Seasonal changes in the hippocampal formation of hoarding and non-hoarding tits.

The hippocampal formation (HF) processes spatial memories for cache locations in food-hoarding birds. Hoarding is a seasonal behavior, and seasonal changes in the HF have been described in some studies, but not in others. One potential reason is that birds may have been sampled during the seasonal hoarding peak in some studies, but not in others. In this study, we investigate the seasonal changes in hoarding and HF in willow tits (Poecile montanus). We compare this to seasonal changes in HF in a closely related non-hoarding bird, the great tit (Parus major). Willow tits near Oulu, Finland, show a seasonal hoarding peak in September and both HF volume and neuron number show a similar peak. HF neuronal density also increases in September, but then remains the same throughout winter. Unexpectedly, the great tit HF also changes seasonally, although in a different pattern: the great tit telencephalon increases in volume from July to August and decreases again in November. Great tit HF volume follows suit, but with a delay. Great tit HF neuron number and density also increase from August to September and stay high throughout winter. We hypothesize that seasonal changes in hoarding birds' HF are driven by food-hoarding experience (e.g., the formation of thousands of memories). The seasonal changes in great tit brains may also be due to experience-dependent plasticity, responding to changes in the social and spatial environment. Large-scale experience-dependent neural plasticity is therefore probably not an adaptation of food-hoarding birds, but a general property of the avian HF and telencephalon.

Effects of ambient temperatures on evolutionary potential of reproductive timing in boreal passerines.

Many populations need to adapt to changing environmental conditions, such as warming climate. Changing conditions generate directional selection for traits critical for fitness. For evolutionary responses to occur, these traits need to be heritable. However, changes in environmental conditions can alter the amount of heritable variation a population expresses, making predictions about expected responses difficult. The aim of this study was to evaluate the effects of ambient temperatures on evolutionary potential and strength of natural selection on the timing of reproduction in two passerine birds breeding in boreal forests. Long-term data on individually marked Willow Tits Poecile montanus (1975-2018) and Great Tits Parus major (1969-2018) were analysed with random regression animal models to assess if spring temperatures affect the expressed amount of additive genetic variation (VA ) and heritability (h2 ) in the timing of breeding. We assessed if ambient temperatures of different seasons influenced the direction and strength of selection on breeding time. We also evaluated if the strength of selection covaried with evolutionary potential. Levels of VA or h2 expressed in laying date were unaffected by spring temperatures in both study species. Selection for earlier breeding was found in the Willow Tit, but not in the Great Tit. In the Willow Tit, selection for earlier breeding was more intense when the temperatures of following autumns and winters were low. Different measures of evolutionary potential did not covary strongly with the strength of selection in either species. We conclude that there is no or little evidence that climate warming would either constrain or promote evolutionary potential in timing of breeding through changes in amount of genetic variance expressed in boreal Willow and Great Tits. However, selection on the timing of breeding, a life-history event taking place in springtime, is regulated by temperatures of autumns and winters. Rapid warming of these periods have thus potential to reduce the rate of expected evolutionary response in reproductive timing.

Climate change-driven elevational changes among boreal nocturnal moths.

Climate change has shifted geographical ranges of species northwards or to higher altitudes on elevational gradients. These changes have been associated with increases in ambient temperatures. For ectotherms in seasonal environments, however, life history theory relies largely on the length of summer, which varies somewhat independently of ambient temperature per se. Extension of summer reduces seasonal time constraints and enables species to establish in new areas as a result of over-wintering stage reaching in due time. The reduction of time constraints is also predicted to prolong organisms' breeding season when reproductive potential is under selection. We studied temporal change in the summer length and its effect on species' performance by combining long-term data on the occurrence and abundance of nocturnal moths with weather conditions in a boreal location at Värriötunturi fell in NE Finland. We found that summers have lengthened on average 5 days per decade from the late 1970s, profoundly due to increasing delays in the onset of winters. Moth abundance increased with increasing season length a year before. Most of the species occurrences expanded upwards in elevation. Moth communities in low elevation pine heath forest and middle elevation mountain birch forest have become inseparable. Yet, the flight periods have remained unchanged, probably due to unpredictable variation in proximate conditions (weather) that hinders life histories from selection. We conclude that climate change-driven changes in the season length have potential to affect species' ranges and affect the structure of insect assemblages, which may contribute to alteration of ecosystem-level processes.

The relevance of food peak architecture in trophic interactions.

Phenological shifts and associated changes in the temporal match between trophic levels have been a major focus of the study of ecological consequences of climate change. Previously, the food peak has been thought to respond as an entity to warming temperatures. However, food peak architecture, that is, timings and abundances of prey species and the level of synchrony between them, determines the timing and shape of the food peak. We demonstrate this with a case example of three passerine prey species and their predator. We explored temporal trends in the timing, height, width, and peakedness of prey availabilities and explained their variation with food peak architecture and ambient temperatures of prebreeding and breeding seasons. We found a temporal match between the predator's breeding schedule and food availability. Temporal trends in the timing of the food peak or in the synchrony between the prey species were not found. However, the food peak has become wider and more peaked over time. With more peaked food availabilities, predator's breeding success will depend more on the temporal match between its breeding schedule and the food peak, ultimately affecting the timing of breeding in the predator population. The height and width of the food peak depended on the abundances and breeding season lengths of individual prey species and their reciprocal synchronies. Peakednesses of separate prey species' availability distributions alone explained the peakedness of the food peak. Timing and quantity of food production were associated with temperatures of various time periods with variable relevance in different prey species. Alternating abundances of early and late breeding prey species caused high annual fluctuation in the timing of the food peak. Interestingly, the food peak may become later even when prey species' schedules are advanced. Climate warming can thus produce unexpected changes in the food availabilities, intervening in trophic interactions.

Different Seasonal Patterns in Song System Volume in Willow Tits and Great Tits.

In most species of seasonally breeding songbirds studied to date, the brain areas that control singing (i.e. the song control system, SCS) are larger during the breeding season than at other times of the year. In the family of titmice and chickadees (Paridae), one species, the blue tit (Cyanistes caeruleus), shows the typical pattern of seasonal changes, while another species, the black-capped chickadee (Poecile atricapillus), shows, at best, very reduced seasonal changes in the SCS. To test whether this pattern holds up in the two Parid lineages to which these two species belong, and to rule out that the differences in seasonal patterns observed were due to differences in geography or laboratory, we compared the seasonal patterns in two song system nuclei volumes (HVC and Area X) in willow tits (Poecile montanus), closely related to black-capped chickadees, and in great tits (Parus major), more closely related to blue tits, from the same area around Oulu, Finland. Both species had larger gonads in spring than during the rest of the year. Great tit males had a larger HVC in spring than at other times of the year, but their Area X did not change in size. Willow tits showed no seasonal change in HVC or Area X size, despite having much larger gonads in spring than the great tits. Our findings suggest that the song system of willow tits and their relatives may be involved in learning and producing nonsong social vocalizations. Since these vocalizations are used year-round, there may be a year-round demand on the song system. The great tit and blue tit HVC may change seasonally because the demand is only placed on the song system during the breeding season, since they only produce learned vocalizations during this time. We suggest that changes were not observed in Area X because its main role is in song learning, and there is evidence that great tits do not learn new songs after their first year of life. Further study is required to determine whether our hypothesis about the role of the song system in the learned, nonsong vocalizations of the willow tit and chickadee is correct, and to test our hypothesis about the role of Area X in the great tit song system.

Does the temporal mismatch hypothesis match in boreal populations?

The temporal mismatch hypothesis suggests that fitness is related to the degree of temporal synchrony between the energetic needs of the offspring and their food supply. The hypothesis has been a basis in studying the influence of climate warming on nature. This study enhances the knowledge on prevalence of temporal mismatches and their consequences in boreal populations, and questions the role of the temporal mismatch hypothesis as the principal explanation for the evolution of timing of breeding. To test this, we examined if synchrony with caterpillar prey or timing of breeding per se better explains reproductive output in North European parid populations. We compared responses of temperate-origin species, the great tit (Parus major) and the blue tit (Cyanistes caeruleus), and a boreal species, the willow tit (Poecile montanus). We found that phenologies of caterpillars and great tits, but not of blue tits, have advanced during the past decades. Phenologies correlated with spring temperatures that may function as cues about the timing of the food peak for great and blue tits. The breeding of great and blue tits and their caterpillar food remained synchronous. Synchrony explained breeding success better than timing of breeding alone. However, the synchrony effect arose only in certain conditions, such as with high caterpillar abundances or high breeding densities. Breeding before good synchrony seems advantageous at high latitudes, especially in the willow tit. Thus, the temporal mismatch hypothesis appears insufficient in explaining the evolution of timing of breeding.

Divergence in evolutionary potential of life history traits among wild populations is predicted by differences in climatic conditions.

Short-term adaptive evolution represents one of the primary mechanisms allowing species to persist in the face of global change. Predicting the adaptive response at the species level requires reliable estimates of the evolutionary potential of traits involved in adaptive responses, as well as understanding how evolutionary potential varies across a species' range. Theory suggests that spatial variation in the fitness landscape due to environmental variation will directly impact the evolutionary potential of traits. However, empirical evidence on the link between environmental variation and evolutionary potential across a species range in the wild is lacking. In this study, we estimate multivariate evolutionary potential (via the genetic variance-covariance matrix, or G-matrix) for six morphological and life history traits in 10 wild populations of great tits (Parus major) distributed across Europe. The G-matrix significantly varies in size, shape, and orientation across populations for both types of traits. For life history traits, the differences in G-matrix are larger when populations are more distant in their climatic niche. This suggests that local climates contribute to shaping the evolutionary potential of phenotypic traits that are strongly related to fitness. However, we found no difference in the overall evolutionary potential (i.e., G-matrix size) between populations closer to the core or the edge of the distribution area. This large-scale comparison of G-matrices across wild populations emphasizes that integrating variation in multivariate evolutionary potential is important to understand and predict species' adaptive responses to new selective pressures.

The great tit HapMap project: A continental-scale analysis of genomic variation in a songbird.

A major aim of evolutionary biology is to understand why patterns of genomic diversity vary within taxa and space. Large-scale genomic studies of widespread species are useful for studying how environment and demography shape patterns of genomic divergence. Here, we describe one of the most geographically comprehensive surveys of genomic variation in a wild vertebrate to date; the great tit (Parus major) HapMap project. We screened ca 500,000 SNP markers across 647 individuals from 29 populations, spanning ~30 degrees of latitude and 40 degrees of longitude - almost the entire geographical range of the European subspecies. Genome-wide variation was consistent with a recent colonisation across Europe from a South-East European refugium, with bottlenecks and reduced genetic diversity in island populations. Differentiation across the genome was highly heterogeneous, with clear 'islands of differentiation', even among populations with very low levels of genome-wide differentiation. Low local recombination rates were a strong predictor of high local genomic differentiation (FST), especially in island and peripheral mainland populations, suggesting that the interplay between genetic drift and recombination causes highly heterogeneous differentiation landscapes. We also detected genomic outlier regions that were confined to one or more peripheral great tit populations, probably as a result of recent directional selection at the species' range edges. Haplotype-based measures of selection were related to recombination rate, albeit less strongly, and highlighted population-specific sweeps that likely resulted from positive selection. Our study highlights how comprehensive screens of genomic variation in wild organisms can provide unique insights into spatio-temporal evolutionary dynamics.

Temperature synchronizes temporal variation in laying dates across European hole-nesting passerines.

Identifying the environmental drivers of variation in fitness-related traits is a central objective in ecology and evolutionary biology. Temporal fluctuations of these environmental drivers are often synchronized at large spatial scales. Yet, whether synchronous environmental conditions can generate spatial synchrony in fitness-related trait values (i.e., correlated temporal trait fluctuations across populations) is poorly understood. Using data from long-term monitored populations of blue tits (Cyanistes caeruleus, n = 31), great tits (Parus major, n = 35), and pied flycatchers (Ficedula hypoleuca, n = 20) across Europe, we assessed the influence of two local climatic variables (mean temperature and mean precipitation in February-May) on spatial synchrony in three fitness-related traits: laying date, clutch size, and fledgling number. We found a high degree of spatial synchrony in laying date but a lower degree in clutch size and fledgling number for each species. Temperature strongly influenced spatial synchrony in laying date for resident blue tits and great tits but not for migratory pied flycatchers. This is a relevant finding in the context of environmental impacts on populations because spatial synchrony in fitness-related trait values among populations may influence fluctuations in vital rates or population abundances. If environmentally induced spatial synchrony in fitness-related traits increases the spatial synchrony in vital rates or population abundances, this will ultimately increase the risk of extinction for populations and species. Assessing how environmental conditions influence spatiotemporal variation in trait values improves our mechanistic understanding of environmental impacts on populations.

Bird populations most exposed to climate change are less sensitive to climatic variation.

The phenology of many species shows strong sensitivity to climate change; however, with few large scale intra-specific studies it is unclear how such sensitivity varies over a species' range. We document large intra-specific variation in phenological sensitivity to temperature using laying date information from 67 populations of two co-familial European songbirds, the great tit (Parus major) and blue tit (Cyanistes caeruleus), covering a large part of their breeding range. Populations inhabiting deciduous habitats showed stronger phenological sensitivity than those in evergreen and mixed habitats. However, populations with higher sensitivity tended to have experienced less rapid change in climate over the past decades, such that populations with high phenological sensitivity will not necessarily exhibit the strongest phenological advancement. Our results show that to effectively assess the impact of climate change on phenology across a species' range it will be necessary to account for intra-specific variation in phenological sensitivity, climate change exposure, and the ecological characteristics of a population.

Geographical trends in the yolk carotenoid composition of the pied flycatcher (Ficedula hypoleuca).

Carotenoids in the egg yolks of birds are considered to be important antioxidants and immune stimulants during the rapid growth of embryos. Yolk carotenoid composition is strongly affected by the carotenoid composition of the female's diet at the time of egg formation. Spatial and temporal differences in carotenoid availability may thus be reflected in yolk concentrations. To assess whether yolk carotenoid concentrations or carotenoid profiles show any large-scale geographical trends or differences among habitats, we collected yolk samples from 16 European populations of the pied flycatcher, Ficedula hypoleuca. We found that the concentrations and proportions of lutein and some other xanthophylls in the egg yolks decreased from Central Europe northwards. The most southern population (which is also the one found at the highest altitude) also showed relatively low carotenoid levels. Concentrations of ß-carotene and zeaxanthin did not show any obvious geographical gradients. Egg yolks also contained proportionally more lutein and other xanthophylls in deciduous than in mixed or coniferous habitats. We suggest that latitudinal gradients in lutein and xanthophylls reflect the lower availability of lutein-rich food items in the northern F. hypoleuca populations and in montane southern populations, which start egg-laying earlier relative to tree phenology than the Central European populations. Similarly, among-habitat variation is likely to reflect the better availability of lutein-rich food in deciduous forests. Our study is the first to indicate that the concentration and profile of yolk carotenoids may show large-scale spatial variation among populations in different parts of the species' geographical range. Further studies are needed to test the fitness effects of this geographical variation.

From feces to data: A metabarcoding method for analyzing consumed and available prey in a bird-insect food web.

Diets play a key role in understanding trophic interactions. Knowing the actual structure of food webs contributes greatly to our understanding of biodiversity and ecosystem functioning. The research of prey preferences of different predators requires knowledge not only of the prey consumed, but also of what is available. In this study, we applied DNA metabarcoding to analyze the diet of 4 bird species (willow tits Poecile montanus, Siberian tits Poecile cinctus, great tits Parus major and blue tits Cyanistes caeruleus) by using the feces of nestlings. The availability of their assumed prey (Lepidoptera) was determined from feces of larvae (frass) collected from the main foraging habitat, birch (Betula spp.) canopy. We identified 53 prey species from the nestling feces, of which 11 (21%) were also detected from the frass samples (eight lepidopterans). Approximately 80% of identified prey species in the nestling feces represented lepidopterans, which is in line with the earlier studies on the parids' diet. A subsequent laboratory experiment showed a threshold for fecal sample size and the barcoding success, suggesting that the smallest frass samples do not contain enough larval DNA to be detected by high-throughput sequencing. To summarize, we apply metabarcoding for the first time in a combined approach to identify available prey (through frass) and consumed prey (via nestling feces), expanding the scope and precision for future dietary studies on insectivorous birds.

Different Ultimate Factors Define Timing of Breeding in Two Related Species.

Correct reproductive timing is crucial for fitness. Breeding phenology even in similar species can differ due to different selective pressures on the timing of reproduction. These selection pressures define species' responses to warming springs. The temporal match-mismatch hypothesis suggests that timing of breeding in animals is selected to match with food availability (synchrony). Alternatively, time-dependent breeding success (the date hypothesis) can result from other seasonally deteriorating ecological conditions such as intra- or interspecific competition or predation. We studied the effects of two ultimate factors on the timing of breeding, synchrony and other time-dependent factors (time-dependence), in sympatric populations of two related forest-dwelling passerine species, the great tit (Parus major) and the willow tit (Poecile montanus) by modelling recruitment with long-term capture-recapture data. We hypothesized that these two factors have different relevance for fitness in these species. We found that local recruitment in both species showed quadratic relationships with both time-dependence and synchrony. However, the importance of these factors was markedly different between the studied species. Caterpillar food played a predominant role in predicting the timing of breeding of the great tit. In contrast, for the willow tit time-dependence modelled as timing in relation to conspecifics was more important for local recruitment than synchrony. High caterpillar biomass experienced during the pre- and post-fledging periods increased local recruitment of both species. These contrasting results confirm that these species experience different selective pressures upon the timing of breeding, and hence responses to climate change may differ. Detailed information about life-history strategies is required to understand the effects of climate change, even in closely related taxa. The temporal match-mismatch hypothesis should be extended to consider subsequent critical periods when food needs to be abundantly available.

Local adaptation to winter conditions in a passerine spreading north: a common-garden approach.

Sedentary passerine birds living in temperate and boreal regions need a high metabolic capacity for thermogenesis to survive winter conditions. As a consequence of the increased thermogenic capacity, basal energetic demands rise at a time when resources and time to acquire them decrease. In a previous study, great tits (Parus major) from two localities in Fennoscandia with contrasting winter conditions differed in their metabolic response to ambient temperature. To investigate the physiological basis underlying interpopulation differences we performed a common-garden experiment to test whether these differences were genetically based. We found basal metabolic rate to be higher in birds originating from transferred eggs from the southern population compared to the ones from the northern population, contrary to the relationship among birds living in their region of origin. Despite previous evidence suggesting that gene flow prevents local adaptation at the northern range limits of a species expanding northward, we found that great tits differ in their reaction norm to winter conditions according to the population of origin.

Evolution and genetic structure of the great tit (Parus major) complex.

The great tit complex is divided into four groups, each containing several subspecies. Even though the groups are known to differ markedly on morphological, vocal and behavioural characters, some hybridization occurs in the regions where they meet. The great tit has often been referred to as an example of a ring species, although this has later been questioned. Here, we have studied the genetic structure and phylogenetic relationships of the subspecies groups to clarify the evolutionary history of the complex using control region sequences of the mitochondrial DNA. The subspecies groups were found to be monophyletic and clearly distinct in mitochondrial haplotypes, and therefore must have had long-independent evolutionary histories. This conflicts with the ring species assignment and supports the formation of secondary contact zones of previously temporarily isolated groups. According to the phylogenetic species concept, all the subspecies groups could be considered as separate species, but if the definition of the biological species concept is followed, none of the subspecies groups is a true species because hybridization still occurs.

Variable responses to large-scale climate change in European Parus populations.

Spring temperatures in temperate regions have increased over the past 20 years and many organisms have responded to this increase by advancing the timing of their growth and reproduction. However, not all populations show an advancement of phenology. Understanding why some populations advance and others do not will give us insight into the possible constraints and selection pressures on the advancement of phenology. By combining two decades of data on 24 populations of tits (Parus sp.) from six European countries, we show that the phenological response to large-scale changes in spring temperature varies across a species' range, even between populations situated close to each other. We show that this variation cannot be fully explained by variation in the temperature change during the pre- and post-laying periods, as recently suggested. Instead, we find evidence for a link between rising temperatures and the frequency of second broods, which results in complex shifts in the laying dates of first clutches. Our results emphasize the need to consider links between different life-history parameters in order to predict the ecological consequences of large-scale climate changes.

Cost of reproduction: parental survival and production of recruits in the Willow Tit Parus montanus.

Brood sizes of the Willow Tit were altered experimentally by subtracting or adding two nestlings in 1986 and 1987 in the vicinity of Oulu, northern Finland. The manipulated broods were within the normal range observed in natural conditions. Unaltered broods were used as controls. Data from natural broods from 1978-1985 were available for comparison. When the nestlings were 13 days old they were ringed and weighed and their tarsus, wing, and tail lengths were measured. On the same day the parents were caught, weighed, and measured. In 1986 there were no differences in nestling mortality between the reduced, control, or enlarged broods; i.e. parents were able to fledge the two extra young. In 1987 starvation was most pronounced in the enlarged broods. This resulted in the number of fledglings being practically the same in each manipulation category. Especially the body weight, but also the other indices of body size, decreased as a function of the brood size category, suggesting that there may be quality differences between the young reared in different experimental groups. In 1986 there was a non-significant trend towards lower body weight of the parents attending reduced, control, or enlarged broods, in that order. In 1987 the differences were much smaller. These results were not due to size differences between the groups, so possibly the increased reproductive effort of raising extra young was responsible for the trend observed in 1986. There were no significant differences in parental survival associated with the manipulation category, although the trend in the females was consistent with the hypothesis of reproductive cost. It is possible that environmental conditions in 1986 were so favourable that the tits were not unduly stressed even when attending two extra young. Correlative data from 1978-1985 did not support the cost hypothesis either. A non-significant trend towards reduced post-fledging survival and recruitment of the young was observed with increased brood size. The average fitness value of parents, incorporating parental survival and number of recruits, showed that the success of the adults raising enlarged broods may be lower than that of others. It seems that the reproductive cost, if it exists, decreases individual fitness value by reducing the chances of recruiting descendants into the next generation. The reproductive stress may be insufficient to reduce the subsequent survival of parents. More data are however needed to confirm these results.

Correlation between two components of parental investment: nest defence intensity and nestling provisioning effort of willow tits.

Nest defence intensity and nestling provisioning effort of female willow tits (Parus montanus) were significantly correlated at the end of nestling period: well-fed young were defended most intensely. Increased effort was rewarded, since broods with the highest female per-offspring provisioning rates were the most likely to produce local recruits. This suggests that the feeding ability is an important cue for parental investment decisions, at least in a species like the willow tit which has adopted the clutch adjustment strategy. Thus, the most valuable broods would not necessarily be the largest ones, but the ones in which the original number of young could be fed most adequately. However, no associations were found between the level of parental effort and offspring weight, size or condition, nor did the broods producing recruits differ from other broods in timing of breeding or number and size of offspring. The female behaviour may suggest that they invest the most time, energy and risk in the young whose chances of joining the winter flock are the best. The first well-fed young also gain an advantage of prior residency in joining the flock. The first to join normally obtain higher social status, and hence better winter survival, than latecomers. The corresponding patterns in male parental investment behaviour were weak or absent, which suggested that the male effort was affected by the female behaviour. Males seemed to invest in nestling provisioning in such a way as to supplement the female effort. During nest defence action males also seemed to invest in protection of females against predation.

To breed or not to breed: causes and implications of non-breeding habit in the willow tit Parus montanus.

Causes and consequences of non-breeding in willow tits were studied in northern Finland during 1986-1992. The breeding status was sex and age biased; males and yearling birds were in excess among the non-reproducers. Due to sex bias in the population it appeared detrimental for males to lose a mate, especially shortly before breeding. Lack of a mate was a important factor for males not reproducing (37% of non-breeding males) than for females (14%). Most of the non-breeding birds maintained a pair bond which only rarely broke up for the next breeding season (divorce rate 5.5%). This implies that parental incompatibility is not a possible explanation for pairs not reproducing. Males that did not breed tended to survive better than reproducing ones, whereas such a relationship was not found for females. It is possible that this sex-related difference in survival cost is attributable to quality differences among non-breeding individuals. It was especially low-quality yearling females, with low survival prospects, that were responsible for the discrepancy. The proportion of non-breeding females in the population correlated highly with clutch size and subsequent juvenile survival. It is therefore suggested that for most of these females non-breeding is a phenotypic response to low offspring value in the prevailing circumstances (inter-generational tradeoff). However, it is uncertain whether willow tits in a northern population can use breeding density as an indicator of changing survival prospects of their descendants, as suggested by Ekman and Askenmo (1986) for southern Sweden.

Large-scale geographical variation in eggshell metal and calcium content in a passerine bird (Ficedula hypoleuca).

Birds have been used as bioindicators of pollution, such as toxic metals. Levels of pollutants in eggs are especially interesting, as developing birds are more sensitive to detrimental effects of pollutants than adults. Only very few studies have monitored intraspecific, large-scale variation in metal pollution across a species' breeding range. We studied large-scale geographic variation in metal levels in the eggs of a small passerine, the pied flycatcher (Ficedula hypoleuca), sampled from 15 populations across Europe. We measured 10 eggshell elements (As, Cd, Cr, Cu, Ni, Pb, Zn, Se, Sr, and Ca) and several shell characteristics (mass, thickness, porosity, and color). We found significant variation among populations in eggshell metal levels for all metals except copper. Eggshell lead, zinc, and chromium levels decreased from central Europe to the north, in line with the gradient in pollution levels over Europe, thus suggesting that eggshell can be used as an indicator of pollution levels. Eggshell lead levels were also correlated with soil lead levels and pH. Most of the metals were not correlated with eggshell characteristics, with the exception of shell mass, or with breeding success, which may suggest that birds can cope well with the current background exposure levels across Europe.