Identifying the paths of climate effects on population dynamics: dynamic and multilevel structural equation model around the annual cycle.

How environmental factors influence population dynamics in long-distance migrants is complicated by the spatiotemporal diversity of the environment the individuals experience during the annual cycle. The effects of weather on several different aspects of life history have been well studied, but a better understanding is needed on how weather affects population dynamics through the different associated traits. We utilise 77 years of data from pied flycatcher (Ficedula hypoleuca), to identify the most relevant climate signals associated with population growth rate. The strongest signals on population growth were observed from climate during periods when the birds were not present in the focal location. The population decline was associated with increasing precipitation in the African non-breeding quarters in the autumn (near the arrival of migrants) and with increasing winter temperature along the migration route (before migration). The number of fledglings was associated positively with increasing winter temperature in non-breeding area and negatively with increasing winter temperature in Europe. These possible carry-over effects did not arise via timing of breeding or clutch size but the exact mechanism remains to be revealed in future studies. High population density and low fledgling production were the intrinsic factors reducing the breeding population. We conclude that weather during all seasons has the potential to affect the reproductive success or population growth rate of this species. Our results show how weather can influence the population dynamics of a migratory species through multiple pathways, even at times of the annual cycle when the birds are in a different location than the climate signal.

Interspecific variation in the relationship between clutch size, laying date and intensity of urbanization in four species of hole-nesting birds.

The increase in size of human populations in urban and agricultural areas has resulted in considerable habitat conversion globally. Such anthropogenic areas have specific environmental characteristics, which influence the physiology, life history, and population dynamics of plants and animals. For example, the date of bud burst is advanced in urban compared to nearby natural areas. In some birds, breeding success is determined by synchrony between timing of breeding and peak food abundance. Pertinently, caterpillars are an important food source for the nestlings of many bird species, and their abundance is influenced by environmental factors such as temperature and date of bud burst. Higher temperatures and advanced date of bud burst in urban areas could advance peak caterpillar abundance and thus affect breeding phenology of birds. In order to test whether laying date advance and clutch sizes decrease with the intensity of urbanization, we analyzed the timing of breeding and clutch size in relation to intensity of urbanization as a measure of human impact in 199 nest box plots across Europe, North Africa, and the Middle East (i.e., the Western Palearctic) for four species of hole-nesters: blue tits (Cyanistes caeruleus), great tits (Parus major), collared flycatchers (Ficedula albicollis), and pied flycatchers (Ficedula hypoleuca). Meanwhile, we estimated the intensity of urbanization as the density of buildings surrounding study plots measured on orthophotographs. For the four study species, the intensity of urbanization was not correlated with laying date. Clutch size in blue and great tits does not seem affected by the intensity of urbanization, while in collared and pied flycatchers it decreased with increasing intensity of urbanization. This is the first large-scale study showing a species-specific major correlation between intensity of urbanization and the ecology of breeding. The underlying mechanisms for the relationships between life history and urbanization remain to be determined. We propose that effects of food abundance or quality, temperature, noise, pollution, or disturbance by humans may on their own or in combination affect laying date and/or clutch size.

Natural selection for earlier male arrival to breeding grounds through direct and indirect effects in a migratory songbird.

For migratory birds, the earlier arrival of males to breeding grounds is often expected to have fitness benefits. However, the selection differential on male arrival time has rarely been decomposed into the direct effect of male arrival and potential indirect effects through female traits. We measured the directional selection differential on male arrival time in the pied flycatcher (Ficedula hypoleuca) using data from 6 years and annual number of fledglings as the fitness proxy. Using structural equation modeling, we were able to take into account the temporal structure of the breeding cycle and the hierarchy between the examined traits. We found directional selection differentials for earlier male arrival date and earlier female laying date, as well as strong selection differential for larger clutch size. These selection differentials were due to direct selection only as indirect selection for these traits was nonsignificant. When decomposing the direct selection for earlier male arrival into direct and indirect effects, we discovered that it was almost exclusively due to the direct effect of male arrival date on fitness and not due to its indirect effects via female traits. In other words, we showed for the first time that there is a direct effect of male arrival date on fitness while accounting for those effects that are mediated by effects of the social partner. Our study thus indicates that natural selection directly favored earlier male arrival in this flycatcher population.

Long-term recovery of clutch size and egg shell quality of the pied flycatcher (Ficedula hypoleuca) in a metal polluted area.

We explored if breeding parameters and egg shell quality of an insectivorous passerine, pied flycatcher Ficedula hypoleuca, have fully recovered after c.a. 99% decrease in dust emissions from a non-ferrous smelter in the course of the 23 year study period. Some potentially important population characteristics (density, phenology, age) and environmental variables (habitat, inter-specific competition) were taken into account in the analysis. We found marked increase in reproductive parameters (egg shell quality, clutch size, hatchability, and fledgling number) in the metal polluted area especially in 1990's when metal-rich dust emissions from the smelter were markedly reduced. Still clutch sizes and fledgling numbers remain below the levels of the reference area. There is currently very little evidence of direct toxic effects of metals in our study population but full recovery of breeding parameters may not be reached until the full recovery of food chains, which is likely to be a slow process.

Variation in clutch size in relation to nest size in birds.

Nests are structures built to support and protect eggs and/or offspring from predators, parasites, and adverse weather conditions. Nests are mainly constructed prior to egg laying, meaning that parent birds must make decisions about nest site choice and nest building behavior before the start of egg-laying. Parent birds should be selected to choose nest sites and to build optimally sized nests, yet our current understanding of clutch size-nest size relationships is limited to small-scale studies performed over short time periods. Here, we quantified the relationship between clutch size and nest size, using an exhaustive database of 116 slope estimates based on 17,472 nests of 21 species of hole and non-hole-nesting birds. There was a significant, positive relationship between clutch size and the base area of the nest box or the nest, and this relationship did not differ significantly between open nesting and hole-nesting species. The slope of the relationship showed significant intraspecific and interspecific heterogeneity among four species of secondary hole-nesting species, but also among all 116 slope estimates. The estimated relationship between clutch size and nest box base area in study sites with more than a single size of nest box was not significantly different from the relationship using studies with only a single size of nest box. The slope of the relationship between clutch size and nest base area in different species of birds was significantly negatively related to minimum base area, and less so to maximum base area in a given study. These findings are consistent with the hypothesis that bird species have a general reaction norm reflecting the relationship between nest size and clutch size. Further, they suggest that scientists may influence the clutch size decisions of hole-nesting birds through the provisioning of nest boxes of varying sizes.

Density effect on great tit (Parus major) clutch size intensifies in a polluted environment.

Long-term data on a great tit (Parus major) population breeding in a metal-polluted zone around a copper-nickel smelter indicate that, against expectations, the clutch size of this species is decreasing even though metal emissions in the area have decreased considerably over the past two decades. Here, we document long-term population-level changes in the clutch size of P. major and explore if changes in population density, population numbers of competing species, timing of breeding, breeding habitat, or female age distribution can explain decreasing clutch sizes. Clutch size of P. major decreased by one egg in the polluted zone during the past 21 years, while there was no significant change in clutch size in the unpolluted reference zone over this time period. Density of P. major nests was similar in both environments but increased threefold during the study period in both areas (from 0.8 to 2.4 nest/ha). In the polluted zone, clutch size has decreased as a response to a considerable increase in population density, while a corresponding density change in the unpolluted zone did not have such an effect. The other factors studied did not explain the clutch size trend. Fledgling numbers in the polluted environment have been relatively low since the beginning of the study period, and they do not show a corresponding decrease to that noted for the clutch size over the same time period. Our study shows that responses of commonly measured life-history parameters to anthropogenic pollution depend on the structure of the breeding population. Interactions between pollution and intrinsic population characters should therefore be taken into account in environmental studies.

Corticosterone secretion patterns prior to spring and autumn migration differ in free-living barn swallows (Hirundo rustica L.).

Recent studies of long-distance migratory birds show that behavioural and physiological changes associated with predictable or unpredictable challenges during the annual cycle are distinctively regulated by hormones. Corticosterone is the primary energy regulating hormone in birds. Corticosterone levels are elevated during stresses but they are also modulated seasonally according to environmental conditions and life-history demands. We measured the baseline and stress-induced levels of corticosterone in the barn swallow (Hirundo rustica L.) just before spring and autumn migrations in South Africa and Finland, respectively. Barn swallows completing their pre-breeding moult had low body condition (residual body mass) and high baseline corticosterone levels in the wintering grounds. In contrast, baseline corticosterone levels in Finland were low and not related to residual mass. These data contradict the first prediction of the migration modulation hypothesis (MMH) by showing no association with baseline corticosterone levels and pre-migratory fuelling. Yet, the adrenocortical response to the capture and handling stress was notably blunted in South Africa compared to a strong response in Finland. Further, individuals that had started fuelling in Finland showed a reduced response to the handling stress. Taken together, elevated baseline corticosterone levels and high residual mass may blunt the adrenocortical response in long-distance migrants and aerial feeders such as the barn swallow. This observation lends support to the second prediction of the MMH.

Seasonal variation in the regulation of redox state and some biotransformation enzyme activities in the barn swallow (Hirundo rustica L.).

Little is known of the normal seasonal variation in redox state and biotransformation activities in birds. In long-distance migratory birds, in particular, seasonal changes could be expected to occur because of the demands of migration and reproduction. In this study, we measured several redox parameters in the barn swallow (Hirundo rustica L.) during the annual cycle. We captured the wintering barn swallows before spring migration in South Africa, and we captured the barn swallows that arrived in spring, bred in summer, and migrated in autumn in Finland. The redox status and biotransformation activities of barn swallows varied seasonally. Wintering birds in South Africa had high biotransformation activities and appeared to experience oxidative stress, whereas in spring and summer, they showed relatively low redox (superoxide dismutase [SOD], catalase [CAT], and glutathione reductase [GR]) and biotransformation enzyme activities. Autumn birds had very low biotransformation enzyme activities and low indication of oxidative stress but high activity of some redox enzymes (GR and glucose 6-phosphate dehydrogenase [G6PDH]). High activities of some redox enzymes (SOD, GR, and G6PDH) seem to be related to migration, whereas low activities of some redox enzymes (SOD, CAT, and GR) may be associated with breeding. Barn swallows in South Africa may experience pollution-related oxidative stress, which may hamper interpretation of normal seasonal variation in redox parameters.

Selection on laying date is connected to breeding density in the pied flycatcher.

Timing of reproduction and clutch size are important determinants of breeding success, especially in seasonal environments. Several recent bird population studies have shown changes in breeding time and in natural selection on it. These changes have often been linked with climate change, but few studies have investigated how the traits or natural selection are actually connected with climatic factors. Furthermore, the effect of population density on selection has been rarely considered, despite the potential importance of density in demographic processes. We studied variation in natural selection on laying date and on clutch size in relation to measures of spring phenology and population density in a long-term study of pied flycatchers in SW Finland. The phenological stage of the environment at mean egg-laying did not affect the direction of selection on either laying date or on clutch size. There was, however, stronger selection for earlier laying date when the breeding density of the population was high, suggesting that early breeding is not necessarily beneficial as such, but that its importance is emphasized when high population density increases competition. In addition, early breeding was favoured when the pre-breeding period was cool, which may indicate an increased advantage for the fittest individuals in harsher conditions. In the middle of the twentieth century, there was selection for large clutch size, which subsequently ceased, along with an overall decrease in recruit production. Our results indicate that attention should be paid to demographic factors such as breeding density when studying natural selection and temporal changes in it.

Challenging claims in the study of migratory birds and climate change.

Recent shifts in phenology in response to climate change are well established but often poorly understood. Many animals integrate climate change across a spatially and temporally dispersed annual life cycle, and effects are modulated by ecological interactions, evolutionary change and endogenous control mechanisms. Here we assess and discuss key statements emerging from the rapidly developing study of changing spring phenology in migratory birds. These well-studied organisms have been instrumental for understanding climate-change effects, but research is developing rapidly and there is a need to attack the big issues rather than risking affirmative science. Although we agree poorly on the support for most claims, agreement regarding the knowledge basis enables consensus regarding broad patterns and likely causes. Empirical data needed for disentangling mechanisms are still scarce, and consequences at a population level and on community composition remain unclear. With increasing knowledge, the overall support ('consensus view') for a claim increased and between-researcher variability in support ('expert opinions') decreased, indicating the importance of assessing and communicating the knowledge basis. A proper integration across biological disciplines seems essential for the field's transition from affirming patterns to understanding mechanisms and making robust predictions regarding future consequences of shifting phenologies.

Climate warming, ecological mismatch at arrival and population decline in migratory birds.

Climate is changing at a fast pace, causing widespread, profound consequences for living organisms. Failure to adjust the timing of life-cycle events to climate may jeopardize populations by causing ecological mismatches to the life cycle of other species and abiotic factors. Population declines of some migratory birds breeding in Europe have been suggested to depend on their inability to adjust migration phenology so as to keep track of advancement of spring events at their breeding grounds. In fact, several migrants have advanced their spring arrival date, but whether such advancement has been sufficient to compensate for temporal shift in spring phenophases or, conversely, birds have become ecologically mismatched, is still an unanswered question, with very few exceptions. We used a novel approach based on accumulated winter and spring temperatures (degree-days) as a proxy for timing of spring biological events to test if the progress of spring at arrival to the breeding areas by 117 European migratory bird species has changed over the past five decades. Migrants, and particularly those wintering in sub-Saharan Africa, now arrive at higher degree-days and may have therefore accumulated a 'thermal delay', thus possibly becoming increasingly mismatched to spring phenology. Species with greater 'thermal delay' have shown larger population decline, and this evidence was not confounded by concomitant ecological factors or by phylogenetic effects. These findings provide general support to the largely untested hypotheses that migratory birds are becoming ecologically mismatched and that failure to respond to climate change can have severe negative impacts on their populations. The novel approach we adopted can be extended to the analysis of ecological consequences of phenological response to climate change by other taxa.

Metal pollution does not bias offspring sex ratio in great tit (Parus major).

PURPOSE: We measured offspring sex ratios in a resident insectivorous passerine bird, the Great tit Parus major, to find out whether metal pollution around a Finnish copper smelter would bias sex ratios in this territorial forest bird species. Besides a direct effect of pollutants, we were interested in possible indirect effects of pollution-related resource limitation, i.e., changes in food quality and quantity, on sex ratio. METHODS: We manipulated the diet quantity and quality (carotenoid levels) of P. major nestlings in polluted and unpolluted environments to find out if variation in dietary resource levels has any effect on fledgling sex ratios. Faecal metal levels of nestlings were analysed to monitor the level of pollution exposure at each territory. Near the time of fledging (at the age of 15 days), feather samples were collected for molecular sex determination. RESULTS: We did not find any association between metal exposure levels and offspring sex ratio, and thus, our results do not support the hypothesis that metal pollution would have direct effects on P. major offspring sex ratio. We found that, irrespective of pollution level, high nestling mortality was associated with male-biased sex ratio, most likely due to sex-specific mortality. Our feeding treatment, however, showed no significant effects on offspring sex ratio, possibly because natural variation in food availability outweighed the effect of our food manipulation on nestling mortality. CONCLUSIONS: We conclude that metal exposure at the levels found in our study area does not bias fledgling sex ratios in this species, but biased sex ratios may follow under natural or pollution-related resource limitation.

Body condition is associated with adrenocortical response in the barn swallow (Hirundo rustica L.) during early stages of autumn migration.

Migration is an energy-demanding life-history period and also a significant population-limiting factor of long-distance migratory birds. It is important to understand how corticosterone, the main energy regulating hormone in birds, is associated with behavioural and physiological changes during migration. According to the migration modulation hypothesis (MMH), individual birds may express elevated levels of baseline corticosterone to facilitate fuelling, but down-regulate the adrenocortical response in order to protect skeletal muscles from the catabolic effects of the hormone. We measured the baseline and stress-induced levels of corticosterone in barn swallows (Hirundo rustica L.) during early stages of autumn migration. Here, we show that, while barn swallows clearly responded to the capture and handling stress by increasing the corticosterone level, the strength of this acute response was related to their energetic condition: birds with high body mass responded more rapidly and had lower peak values of corticosterone than lighter birds. Further, the baseline levels of corticosterone correlated negatively with the magnitude of the adrenocortical response. Barn swallows did not show elevated baseline levels of corticosterone in the course of autumn, which suggests that, instead of fuelling, the birds were actively migrating. Our results indicate that MMH also applies to aerial feeders, whose foraging habits differ from model birds of previous studies.

Local temperature fine-tunes the timing of spring migration in birds.

Evidence for climate-driven phenological changes is rapidly increasing at all trophic levels. Our current poor knowledge of the detailed control of bird migration from the level of genes and hormonal control to direct physiological and behavioral responses hampers our ability to understand and predict consequences of climatic change for migratory birds. In order to better understand migration phenology and adaptation in environmental changes, we here assess the scale at which weather affects timing of spring migration in passerine birds. We use three commonly used proxies of spring-time climatic conditions: (1) vegetation "greenness" (NDVI) in Europe, (2) local spring temperatures in northern Europe, and (3) the North Atlantic Oscillation Index (NAO) as predictors of the phenology of avian migration as well as the strength of their effect on different subsets of populations and the dependence of correlations on species-specific migratory strategy. We analyze phenological patterns of the entire spring migration period in 12 Palaearctic passerine species, drawing on long-term data collected at three locations along a longitudinal gradient situated close to their northern European breeding area. Local temperature was the best single predictor of phenology with the highest explanatory power achieved in combination with NAO. Furthermore, early individuals are more affected by climatic variation compared to individuals on later passage, indicating that climatic change affects subsets of migratory populations differentially. Species wintering closer to the breeding areas were affected more than were those travelling longer distances and this pattern was strongest for the earliest subsets of the population. Overall, our results suggest that at least early subsets of the population are affected by local conditions and early birds use local conditions to fine-tune the date of their spring arrival while individuals arriving later are driven by other factors than local conditions e.g. endogenous control. Understanding what cues migratory organisms use to arrive at an optimum time is important for increasing our knowledge of fundamental issues like decision making in organisms during migration and is crucial for future protection of migratory organisms.

Polluted environment and cold weather induce laying gaps in great tit and pied flycatcher.

We studied the occurrence of laying gaps in free-living populations of the pied flycatcher, Ficedula hypoleuca, and the great tit, Parus major, in a pollution gradient of a copper smelter in south-west Finland. Laying gaps were 2.8 times more common in F. hypoleuca than in P. major. The probability of laying gaps was highest in the heavily polluted zone and lowest in the unpolluted zone for both bird species. Cold weather at the time of laying increased the number of laying gaps in both species, but in P. major this effect was most pronounced in the heavily polluted environment. In the most heavily polluted environment the laying gaps were more likely to occur near the beginning of the laying sequence in both species. The laying gap probability increased with increasing laying date in P. major but not in F. hypoleuca. We suggest that the increased number of laying gaps in the polluted environment results from limited Ca availability and the interference of heavy metals with Ca metabolism in laying females.

Great tits lay increasingly smaller clutches than selected for: a study of climate- and density-related changes in reproductive traits.

1. The phenology of temperate environments and therefore timing of breeding has advanced in a number of bird species due to climate warming. Few studies, however, have examined the mechanisms behind the observed changes, the role of natural selection in them or the determinants of the selection. In other traits such as clutch size, even changes over years have been rarely studied. 2. We studied patterns and trends in timing of breeding, clutch size and fledgling production in the great tit Parus major in South-West Finland during 1953-2008, as well as natural selection on the timing and clutch size, based on fledgling production. We also examined connections between these parameters and a number of climatic and population intrinsic factors. 3. Laying date was earlier when the pre-breeding period was warm and tended to be earlier when breeding density was high, but it did not show any temporal change during the study period despite temporal increases in both explanatory factors. Number of fledglings decreased through declines in both mean clutch size and fledging success. Fledging success was better with higher breeding-time temperature and larger clutch size. Both the clutch size and fledging success were lower at higher breeding density. 4. Selection on laying date did not change through time, but there was a selection for early laying with high breeding-time temperature and high breeding density. Interestingly, in contrast to the decrease in reproductive output, the selection for larger than average clutch size strengthened with time, which was not explained by any tested factor. 5. We suggest that increasingly favourable conditions in winters have enhanced the survival and resulted in the observed increase in great tit breeding density. This may have most concerned young and otherwise low-quality individuals, which also most likely end up breeding in the increasingly occupied low-quality territories. This hypothesis was indicatively supported by increased within-year variation in both laying date and clutch size. The changes could also explain the lack of advancement in laying date as well as the increasing selection for large clutch sizes as the fittest individuals most likely occupy the best territories and lay largest clutches.

Climate change effects on migration phenology may mismatch brood parasitic cuckoos and their hosts.

Phenological responses to climate change vary among taxa and across trophic levels. This can lead to a mismatch between the life cycles of ecologically interrelated populations (e.g. predators and prey), with negative consequences for population dynamics of some of the interacting species. Here we provide, to our knowledge, the first evidence that climate change might disrupt the association between the life cycles of the common cuckoo (Cuculus canorus), a migratory brood parasitic bird, and its hosts. We investigated changes in timing of spring arrival of the cuckoo and its hosts throughout Europe over six decades, and found that short-distance, but not long-distance, migratory hosts have advanced their arrival more than the cuckoo. Hence, cuckoos may keep track of phenological changes of long-distance, but not short-distance migrant hosts, with potential consequences for breeding of both cuckoo and hosts. The mismatch to some of the important hosts may contribute to the decline of cuckoo populations and explain some of the observed local changes in parasitism rates of migratory hosts.

Populations of migratory bird species that did not show a phenological response to climate change are declining.

Recent rapid climatic changes are associated with dramatic changes in phenology of plants and animals, with optimal timing of reproduction advancing considerably in the northern hemisphere. However, some species may not have advanced their timing of breeding sufficiently to continue reproducing optimally relative to the occurrence of peak food availability, thus becoming mismatched compared with their food sources. The degree of mismatch may differ among species, and species with greater mismatch may be characterized by declining populations. Here we relate changes in spring migration timing by 100 European bird species since 1960, considered as an index of the phenological response of bird species to recent climate change, to their population trends. Species that declined in the period 1990-2000 did not advance their spring migration, whereas those with stable or increasing populations advanced their migration considerably. On the other hand, population trends during 1970-1990 were predicted by breeding habitat type, northernmost breeding latitude, and winter range (with species of agricultural habitat, breeding at northern latitudes, and wintering in Africa showing an unfavorable conservation status), but not by change in migration timing. The association between population trend in 1990-2000 and change in migration phenology was not confounded by any of the previously identified predictors of population trends in birds, or by similarity in phenotype among taxa due to common descent. Our findings imply that ecological factors affecting population trends can change over time and suggest that ongoing climatic changes will increasingly threaten vulnerable migratory bird species, augmenting their extinction risk.

Carotenoids in a food chain along a pollution gradient.

Carotenoids are synthesized by plants, therefore insects and birds must obtain them from their diet. They function in pigmentation and as antioxidants. We studied the carotenoid profiles in a model food chain (plant-insect-bird) in an air pollution gradient to find out whether heavy metal pollution affects the transfer of carotenoids across the trophic levels. Birch leaves showed higher beta-carotene and, one of the birch species (Betula pendula), higher total carotenoids levels in the polluted area. There was no difference in the lutein concentration of caterpillars' food source, birch leaves, between the study areas. Autumnal moth larvae accumulated lutein more efficiently than beta-carotene while sawfly larvae accumulated beta-carotene over lutein. Because of different antioxidant profiles in different leaf chewing insects their sensitivity to pollution stress may differ. The lutein concentration of plasma and feathers of Great tit nestlings did not differ along the pollution gradient. The lack of difference in lutein concentration of autumnal moth larvae along pollution gradient may partly explain the lutein concentrations of Great tit nestlings, since the abundance of autumnal moth larvae peak during the nestling phase of Great tit. The lutein concentration of autumnal moth larvae was positively associated to circulating plasma lutein level of Great tit indicating the importance of carotenoid rich diet during the nestling phase. In addition, the higher the plasma lutein concentration the more lutein was deposited to feathers, irrespective of the other possible functions of lutein in nestlings. We found that carotenoid levels differed between the polluted and the unpolluted area especially at lower levels of food chain: in birches and in caterpillars.

Environmental pollution affects the plumage color of Great tit nestlings through carotenoid availability.

Birds need to acquire carotenoids for their feather pigmentation from their diet, which means that their plumage color may change as a consequence of human impact on their environment. For example, the carotenoid-based plumage coloration of Great tit, Parus major, nestlings is associated with the degree of environmental pollution. Breast feathers of birds in territories exposed to heavy metals are less yellow than those in unpolluted environments. Here we tested two hypotheses that could explain the observed pattern: (I) deficiency of carotenoids in diet, and (II) pollution-related changes in transfer of carotenoids to feathers. We manipulated dietary carotenoid levels of nestlings and measured the responses in plumage color and tissue concentrations. Our carotenoid supplementation produced the same response in tissue carotenoid concentrations and plumage color in polluted and unpolluted environments. Variation in heavy metal levels did not explain the variation in tissue (yolk, plasma, and feathers) carotenoid concentrations and was not related to plumage coloration. Instead, the variation in plumage yellowness was associated with the availability of carotenoid-rich caterpillars in territories. Our results support the hypothesis that the primary reason for pollution-related variation in plumage color is carotenoid deficiency in the diet.