induced CORT concentrations were higher during breeding and lower during molt. In addition, for both species, the overall pattern of seasonal modulation of stress response was heavily influenced by the migration strategy, with long-distance migrants showing significantly higher stress-induced CORT levels. Our results highlight the need for more data collection in the Neotropics. Comparative data would shed further light on the sensitivity of the adrenocortical response to stress under different scenarios of environmental seasonality and unpredictability.

Unexpected long-term retention of subcutaneous beeswax implants and additional notes on dose and composition from four testosterone implant studies.

Experimental manipulations of testosterone have advanced our understanding of the hormonal control of traits across vertebrates. Implants are commonly used to supplement testosterone and other hormones to organisms, as they can be readily scaled to produce desired hormone levels in circulation. Concerns about pharmacological (i.e. unnatural) doses of traditional silastic implants led to innovation in implant methods, with time-release pellets and beeswax implants proposed as solutions. A study comparing silastic, time-release pellets, and beeswax implants found the latter to be most effective in delivering a physiologically relevant dose. One proposed advantage to subcutaneous beeswax implants is that they are expected to degrade within the body, thus removing the obligation to recapture implanted individuals in the field. However, few studies have reported on dosage and no published literature has examined the assumption that beeswax implants readily degrade as expected. Here we present time-release androgen data in relation to implants containing varying levels of testosterone from four separate implant studies. In addition, we report long-term persistence of subcutaneous implants, including two cases of implants being retained for > 2 years. Finally, we offer recommendations on the composition and implementation of beeswax implants to aid the pursuit of minimally invasive and physiologically relevant manipulations of circulating hormones.

Austral birds offer insightful complementary models in ecology and evolution.

The Southern Hemisphere differs from the Northern Hemisphere in many aspects. However, most ecological and evolutionary research is conducted in the Northern Hemisphere and its conclusions are extrapolated to the entire globe. Therefore, unique organismal and evolutionary characteristics of the south are overlooked. We use ornithology to show the importance of including a southern perspective. We present examples of plumage pigmentation, brood-parasitic nestling ejection, flightlessness, female song, and female aggression modulated by progesterone as complementary models for investigating fundamental biological questions. More research in the Southern Hemisphere, together with increased cooperation among researchers across the hemispheres and within the Southern Hemisphere, will provide a greater global outlook into ecology and evolution.

Neuroendocrine patterns underlying seasonal song and year-round territoriality in male black redstarts.

BACKGROUND: The connection between testosterone and territoriality in free-living songbirds has been well studied in a reproductive context, but less so outside the breeding season. To assess the effects of seasonal androgenic action on territorial behavior, we analyzed vocal and non-vocal territorial behavior in response to simulated territorial intrusions (STIs) during three life-cycle stages in free-living male black redstarts: breeding, molt and nonbreeding. Concurrently, we measured changes in circulating testosterone levels, as well as the mRNA expression of androgen and estrogen receptors and aromatase in the preoptic, hypothalamic and song control brain areas that are associated with social and vocal behaviors. RESULTS: Territorial behavior and estrogen receptor expression in hypothalamic areas did not differ between stages. But plasma testosterone was higher during breeding than during the other stages, similar to androgen receptor and aromatase expression in the preoptic area. The expression of androgen receptors in the song control nucleus HVC was lower during molt when birds do not sing or sing rarely, but similar between the breeding and the nonbreeding stage. Nevertheless, some song spectral features and the song repertoire differed between breeding and nonbreeding. Territorial behavior and song rate correlated with the expression of steroid receptors in hypothalamic areas, and in the song control nucleus lMAN. CONCLUSIONS: Our results demonstrate seasonal modulation of song, circulating testosterone levels, and brain sensitivity to androgens, but a year-round persistency of territorial behavior and estrogen receptor expression in all life-cycle stages. This suggests that seasonal variations in circulating testosterone concentrations and brain sensitivity to androgens is widely uncoupled from territorial behavior and song activity but might still affect song pattern. Our study contributes to the understanding of the complex comparative neuroendocrinology of song birds in the wild.

Seasonality at the equator: isotope signatures and hormonal correlates of molt phenology in a non-migratory Amazonian songbird.

BACKGROUND: Birds, across their annual cycle, progress through sequences of life-history stages such as reproduction and molt. The mechanisms that control annual avian itineraries involve endocrine responses triggered by seasonal environmental factors, including changes in resource availability and/or photoperiod. However, at equatorial latitudes birds are exposed to different degrees of seasonality, and the mechanisms underlying phenology of birds near the equator remain less explored. We studied the silver-beaked tanager, an endemic Amazonian songbird, from an equatorial lowland population. Remarkably, in this species, song behavior has been shown to be seasonally aligned to minimal changes in day length near the equator. Here, we aimed to further explore the phenology of silver-beaked tanagers by assessing shifts of food sources utilization as potential ultimate factors. We measured triple isotopic tracers of carbon (δ13C), nitrogen (δ15N) and sulphur (δ34S) in blood and feathers of birds throughout a whole year. In addition, we assessed the degree of seasonality in the molting activity, in relation to circulating levels of corticosterone, as well as to testosterone as a proxy of the reproductive condition of males. RESULTS: There was important seasonal variation of δ34S values in relation to rainfall patterns and changes in estuarine water composition. Despite the seasonal rainfall, we found no substantial variation in the foraging ecology of birds over seasons. This was accompanied by uniform levels of corticosterone throughout the year, probably associated with the absence of drastic seasonal resource shortages. Even so, silver-beaked tanagers showed a marked seasonal molting schedule, which was related to variation in the circulating levels of both corticosterone and testosterone. CONCLUSIONS: These findings suggest that foraging niche is not life history stage-dependent in silver-beaked tanagers, and highlight rainfall as an important environmental cue for bird phenology. Our stable isotope results encourage further studies addressing the influence of estuarine water dynamics on bird timing. In addition, the results suggest a primary role of steroid hormones in regulating seasonal life history stages under the absence of a marked photoperiod. Contrary to what might be expected for a tropical songbird, our physiological data in silver-beaked tanagers do not support reproduction-molt overlapping.

Seasonal singing of a songbird living near the equator correlates with minimal changes in day length.

Behaving in accordance with natural cycles is essential for survival. Birds in the temperate regions use the changes of day length to time their behavior. However, at equatorial latitudes the photoperiod remains almost constant throughout the year, and it is unclear which cues songbirds use to regulate behaviors, such as singing. Here, we investigated the timing of dawn-song of male silver-beaked tanagers in the equatorial lowland Amazonas over two years. In this region, birds experience around nine minutes of annual day length variation, with sunrise times varying by 32 minutes over the year. We show that the seasonal timing of dawn-song was highly regular between years, and was strongly correlated with slight increases in day length. During the singing season the daily dawn-song onset was precisely aligned to variations in twilight time. Thus, although photoperiodic changes near the equator are minimal, songbirds can use day length variation to time singing.

Dawn-song onset coincides with increased HVC androgen receptor expression but is decoupled from high circulating testosterone in an equatorial songbird.

The song of songbirds is a testosterone-sensitive behavior that is controlled by brain regions expressing androgen receptors. At higher latitudes, seasonal singing is stimulated by increasing day-length and elevated circulating testosterone. However, a large number of songbird species inhabit equatorial regions under a nearly constant photoperiod, and the neuroendocrine mechanisms of seasonal song in these species have rarely been investigated. We studied males from an equatorial population of the silver-beaked tanager (Ramphocelus carbo), an Amazonian songbird. We found seasonality in dawn-song behavior, which was displayed continuously for more than half a year throughout an extended breeding territoriality stage. The seasonal activation of dawn-song was correlated with an increased area of androgen receptor expression in HVC, a major brain area of song control. However, testosterone levels remained low for several weeks after activation of dawn-song. Circulating levels of testosterone were elevated only later in the breeding season, coinciding with a higher dawn-song output and with the mating period. Our results suggest that the seasonal activation of dawn-song and territoriality involves an increase of androgen target cells in HVC. This mechanism could potentially function to circumvent adverse effects of high testosterone levels in a species with an extended breeding season.

Towards more physiological manipulations of hormones in field studies: comparing the release dynamics of three kinds of testosterone implants, silastic tubing, time-release pellets and beeswax.

Hormone manipulations are of increasing interest in the areas of physiological ecology and evolution, because hormones are mediators of complex phenotypic changes. Often, however, hormone manipulations in field settings follow the approaches that have been used in classical endocrinology, potentially using supra-physiological doses. To answer ecological and evolutionary questions, it may be important to manipulate hormones within their physiological range. We compare the release dynamics of three kinds of implants, silastic tubing, time-release pellets, and beeswax pellets, each containing 3mg of testosterone. These implants were placed into female Japanese quail, and plasma levels of testosterone measured over a period of 30 days. Testosterone in silastic tubing led to supraphysiological levels. Also, testosterone concentrations were highly variable between individuals. Time-release pellets led to levels of testosterone that were slightly supraphysiological during the first days. Over the period of 30 days, however, testosterone concentrations were more consistent. Beeswax implants led to a physiological increase in testosterone and a relatively constant release. The study demonstrated that hormone implants in 10mm silastic tubing led to a supraphysiological peak in female quail. Thus, the use of similar-sized or even larger silastic implants in males or in other smaller vertebrates needs careful assessment. Time-release pellets and beeswax implants provide a more controlled release and degrade within the body. Thus, it is not necessary to recapture the animal to remove the implant. We propose beeswax implants as an appropriate procedure to manipulate testosterone levels within the physiological range. Hence, such implants may be an effective alternative for field studies.

Seasonal variations of basal cortisol and high stress response to captivity in Octodon degus, a mammalian model species.

Across vertebrates, the hypothalamic-pituitary-adrenal axis is a conserved neuroendocrine network that responds to changing environments and involves the release of glucocorticoids into the blood. Few studies have been carried out concerning mammalian adrenal regulation in wild species either in the laboratory or field, and even fewer have been able to determine true glucocorticoid baselines. We studied the South-American caviomorph rodent Octodon degus, a diurnal and social mammal that has become an important species in the biological research. First, we determined the plasma cortisol baseline and the acute stress concentrations during the non-reproductive and mating seasons in free-living individuals. Second, using the same protocol we assessed the impact of long-term captivity on the adrenal function in wild-caught degus and degus born in laboratory. Third, we examined laboratory groups formed with degus taken from two distant natural populations; one of them originally occurs at the Andes Mountains in high altitude conditions. The data revealed seasonal modulation of basal cortisol in the wild associated with mating. In laboratory, degus presented higher cortisol stress responses, with greater magnitudes shown in degus born and reared in captivity. No differences between populations were found. The results suggest differential regulatory mechanisms between basal and stress-induced cortisol levels, and context dependence of cortisol modulation in a mammalian species.

Ecological drivers of group living in two populations of the communally rearing rodent, Octodon degus.

Intraspecific variation in sociality is thought to reflect a trade-off between current fitness benefits and costs that emerge from individuals' decision to join or leave groups. Since those benefits and costs may be influenced by ecological conditions, ecological variation remains a major, ultimate cause of intraspecific variation in sociality. Intraspecific comparisons of mammalian sociality across populations facing different environmental conditions have not provided a consistent relationship between ecological variation and group-living. Thus, we studied two populations of the communally rearing rodent Octodon degus to determine how co-variation between sociality and ecology supports alternative ecological causes of group living. In particular, we examined how variables linked to predation risk, thermal conditions, burrowing costs, and food availability predicted temporal and population variation in sociality. Our study revealed population and temporal variation in total group size and group composition that covaried with population and yearly differences in ecology. In particular, predation risk and burrowing costs are supported as drivers of this social variation in degus. Thermal differences, food quantity and quality were not significant predictors of social group size. In contrast to between populations, social variation within populations was largely uncoupled from ecological differences.