Free-living Tree Swallows Research Articles

Heat shock protein gene expression varies among tissues and populations in free-living birds

Abstract Climate change is dramatically altering our planet, yet our understanding of mechanisms of thermal tolerance is limited in wild birds. We characterized natural variation in heat shock protein (HSP) gene expression among tissues and populations of free-living Tree Swallows (Tachycineta bicolor). We focused on HSPs because they prevent cellular damage and promote recovery from heat stress. We used quantitative PCR to measure gene expression of 3 HSPs, including those in the HSP70 and HSP90 families that have robust experimental connections to heat in past literature. First, to evaluate how tissues and, by extension, the functions that they mediate, may vary in their thermal protection, we compared HSP gene expression among neural and peripheral tissues. We hypothesized that tissues with particularly vital functions would be more protected from heat as indicated by higher HSP gene expression. We found that brain tissues had consistently higher HSP gene expression compared to the pectoral muscle. Next, we compared HSP gene expression across 4 distinct populations that span over 20° of latitude (>2,300 km). We hypothesized that the more southern populations would have higher HSP gene expression, suggesting greater tolerance of, or experience with, warmer local conditions. We observed largely higher HSP gene expression in more southern populations than northern populations, although this pattern was more striking at the extremes (southern Indiana vs. Alaska), and it was stronger in some brain areas than others (ventromedial telencephalon vs. hypothalamus). These results shed light on the potential mechanisms that may underlie thermal tolerance differences among populations or among tissues.

The telomere regulatory gene POT1 responds to stress and predicts performance in nature: Implications for telomeres and life history evolution.

Telomeres are emerging as correlates of fitness-related traits and may be important mediators of ecologically relevant variation in life history strategies. Growing evidence suggests that telomere dynamics can be more predictive of performance than length itself, but very little work considers how telomere regulatory mechanisms respond to environmental challenges or influence performance in nature. Here, we combine observational and experimental data sets from free-living tree swallows (Tachycinetabicolor) to assess how performance is predicted by the telomere regulatory gene POT1, which encodes a shelterin protein that sterically blocks telomerase from repairing the telomere. First, we show that lower POT1 gene expression in the blood was associated with higher female quality, that is, earlier breeding and heavier body mass. We next challenged mothers with an immune stressor (lipopolysaccharide injection) that led to "sickness" in mothers and 24h of food restriction in their offspring. While POT1 did not respond to maternal injection, females with lower constitutive POT1 gene expression were better able to maintain feeding rates following treatment. Maternal injection also generated a 1-day stressor for chicks, which responded with lower POT1 gene expression and elongated telomeres. Other putatively stress-responsive mechanisms (i.e., glucocorticoids, antioxidants) showed marginal responses in stress-exposed chicks. Model comparisons indicated that POT1 mRNA abundance was a largely better predictor of performance than telomere dynamics, indicating that telomere regulators may be powerful modulators of variation in life history strategies.

Brief Increases in Corticosterone Affect Morphology, Stress Responses, and Telomere Length but Not Postfledging Movements in a Wild Songbird.

Organisms are frequently exposed to challenges during development, such as poor weather and food shortage. Such challenges can initiate the hormonal stress response, which involves secretion of glucocorticoids. Although the hormonal stress response helps organisms deal with challenges, long-term exposure to high levels of glucocorticoids can have morphological, behavioral, and physiological consequences, especially during development. Glucocorticoids are also associated with telomere shortening, and they have a complex relationship with survival. To investigate whether brief, acute exposures to glucocorticoids can also produce these phenotypic effects in free-living birds, we exposed wild tree swallow (Tachycineta bicolor) nestlings to a brief exogenous dose of corticosterone once per day for 5 d and then measured their morphology, baseline and stress-induced corticosterone levels, and telomere length. We also deployed radio tags on a subset of nestlings, which allowed us to determine the age at which tagged nestlings left the nest (fledged) and their pattern of presence and absence at the natal site during the postbreeding period. Corticosterone-treated nestlings had lower mass, higher baseline and stress-induced corticosterone, and reduced telomeres; other metrics of morphology were affected weakly or not at all. Our treatment resulted in no significant effect on survival to fledging, fledge age, or age at first departure from the natal site, and we found no negative effect of corticosterone on interannual return rate. These results show that brief acute corticosterone exposure during development can have measurable effects on phenotype in free-living tree swallows. Corticosterone may therefore mediate correlations between rearing environment and phenotype in developing organisms, even in the absence of prolonged stressors.

Mercury Exposure and Altered Parental Nesting Behavior in a Wild Songbird.

Methylmercury is a neurotoxin and endocrine disruptor and may impair avian reproduction directly through embryotoxicity or by altering parental care behaviors. We studied mercury exposure and incubation behavior of free-living tree swallows ( Tachycineta bicolor) nesting in artificial nest boxes. Using small temperature dataloggers, we measured incubation constancy (the proportion of each day the female spent incubating eggs), the number of incubation recesses taken per day, and the duration of incubation recesses. We also assessed maternal mercury exposure by measuring mercury concentrations in both blood and eggs. Females with higher mercury concentrations exhibited lower incubation constancy, took more frequent and shorter incubation recesses, and were more likely to take incubation recesses that caused nest temperature decreases that were likely to slow embryonic development. Overall, females that laid eggs with the highest observed mercury concentration (0.53 μg/g fww) spent an average of 12% less time incubating their eggs over the 14-day incubation period than females that laid eggs with the lowest mercury concentration (0.07 μg/g fww). Because less time spent incubating can lower egg temperatures, slow embryonic development, and potentially lengthen the incubation period, these results suggest that environmentally relevant mercury concentrations may negatively influence reproduction by altering parental nesting behaviors of wild songbirds.

Do Hormones, Telomere Lengths, and Oxidative Stress form an Integrated Phenotype? A Case Study in Free-Living Tree Swallows.

Synopsis All organisms must anticipate and balance energetic demands and available resources in order to maximize fitness. As hormones coordinate many interactions between an organism's internal condition and the external environment, they may be key in mediating the allocation of resources to meet these demands. However, given that individuals differ considerably in how they react to changes in energetic demand, we asked whether variations in endocrine traits also correspond with life history variation. We tested whether natural variation in glucocorticoid hormone levels, oxidative stress measurements, and condition related to reproductive effort in a free-living songbird, the tree swallow, Tachycineta bicolor We then tested whether any of these traits predicted the probability of a particular individual's return to the local population in the following two years, an indicator of survival in this philopatric species. We found that males and females with longer telomeres had lighter nestlings. Moreover, individuals with lower plasma antioxidant capacity and higher reactive oxygen metabolites (i.e., greater oxidative stress) were less likely to return to the population. However, none of these traits were related to glucocorticoid levels. Our findings suggest a trade-off between reproduction and survival, with individuals with shorter telomeres having heavier nestlings but potentially paying a cost in terms of higher oxidative stress and lower survival. Interestingly, the evidence of this trade-off was unrelated to natural variation in glucocorticoids.

Interrelations among Immune Defense Indexes Reflect Major Components of the Immune System in a Free-Living Vertebrate

Understanding the relationships among immune components in free-living animals is a challenge in ecoimmunology, and it is important not only for selecting the immune assays to be used but also for more knowledgeable interpretation of results. In this study, we investigated the relationships among six immune defense indexes commonly used by ecoimmunologists and measured simultaneously in individual free-living tree swallows. Three main axes of variation in immune function were identified using a principal components analysis, representing variation in T-cell, B-cell, and innate immunity. Measures within each axis tended to be positively correlated among individuals, while measures in different axes were uncorrelated. A trade-off between T-cell function and B-cell function became apparent only when variation among individuals in body condition, age, and general quality was taken into account. Interestingly, the level of natural antibodies, a component of innate immunity, showed the strongest association with components of acquired B-cell function, possibly reflecting a common underlying genetic mechanism, as has been documented in poultry. Our results indicate that despite the complexity of the immune system, important insights can be gained by using the currently available assays but in a more comprehensive approach than has generally been used in the field of ecoimmunology.

Consequences of immune system aging in nature: a study of immunosenescence costs in free-living Tree Swallows

Immunosenescence, the aging of the immune system, is well documented in humans and laboratory models and is known to increase infection risk, morbidity, and mortality among the old. Immunosenescence patterns have recently been unveiled in various free-living populations, but their consequences in the wild have not been explored. We investigated the consequences of immunosenescence in free-living Tree Swallows Tachycineta bicolor through a field experiment simulating a bacterial infection (challenge with lipopolysaccharide, LPS) in females of different ages during the nestling rearing period. We assessed behavioral and physiological responses of females, as well as growth and quality of their offspring, to determine the costs associated with the simulated infection. Results of the experiment differed between the two years of study. In the first year, old females challenged with LPS lost more body mass and reduced their nest visitation rates more, and their offspring tended to grow slower compared to similarly challenged younger females. In contrast, in the second year, old females did not appear to suffer larger costs than younger ones. Interestingly, immunosenescence was only detected during the first year of the study, suggesting that it is the dysregulated immune function characteristic of immunosenescent individuals rather than age per se that can lead to higher costs of immune defense in old individuals. These results provide the first evidence of costs of immunosenescence in free-living animals and support the hypothesis that old, immunosenescent individuals pay higher costs than younger ones when faced with a challenge to their immune system. Our results also suggest that these costs are mediated by an exaggerated sickness behavior, as seen in laboratory models, and can be modulated by ecological factors such as weather conditions and food availability.

Mercury exposure and survival in free-living tree swallows (Tachycineta bicolor)

Mercury has become a ubiquitous contaminant in food chains worldwide. A large body of literature detailing bioaccumulation and effects on birds has revealed the potential for mercury to adversely impact avian physiology and reproduction. However, the extent to which these effects impair survival remains poorly understood. The objective of this study was to determine whether mercury exposure was associated with reduced annual survivorship in tree swallows (Tachycineta bicolor) breeding at a site with legacy industrial contamination. From 2005 to 2008, we captured and marked 932 adult swallows. We used Cormack-Jolly-Seber models and an information-theoretic approach to test our hypotheses that adult survival varied by sex, breeding location, and cumulative individual mercury exposure. Blood mercury was significantly elevated on contaminated sites (2005-2007 combined mean ± SE: 2.84 ± 0.09 μg/g; reference: 0.17 ± 0.01 μg/g). Model-averaged estimates of female apparent survival ranged from 0.483 to 0.488 on reference sites and 0.473 to 0.477 on contaminated sites. For males, apparent survival ranged from 0.451 to 0.457 on reference sites and 0.444 to 0.448 on contaminated sites. Thus, we observed approximately a 1% difference in survival between mercury-contaminated and reference sites. Such a small difference is unlikely to impact population viability in this short-lived species; however, some songbirds accumulate mercury to a greater degree than tree swallows and do not possess the migratory behavior that removes swallows to less contaminated areas for the majority of the year. Identifying whether such species are at risk of suffering biologically significant reductions in survival should become a focus of future research.

The corticosterone stress response and mercury contamination in free-living tree swallows, Tachycineta bicolor

We determined mercury concentrations in tree swallows, Tachycineta bicolor, from Massachusetts and Maine with different levels of contamination. Baseline and stress-induced plasma corticosterone concentrations from adults and nestlings (Massachusetts only) were compared with mercury concentrations. In Massachusetts, adult baseline corticosterone was negatively correlated with blood mercury, but showed a nearly-significant positive correlation with feather mercury. There was a negative relationship between baseline corticosterone and blood mercury in nestlings and between baseline corticosterone and egg mercury. There was no relationship between mercury and stress-induced corticosterone in any of the groups, or with baseline corticosterone in Maine sites where mercury levels were lower. The findings suggest blood and egg mercury may be a better indicator of current condition than feather mercury. Further, mercury contamination may not alter stress-induced corticosterone concentrations in tree swallows but appears to have a significant impact on baseline circulating corticosterone.

Compromised immune competence in free-living tree swallows exposed to mercury

Mercury is a pervasive environmental contaminant and a well-documented immunosuppressor. However, little is known about the effects of mercury contamination on health of free-living vertebrate populations. The South River in Virginia, USA was heavily contaminated with industrial mercury from 1929 to 1950, and recent studies have documented high levels of circulating mercury in riparian songbirds breeding below the site of contamination. Here we used two standardized immune assays, mitogen-induced swelling in response to phytohaemagglutinin (PHA) and antibody response to sheep red blood cells (SRBCs), to test for effects of mercury toxicity on the immune system of female tree swallows (Tachycineta bicolor) which feed on terrestrial and aquatic insects along the contaminated waterway. We found that females breeding at mercury-contaminated sites mounted significantly weaker PHA-induced swelling responses than those at reference sites in both years of study. However, among females on the contaminated sites, individual bloodstream mercury concentration did not predict the extent of mitogen-induced swelling. We did not detect any differences between reference and contaminated females in the strength of antibody responses to SRBCs, but sample sizes for this assay were significantly smaller. Overall, our results suggest that mercury toxicity can exert sub-lethal immunosuppression in free-living, insectivorous songbirds. The potential fitness consequences of the detected differences in immunocompetence caused by mercury toxicity warrant further study.

Ontogeny of innate and adaptive immune defense components in free-living tree swallows, Tachycineta bicolor

Little is known about the development of immune function in wild animals. We investigated the ontogeny of immune defense in a free-living bird, the tree swallow. We assessed total and differential leukocyte counts, natural antibodies, complement activity, in vivo skin swelling response, and in vitro lymphocyte proliferation and compared the levels of development between nestlings and young adults. We also assessed whether body condition explained variation in these immune components. We found some support for the prediction that innate defenses, which do not need to generate a broad repertoire of specific receptors, would reach adult levels earlier than adaptive defenses. In contrast, we found limited support for the prediction that adaptive defenses, which are thought to be more costly to develop, would be more related to body condition than innate defenses. We discuss our findings in the context of other studies on the ontogeny of immune function.

Seasonal trade-offs in cell-mediated immunosenescence in ruffs (Philomachus pugnax).

The immune system is an energetically expensive self-maintenance complex that, given the risks of parasitism, cannot be carelessly compromised. Life-history theory posits that trade-offs between fitness components, such as self-maintenance and reproduction, vary between genders and age classes depending on their expected residual lifetime reproductive success, and seasonally as energetic requirements change. Using ruff (Philomachus pugnax), a bird with two genetically distinct male morphs, we demonstrate here a decrease in male immunocompetence during the breeding season, greater variance in immune response among males than females, immunosenescence in both sexes and male morphs, and a seasonal shift in the age range required to detect senescence. Using a phytohaemagglutinin delayed hypersensitivity assay, we assessed cell-mediated immunity (CMI) of males of typical breeding age during the breeding and nonbreeding seasons, and of a larger sample that included females and birds of a greater age range during the non-breeding period. CMI was higher for breeding-aged males in May than in November, but the increase was not related to age or male morph. In November, mean CMI did not differ between the sexes, but the variance was higher for males than for females, and there were no differences in mean or variance between the two male morphs. For both sexes and male morphs, CMI was lower for young birds than for birds of typical breeding ages, and it declined again for older birds. In males, senescence was detected in the non-breeding season only when very old birds were included. These results, generally consistent with expectations from life-history theory, indicate that the immune system can be involved in multifarious trade-offs within a yearly cycle and along an individual's lifetime, and that specific predictions about means and variances in immune response should be considered in future immunoecological research.