The effect of air pollution on immunological, antioxidative and hematological parameters, and body condition of Eurasian tree sparrows.

Air pollution constitutes potential threats to wildlife and human health; therefore, it must be monitored accurately. However, little attention has been given to understanding the toxicological effects induced by air pollution and the suitability of bird species as bioindicators. The Eurasian tree sparrow (Passer montanus), a human commensal species, was used as a study model to examine toxic metal accumulation, retention of particulate matter (PM), immunological and antioxidant capacities, and hematological parameters in birds inhabiting those areas with relatively higher (Shijiazhuang city) or lower (Chengde city) levels of PM2.5 and PM10 in China. Our results showed that Shijiazhuang birds had significantly more particle retention in the lungs and toxic metal (including aluminum, arsenic, cadmium, iron, manganese, and lead) accumulation in the feathers relative to Chengde birds. They also had lower superoxide dismutase, albumin, immunoglobulin M concentrations in the lung lavage fluid, and total antioxidant capacity (T-AOC) in the lungs and hearts. Furthermore, although they had higher proportions of microcytes, hypochromia, and polychromatic erythrocytes in the peripheral blood (a symptom of anemia), both populations exhibited comparable body conditions, white cell counts, heterophil and lymphocyte ratios, and plasma T-AOC and corticosterone levels. Therefore, our results not only confirmed that Shijiazhuang birds experienced a greater burden from environmental PM and toxic metals but also identified a suite of adverse effects of environmental pollution on immunological, antioxidative, and hematological parameters in multiple tissues. These findings contribute to our understanding of the physiological health consequences induced by PM exposure in wild animals. They suggest that free-living birds inhabiting urban areas could be used as bioindicators for evaluating the adverse effects induced by environmental pollution.

A comparison of the nutritional physiology and gut microbiome of urban and rural house sparrows (Passer domesticus).

Urbanization influences food quality and availability for many wild species, but our knowledge of the consequences urbanization has on the nutritional physiology of these animals is currently limited. To fill this gap, we captured House Sparrows (Passer domesticus) from rural and urban environments and hypothesized that increased access to human refuse in urban areas may significantly alter the gut microbiome and nutritional physiology of Sparrows. While there were no significant differences in circulating triglycerides or free glycerol concentrations between populations, urban birds had significantly greater blood glucose concentrations, which suggests greater circulating glucagon concentrations, accessibility to carbohydrates, and/or higher rates of gluconeogenesis in an urban setting. Rural birds had significantly more plasma uric acid, suggesting that they may metabolize more proteins or experience lower inflammation than urban birds. Rural birds also had significantly higher liver free glycerol concentrations, indicating that they metabolize more fat than urban birds. There were no significant differences in the relative abundance of gut microbial taxa at the phyla level between the two populations, but linear discriminant analysis effect size (LEfSe) showed that urban House Sparrows were more enriched with class- and order-level microbes from the phylum Proteobacteria, which are implicated in several mammalian intestinal and extra-intestinal diseases. These findings demonstrate that urbanization significantly alters the nutritional physiology and the composition of the gut microbiome of House Sparrows.

Recovery periods during repeated stress impact corticosterone and behavioral responses differently in house sparrows.

A number of studies have shown that chronic stress can negatively impact both physiology and behavior in a variety of organisms. What has yet to be extensively explored is whether these changes permanently alter an animal's functioning, or if they can be reversed. In this study, we used wild-caught house sparrows (Passer domesticus) to assess how recovery periods influence the physiological and behavioral impacts of an initial four days and subsequent four days of repeated stressors. Birds were randomly assigned to a recovery group and either experienced 0, 24, or 72 h of recovery between the two sets of stressors (cage rolling and cage tapping). We measured the regulation of the hypothalamic pituitary adrenal (HPA) axis by quantifying baseline and stress-induced corticosterone as well as negative feedback strength. We also assessed behavior using neophobia trials to measure how birds altered their approach towards novel objects and their overall activity. Both behavior and corticosterone responses were assessed before the experiment, after the recovery time, and following the final 4 days of stressors. We found that birds that experienced 24 h of recovery had reduced stress-induced corticosterone, but enhanced negative feedback relative to the pre-experiment sample. Additionally, 4 days of stressors was enough to significantly reduce approach latency towards novel objects; however, pre-experiment levels returned with longer periods of recovery. Finally, recovery time did not significantly influence responses to the second 4 days of stressors. Our results indicate that brief recovery periods partially ameliorate the hormonal and behavioral effects of repeated stress.

DNA damage as an indicator of chronic stress: Correlations with corticosterone and uric acid.

Corticosterone does not change in consistent ways across species and contexts, making it challenging to use as an indicator of chronic stress. We assessed DNA damage as a potential metric that could be a more integrative stress measurement with direct links to health. We captured free-living house sparrows, took an immediate blood sample, and transferred them to the laboratory, exposing them to the chronic stress of captivity. Biweekly blood and weight samples were then taken for 4 weeks. We immediately assessed DNA damage in red blood cells using the comet assay and later quantified corticosterone. Uric acid was analyzed in a separate group of birds. We found that birds initially lost, but began to regain weight over the course of captivity. DNA damage peaked within the first 10 days of captivity, and mostly remained elevated. However, the cellular distribution of damage changed considerably over time; most cells showed low levels of damage early, a bimodal distribution of high and low DNA damage during the peak of damage, and a wide unimodal distribution of damage at the end of the 4 weeks. Furthermore, corticosterone increased and remained elevated and uric acid decreased and remained depleted over the same period. Although both a molecular (DNA damage) and an endocrine (corticosterone) marker showed similar response profiles over the 4 weeks, they were not correlated, suggesting they reflect different aspects of the underlying physiology. These data provide convincing preliminary evidence that DNA damage has potential to be an additional indicator of chronic stress.

Preparing to migrate: expression of androgen signaling molecules and insulin-like growth factor-1 in skeletal muscles of Gambel's white-crowned sparrows.

Migratory birds, including Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii), exhibit profound modifications of skeletal muscles prior to migration, notably hypertrophy of the pectoralis muscle required for powered flight. Muscle growth may be influenced by anabolic effects of androgens; however, prior to spring departure, circulating androgens are low in sparrows. A seasonal increase in local androgen signaling may occur within muscle to promote remodeling. We measured morphological parameters, plasma and tissue levels of testosterone, as well as mRNA expression levels of androgen receptor, 5α-reductase (converts testosterone to 5α-dihydrotestosterone), and the androgen-dependent myotrophic factor insulin-like growth factor-1. We studied the pectoralis muscle as well as the gastrocnemius (leg) muscle of male sparrows across three stages on the wintering grounds: winter (February), pre-nuptial molt (March), and pre-departure (April). Testosterone levels were low, but detectable, in plasma and muscles at all three stages. Androgen receptor mRNA and 5α-reductase Type 1 mRNA increased at pre-departure, but did so in both muscles. Notably, mRNA levels of insulin-like growth factor-1, an androgen-dependent gene critical for muscle remodeling, increased at pre-departure in the pectoralis but decreased in the gastrocnemius. Taken together, these data suggest a site-specific molecular basis for muscle remodeling that may serve to enable long-distance flight.

House sparrows (Passer domesticus) adjusted hypothalamic-pituitary-adrenal axis negative feedback and perch hopping activities in response to a single repeated stimulus.

Chronic stress has been extensively studied in both laboratory and field settings; however, a conclusive and consistent phenotype has not been reached. Several studies have reported attenuation of the hypothalamic-pituitary-adrenal axis during experiments intended to cause chronic stress. We sought to determine whether this attenuation could be indicative of habituation. Importantly, we were not investigating habituation to a specific stimulus-as many stress physiology studies do-but rather we assessed how the underlying physiology and behavior changed in response to repeated stressor presentation. We exposed house sparrows (Passer domesticus) to a single stimulus twice per day at random times for 8 consecutive days. We predicted that this period of time would be long enough for the birds to determine that these acute stressors were not, in fact, dangerous and they would, therefore, acclimate. A second control group remained undisturbed for the same period of time. We measured baseline, stress-induced, negative feedback strength, and maximum production of corticosterone as well as neophobic behavior before, during, and after this 8-day experiment. When birds experienced a stimulus for 4 days, their negative feedback strength was significantly diminished, but recovered after the second 4 days. Additionally, perch hopping decreased and recovered in this same time frame. These data suggest that distinct physiological and behavioral responses arise when house sparrows are exposed to the same stressor for several consecutive days as opposed to many stressors layered on top of one another. Furthermore, they indicate that habituation-as with chronic stress-can appear differently depending on the metric being examined.

Stress-Induced Hyperglycemia in White-Throated and White-Crowned Sparrows: A New Technique for Rapid Glucose Measurement in the Field.

Organisms experience stressors, and the physiological response to these stressors is highly conserved. Acute stress activates both the sympathetic nervous system and the hypothalamic-pituitary-adrenal axis, increasing epinephrine, norepinephrine, and glucocorticoids, collectively promoting glucose mobilization. While this is well characterized in mammals, the hyperglycemic response to stress in avian and nonavian reptiles has received less attention. A number of factors, ranging from time of day to blood loss, are reported to influence the extent to which acute stress leads to hyperglycemia in birds. Here we characterized the glycemic response to acute handling stress in two species of free-living sparrows: white-throated sparrows (WTSPs: Zonotrichia albicollis) in St. Mary's County, Maryland, and white-crowned sparrows (WCSPs: Zonotrichia leucophrys) in Tioga Pass Meadow, California. We validated a novel technique for rapid field measurement of glucose using a human blood glucose meter, FreeStyle Lite. As expected, acute handling stress elevated blood glucose at both 15 and 30 min postcapture as compared to baseline for both WTSPs and WCSPs. In addition, handling for 30 min without bleeding had the same hyperglycemic effect as handling with serial bleeds in WCSPs. Finally, body condition that was measured as abdominal fat score predicted stress-induced blood glucose in WTSPs but not in WCSPs. Our results are consistent with the mammalian literature on acute stress and energy mobilization, and we introduce a new field technique for avian field biologists.

House sparrow biomarkers as lead pollution bioindicators. Evaluation of dose and exposition length on hematological and oxidative stress parameters.

House sparrows (Passer domesticus) have been proposed as a key ecological indicator of urban pollution. Remarkably, we lack knowledge about the physiological effects of lead on this bird species. Therefore, this study was aimed to evaluate the effect of Pb on several physiological parameters in house sparrows exposed to environmental Pb concentrations. In a first experiment, birds were exposed to Pb sub-lethal doses (from 1.3 to 14.0 µg of Pb/g animal/day) during 5 days, which resulted in a dose response increase of blood Pb levels and decrease of blood ALAD activity. However, at the higher doses tested (> 7 µg of Pb/g animal/day) the blood ALAD activity inhibition (~82%) remained constant. Hematocrit and hemoglobin were significantly reduced only at the highest-doses, and the stress indicator, heterophils to lymphocyte (H/L) ratio, did not show apparent changes. In a second experiment, house sparrows were exposed to Pb in drinking water (12.3 ppm) during either 15 or 30 days. Pb concentration used in this study was enough to produce blood lead levels equivalents to those found recently in house sparrows inhabiting urban areas, reduced blood ALAD activity and inversion of the H/L ratio. Decreasing blood ALAD activities were correlated with increasing blood Pb levels. In addition, Pb exposure produced modification in the levels of hepatic antioxidant enzymes, increased GST activity and decreased CAT activity, without lipid peroxidation. In conclusion, our results suggest that blood ALAD activity is a reliable and sensitive biomarker for environmental Pb exposure in house sparrows, additionally chronic exposure produce physiological stress (H/L inversion) and small changes in antioxidant enzyme activity. Finally, this specie could be considered a bioindicator for monitoring the urban Pb contamination.

Territorial aggression in urban and rural Song Sparrows is correlated with corticosterone, but not testosterone.

Urban songbirds of several species more vigorously defend their territories in response to conspecific song playback than do their rural counterparts, but the hormonal basis of this behavioral difference is unclear. It is well established in vertebrates that both testosterone and corticosterone affect the intensity of territoriality. Previous studies have found no evidence that initial (i.e., immediately following territorial challenge, but prior to restraint) plasma testosterone accounts for the elevated territorial aggression of urban birds. Determining if testosterone still contributes to urban-rural differences in territoriality requires also assessing males' abilities to transiently increase plasma testosterone (in response to an injection of gonadotropin-releasing hormone). We tested whether these hormones are correlated with the territorial response to conspecific song playback in urban and rural male Song Sparrows (Melospiza melodia) in Montgomery County, Virginia. We found that the elevated territorial aggression of urban sparrows was not related to variation in either initial plasma testosterone or the ability to transiently increase testosterone. In contrast, despite no overall habitat difference in initial corticosterone, levels of this hormone were positively correlated with territoriality in urban and rural sparrows. Furthermore, for a given level of corticosterone, urban sparrows were more territorially aggressive. Our findings suggest that initial corticosterone may either play a role in the regulation of persistent differences in territorial behavior between free-ranging urban and rural male Song Sparrows or be affected by the intensity of behavioral response to territorial challenge.

Chronic stress and the introduction to captivity: How wild house sparrows (Passer domesticus) adjust to laboratory conditions.

The conditions of captivity can cause chronic stress in wild animals. Newly-captured animals may experience weight loss, elevated glucocorticoid hormones, increased heart rate, increased resting adrenomedullary activation, and an altered heart rate response to acute stressors. As captivity conditions persist, chronic stress may decrease as animals adjust to the stressors of captivity. In this study, house sparrows (Passer domesticus) were captured from the wild, fitted with heart rate transmitters in a minor surgical process, and individually housed in an indoor bird facility. Mass, baseline corticosterone, resting heart rate, resting adrenomedullary activation, and the acute heart rate response to a sudden noise were measured over the course of the first 6 weeks of captivity. Birds lost weight during the first weeks of captivity, which was regained by week 5. Baseline corticosterone peaked at day 7, decreased sharply by day 11, and continued to decrease throughout the 6 weeks. Although heart rate in the first 24 h could not be collected, daytime heart rate decreased from day 1 through day 20, where it reached a stable plateau. Daytime heart rate variability decreased through the entire 6 weeks, which may indicate a gradual shift from sympathetic to parasympathetic nervous system regulation of heart rate. The acute heart rate response to a sudden noise lasted longer at day 6 than earlier or later in captivity. In conclusion, the data indicate that the different physiological systems associated with chronic stress adjust to captivity over different timelines.

Effects of thyroid hormone manipulation on pre-nuptial molt, luteinizing hormone and testicular growth in male white-crowned sparrows (Zonotrichia leuchophrys gambelii).

Most seasonal species rely on the annual change in day length as the primary cue to appropriately time major spring events such as pre-nuptial molt and breeding. Thyroid hormones are thought to be involved in the regulation of both of these spring life history stages. Here we investigated the effects of chemical inhibition of thyroid hormone production using methimazole, subsequently coupled with either triiodothyronine (T3) or thyroxine (T4) replacement, on the photostimulation of pre-nuptial molt and breeding in Gambel's white-crowned sparrows (Zonotrichia leuchophrys gambelii). Suppression of thyroid hormones completely prevented pre-nuptial molt, while both T3 and T4 treatment restored normal patterns of molt in thyroid hormone-suppressed birds. Testicular recrudescence was blocked by methimazole, and restored by T4 but not T3, in contrast to previous findings demonstrating central action of T3 in the photostimulation of breeding. Methimazole and replacement treatments elevated plasma luteinizing hormone levels compared to controls. These data are partially consistent with existing theories on the role of thyroid hormones in the photostimulation of breeding, while highlighting the possibility of additional feedback pathways. Thus we suggest that regulation of the hypothalamic pituitary gonad axis that controls breeding may be more complex than previously considered.

House sparrows mitigate growth effects of post-natal glucocorticoid exposure at the expense of longevity.

Acute, short-term effects of early-life stress and associated glucocorticoid upregulation on physiology and survival are widely documented across vertebrates. However, the persistence and severity of these effects are largely unknown, especially through the adult stage and for natural systems. Here, we investigate physiological, morphological, and survival effects of post-natal glucocorticoid upregulation across the nestling, juvenile, and adult life stages in house sparrows (Passer domesticus). We manipulate circulating corticosterone concentration in wild, free-living house sparrow nestlings and monitor body size, size-corrected mass, two measures of health (hematocrit and phytohemagglutinin-induced skin swelling), and survival in a captive environment until adulthood. We find that early-life corticosterone exposure depresses nestling size-corrected mass in both sexes, with no strong effect of the treatment on body size or our two measures of health. Birds are able to compensate for negative effects of high early-life corticosterone exposure in the long-term and this effect largely disappears by the juvenile and adult stages. However, treatment has a negative effect on survival through one year of age, suggesting that long-term compensation comes at a price.

Captivity influences immune responses, stress endocrinology, and organ size in house sparrows (Passer domesticus).

Studies using wild animals in laboratory-based research require bringing wild-captured organisms into a novel setting, which can have long-lasting impacts on physiology and behavior. In several species, captivity stimulates stress hormone production and can alter immune function. Despite this, there is little consensus on how captivity influences stress hormone regulation, or if captivity-induced changes in stress hormone production and regulation mediate changes in immune function. In this study, we investigate the influence of captivity on the physiology of a wild bird commonly-used in laboratory-based research, the house sparrow (Passer domesticus). We tested how captivity influences stress endocrinology, immune responses, and organ mass, and also investigated if the production or regulation of corticosterone, the main stress hormone in birds, correlated with changes in immunity. We found that baseline corticosterone concentrations and maximum capacity of the adrenals to secrete corticosterone increase following captivity and remain elevated after 9weeks of captivity. A measure of innate immune function, the bactericidal ability of plasma, also increased with time spent in captivity. Wound healing was also influenced by time spent in captivity, with birds taking almost 2days longer to heal if they were wounded after 3weeks in captivity when compared with birds that were wounded immediately upon capture. Additionally, captivity caused notable reductions in spleen and liver mass. Together, these results imply that captivity can have long-lasting effects on house sparrow corticosterone release and immune function, and suggest that even after 9weeks house sparrows do not acclimate physiologically to life in captivity.

Time as tyrant: The minute, hour and day make a difference for corticosterone concentrations in wild nestlings.

The hypothalamic-pituitary-adrenal (HPA) axis has been studied extensively in adults, but the HPA axis in early life is not well characterized, and there is an immense amount of unexplained variation in glucocorticoid levels during early life, especially in wild animals. To characterize population-wide natural variation in early-life HPA axis function, we compared plasma corticosterone levels (at baseline and after 30min acute restraint-stress) from seven-day-old nestlings (n=123) from a free-living, marked population of Savannah sparrows (Passerculus sandwichensis). We found a surprising sensitivity of the HPA axis to timing of sample collection across time scales. Even within the accepted 3-min framework to collect baseline samples, time to collect blood had a significant effect on baseline corticosterone concentrations. Daily rhythms also influenced baseline levels, which increased significantly during the relatively short window of sample collection (1100 and 1600). On a broader timeframe, there was a strong effect of hatch date (over a 2month period) on HPA axis responsiveness, where nestlings hatched later in the breeding season had lower stress-induced corticosterone levels than those hatched earlier. The ecophysiological mechanisms and implications of these patterns warrant future investigation; meanwhile this study highlights the critical need to consider, and potentially restrict, time across scales when collecting blood samples from wild birds to assess stress physiology.

Male song sparrows have elevated testosterone in response to neighbors versus strangers.

Upon hearing a conspecific signal, animals must assess their relationship with the signaller and respond appropriately. Territorial animals usually respond more aggressively to strangers than neighbors in a phenomenon known as the "dear enemy effect". This phenomenon likely evolved because strangers represent a threat to an animal's territory tenure and parentage, whereas neighbors only represent a threat to an animal's parentage because they already possess a territory (providing territory boundaries are established and stable). Although the dear enemy effect has been widely documented using behavioral response variables, little research has been conducted on the physiological responses of animals to neighbors versus strangers. We sought to investigate whether the dear enemy effect is observed physiologically by exposing territorial male song sparrows (Melospiza melodia) to playback simulating a neighbor or a stranger, and then collecting blood samples to measure plasma testosterone levels. We predicted that song sparrows would exhibit increased testosterone levels after exposure to stranger playback compared to neighbor playback, due to the role testosterone plays in regulating aggression. Contrary to our prediction, we found that song sparrows had higher testosterone levels after exposure to neighbor playback compared to stranger playback. We discuss several explanations for our result, notably that corticosterone may regulate the dear enemy effect in male song sparrows and this may inhibit plasma testosterone. Future studies will benefit from examining corticosterone in addition to testosterone, to better understand the hormonal underpinnings of the dear enemy effect.

Behavioral and physiological traits of migrant and resident white-crowned sparrows: a common garden approach.

To accommodate a migratory life history, migrants express a greater number of physiological and behavioral stages per annum than residents and are thus considered to have higher finite state diversity (FSD). To investigate the physiological mechanisms and constraints associated with migration, direct comparison of two subspecies of white-crowned sparrow - migrant, Zonotrichia leucophrys gambelii, and resident, Z. l. nuttalli - were made under common garden conditions of photoperiod and housing, as birds progressed from winter through the vernal life history stages. We tested the hypothesis that migrants (higher FSD) respond differently than residents (lower FSD) to the initial predictive cue, photoperiod, to initiate and integrate the progression of vernal stages of prenuptial molt, migration and development of breeding. If differences in vernal phenology were noted, then the basis for the distinctions was considered genetic. Results indicate that (1) residents had a lower threshold to vernal photoperiod with elevations of plasma androgen, growth and development of reproductive structures preceding those of migrants; (2) only migrants displayed prenuptial molt, preparations for migration and migratory restlessness; and (3) neither baseline nor stress-induced plasma corticosterone differed across subspecies, suggesting energetic demands of the common garden were insufficient to induce a differential adrenocortical response in either subspecies, highlighting the impact of environmental conditions on corticosterone secretion. Thus, in a common garden experiment, Z. l. gambelii responds differently to the initial predictive cue, photoperiod, to initiate and execute the vernal stages of molt, migration and development of breeding in comparison to the shared stage of breeding with Z. l. nuttalli, confirming a genetic basis for the subspecies differences.

Repeated immune challenges affect testosterone but not sperm quality.

Mounting an immunological response is energetically demanding and necessarily redirects allocation of resources toward immune system activation and away from other energetically expensive processes, such as reproduction. Lipopolysaccharide (LPS), a major component of the outer membrane of the cell wall of Gram-negative bacteria Escherichia coli, mimics a bacterial infection without producing the cost of replicating the pathogen and is one of the most commonly used agents to induce an acute phase immune response. Here, we ask if a trade-off can be induced between activation of the acute phase immune response and sperm function, a key indicator of sperm competitive ability. Further, we ask whether repeated exposure to this endotoxin in a social species such as the house sparrow (Passer domesticus), where repeated pathogen exposure may be common, may have a more pronounced effect. To address our questions, we exposed individuals to two rounds of LPS treatment or control, to mimic a repeated pathogen exposure in the wild. We predicted that repeated pathogen exposure would have detrimental effects on sperm quality, and therefore, reproductive success. We compared a measure of sperm quality (straight-line velocity) in captive male house sparrows between LPS-treated and control individuals. We found that although LPS treatment impaired circulating testosterone and induced a hypothermic state when compared with controls, it did not affect sperm quality within days or weeks following a single or repeated LPS exposure.

Dynamic interactions between corticosterone, corticosteroid binding globulin and testosterone in response to capture stress in male breeding Eurasian tree sparrows.

In birds, corticosterone (CORT), testosterone (T), and corticosteroid binding globulin (CBG) are involved in modulating the trade-off between reproduction and survival. In response to acute stress, increased total plasma CORT is a ubiquitous phenomenon while T levels can decrease, or remain unchanged. Since CORT and T bind competitively with CBG in birds, the underlying regulatory mechanisms and consequences of their dynamic interactions remain largely unknown. Here, we studied the dynamic changes of total CORT, T, and CBG, and estimated free and bound CORT and T in response to capture stress in male Eurasian tree sparrows (Passer montanus) across the nest building, egg-laying, and nestling stages. We predicted that free, bound and total hormone concentrations would increase for CORT and decrease for T in response to acute stress, and the relative magnitude of these changes would vary with life history stage. We found that baseline and stressed-induced CORT values did not vary across breeding sub-stages. However, total and bound CORT increased with stress while free remained unchanged. Baseline levels of total, free and bound T were highest during the nest building and it was the only stage in which all measures of T were affected by stress. Regardless of breeding stage or restraint stress, we did not detect a significant correlation between CORT and T. CBG was found to be mostly unoccupied by steroid hormones under stress and stress-free conditions and this likely provided an adequate buffer for changes in free levels of CORT and T during unpredictable environmental perturbations.

An experimental examination of interindividual variation in feather corticosterone content in the house sparrow, Passer domesticus in southeast Australia.

Non-invasive techniques for measuring glucocorticoids (GCs) have become more prevalent, due to the advantage of eliminating the effects of animal disturbance on GC levels and their potential to provide an integrated, historic estimate of circulating GC levels. In the case of birds, corticosterone (CORT) is deposited in feathers, and may reflect a bird's GC status over the period of feather synthesis. This technique thus permits a retrospective view of the average circulating GC levels during the moult period. While it is generally assumed that differences in feather CORT content (CORTf) between individuals reflects their different stress histories during either natural or induced moult, it is not clear how much of this variation is due to extrinsic versus intrinsic factors. We examined this question by determining CORTf in free-living house sparrows (Passer domesticus) from two populations, one urban and the other rural, that were plucked before and after exposure to different plasma CORT levels while held captive. We experimentally manipulated plasma CORT by implanting birds with either a corticosterone-filled, metyrapone-filled, or empty ('sham') silastic capsule as replacement feathers first emerged. The pattern of post-treatment CORTf was consistent with our expectations, based on plasma CORT levels of an experimentally implanted reference group. However, there was no statistically significant difference in CORTf between these treatment groups unless sex, population origin, and CORTf of original feathers for each individual were included in a model. Thus, birds with higher CORTf in feathers removed for this experiment tended to have higher CORTf in post-treatment replacement feathers, irrespective of treatment. In addition, we found that feather fault bar scores were significantly higher in CORT-treated birds than in the other two treatment groups, but did not vary directly with CORTf level. Our study therefore broadly confirms the use of feathers as a non-invasive tool to estimate plasma CORT during moult in birds, but importantly demonstrates the potential for intrinsic differences in stress characteristics between populations and individuals to obscure the effects extrinsic stressors might have on CORTf.

Testosterone and Haemosporidian Parasites Along a Tropical Elevational Gradient in Rufous-Collared Sparrows (Zonotrichia capensis).

Elevation has been proposed as a dominant ecological variable shaping life history traits and subsequently their underlying hormonal mechanisms. In an earlier meta-analysis of tropical birds, elevation was positively related to testosterone levels. Furthermore, parasitism by avian haemosporidians should vary with elevation as environmental conditions affect vector abundance, and while testosterone is needed for breeding, it is hypothesized to be immunosuppressive and thus could exacerbate haemosporidian infection. Our objective in this study was to examine the relationships between elevation, testosterone levels, and parasitism by avian haemosporidians. We surveyed breeding male rufous-collared sparrows (Zonotrichia capensis) across a wide elevational range along the equator. We measured baseline testosterone levels, haemosporidian infection at four elevations spanning the species' natural range in the Ecuadorian Andes (600, 1500, 2100, 3300 m). Testosterone levels from breeding males were not related to elevation, but there was high intrapopulation variability. Testosterone levels were not related to the probability of parasitism, but our results from one population suggested that the likelihood of being infected by haemosporidian parasites was greater when in breeding condition. In conclusion, even though there is variation in life history strategies among the studied populations, wider divergence in seasonality and life history traits would probably be needed to detect an effect of elevation on testosterone if one exists. Additionally, our results show that variation in testosterone is not related to infection risk of haemosporidians, thus other factors that take a toll on energetic resources, such as reproduction, should be looked at more closely.