Eggshell Maculation Research Articles

Eggshell Maculation is Not a Signal of Maternal Condition in Two Species of Rail

Eggshell Maculation is Not a Signal of Maternal Condition in Two Species of Rail

Correlated evolution of eggshell maculation with social breeding and nest type in Hirundinidae

Maculation on avian eggshells has the potential to serve as an identity signal, and this information may help females recognize their eggs/nest or reject foreign eggs laid by hetero‐ or conspecific brood parasites. Recognizing eggs could be adaptive in cases where birds nest in dense colonies, as reports of conspecific brood parasitism are over‐represented in colony‐nesting species. We utilized the variation in breeding biology (solitary vs. colonial breeding) and eggshell phenotype in swallows and martins (Hirundinidae) to test for correlated evolution between these traits, while also accounting for nest type, as maculation may camouflage eggs in open‐cup nests. We found that maculated eggs were more likely to be laid by species that breed socially and build open‐cup nests where maculation would be more visible than in dark cavity nests.

Ecological drivers of eggshell wettability in birds.

Complex and at times extreme environments have pushed many bird species to develop unique eggshell surface properties to protect the embryo from external threats. Because microbes are usually transmitted into eggs by moisture, some species have evolved hydrophobic shell surfaces that resist water absorption, while also regulating heat loss and the exchange of gases. Here, we investigate the relationship between the wettability of eggshells from 441 bird species and their life-history traits. We measured the initial contact angle between sessile water droplets and the shell surface, and how far the droplet spread. Using phylogenetic comparative methods, we show that body mass, annual temperature and eggshell maculation primarily explained variance in water contact angle across eggshells. Species nesting in warm climates were more likely to exhibit highly hydrophobic eggshells than those nesting in cold climates, potentially to reduce microbial colonization. In non-passerines, immaculate eggs were found to have more hydrophobic surfaces than maculate eggshells. Droplets spread more quickly on eggshells incubated in open nests compared to domed nests, likely to decrease heat transfer from the egg. Here, we identify clear adaptations of eggshell wettability across a diverse range of nesting environments, driven by the need to retain heat and prevent microbial adhesion.

The blunt pole is not a source of more salient recognition cues than the sharp pole for the rejection of model eggs by American robins (Turdus migratorius)

Hosts of obligate avian brood parasites can reduce the costs of raising parasitic offspring by rejecting foreign eggs from their nests. Rejecter hosts use various visual and tactile cues to discriminate between own and foreign eggs. The blunt pole hypothesis specifically states that avian-perceivable visual information at and around the broader pole of the eggshell contains more salient recognition cues than does the sharp pole of the same egg. The directional prediction is, therefore, that eggs painted non-mimetically on their blunt pole should more likely be rejected relative to those similarly painted on their sharp pole. This hypothesis had been experimentally tested and its predictions supported solely in mimetic avian host-parasite systems, with hosts producing denser and more variable eggshell maculation patterns at the blunt pole, and in one species with immaculate eggs but still with distinctly discernible blunt-pole specific colouration. Here we aimed to expand upon these previous works and assessed whether the blunt pole of model eggs contains more salient egg rejection cues, relative to the sharp pole, for the American robin (Turdus migratorius), a robust rejecter of non-mimetic brown-headed cowbird (Molothrus ater) eggs. In this system host eggs are uniformly immaculate whereas the brood parasitic shell is maculated. We painted model cowbird-sized eggs on either the blunt or the sharp half to mimic the immaculate robin egg colours and the other half to resemble non-mimetic egg colours and patterns. There was no statistical support for the predicted outcomes of the blunt pole hypothesis in our trials as rejection rates were similar regardless of whether eggs were painted with non-mimetic colours on the blunt or sharp poles. Future work should test the role of asymmetrical signalling content for anti-parasitic rejection of eggs in additional host species, especially those with both immaculate own and mimetic parasitic eggs.

Variations in tree sparrow (Passer montanus) egg characteristics under environmental metal pollution

Environmental metal pollution is known to adversely affect bird reproduction, for which the variations of egg characteristics are considered very important. Our study explored whether variations in egg characteristics, such as egg volume, eggshell spotting pattern, eggshell coloration, and eggshell thickness, were correlated with heavy metal levels (Cu, Zn, Pb, and Cd) and Ca levels in the eggshells of tree sparrows (Passer montanus), a widespread passerine species. Eggs were collected from a long-term heavy metal polluted area (Baiyin, BY, northwest China) and a relatively unpolluted area (Liujiaxia, LJX, northwest China). Our results showed that the embryonated (eggshell: Cu: p = 0.003, Pb: p = 0.002) and non-embryonated (egg contents: Pb: p = 0.044, Ca: p = 0.045) eggs collected from BY contained relatively higher metal concentrations than those from LJX. Eggs from BY were smaller in volume (p < 0.01) and thinner in eggshell thickness (p < 0.01) than those from LJX. Mean egg volume increased with clutch size in BY (p = 0.017), which was also accompanied by an increase in the within-clutch coefficient of variation (CV) for egg volume (p = 0.045). Clutches with a higher CV for egg volume tended to contain higher concentrations of Zn and Pb (Zn: p = 0.084; Pb: p = 0.081) in the eggshells from BY. No differences were found in the eggshell spotting coverage ratio of eggs; however, eggshells were much darker in BY than in LJX. A more aggregated eggshell spotting distribution indicated higher eggshell Zn and Pb levels (BY: Zn: p = 0.040, Pb: p = 0.076; LJX: Pb: p = 0.066). The results demonstrate that the egg characteristics of tree sparrows can be used as indicators of metal pollution, especially for the within-clutch CV for egg volume, eggshell spotting pattern and eggshell coloration.

Quantifying the maculation of avian eggs using eggshell geometry

Birds’ eggs often exhibit considerable variation in the presence and pattern of superficial pigmentation, or maculation. However, despite long appreciation of the nature of this variation both between and within species, there is still poor understanding of why such a diversity of patterns has evolved. Accurately quantifying eggshell maculation is therefore key to elucidating its functional significance. Maculation is typically quantified from photographs, and so measurements may be biased by the perceived distortion introduced by the curvature of the eggshell. Here, I consider egg geometry in order to derive measures of curved distance and surface area that can be used to characterize eggshell maculation patterns more accurately, and demonstrate the utility of this approach for quantifying and visualizing eggshell maculation from standard photographs.

Eggshell spotting pattern is related to hatching asynchrony, hematocrit value and growth of nestling great titsParus major

Eggshell pigmentation may signal the quality of the egg, that of the female and the environment, and thus nestling development may be related to this egg trait. However, so far, few studies have investigated the relationship between eggshell pigmentation and nestling development. Our aim was to study in a partial cross‐fostering experiment whether the protoporphyrin‐based eggshell pigmentation (spot intensity and spot distribution) showed any significant associations with several traits related to the development of nestling great titsParus major. We found that nestlings from clutches with darker spotted eggshells hatched more synchronously, had higher hematocrit value and shorter wings. Females under adverse environmental circumstances were previously shown to lay darker‐spotted eggs, and these females probably delayed the start of incubation due to energetic constraints, resulting in a more synchronous hatching. The shorter wings of fledglings originating from clutches with darker‐spotted eggs may be caused by the lower general quality of their mothers, mediated through either genetic factors or early maternal effects. Darker spotted eggs may contain different levels of some biomolecules than paler spotted eggs, which could have an effect on nestling metabolism and activity, resulting in an elevated hematocrit value. More aggregated eggshell spotting was related to faster bone growth and slightly longer fledging tarsus length. Eggs with aggregated spotting may have thinner eggshells that could lead to hatchlings with smaller bones due to the lower availability of calcium during development. Nestlings may increase bone growth rate to compensate for their smaller initial size. Overall, our results indirectly suggest lower fitness for nestlings hatched from darker spotted eggs and for nestlings hatched from eggs with more aggregated spotting distribution due to the possible long‐term costs of synchronous hatching, shorter wings, and accelerated tarsus growth rate.

Effects of environmental conditions on the egg mass, yolk antioxidant level, eggshell thickness and eggshell spotting patterns of Great Tits (Parus major)

Avian mothers can influence the fitness of their offspring by resource investment into the egg. Allocation of macro- and micronutrients into the eggs may be costly for the female, therefore, we expect that resource investment may be affected by the environmental and social conditions the mother experiences during egg formation. Here, we investigated whether environmental circumstances experienced by the reproducing female exert an influence on egg mass, yolk antioxidant (lutein and tocopherol) concentration, eggshell thickness, and the eggshell spotting patterns of Great Tits (Parus major). Our study showed that when caterpillars were less abundant, in colder weather and when breeding in an area of higher local breeding density, female Great Tits laid eggs of lower mass, suggesting that adverse environmental circumstances constrain the macronutrient investment into the eggs. However, we found no evidence that yolk antioxidant concentration and eggshell thickness were affected by environmental factors. Female Great Tits may use their endogenous stores or spend more time and energy in finding sufficient amounts of dietary antioxidants and calcium under unfavourable environmental conditions, which may have a cost effect on their own conditions. We found that birds that bred in the beginning of the season laid eggs with darker eggshell spots. Moreover, females laid more spotted eggs in colder weather and when breeding in higher density areas. Our results suggest that Great Tits deposit more of the potentially harmful pro-oxidant protoporphyrin pigment into the eggshell under unfavourable environmental conditions.

Darker eggshell spotting indicates lower yolk antioxidant level and poorer female quality in the Eurasian Great Tit (Parus major)

ABSTRACT Protoporphyrin pigment causes the red-brown eggshell colors; however, for many species, the function of this pigment is unknown. It has been proposed that eggshell pigmentation may strengthen the shell or that it may be a sexually selected signal, which advertises the quality of the female and that of her offspring to the male parent. In this study, we aimed to discover whether protoporphyrin-based eggshell pigmentation patterns of Eurasian Great Tits (Parus major) were related to female or egg quality. Additionally, we tested whether different methods of eggshell pigmentation estimation could be reliable predictors of eggshell protoporphyrin levels. We found that spot intensity, spot size, spotting coverage, and brown spot chroma indicated the protoporphyrin pigment concentration of the eggshell. Our results revealed that Eurasian Great Tit females that laid eggs with darker pigmentation had more lymphocytes in their circulation and had paler yellow breast and lower UV plumage reflectance, possi...

Effects of breeding habitat (woodland versus urban) and metal pollution on the egg characteristics of great tits (Parus major)

In an urban environment, birds are exposed to metals, which may accumulate in their tissues and cause oxidative stress. Female birds may eliminate these pollutants through depositing them into eggs, thus eggs become suitable bioindicators of pollution. In this study, we aimed to analyse whether eggshell spotting pattern, egg volume, eggshell thickness and egg yolk antioxidant (lutein, tocopherol, retinol and selenium) levels were related to the breeding area (woodland versus urban) and the metal levels in the eggshell of a small passerine species, the great tit (Parus major). In the urban habitat, soil and eggshells contained higher concentrations of metals, and soil calcium level was also higher than that in the woodland. Eggshell spotting intensity and egg volume did not differ between eggs laid in the woodland and the urban park, and these traits were not related to the metal levels of the eggshell, suggesting that these egg characteristics are not sensitive indicators of metal pollution. A more aggregated eggshell spotting distribution indicated a higher Cu concentration of the eggshell. We found that eggshells were thinner in the less polluted woodland habitat, which is likely due to the limited Ca availability of the woodland area. Great tit eggs laid in the urban environment had lower yolk lutein, retinol and selenium concentrations, however, as a possible compensation for these lower antioxidant levels, urban females deposited more tocopherol into the egg yolk. It appears that females from different breeding habitats may provide similar antioxidant protection for their offspring against oxidative damage by depositing different specific dietary antioxidants. Egg yolk lutein and retinol levels showed a negative relationship with lead concentration of the eggshell, which may suggest that lead had a negative impact on the amount of antioxidants available for embryos during development in great tits.

Modelling Eggshell Maculation

The eggshells of many avian species are characterised by distinctive patterns of maculation, consisting of speckles, spots, blotches or streaks, the spatial-statistical properties of which vary considerably between (and often within) species. Understanding the mechanisms underlying the production of eggshell maculation would enable us to explore the costs and constraints on the evolution of maculation patterns, but as yet this area is surprisingly understudied. Here I present a simple model of eggshell maculation, which is based on the known biology of pigment deposition, and which can produce a range of realistic maculation patterns. In particular, it provides an explanation for previous observations of maculation heterogeneity and diversity, and allows testable predictions to be made regarding maculation patterns, including a possible signalling role.

Maternal influence on eggshell maculation: implications for cryptic camouflaged eggs

Egg camouflage may explain the adaptive significance of avian eggshell pigmentation in ground-nesting species. Eggshell maculation (spots) is predominantly due to protoporphyrin, but both biliverdin (antioxidant) and protoporphyrin (pro-oxidant) may be present in spotted eggshells. Because of their role in oxidative stress, the deposition of eggshell pigments might be condition-dependent. However, because of the fitness benefits of eggshell coloration, cryptic eggshell appearance should be strongly conserved in ground-nesting species regardless of female condition and eggshell pigment concentrations. We investigated whether Japanese quails (Coturnix coturnix japonica) maintained eggshell maculation under food restriction. We quantified eggshell maculation (i.e., percentage of spot coverage) using digital photography, and both protoporphyrin and biliverdin concentrations of eggs laid by females either on a food-restricted or an ad libitum diet. Females on a high quality diet, which are known to decrease the deposition of eggshell protoporphyrin, decreased eggshell maculation compared with food-restricted females that maintained it. For the first time, we propose an experimental study which suggests that eggshell maculation depends on female body condition and that manipulating eggshell maculation may be the strategy used by females to potentially optimize egg camouflage.

Female oxidative status, egg antioxidant protection and eggshell pigmentation: a supplemental feeding experiment in great tits

Oxidative stress has been suggested as a mechanism underlying the costs of reproduction and life history trade-offs. Reproductive activities may lead to high production of pro-oxidants, whose activity can generate oxidative damage when not countered by adequate antioxidant defenses. Because inter-individual differences in the efficiency of the antioxidant system are influenced by an individual’s diet, food availability experienced during reproduction may affect the females’ antioxidant status and, in birds, their ability to transfer antioxidants into their eggs. Moreover, a female’s ability to cope with oxidative stress has been suggested to influence pigment deposition in the eggshell, suggesting a possible signaling function of eggshell maculation. Here we performed a food supplementation experiment in a natural population of great tits (Parus major) in order to investigate how nutritional conditions experienced during the egg laying period affect the female’s oxidative status and egg investment and how maternal oxidative status and egg antioxidant protection relate to eggshell pigmentation. We show that food-supplemented females had lower oxidative damage levels (ROMs) than non-food-supplemented females. Furthermore, a female’s ROMs levels were negatively associated with the levels of yolk antioxidant protection in her eggs, but this negative association was only significant in non-food-supplemented females. This suggests that oxidative stress experienced during reproduction influences the allocation of antioxidants into the eggs. Moreover, we observed a positive relationship between eggshell pigment distribution and maternal and yolk antioxidant protection, suggesting that eggshell pigmentation is a cue of female (and offspring) quality.

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.

Effects of experimental calcium availability, egg parameters and laying order on Great TitParus majoreggshell 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.

Asymmetrical signal content of egg shape as predictor of egg rejection by great reed warblers, hosts of the common cuckoo

The size, patterning and coloration of bird eggs may signal different information content to nest owners, mates, predators, hosts, or brood parasites. Recent studies suggested that the pigmentation at one pole of the typically asymmetrical avian egg plays a critical role in the discrimination of own and foreign eggs by several host species parasitized by the common cuckoo (Cuculus canorus). Typically, both eggshell maculation and background colour are more consistent on the blunt pole, and hosts react more strongly to experimental changes in coloration of the blunt pole compared to the sharp pole. However, it remains unclear whether the asymmetrical shape of natural eggs per se enhances the behavioural responses of hosts to foreign eggs. To evaluate the salience of asymmetrical egg shape, we studied reactions of a rejecter cuckoo host, the great reed warbler (Acrocephalus arundinaceus), to artificial shapes of model eggs painted a non-mimetic blue colour. Artificial eggs with two blunt poles were rejected significantly more often than those with a single blunt pole or two sharp poles. These results corroborate the hypothesis that the different egg poles have different signal salience and may have implications for the evolution of diversity of not only egg coloration but also of egg shape in the arms race between hosts and brood parasitic birds.

Eggshell spotting does not predict male incubation but marks thinner areas of a shorebird's shells

Many birds lay eggs speckled with black or reddish-brown spots of protoporphyrin pigment, but the function of these spots is debated. Two recent hypotheses have received considerable attention. Under the “signaling-function hypothesis,” speckling reflects female quality and influences allocation of male parental care; under the “structural-function hypothesis,” the speckles strengthen the eggshell when calcium is scarce. The evidence for each is taxonomically uneven. For instance, evidence for the signaling-function hypothesis comes from cavity-nesting and uniparentally incubating species in which the viewing conditions and possibilities for males to assess the speckling are limited, and male response to speckling is indirect (i.e., female-feeding) or postponed (i.e., chickfeeding). We tested the predictions of both hypotheses in a biparentally incubating ground-nesting shorebird, the Northern Lapwing (Vanellus vanellus), in which the potential for a direct male response to eggshell speckling exists and visual inspection of the eggs is not hindered by light availability in a nest cavity. The speckling parameters (spot intensity, distribution, and cover) were repeatable within Northern Lapwing clutches, a requirement for a sexually selected signal, but we found no relationship between male incubation and speckling. However, the spots were associated with thinner areas of eggshell, which strongly supports the suggestion of a structural (strengthening) function. Our results do not support the signaling-function hypothesis of eggshell speckling, but extend the structural-function hypothesis and prior findings (that speckling occurs primarily at thinner areas of the shell) to other avian taxa. Furthermore, if the generally accepted view that Northern Lapwing eggs are cryptically marked is correct, our findings suggest that more than one significant function can drive the evolution of avian eggshell pigmentation.

Testing the structural–function hypothesis of eggshell maculation in the Great Tit: an experimental approach

The function of eggshell pigmentation has long been the subject of much speculation. Most of the proposed hypotheses have been based on a signalling function. According to the recently proposed structural–function hypothesis, birds use the pigment protoporphyrin to strengthen the eggshell in response to dietary calcium (Ca) shortage. The aim of the study reported here was to test this hypothesis in a Ca-provisioning experiment involving Great Tit Parus major populations breeding in Ca-poor coniferous and Ca-rich deciduous Estonian forests, respectively, over a 2-year period. Ca-supplemented birds laid eggs with ‘darker’ pigmentation than birds in the unsupplemented (control) group, which is at odds with the structural–function hypothesis. Although eggs were smaller and had thinner shells in Ca-poor habitat, eggshell maculation did not differ between the habitats. Eggshells were more pigmented in the year when they were also thinner, but their thickness was not related to their maculation within each year. Although the results of our study neither prove nor disprove the structural–function hypothesis, they do suggest that the relationship between eggshell pigmentation and Ca availability cannot be explained simply by this hypothesis.

Experimental evidence for the role of calcium in eggshell pigmentation pattern and breeding performance in Blue Tits Cyanistes caeruleus

Previous studies have found strong relationships between calcium availability and eggshell pigmentation in the Great Tit (Parus major). According to the “structural function hypothesis”, protoporphyrins, the pigments responsible for reddish spots on speckled eggs, are deposited in those areas of the shell where calcium deposition is less intense. In the study reported here, which was carried out in three Blue Tit (Cyanistes caeruleus) populations in central Spain, we provide partial experimental support for the association between protoporphyrin eggshell pigmentation and shell thinning. Contrary to our expectations, we did not observe a decrease in the size and intensity of pigment spots for those eggs from calcium-supplemented nests. However, we did find that the provision of calcium-rich material during the egg-laying period led to a more wide distribution of pigment spots and reduced the proportion of eggs with defective shells (deviant pigmentation, dull and rough surface). When only the supplemented nests were considered in the analysis, within the same clutch we also detected differences in the spotting distribution between those eggs laid by female Blue Tits observed to have consumed calcium consumption on the day prior to laying and those observed not to have consumed calcium on the day prior to laying. Clutch size was not affected by the calcium supplementation. Female Blue Tits experimentally supplied with calcium-rich material had a shorter incubation period than control females, and they laid eggs with thicker shells. Eggshell thickness markedly affects the probability of hatching and could explain the lower proportion of unhatched late eggs found in supplemented nests in comparison with control ones. This study highlights the role of calcium in eggshell maculation and its effects on breeding performance of small passerines. We found the spotting distribution to be a good predictor of calcium deficiency. However, our results provide only mixed support for the “structural function hypothesis”: in our study populations, the thickness of the eggshell was intimately associated with calcium availability, but the relationship between calcium and protoporphyrin deposition remains far from clear.

Eggshell spotting in brood parasitic shiny cowbirds (Molothrus bonariensis) is not linked to the female sex chromosome

For avian brood parasites in which individual females are host-specialists, the arms race between hosts and parasites has favored egg color polymorphism in the parasite, with female lineages laying mimetic eggs that resemble those of the host species they parasitize. Female sex-linked inheritance of egg color fosters evolutionary stability of egg polymorphism if female lineages show both consistent eggshell color and host use. This co-evolutionary relationship is unlikely to occur if individual brood parasites use different hosts or if egg color is not maternally inherited. The shiny cowbird (Molothrus bonariensis) is an extreme generalist brood parasite that shows a very high degree of egg polymorphism. We tested whether egg spotting in this species has female sex-linked inheritance. If genetic factors controlling the expression of egg spotting were present on the female-specific W chromosome, we expected co-segregation between spotting patterns and mtDNA haplotypes, as both W and mtDNA are maternally inherited. In contrast to the known maternal inheritance of spotting patterns in great tits, we found no associations between eggshell spotting and mtDNA haplotypes, which suggests that eggshell spotting is not maternally inherited in this cowbird species.