Speckled eggs: water-loss and incubation behaviour in the great tit Parus major

Many small passerine birds worldwide lay white eggs speckled with red, brown and black protoporphyrin pigment spots (maculation). Unlike some patterns of avian eggshell pigmentation which clearly serve a crypsis or signalling function, the ubiquity of maculation among passerines suggests that its origins lie in another function, not specific to any particular ecological or behavioural group. Elsewhere, we have presented evidence that protoporphyrin pigments serve a structural function related to eggshell thickness and calcium availability: eggshell maculation in the great tit Parus major increases with decreasing soil calcium levels, pigments demarcate thinner areas of shell, and both the pigment intensity and distribution are related to shell thickness. Here we show that maculation also affects the rate of water loss from the egg during incubation (≈ Mass Loss per Day or MLD, which is critical to egg viability), but not that of unincubated eggs. We also demonstrate, both by observation and experiment, that the effect of female incubation behaviour on MLD compensates in some way for variation in egg characteristics, and that differences between females in the degree of such compensation are related to differences in clutch maculation. Our results suggest that, while a principal function of maculation in this species may be to strengthen the eggshell, it may also reduce eggshell permeability when large amounts of pigment are used, and that this necessitates a behavioural adjustment from the female during incubation. We discuss these findings and make further testable predictions from our model.     

An evaluation of different methods for assessing eggshell pigmentation and pigment concentration using great tit eggs

The striking variation in colour and maculation of bird eggs has fascinated biologists since centuries, and many hypotheses based on mechanical, physiological or signalling functions have been proposed for its evolution by natural and sexual selection. Protoporphyrin is the main eggshell pigment found in brown maculated eggs, and is assumed to function as a mediator of these selection processes. It is a precursor of heme with pro‐oxidant properties, and hence a link between brown maculation and female condition has been proposed and tested in a number of studies, albeit with contrasting results. A variety of different visual methods have been used to quantify outer eggshell pigmentation, which has been assumed to correspond to overall quantity of protoporphyrin in the shell. Yet, this relationship has rarely been tested. The aim of this study was to apply four commonly used methods to assess pigmentation in great tit eggs with protoporphyrin as the predominant eggshell pigment, and to compare the results of these methods. Specifically, we 1) ranked eggshell pigmentation by human naked eye, 2) applied a granularity approach and 3) measured spectrophotometric reflectance of eggshell pigments. Second, we estimated the relationship between outer eggshell pigmentation (i.e. estimated by three different methods above) and true protoporphyrin concentration deposited in the entire shell measured by HPLC. Among‐method estimates were significantly correlated for the traits describing pigment ‘darkness’ only. While the model including scores based on human naked eyes explained 16% of the variance of pigment concentration in the entire shell, spectrometry explained 27%, and the granularity approach explained 40%. Thus, the estimation of true pigment concentration in the entire shell from the visible outer side of the shell is most reliable with the granularity approach. It is relevant for studies where the maintenance of the integrity of the eggs is essential.     

Effects of experimental calcium availability,egg parameters and laying order on Great Tit Parus major eggshell pigmentation patterns

Many bird species lay eggs speckled with protoporphyrin‐based spots, however, for most of them the function of eggshell spotting is unknown. A plausible hypothesis is that protoporphyrin might have a structural function in strengthening the eggshell and is therefore deposited when calcium is scarce. In this study, we experimentally provided Great Tit Parus major females with supplemental calcium to examine its effect on the protoporphyrin‐based maculation of their eggs. In addition, we studied variation in eggshell pigmentation patterns in relation to other egg parameters and laying order. Calcium‐supplemented females laid larger eggs but shell thickness was not significantly affected by the treatment. Calcium supplementation may reduce the time and energy females devote to searching for calcium‐rich material, so that they can collect more nutrients and so lay larger eggs. Furthermore, pigment darkness was associated with egg volume and shape, which suggests that female quality and environmental food availability may also influence the shell pigmentation pattern. Within clutches, later‐laid eggs had larger and darker spots that were distributed more unevenly on the shell surface. This within‐clutch pattern could be explained by the increase in egg volume and egg shape and a decline in shell thickness with egg‐laying order, which characteristics were all related to shell‐spotting pattern. Eggs with a coronal ring had thinner shells, but pigment intensity and spot size were not related to shell thickness. Thus, our results suggest that concentrated spotting distribution may have a mechanical function, supporting the structural‐function hypothesis.     

Eggshell Appearance Does Not Signal Maternal Corticosterone Exposure in Japanese Quail: An Experimental Study with Brown-Spotted Eggs

Reproduction is a critical period for birds as they have to cope with many stressful events. One consequence of an acute exposure to stress is the release of corticosterone, the avian stress hormone. Prolonged stress can have negative impacts on the immune system, resulting in, for example, increased oxidative stress. Through maternal effects, females are known to modulate their investment in eggs content according to their own physiological condition. Less is known about maternal investment in eggshells, especially in pigments. The two main eggshell pigments may possess opposite antioxidant properties: protoporphyrin (brown) is a pro-oxidant, whereas biliverdin (blue-green) is an antioxidant. In Japanese quail, we know that the deposition of both pigments is related to female body condition. Thus, a chronic stress response may be reflected in eggshell coloration. Using female Japanese quails that lay brown-spotted eggs, we explored whether physiological exposure to corticosterone induces a change in female basal stress and antioxidant factors, and eggshell pigment concentration, spectrophotometric reflectance, and maculation coverage. We supplemented adult females over a 2 week period with either peanut oil (control) or corticosterone (treatment). We collected pre- and post-supplementation eggs and analysed the effect of corticosterone treatment on female physiology and eggshell appearance parameters. Except for corticosterone-fed birds which laid eggs with brighter spots, supplementation had no significant effect on female physiology or eggshell pigment concentration, reflectance and maculation. The change in eggshell spot brightness was not detected by a photoreceptor noise-limited color opponent model of avian visual perception. Our data confirms that eggshell reflectance in spotted eggs varies over the laying sequence, and spot reflectance may be a key factor that is affected by females CORT exposure, even if the changes are not detected by an avian visual model.     

Avian eggshell coloration: new perspectives on adaptive explanations

Recent work suggests that the evolution of egg coloration may have been constrained in three important ways that have not yet been critically synthesized in any review. First, on account of birds being able to see in the ultraviolet spectrum, the interaction between the properties of avian vision and the light environment of nests imply different perceptions of egg coloration from those experienced by humans. Second, a new hypothesis to explain blue–green egg coloration interprets it as a sexually selected signal to males of the laying female's genetic quality. Third, evidence from taxa as divergent as sparrowhawks and great tits indicates that protoporphyrin pigments responsible for maculation (spotting patterns) have a structural function in compensating for eggshell thinning, as caused by calcium stress, and, more recently, dichlorodiphenyltrichloroethane. We consider this to be the most convincing explanation for the primary function of spotting, although an important secondary function might arise through the fact that individual patterns of maculation may allow birds to identify their own eggs, effectively serving as signatures in the face of inter‐ or intra‐specific brood parasitism. These constraints or hypotheses are not mutually exclusive, and should not be taken to imply that one, but not other, agents of selection might apply to any one species. However, the sexually‐selected eggshell coloration hypothesis is least plausible for hole‐nesting birds because of the poor light quality available, although such species have been the focus of research in this area, and only a single experimental study has shown a link between egg coloration and male provisioning. Furthermore, the observed relationships between female phenotypic quality and egg traits do not necessarily imply that they have signalling functions. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 100 , 753–762.     

Eggshell spot scoring methods cannot be used as a reliable proxy to determine pigment quantity

Eggshell maculation of most passerines is due to the deposition of the pigment protoporphyrin which is produced during biosynthesis of blood haem. Its functional significance has only received empirical attention in recent years. This interest has generated a number of hypotheses of which some remain untested partly because the quantification of protoporphyrin is analytically challenging and can be prohibitively expensive. Many studies have therefore used the extent of eggshell spotting as a proxy for total eggshell protoporphyrin concentration, although this has not been formally tested. Pigment scoring involves recording visible eggshell pigment attributes, such as spot intensity, distribution and size. Since even immaculate eggs can contain some protoporphyrin, there remains doubt over the degree to which visible pigment correlates with total pigment content of the shell. In this study, we test whether visible pigment scoring can be used as a proxy for protoporphyrin concentration of an eggshell. We use pigmented eggshells of two common British passerine species to compare eggshell spot intensity, distribution and spot size (as used by the visual pigment scoring method) with direct measures of eggshell protoporphyrin concentration. In addition, we compared an alternative method of pigment scoring, the pixel pigment scoring method, using a computer programme to quantify the number of pixels exceeding a specified colour threshold. We demonstrate that although results from both scoring methods were positively correlated with eggshell protoporphyrin concentrations, the correlations were not sufficiently strong to be used as surrogates in studies where actual pigment concentrations are required.     

A standardized method for quantifying eggshell spot patterns

Spottiness is an important component of the morphology of bird eggs and a number of methods have been developed for characterizing variation in spottiness. We developed a quantitative method for measuring and comparing eggs to determine if female European Cranes (Grus grus) lay eggs with individually distinct color and spotting patterns. We used photographs taken under standard conditions and developed a computer program (ESPANA) to quantify egg‐spot patterns. The goal of the analysis was to create a “fingerprint” of each eggshell by measuring reflection along virtually drawn lines (transects) on an egg's image. Values measured in the same positions along transects can be compared among eggs by considering them as separate variables defining the pattern. Data were analyzed using cluster analyses and by performing analyses of similarity (ANOSIM). We found that the eggs of female European Cranes (N = 11) had individually distinct color patterns, with eggs laid by a given female more similar to each other than to the eggs of other females. Beyond its potential use for identifying the eggs of specific females, we believe our method could also be useful for investigators quantifying differences in egg‐spot patterns for other reasons, for example, examining possible relationships between egg‐spot patterns and female quality. The program ESPANA is implemented using the Java programming language and is available as supporting information on the Journal of Field Ornithology website.     

The quantification of local substrate heterogeneity in streams and its significance for macroinvertebrate assemblages

The effect of substrate heterogeneity on the structure of stream macroinvertebrate assemblages (total abundance, taxon richness, and evenness) is still not clear, but this could be due to the lack of standard methods for quantifying substrate heterogeneity. An accurate quantification of substrate heterogeneity was obtained from photographs of sampled areas (each 225 cm²), which were used to create maps that were subsequently digitized and analyzed using image analysis software. These maps allowed the calculation of multiple metrics quantifying two aspects of substrate heterogeneity: composition and spatial configuration of substrate patches. The diversity of substrate types (calculated as the Shannon diversity index), and the heterogeneity of patch compactness (calculated as the coefficient of variation of the relationship between patch dimensions) were the metrics explaining more biotic variance at the sample scale, but at higher scales there were no relationships between assemblage structure and substrate heterogeneity. Most variation in substrate heterogeneity occurred at the sample scale, while some metrics varied significantly at riffle or segment scales; these patterns of variation match those of macroinvertebrate assemblages, which had been previously studied. The importance of quantifying substrate heterogeneity and considering the spatial scales of its study are discussed.     

Review: an embryo's eye view of avian eggshell pigmentation

Current research into the evolution and adaptive function of avian eggshell pigmentation, including maculation, has focused mostly on signalling‐based and structural function hypotheses but ignored the potential consequences of shell pigmentation for the developing avian embryo, especially in moderating the embryo's interaction with its light environment. The exposure of the eggs to sunlight that frequently accompanies avian incubation behaviour is one of the major evolutionary steps setting apart birds and reptiles, and coincides with the appearance of eggshell pigmentation. This suggests that shell pigments could play a major role in ensuring the successful development of the avian embryo. We propose that the effects of shell pigments on the egg contents should be considered in addition to established hypotheses of shell pigmentation such as crypsis, egg recognition or a possible structural function. This approach has the potential to identify trade‐offs between different pigment functions and to resolve some of the long standing paradoxa in the evolution of eggshell colour, such as the occurrence of conspicuous blue eggs in passerines or the secondary evolution of white eggshells in cavity nesters. In particular, we identify seven hypotheses, which address how the interaction of eggshell pigments and the light environment may influence embryonic development. These hypotheses are the: thermo‐regulation; UV‐B protection; photo‐acceleration; lateralization; circadian rhythm; photo‐reactivation; and antimicrobial defence. We believe that the understanding of eggshell pigmentation will greatly benefit from taking these hypotheses into consideration when studying the functional significance of eggshell pigmentation and suggest a number of promising directions for future experimental and comparative research.     

Protoporphyrin‐based eggshell pigmentation predicts hatching success and offspring sex ratio in the barn swallow

Inter‐ and intraspecific variation in eggshell colouration has long fascinated evolutionary biologists. Among species, such variation may accomplish different functions, the most obvious of which is camouflage and background matching. Within species, it has been proposed that inter‐female variation in eggshell pigmentation patterns can reflect egg, maternal or paternal traits and hence may provide cues to conspecifics about egg, maternal or paternal phenotypic quality. However, the relationship between protoporphyrin‐based eggshell pigmentation and egg or maternal/paternal traits appears to be highly variable among species. We investigated patterns of intraspecific variation in Eurasian barn swallow Hirundo r. rustica protoporphyrin‐based eggshell pigmentation, and analysed its association with egg and clutch characteristics, maternal/paternal phenotypic traits and parental feeding effort. Eggshell pigmentation pattern significantly varied between breeding colonies, was significantly repeatable in first clutches laid by the same females in different years (intraclass correlation coefficient ranging between 0.56 and 0.63), but it was not significantly associated with egg traits, such as position in the laying sequence, egg mass, yolk testosterone concentration and antioxidant capacity. It was weakly or non‐significantly associated with female and male traits (sexual ornaments), but females laying darker (higher pigment intensity) first clutches had higher hatching success, suggesting that eggshell pigment intensity may predict fitness. Male nestling feeding effort was not predicted by eggshell pigmentation. In addition, females with darker breast plumage colouration (a melanin‐based trait related to fitness) laid highly protoporphyrin‐covered eggs, suggesting the presence of a previously unappreciated link between protoporphyrin biosynthesis and plumage melanisation. Moreover, the proportion of male offspring increased in clutches originating from highly protoporphyrin‐covered eggs, suggesting that parents could acquire visual cues about their future brood sex composition before egg hatching. Our results support the idea that intraspecific signalling via eggshell pigmentation is a species‐specific rather than a general feature of avian taxa.     

Multicomponent shell traits are consistent with an individual recognition function of the appearance of common murre (Uria aalge) eggs: A biological replication study

In dense breeding colonies, and despite having no nest structure, common murres (or guillemots: Uria aalge) are still able to identify their own eggs. Each female murre''s egg is thought to be recognized individually by the shell''s avian‐perceivable traits. This is because the eggshells’ visible traits conform to expectations of the identity‐signaling hypothesis in that they show both high intraindividual repeatability and high interindividual variability. Identity signaling also predicts a lack of correlation between each of the putative multicomponent recognition traits, yielding no significant relationships between those eggshell traits that are generated by mutually exclusive physiological factors. Using a multivariate analysis across eggshell size and shape, avian‐perceivable background coloration, spot (maculation) shape, and spot density, we detected no unexpected statistical correlations between Icelandic common murre egg traits lacking known physiological or mathematical relationships with one another. These results biologically replicate the conclusions of a recent eggshell trait study of Canadian common murres using similar methodology. We also demonstrate the use of static correlations to infer identity signaling function without direct behavioral observations, which in turn may also be applied to rare or extinct species and provide valuable insight into otherwise unknown communicative and behavioral functions.     

Eggshell pigmentation pattern in relation to breeding performance of blue tits Cyanistes caeruleus

1. We test the consequences, in terms of breeding success and parental effort, of eggshell pigmentation pattern in a hole-nesting bird, the blue tit Cyanistes caeruleus that lays eggs asymmetrically speckled with reddish spots (maculated eggs). 2. We assess the effect of distribution of spots (pigment 'spread') and spot size and pigment intensity (pigment 'darkness') on eggshell physical properties and breeding parameters concerning nestling condition, investment of parents in offspring care and reproductive output in two different habitat types: a deciduous oakwoodland and an evergreen forest. 3. Blue tit clutches with more widely distributed spots showed a thicker eggshell, a shorter incubation period, a lesser amount of mass loss per day and a higher hatching probability than those with spots forming a 'corona' ring. While eggs with larger and darker (more pigment intensity) spots showed a thicker eggshell and a shorter incubation period. In the light of 'signal function hypothesis', these egg traits may reflect female health status and, consequently, this could affect male parental effort. 4. Here we show supports for some of the necessary assumptions of this hypothesis. We found a positive relationship between egg pigment 'spread' and male but not female provisioning rates per day. On the other hand, pigment 'darkness' of blue tits' clutches was positively related to female tarsus length, while pigment 'spread' was positively related to clutch size, male body mass and nestling tarsus length. Our study shows that eggshell pigment 'spread' can be used as an indicator of clutch quality. Further investigations are needed to understand the role of calcium availability as possible causal agent of deviant eggs and its relation to the maculation phenomenon.     

Landscape-mediated variation in diet is associated with egg size and maculation in a generalist forager

Human impacts alter landscapes with consequences for the distribution and availability of high-quality food resources to populations inhabiting those landscapes, which may impact on the reproductive output of individuals in those populations. The sensitivity of wild populations to changes in food resources may vary among stages of the annual cycle. For example, in birds, effects are likely to be greater during costly stages such as egg production. Here we compare assimilated diet (from stable isotope analysis of chick feathers) and egg traits (egg size, shape, eggshell colour and maculation, using pattern-analysis software) in Herring Gulls Larus argentatus, across seven colonies in southwest Scotland and Northern Ireland. The Herring Gull is an opportunistic, generalist forager on both marine and terrestrial resources which frequently exploits anthropogenic food sources such as fishery discards and human refuse. We found that larger eggs were laid in colonies where females consumed either a higher proportion of marine resources or terrestrial resources; smaller eggs were laid in colonies where females had an intermediate diet. In colonies where females consumed more marine items, they also laid eggs with higher maculation (intensity and size of spots) compared with colonies where females mainly consumed terrestrial food. We also found smaller and more pointed eggs, suggestive of resource shortages, in larger colonies. Generalist foragers are often thought to have the capacity to buffer themselves against changes in the food web, provided that enough alternative food is available. However, this study highlights that specializing on the most profitable or available resources has consequences for egg traits even in an opportunistic generalist forager exploiting a large range of habitats. If variation in egg traits is related to reproductive output, then understanding the impact of assimilated diet on reproduction early in the breeding season can provide important insights into how populations will respond to landscapes altered by human impact.     

Variability in Avian Eggshell Colour: A Comparative Study of Museum Eggshells

Background

The exceptional diversity of coloration found in avian eggshells has long fascinated biologists and inspired a broad range of adaptive hypotheses to explain its evolution. Three main impediments to understanding the variability of eggshell appearance are: (1) the reliable quantification of the variation in eggshell colours; (2) its perception by birds themselves, and (3) its relation to avian phylogeny. Here we use an extensive museum collection to address these problems directly, and to test how diversity in eggshell coloration is distributed among different phylogenetic levels of the class Aves.

Methodology and Results

Spectrophotometric data on eggshell coloration were collected from a taxonomically representative sample of 251 bird species to determine the change in reflectance across different wavelengths and the taxonomic level where the variation resides. As many hypotheses for the evolution of eggshell coloration assume that egg colours provide a communication signal for an avian receiver, we also modelled reflectance spectra of shell coloration for the avian visual system. We found that a majority of species have eggs with similar background colour (long wavelengths) but that striking differences are just as likely to occur between congeners as between members of different families. The region of greatest variability in eggshell colour among closely related species coincided with the medium-wavelength sensitive region around 500 nm.

Conclusions

The majority of bird species share similar background eggshell colours, while the greatest variability among species aligns with differences along a red-brown to blue axis that most likely corresponds with variation in the presence and concentration of two tetrapyrrole pigments responsible for eggshell coloration. Additionally, our results confirm previous findings of temporal changes in museum collections, and this will be of particular concern for studies testing intraspecific hypotheses relating temporal patterns to adaptation of eggshell colour. We suggest that future studies investigating the phylogenetic association between the composition and concentration of eggshell pigments, and between the evolutionary drivers and functional impacts of eggshell colour variability will be most rewarding.     

Why are birds’ eggs speckled?

Birds are unique in laying eggs with pigmented shells, but for most species (e.g. most passerines, which lay white eggs speckled with reddish spots of protoporphyrin) the pigments’ function is unknown. We studied a bird population at a geologically variable site, and considered a hitherto untested hypothesis: that protoporphyrin pigments might compensate for reduced eggshell‐thickness (caused partly by calcium deficiency), which is known to reduce eggshell‐strength and increase eggshell‐permeability. We found that pigment spots specifically demarcated thinner areas of shell, with darker spots marking yet thinner shell than paler spots. Variation in pigmentation was thus associated with variation in shell thickness both within and between clutches, so accounting for the eggshell's characteristic spot patterns. Geological variability at this site has resulted in a great range of calcium availability and, as predicted by the hypothesis, variation in calcium availability was found to affect between‐clutch variation in both eggshell‐mass (+) and pigmentation characteristics (?). We suggest a physiological mechanism and some important implications of these findings.     

A comparison of indices and measured values of eggshell thickness of different shell regions using museum eggs of 230 European bird species

The thickness of avian eggshells is used to assess shell quality in wild and domestic species, as an indicator of environmental pollution and as an adaptive explanation for shell maculation. Both direct measurements and calculated eggshell thickness indices (ETI) are used in such research, yet this is the first study to quantify, across a large spectrum of bird families (and thus egg shapes), the correlation between measured thicknesses and ETI. Furthermore, few studies have quantified thickness variation across the entire length of the shell, although this variation may influence both gas transfer and embryonic development. We measured the thickness of 942 eggshells of 230 European bird species from the Class II material at the Natural History Museum, Tring, UK, both in the conventional manner, at the equator through the blowhole and, uniquely, after a single longitudinal, cut at its equator at the blunt and pointed ends. Over half of the samples revealed shell defects, cautioning against the indiscriminate use of museum specimens. Strong positive associations were found between species‐specific means of shell thickness with each other and also with ETI, especially those derived from Schönwetter's ‘Handbuch der Oologie’ method, validating the interspecific comparative use of ETI. Thickness measurements and ETI factors are provided for all 230 species. Eggshells were usually thinner at the blunt end (the location of the air sac) than at the equator, but of equal thickness in passerine eggs. This difference was greatest in species producing elongate eggs and suggests that there is a functional significance of shell thickness variation among species that requires further investigation.     

Test of an adaptive hypothesis for egg speckling along an elevational gradient in a population of Mexican jays Aphelocoma ultramarina

The adaptive significance of avian egg speckling patterns has been a subject of ongoing debate. We examined speckling in a population of Mexican jays Aphelocoma ultramarina exhibiting extreme eggshell variability. We sampled 167 eggs at 55 nests from sites ranging across a steep elevation gradient within the Sierra del Carmen mountain range in Coahuila, Mexico, in order to test the recent hypothesis that egg speckling lends structural support to eggs and should therefore be more prevalent in females subject to reduced environmental calcium. Although we documented high variation in the amount and distribution of eggshell speckling within the Sierra del Carmen jays, we found no relationship between local soil calcium levels and the pattern of speckling. Our results indicate that explanations in addition to soil calcium levels are necessary to explain extreme variation in eggshell speckling in birds.     

Eggshell biliverdin concentration does not sufficiently predict eggshell coloration

Avian eggshell coloration may have arisen due to selection on the biological, chemical, or physical properties of the pigments embedded within the eggshell, or due to selection on the coloration that emerges due to pigment deposition. Within both hypothetical frameworks, pigment‐based eggshell coloration would be related to metrics of egg quality; however, no one has evaluated the relative strength of coloration and pigment concentration in predicting egg quality. Here, we examined 66 European starling Sturnus vulgaris eggs and quantified eggshell biliverdin concentration (an antioxidant that produces the eggshell's blue coloration) and used 28 different coloration metrics derived from both photographic and spectrophotometric data. We also measured egg size, eggshell thickness, concentration of carotenoids in the yolk, and concentration of lysozyme in the albumen to capture variation in egg quality. We found that throughout the laying sequence, biliverdin concentration increased while eggshell thickness, yolk carotenoid concentration, and lysozyme concentration in the albumen all decreased, but this variation was not captured by any eggshell coloration metric. Both eggshell coloration and biliverdin concentration were negatively associated with yolk carotenoid concentration, but only eggshell biliverdin concentration was negatively associated with yolk mass. Lastly, biliverdin concentration explained, at most, only 46% of the variation for all eggshell coloration metrics. Our results suggest biliverdin concentration is a better predictor of egg quality than egg coloration in European starlings, supporting the hypothesis that eggshell pigment concentration per se may be the target of selection.     

Quantitative description and discrimination of butterfly wing patterns using moment invariant analysis

Studies examining and using pattern variation in insects for identification and characterization of individuals and populations have been limited by the methods available for quantifying wing patterns objectively. In this paper, differences in wing pattern are demonstrated statistically using moment invariant data sets generated automatically from digitized images of the speckled wood butterfly, Pararge aegeria (Linnaeus). Studies with other biological subjects have already shown moment invariants to work well with outline shapes and silhouettes. A pilot study with replicated monochrome photographs of a single butterfly showed the method could detect pattern differences between wing surfaces, even in the presence of simulated wing fading and damage. In a further study of the wings of 228 specimens, multivariate analyses of variance using the moment data reliably detected differences between groups of butterflies according to sex, geographical origin and culture history. Potential applications and future improvements of the moment methodology are considered.     

Variation in eggshell traits between geographically distant populations of pied flycatchers Ficedula hypoleuca

The expression and impact of maternal effects may vary greatly between populations and environments. However, little is known about large‐scale geographical patterns of variation in maternal deposition to eggs. In birds, as in other oviparous animals, the outermost maternal component of an egg is the shell, which protects the embryo, provides essential mineral resources and allows its interaction with the environment in the form of gas exchange. In this study, we explored variation of eggshell traits (mass, thickness, pore density and pigmentation) across 15 pied flycatcher populations at a large geographic scale. We found significant between‐population variation in all eggshell traits, except in pore density, suggesting spatial variation in their adaptive benefits or in the females’ physiological limitations during egg laying. Between‐ population variation in shell structure was not due to geographic location (latitude and longitude) or habitat type. However, eggshells were thicker in populations that experienced higher ambient temperature during egg laying. This could be a result of maternal resource allocation to the shell being constrained under low temperatures or of an adaptation to reduce egg water loss under high temperatures. We also found that eggshell colour intensity was positively associated with biliverdin pigment concentration, shell thickness and pore density. To conclude, our findings reveal large‐ scale between‐population variation of eggshell traits, although we found little environmental dependency in their expression. Our findings call for further studies that explore other environmental factors (e.g. calcium availability and pollution levels) and social factors like sexual selection intensity that may account for differences in shell structure between populations.