Liliana D'Alba

Iridescent colors in feathers are some of the brightest in nature, and are produced by coherent light scattering from periodic arrangements of melanosomes (melanin-containing organelles). Hollow melanosomes, an evolutionary innovation largely restricted to birds, contain an optically powerful combination of high and low refractive indices (from the melanin and air, respectively) that enables production of brighter and more saturated colors than solid melanosomes. However, despite their significance to avian color and potential utility as optical biomaterials, little is known about the ontogeny of either the melanosomes themselves or the nanostructures they comprise. We used light and electron microscopy to characterize nanostructural development in regenerating feathers of wild turkeys, a species with iridescent color produced by a hexagonally close-packed array of hollow melanosomes. We found that melanosomes form as solid bodies in melanocytes. Later in development, largely after placement in developing barbules, their interiors dissolve and leave hollow cores. These now hollow melanosomes are initially disorganized in the barbule, but become close-packed as they are pulled to the edge of the barbule, likely through a combination of forces including depletion-attraction. These data suggest that these structurally colored tissues are self-assembled and represent novel pathways of development. J. Morphol. 276:378-384, 2015. © 2014 Wiley Periodicals, Inc.

One of the greatest threats to the survival of avian eggs is the risk of infection by microbes; as such, a large number of parental defense mechanisms have evolved in response to the decreased fitness imposed by microbial infection. The existing literature on this topic has focused largely on the mechanisms of microbial invasion through eggshells and the identification of molecules with antimicrobial properties in eggs of commercial species. However, little is still known about antimicrobial mechanisms in wild birds or how they vary with environmental pressures. This review concentrates on recent findings that shed new light on the role of parental behaviors (including incubation and placement of vegetation with antifungal activity in the nest) and the physical properties of eggshells (including nanometer-scale spheres that prevent microbial attachment) that protect eggs from contamination in high-risk environments. In addition to presenting a summary of current information, we identify evident gaps in knowledge and highlight research avenues for the future.

Understanding the mechanistic bases of natural color diversity can provide insight into its evolution and inspiration for biomimetic optical structures. Metazoans can be colored by absorption of light from pigments or by scattering of light from biophotonic nanostructures, and these mechanisms have largely been treated as distinct. However, the interactions between them have rarely been examined. Captive breeding of budgerigars (Aves, Psittacidae, Melopsittacus undulatus) has produced a wide variety of color morphs spanning the majority of the spectrum visible to birds, including the ultraviolet, and thus they have been used as examples of hypothesized structure–pigment interactions. However, empirical data testing these interactions in this excellent model system are lacking. Here we used ultraviolet–visible spectrometry, light and electron microscopy, pigment extraction experiments and optical modeling to examine the physical bases of color production in seven budgerigar morphs, including grey and chromatic (purple to yellow) colors. Feathers from all morphs contained quasi-ordered air–keratin ʻspongy layerʼ matrices, but these were highly reduced and irregular in grey and yellow feathers. Similarly, all feathers but yellow and grey had a layer of melanin-containing melanosomes basal to the spongy layer. The presence of melanosomes likely increases color saturation produced by spongy layers whereas their absence may allow increased expression of yellow colors. Finally, extraction of yellow pigments caused some degree of color change in all feathers except purple and grey, suggesting that their presence and contribution to color production is more widespread than previously thought. These data illustrate how interactions between
structures and pigments can increase the range of colors attainable in birds and potentially in synthetic systems.

Relative to other metazoans, the mammalian integument is thought to be limited in colour. In particular, while iridescence is widespread among birds and arthropods, it has only rarely been reported in mammals. Here, we examine the colour, morphology and optical mechanisms in hairs from four species of golden mole (Mam- malia: Chrysochloridae) that are characterized by sheens ranging from purple to green. Micro- spectrophotometry reveals that this colour is weak and variable. Iridescent hairs are flattened and have highly reduced cuticular scales, pro- viding a broad and smooth surface for light reflection. These scales form multiple layers of light and dark materials of consistent thickness, strikingly similar to those in the elytra of irides- cent beetles. Optical modelling suggests that the multi-layers produce colour through thin-film interference, and that the sensitivity of this mechanism to slight changes in layer thickness and number explains colour variability. While coloured integumentary structures are typically thought to evolve as sexual ornaments, the blind- ness of golden moles suggests that the colour may be an epiphenomenon resulting from evolution via other selective factors, including the ability to move and keep clean in dirt and sand.

Iridescent feather colors involved in displays of many extant birds are produced by nanoscale arrays of melanin-containing organelles (melanosomes). Data relevant to the evolution of these colors and the properties of melanosomes involved in their generation have been limited. A data set sampling variables of extant avian melanosomes reveals that those forming most iridescent arrays are distinctly narrow. Quantitative comparison of these data with melanosome imprints densely sampled from a previously unknown specimen of the Early Cretaceous feathered Microraptor predicts that its plumage was predominantly iridescent. The capacity for simple iridescent arrays is thus minimally inferred in paravian dinosaurs. This finding and estimation
of Microraptor feathering consistent with an ornamental function for the tail suggest a centrality for signaling in early evolution of plumage and feather color.

Colors of living organisms are produced by selective light absorption from pigments and/or by light scattering from highly ordered nanostructures (i.e., structural color). While the physical bases of metallic colors of arthropods and fish are fairly well-known, those of birds are not. Here we examine structurally based silver color and its production in feathers of the waterbird species Anhinga. This achromatic color is distinguished from grey by high specular reflectance, from white by low diffuse reflectance, and from both by high gloss. Light and electron microscopy revealed three modifications of feathers likely leading to silver color. First, proximal barbules were highly elongated and contained glossy black color at their base and white color at their pennulum. Second, this glossy black portion contained a single outer layer of keratin weakly bounded by melanosomes. Finally, the white portion contained a disordered amorphous matrix of keratin and air. Optical analyzes suggest that these structures produce, respectively, glossy black color through thin-film interference and white color through incoherent light scattering. Silver color likely results from the combined reflectance of these adjacent structures. This represents a distinct mechanism for attaining silver colors that may have been partially derived through selection for display, thermoregulation or decreased hydrophobicity.

Stressful conditions early in life can give rise to exaggerated stress responses, which, while beneficial in the short term, chronically increase lifetime exposure to stress hormones and elevate disease risk later in life. Using zebra finches Taeniopygia guttata, we show here that individuals whose glucocorticoid stress hormones were experimentally increased for only a brief period in early post-natal life, inducing increased stress sensitivity, had reduced adult lifespans. Remarkably, the breeding partners of such exposed individuals also died at a younger age. This negative effect on partner longevity was the same for both sexes; it occurred irrespective of the partner's own early stress exposure and was in addition to any longevity reduction arising from this. Furthermore, this partner effect continued even after the breeding partnership was terminated. Only 5 per cent of control birds with control partners had died after 3 years, compared with over 40 per cent in early stress–early stress pairs. In contrast, reproductive capability appeared unaffected by the early stress treatment, even when breeding in stressful environmental circumstances. Our results clearly show that increased exposure to glucocorticoids early in life can markedly reduce adult life expectancy, and that pairing with such exposed partners carries an additional and substantial lifespan penalty.

Nest shelter in incubating birds is of major importance in providing protection against unfavourable conditions such as harshness of the environment and exposure to predators. We examined the link between nest shelter, baseline corticosterone (CORT) levels and hatching success in common eiders (Somateria mollissima) incubating at nest sites with different levels of shelter. Since more sheltered nest sites could be occupied by better-quality females, we also used an experimental manipulation of nest shelter to separate the effects of the physical attributes of the nest site from those of individual quality. We compared birds with naturally sheltered nests, exposed nests and exposed nests provided with artificial nest shelters and measured clutch size, body condition, CORT levels at the end of incubation and hatching success. If nest shelter reduces CORT levels, we predicted that CORT levels would be highest at the least sheltered sites, and that the provision of artificial shelter would reduce CORT levels. We found that nest shelter was not related to CORT levels in incubating eiders. Nest shelter, however, affected body condition, with females at exposed sites losing more body mass during incubation than females at naturally and artificially sheltered nests. Interestingly however, in those birds nesting at the exposed sites, with and without artificial shelter, those with the highest CORT levels had the lowest hatching success. This relationship was not evident in females nesting at naturally sheltered sites. These results suggest that the level of nest shelter does not directly affect CORT levels in females. Instead, we suggest that the relationship between CORT levels and hatching success is state-dependent. Exposed sites are occupied by individuals that laid smaller clutches, and hence are likely to be of lower quality, and the negative effects of elevated CORT on hatching success are more pronounced in these females.

The colours of living organisms are produced by the differential absorption of light by pigments (e.g. carotenoids, melanins) and/or by the physical interactions of light with biological nanostructures, referred to as structural colours. Only two fundamental morphologies of non-iridescent nanostructures are known in feathers, and recent work has proposed that they self-assemble by intracellular phase separation processes. Here, we report a new biophotonic nanostructure in the non-iridescent blue feather barbs of blue penguins (Eudyptula minor) composed of parallel β-keratin nanofibres organized into densely packed bundles. Synchrotron small angle X-ray scattering and two-dimensional Fourier analysis of electron micrographs of the barb nanostructure revealed short-range order in the organization of fibres at the appropriate size scale needed to produce the observed colour by coherent scattering. These two-dimensional quasi-ordered penguin nanostructures are convergent with similar arrays of parallel collagen fibres in avian and mammalian skin, but constitute a novel morphology for feathers. The identification of a new class of β-keratin nanostructures adds significantly to the known mechanisms of colour production in birds and suggests additional complexity in their self-assembly.

Dynamic changes in integumentary color occur in cases as diverse as the neurologically-controlled iridiphores of cephalopod skin and the humidity-responsive cuticle of longhorn beetles. By contrast, feather colors are generally assumed to be relatively static, changing by small amounts only over periods of months. However, this assumption has rarely been tested even though structural colors of feathers are produced by ordered nanostructures that are analogous to those in the aforementioned dynamic systems. Feathers are neither innervated nor vascularized and therefore any color change must be caused by external stimuli. Thus, we here explore how feathers of iridescent mourning doves Zenaida macroura respond to a simple stimulus: addition and evaporation of water. After three rounds of experimental wetting and subsequent evaporation, iridescent feather color changed hue, became more chromatic and increased in overall reflectance by almost 50%. To understand the mechanistic basis of this change, we used electron microscopy to examine macro- and nanostructure before and after treatment. Transmission electron microscopy and transfer matrix thin film models revealed that color is produced by thin film interference from a single thick (~330 nm) layer of keratin around the edge of feather barbules, beneath which lies a layer of air and melanosomes. After treatment, the most striking morphological difference was a twisting of colored barbules that exposed more of their surface area for reflection following treatment, explaining the observed increase in brightness. These results suggest that some plumage colors may be more malleable than previously thought, leading to new avenues for research on dynamic plumage color.

Colours in feathers are produced by pigments or by nanostructurally organized tissues that interact with light. One of the simplest nanostructures is a single layer of keratin overlying a linearly organized layer of melanosomes that create iridescent colours of feather barbules through thin-film interference. Recently, it has been hypothesized that glossy (i.e. high specular reflectance) black feathers may be evolutionarily intermediate between matte black and iridescent feathers, and thus have a smooth keratin layer that produces gloss, but not the layered organization of melanosomes needed for iridescence. However, the morphological bases of glossiness remain unknown. Here, we use a theoretical approach to generate predictions about morphological differences between matte and glossy feathers that we then empirically test. Thin-film models predicted that glossy spectra would result from a keratin layer 110–180 nm thick and a melanin layer greater than 115 nm thick. Transmission electron microscopy data show that nanostructure of glossy barbules falls well within that range, but that of matte barbules does not. Further, glossy barbules had a thinner and more regular keratin cortex, as well as a more continuous underlying melanin layer, than matte barbules. Thus, their quasi-ordered nanostructures are morphologically intermediate between matte black and iridescent feathers, and perceived gloss may be a form of weakly chromatic iridescence.

Avian incubation dramatically reduces the abundance and diversity of microbial assemblages on eggshells, and this effect has been hypothesized as an adaptive explanation for partial incubation, the bouts of incubation that some birds perform during the egg-laying period. However, the mechanisms for these antimicrobial effects are largely unknown. In this study, we hypothesized that microbial inhibition is partly achieved through removal of liquid water, which generally enhances microbial growth, from eggshells, and experimentally tested this hypothesis in two ways. First, we placed the first- and second-laid eggs of tree swallow (Tachycineta bicolor) clutches in unincubated holding nests with either ambient or increased water on eggshells. Second, we added water to eggshells in naturally partially incubated nests. We compared microbial growth on shells during a 5-day experimental period and found that, as predicted, both unincubated groups had higher microbial growth than naturally partially incubated controls, and that only in the absence of incubation did wetted eggs have higher microbial growth than unwetted eggs. Thus, we have shown that water increases microbial growth on eggshells and that incubation nullifies these effects, suggesting that removal of water from egg surfaces is one proximate mechanism for the antimicrobial effects of incubation.

It is well known that testosterone (T) influences the expression of the behavior and many sexual traits during reproduction in vertebrates. However, patterns of circulating concentrations of T vary tremendously across free-living populations. Here the profiles of plasma T levels in the Magnificent Frigatebird, Fregata magnificens, are presented during the courtship, incubation and chick rearing stages of breeding. In addition, the predicted interrelationship of T and the expression of a sexually selected trait, the red gular pouch of males, is investigated. Plasma levels of corticosterone (Cort) are reported in relation to the demands of breeding conditions in colonies. Blood samples were obtained from 26 males and 32 females in the 1993-1994 breeding season and 41 courting males in 1997. Pouch color and size were also estimated in these males. As expected, T levels changed across the breeding stages: birds showed high levels of T during courtship and much lower circulating levels during incubation and chick rearing. Consistent with the expected effect of T, individual pouch color and size correlated with circulating levels of this hormone. In this highly dimorphic species no correlation was found between T and body condition or tail asymmetry. Cort, in contrast, did not change across the three reproductive stages.

The role of developmental conditions in shaping adult phenotypes has been the focus of a great deal of recent work. However, the effects of early life stress on reproductive performance have been little studied, particularly in avian species. In addition, although there is a large body of evidence to suggest that prevailing environmental conditions are linked to changes in breeding behavior, very little work has investigated the interaction between past and current exposure to environmental stress in determining breeding success. In this study, we examined the effects of early exposure to elevated stress hormone levels (corticosterone, CORT) on parental behavior during incubation in male and female zebra finches (Taeniopygia guttata) breeding in both stressful and nonstressful conditions as adults. We found that female birds fed CORT during postnatal development exhibited reduced incubation effort under both breeding conditions. There were no effects of developmental CORT exposure on male incubation effort; however, males breeding in unpredictable feeding conditions significantly reduced their effort levels compared with control males. There were no effects of either of our experimental treatments on hatching success or length of the incubation period. This may have been due to partial compensation by females paired with males that reduced effort. Our results clearly demonstrate sex-specific responses to developmental and adult environmental conditions in terms of incubation behavior and while in this captive population fitness costs appear to be ameliorated, birds breeding in a less benign environment in the wild may face higher costs having downstream effects on current or future reproduction.

For as long as dinosaurs have been known to exist, there has been speculation about their appearance. Fossil feathers can preserve the morphology of color-imparting melanosomes, which allow color patterns in feathered dinosaurs to be reconstructed. Here, we have mapped feather color patterns in a Late Jurassic basal paravian theropod dinosaur. Quantitative comparisons with melanosome shape and density in extant feathers indicate that the body was gray and dark and the face had rufous speckles. The crown was rufous, and the long limb feathers were white with distal black spangles. The evolution of melanin-based within-feather pigmentation patterns may coincide with that of elongate pennaceous feathers in the common ancestor of Maniraptora, before active powered flight. Feathers may thus have played a role in sexual selection or other communication.

Female birds can influence offspring fitness by varying the relative quantities of egg components they deposit within and between clutches. Antimicrobial proteins (lysozyme, ovotransferrin, and avidin) are significant components of the avian albumen and likely aid in defense of embryos from microbial infection. Within clutches, females may enhance antimicrobial defense of early-laid eggs to protect them from the high risk of infection incurred before the onset of incubation. Among entire clutches, females may invest more resources in young sired by more attractive males because they have higher reproductive value. We tested these hypotheses by quantifying antimicrobial protein distribution within and among clutches in blue tit eggs. Contrary to our hypothesis, clutches showed no differential deposition of lysozyme or avidin within clutches, but eggs laid in the middle of the sequence had higher concentrations of ovotransferrin than eggs in the beginning and end. Consistent with our second hypothesis, we found that females produced eggs with higher concentrations of lysozyme (although not ovotransferrin or avidin) when mated to more attractive (more UV-reflective) males. Furthermore, females mated to polygynous males deposited less lysozyme than those mated to monogamous males. These data suggest that allocation of lysozyme at the clutch level may be a maternal effect mediated by male qualities.

Models of climate change predict that its effects on animal populations will not always be negative, but most studies indicate negative associations between changes in climate and the phenology of animal migration and reproduction. For some populations, however, climate change may render particular environments more favourable, with positive effects on population growth. We used a 30-year population dataset on over 2000 Common Eiders Somateria mollissima at a colony in southwest Iceland to examine the response of this species to climate fluctuations. Eiders are strongly dependent on suitable climatic conditions for successful reproduction and survival. Temperatures in southwest Iceland, in both winter and summer, have generally increased over the past 30 years but have shown considerable fluctuation. We show that females laid earlier following mild winters and that year-to-year variation in the number of nests was related to the temperature during the breeding season 2 years previously. Milder summers could have positive effects on breeding success and offspring survival, producing an increase in nest numbers 2 years later when most Eiders recruit into the breeding population. In this part of their range, Eiders could benefit from a general warming of the climate.

1. In species that actively maintain optimal conditions for offspring development, the quality of the breeding site may have a substantial effect on reproductive effort and breeding success. Here, we studied the effect of nest shelter on reproductive costs for incubating female common eiders (Somateria mollissima) nesting in an arctic environment using both a correlative and an experimental approach.

2. The correlative data showed that females that nested in naturally well-sheltered nests experienced milder temperatures, laid larger clutches, provided a more favourable thermal environment for their eggs and had a higher hatching success than those at exposed nest-sites.

3. We added artificial shelters to exposed nest-sites and compared them to unmanipulated exposed nest-sites to examine the effect of nest shelter in females of similar quality. Costs of nesting in exposed sites were greater as evidenced by the reduced female mass loss and more favourable thermal conditions for the eggs at artificially sheltered nest-sites relative to exposed nest-sites. However, there was no difference in hatchings success between artificially sheltered and exposed nest-sites.

4. This study shows even small changes in the climatic conditions at the nest-site can have substantial consequences for reproductive effort, but the association between nest-site quality and breeding success in eiders is due to better quality individuals occupying better nest-sites.

The physical environment has a strong influence on the lives or organisms by limiting the way energy is gained and expended determining the capacity of organisms to invest in activities like reproduction. The avian nest site and structure, through its effects on the thermal conditions of the proximal environment of the incubating parent can affect several aspects of an individual’s reproductive success. On a larger scale, characteristics of the oviposition site can impact the spatial distribution of a species and consequently several aspects of population dynamics. In this thesis I investigate the importance of the thermal environment and nesting habitat on the reproductive performance of a sea duck, the Common Eider (Somateria mollissima) breeding in a cold environment. First I described the spatial variation in nest distribution in relation to female’s and nest attributes. Females of similar quality formed aggregations of nests and clusters of high productivity were occupied earlier in the season and at higher densities. Eiders seemed to choose to nest sites based on biotic (conspecifics) rather than abiotic (microclimate) cues. By providing females with artificial shelters I tested some of the effects of microclimate on individual physiology and use of energy during incubation. Females experienced improved microclimatic conditions provided by well-sheltered nest-sites. Nest shelter conferred advantages both to incubating females by allowing energy savings and to their clutches by providing more stable incubation conditions. Shelter did not have an appreciable effect on the female’s stress response. However, in exposed areas, females with high levels of corticosterone hatched a lower proportion of eggs than females with low corticosterone. Behavioural and functional aspects nest construction were tested first, by removing down from nests on repeated occasions throughout incubation and then by testing the effects of different amounts of down on the microclimate of incubation. Females did not replace the removed down suggesting the existence of constrains on the allocation of down to the nest. Large amounts of down in the nest contributed to more stable incubation conditions but females were able to counterbalance poor nest insulation and keep incubation temperature constant but the costs of doing this are unclear. Finally, I assessed the influence of environmental variability on the onset of incubation and short-term fluctuations in population size for the study colony with data from 1977 to 2006. I found that after mild winters female Eiders lay earlier in the season perhaps because milder conditions allow them to attain the necessary body condition for reproduction sooner. Summer temperature had a lagged effect (2 yr) on colony size that could be related to the delayed maturity (age at first reproduction) presented by Common Eiders and the influence of climate on influence recruitment rate to the population. The evidence presented here shows that Common Eiders are strongly influenced by their thermal environment on reproduction but by choosing a good nest they can ameliorate those effects. However nest site selection seems to be strongly linked to female quality rather than to nest properties.

Extra-pair reproduction is known to occur in many avian species. However, among passerines, the majority of studies on extra-pair reproduction have been carried out in oscine birds from temperate regions. Conversely, sub-oscines species, and particularly, species that inhabit tropical regions, have been studied to a much lesser extent. Given that a majority of avian species live in the tropics, it is important to study more tropical and sub-oscine species to have a more accurate picture of the rates of extra-pair reproduction among passerines, and a better understanding of the adaptive function of extra-pair reproduction in birds. Tropical species differ from temperate species in several ecological and life history traits, that may influence the occurrence of different modes of extra-pair reproduction and their prevalence. In this study we asked whether extra-pair reproduction occur in a sexually dimorphic and socially monogamous sub-oscine, the vermilion flycatcher (Pyrocephalus rubinus). We report cases of extra-pair paternity, extra-pair maternity and intra-specific brood parasitism, and discuss our results in the view of other studies with passerines.

Females may maximize their lifetime reproductive output by adjusting their investment in each breeding event to the perceived likelihood of success. The Blue-footed Booby (Sula nebouxii) is a long-lived seabird with facultative siblicide. We examined whether there is differential resource allocation to eggs with laying order and whether greater egg mass increases hatching probability, chick survival, hatching interval, and mass and size at hatching. We found that the relative investment in first and second eggs decreased as the season advanced: second eggs were slightly (1.5%) heavier than first eggs in early clutches; by contrast, first eggs were 2% heavier than second eggs in late clutches. Accordingly, hatching probability increased with laying date for first eggs and decreased for second eggs. The mass of the egg increased hatching probability, and no effect on chick survival was detected. Laying interval increased after a heavier egg was laid, and heavier eggs produced heavier hatchlings. Hatching intervals were positively related to laying intervals, but egg mass was unrelated to the length of the incubation period and the hatching interval. Our results suggest that egg mass influences embryo survival and that Blue-footed Booby females may adaptively allocate resources to eggs of different laying order according to breeding conditions.

Abstract  Egg composition, which is under maternal control, can have a profound effect on offspring fitness. The presence of maternal testosterone and carotenoids in avian egg yolk, for example, is thought to enhance the development and competitive ability of the offspring and protect the hatching and growing chick against oxidative stress. Egg quality often differs between females and such variation can be due to differences in maternal social environment, e.g. breeding density. However, this is confounded by the possibility that the quality of individuals breeding in high- or low-density areas may vary. We tested if maternal social environment influences egg composition in a colonial seabird, the lesser black-backed gull (Larus fuscus). To control for confounding effects of female quality, we experimentally manipulated maternal social environment during egg formation. We increased the frequency of intra-specific interactions (i.e. aggressive encounters with conspecifics other than nest mates) in which the females were involved, by placing an elevated platform in their territory. Females that took part in more intra-specific interactions produced a heavier last egg, but the yolk testosterone concentration in eggs laid by control and experimental females did not differ. Differences in yolk testosterone concentration in relation to embryo sex were found neither in the control nor in the experimental group. In contrast, within the control group, eggs with a male embryo contained more carotenoids than eggs with a female embryo. Moreover, experimental females that had been involved in more intra-specific interactions produced female eggs with higher carotenoid levels compared to female eggs of control birds. An experimental increase in carotenoid levels was not observed in eggs containing a male embryo. Our results suggest that intra-specific interactions experienced by female birds during egg formation can influence conditions for embryonic development.

Colors of living organisms are produced by selective light absorption from pigments and/or by light scattering from highly ordered nanostructures (i.e., structural color). While the physical bases of metallic colors of arthropods and fish are fairly well-known, those of birds are not. Here we examine structurally based silver color and its production in feathers of the waterbird species Anhinga. This achromatic color is distinguished from grey by high specular reflectance, from white by low diffuse reflectance, and from both by high gloss. Light and electron microscopy revealed three modifications of feathers likely leading to silver color. First, proximal barbules were highly elongated and contained glossy black color at their base and white color at their pennulum. Second, this glossy black portion contained a single outer layer of keratin weakly bounded by melanosomes. Finally, the white portion contained a disordered amorphous matrix of keratin and air. Optical analyzes suggest that these structures produce, respectively, glossy black color through thin-film interference and white color through incoherent light scattering. Silver color likely results from the combined reflectance of these adjacent structures. This represents a distinct mechanism for attaining silver colors that may have been partially derived through selection for display, thermoregulation or decreased hydrophobicity. J. Morphol., 2011. © 2011 Wiley-Liss, Inc.