Melanin coloration in New World orioles I: carotenoid masking and pigment dichromatism in the orchard oriole complex

Carotenoids produce the brilliant red, orange, and yellow colors of many animals. However, melanin pigments can also confer some of these same hues. Because carotenoid and melanin colors are produced in different ways and may serve different signaling functions, either within or between species, it is important to establish whether one or both types of pigment are responsible for coloration. We have discovered what appears to be an evolutionary switch from carotenoid- to melanin-based color in two sexually dichromatic New World orioles. Using a combination of reflectance spectrometry and chromatographic analyses of plumage pigments, we found that the chestnut plumage of adult male orchard orioles Icterus spurius is produced predominantly by phaeomelanins. Orchard oriole feathers also contain carotenoids, which appear to be masked by the high concentration of phaeomelanins. In contrast, both carotenoids and phaeomelanins appear to contribute to color in adult male Fuertes's orioles I. fuertesi . Moreover, yellow yearling male and female plumage in both species is produced by carotenoids alone. The masking of carotenoids with phaeomelanins in orchard orioles is interesting in light of the signaling roles that carotenoids are thought to play. In addition, these plumage differences produce a unique case of age and sexual pigment dimorphism in orchard and Fuertes's orioles.     

The influence of carotenoid acquisition and utilization on the maintenance of species-typical plumage pigmentation in male American goldfinches (Carduelis tristis) and northern cardinals (Cardinalis cardinalis).

Birds display a tremendous variety of carotenoid-based colors in their plumage, but the mechanisms underlying interspecific variability in carotenoid pigmentation remain poorly understood. Because vertebrates cannot synthesize carotenoids de novo, access to pigments in the diet is one proximate factor that may shape species differences in carotenoid-based plumage coloration. However, some birds metabolize ingested carotenoids and deposit pigments that differ in color from their dietary precursors, indicating that metabolic capabilities may also contribute to the diversity of plumage colors we see in nature. In this study, we investigated how the acquisition and utilization of carotenoids influence the maintenance of species-typical plumage pigmentation in male American goldfinches (Carduelis tristis) and northern cardinals (Cardinalis cardinalis). We supplemented the diet of captive goldfinches with red carotenoids to determine whether males, which are typically yellow in color, were capable of growing red plumage. We also deprived cardinals of red dietary pigments to determine whether they could manufacture red carotenoids from yellow precursors to grow species-typical red plumage. We found that American goldfinches were able to deposit novel pigments in their plumage and develop a striking orange appearance. Thus, dietary access to pigments plays a role in determining the degree to which goldfinches express carotenoid-based plumage coloration. We also found that northern cardinals grew pale red feathers in the absence of red dietary pigments, indicating that their ability to metabolize yellow carotenoids in the diet contributes to the bright red plumage that they display.     

The evolution of carotenoid coloration in estrildid finches: a biochemical analysis

The estrildid finches (Aves: Passeriformes: Estrildidae) of Africa, Asia, and Australia have been the focus of several recent tests of sexual selection theory. Many estrildids display bright red, orange, or yellow colors in the beak or plumage, which typically are generated by the presence of carotenoid pigments. In this study, we used high-performance liquid chromatography to investigate the carotenoid content of feathers and other colorful tissues in seven species of estrildids. Star finches (Neochmia ruficauda) and diamond firetails (Stagonopleura guttata) circulated two main dietary carotenoids (lutein and zeaxanthin) through the blood and liver and used both to acquire a yellow plumage color. However, five other estrildids (common waxbill, Estrilda astrild; black-rumped waxbill, Estrilda troglodytes; zebra waxbill, Amandava subflava; red avadavat, Amandava amandava; and zebra finch, Taeniopygia guttata) circulated these same dietary carotenoids along with two metabolites (dehydrolutein and anhydrolutein) through the blood and/or liver and used all four as yellow plumage colorants. We subsequently tracked the distribution of these pigments using a published phylogeny of estrildid finches to determine the evolutionary pattern of carotenoid metabolism in these birds. We found that finches from the most ancient tribe of estrildids (Estrildini) possessed the ability to metabolize dietary carotenoids. Although carotenoids from the most ancestral extant estrildid species have yet to be analyzed, we hypothesize (based on their relationships with other songbirds known to have such metabolic capabilities) that these finches inherited from their ancestors the capability to metabolize carotenoids. Interestingly, later in estrildid evolution, certain taxa lost the ability to metabolize dietary carotenoids (e.g., in the Poephilini), suggesting that the occurrence of carotenoid metabolism can be labile and is likely shaped by the relative costs and benefits of color signaling across different species.     

Evolutionary innovation and diversification of carotenoid‐based pigmentation in finches

The ornaments used by animals to mediate social interactions are diverse, and by reconstructing their evolutionary pathways we can gain new insights into the mechanisms underlying ornamental innovation and variability. Here, we examine variation in plumage carotenoids among the true finches (Aves: Fringillidae) using biochemical and comparative phylogenetic analyses to reconstruct the evolutionary history of carotenoid states and evaluate competing models of carotenoid evolution. Our comparative analyses reveal that the most likely ancestor of finches used dietary carotenoids as yellow plumage colorants, and that the ability to metabolically modify dietary carotenoids into more complex pigments arose secondarily once finches began to use modified carotenoids to create red plumage. Following the evolutionary “innovation” that enabled modified red carotenoid pigments to be deposited as plumage colorants, many finch species subsequently modified carotenoid biochemical pathways to create yellow plumage. However, no reversions to dietary carotenoids were observed. The finding that ornaments and their underlying mechanisms may be operating under different selection regimes—where ornamental trait colors undergo frequent reversions (e.g., between red and yellow plumage) while carotenoid metabolization mechanisms are more conserved—supports a growing empirical framework suggesting different evolutionary patterns for ornaments and the mechanistic innovations that facilitate their diversification.     

Proximate mechanisms of colour variation in the frillneck lizard: geographical differences in pigment contents of an ornament

Animal coloration has evolved in contexts such as communication, camouflage, and thermoregulation. Most studies of animal coloration focus on its adaptive benefits, whereas its underlying mechanisms have received less attention despite their potential influence on adaptive benefits. In fish and reptiles, for example, colour variation from yellow to red can be produced by carotenoid and/or pteridine pigments, which differ dramatically in the way they are obtained (carotenoids through diet and pteridines synthesized de novo). Hence, potential adaptive benefits could differ greatly depending on the relative contribution to coloration of different pigments. In the present study, we investigate the mechanisms underlying colour variation in the frill of the Australian frillneck lizard (Sauropsida: Chlamydosaurus kingii). Frill colour varies between populations across the species' range (red, orange, yellow or white). We argue that this geographical variation results from different concentrations of carotenoids and pteridines in the frill. Frill carotenoid concentrations were lower in eastern populations (yellow and white forms), and pteridines were present only in the red and orange forms, thereby explaining their redder hues. The observed geographical variation in frill carotenoids suggests variation in carotenoid availability across the species' range, which is backed up by the finding that plasma carotenoid concentrations were higher in the red (western) compared to the yellow (eastern) form. Although no correlations were found between individual colour measurements, frill pigments and plasma carotenoids, our results suggest that selective pressures vary across the species' range and we speculate that predation pressures and/or intrasexual signalling context differ between forms.     

Multiple ways to become red: pigment identification in red feathers using spectrometry

Red hues are a challenge in studies on the evolution of bird coloration, as multiple pigments such as carotenoids, pheomelanin, psittacofulvins, porphyrins, turacin, haemoglobin and even exogenous iron-oxides, may confer red colors. Determining the pigment type is paramount and here we investigate the differences in spectrum reflectance for six pigments resulting in red colorations in feathers of different species, with a focus on discriminating among melanins and carotenoids. Pigment chemical identification was obtained from the literature or using High Performance Liquid Chromatography (HPLC) in our laboratory. We have also derived discriminant formulas for identification of the major known types of pigments based on parameters of the reflectance curves obtained with a portable spectrometer. Our results indicate that the reflectance patterns of coloration perceived as red patches widely differ. The distinction between carotenoid- and melanin-based reflectance curves is relatively straightforward: sigmoid versus straight slope. The spectral reflectance curves of feathers containing red psittacofulvins are sigmoid, whereas iron oxide and porphyrin-containing feathers recall pheomelanins in rendering a straight slope. In the case of turacin-based coloration, the spectral shape is unique. For the pigments with enough number of species sampled (i.e., carotenoids, melanins and psittacofulvins) the differences in reflectance shape are important enough to allow separation of carotenoid and melanin derived colorations based on reflectance curves alone.     

Red-winged blackbirds Agelaius phoeniceus use carotenoid and melanin pigments to color their epaulets

Over the past three decades, the red‐winged blackbird Agelaius phoeniceus has served as a model species for studies of sexual selection and the evolution of ornamental traits. Particular attention has been paid to the role of the colorful red‐and‐yellow epaulets that are striking in males but reduced in females and juveniles. It has been assumed that carotenoid pigments bestow the brilliant red and yellow colors on epaulet feathers, but this has never been tested biochemically. Here, we use high‐performance liquid chromatography (HPLC) to describe the pigments present in these colorful feathers. Two red ketocarotenoids (astaxanthin and canthaxanthin) are responsible for the bright red hue of epaulets. Two yellow dietary precursors pigments (lutein and zeaxanthin) are also present in moderately high concentrations in red feathers. After extracting carotenoids, however, red feathers remained deep brown in color. HPLC tests show that melanin pigments (primarily eumelanin) are also found in the red‐pigmented barbules of epaulet feathers, at an approximately equal concentration to carotenoids. This appears to be an uncommon feature of carotenoid‐based ornamental plumage in birds, as was shown by comparable analyses of melanin in the yellow feathers of male American goldfinches Carduelis tristis and the red feathers of northern cardinals Cardinalis cardinalis, in which we detected virtually no melanins. Furthermore, the yellow bordering feathers of male epaulets are devoid of carotenoids (except when tinged with a carotenoid‐derived pink coloration on occasion) and instead are comprised of a high concentration of primarily phaeomelanin pigments. The dual pigment composition of red epaulet feathers and the melanin‐only basis for yellow coloration may have important implications for the honesty‐reinforcing mechanisms underlying ornamental epaulets in red‐winged blackbirds, and shed light on the difficulties researchers have had to date in characterizing the signaling function of this trait. As in several other birds, the melanic nature of feathers may explain why epaulets are used largely to settle aggressive contests rather than to attract mates.     

CONVERGENT EVOLUTION OF RED CAROTENOID COLORATION IN WIDOWBIRDS AND BISHOPS (EUPLECTES SPP.)

Avian carotenoid‐based signals are classic examples of sexually selected, condition‐dependent threat displays or mate choice cues. In many species, male dominance or mating success is associated with redder (i.e., longer wavelength) color hues, suggesting that red colors are either more efficient or more reliable signals than yellow colors. Few studies, however, have investigated selection for redness in a macroevolutionary context. Here, we phylogenetically reconstruct the evolution of carotenoid coloration in the African widowbirds and bishops (Euplectes spp.), for which agonistic selection for redder hues, as well as pigmentary mechanisms, is well documented. Using reflectance spectrometry for objective color quantification, and accounting for phylogenetic uncertainty, we find that yellow plumage color is a retained ancestral state in Euplectes, and that red color hues have convergently evolved two or three times. Results are discussed in relation to a known diversity in pigment mechanisms, supporting independent origins of red color, and suggesting that agonistic selection and physiological constraints have interacted to generate color diversity in Euplectes.     

Interspecific variation in dietary carotenoid assimilation in birds: links to phylogeny and color ornamentation

Many birds use carotenoid pigments to acquire rich red, orange, and yellow coloration in feathers and bare parts that is used as a signal of mate quality. Because carotenoids are derived from foods, much attention has been paid to the role of diet in generating color variation both within and among avian species. Less consideration has been given to physiological underpinnings of color variability, especially among species. Here, I surveyed published literature (e.g. captive feeding studies) on carotenoid assimilation in six bird species and completed additional controlled carotenoid-supplementation experiments in two others to consider the ability of different taxa to extract carotenoids from the diet in relation to phylogeny and coloration. I found that, for a given level of carotenoids in the diet, passerine birds (zebra finch, Taeniopygia guttata; house finch, Carpodacus mexicanus; American goldfinch, Carduelis tristis; society finch, Lonchura domestica) exhibit higher levels of carotenoids in circulation than non-passerines like gamebirds (domestic chicken, Gallus domesticus; red junglefowl, Gallus gallus; Japanese quail, Coturnix coturnix; red-legged partridge, Alectoris rufa). This difference in carotenoid accumulation is likely due to interspecific variation in micelle, chylomicron, or lipoprotein concentrations or affinities for xanthophyll carotenoids. Passerine birds more commonly develop carotenoid-based colors than do birds from ancient avian lineages such as Galliformes, and the physiological differences I uncover may explain why songbirds especially capitalize on carotenoid pigments for color production. Ultimately, because we can deconstruct color traits into component biochemical, physical, and physiological parts, avian color signals may serve as a valuable model for illuminating the proximate mechanisms behind interspecific variation in signal use in animals.     

EVOLUTION OF CAROTENOID PIGMENTATION IN CACIQUES AND MEADOWLARKS (ICTERIDAE): REPEATED GAINS OF RED PLUMAGE COLORATION BY CAROTENOID C4‐OXYGENATION

Many animals use carotenoid pigments to produce yellow, orange, and red coloration. In birds, at least 10 carotenoid compounds have been documented in red feathers; most of these are produced through metabolic modification of dietary precursor compounds. However, it is poorly understood how lineages have evolved the biochemical mechanisms for producing red coloration. We used high‐performance liquid chromatography to identify the carotenoid compounds present in feathers from 15 species across two clades of blackbirds (the meadowlarks and allies, and the caciques and oropendolas; Icteridae), and mapped their presence or absence on a phylogeny. We found that the red plumage found in meadowlarks includes different carotenoid compounds than the red plumage found in caciques, indicating that these gains of red color are convergent. In contrast, we found that red coloration in two closely related lineages of caciques evolved twice by what appear to be similar biochemical mechanisms. The C4‐oxygenation of dietary carotenoids was responsible for each observed transition from yellow to red plumage coloration, and has been commonly reported by other researchers. This suggests that the C4‐oxygenation pathway may be a readily evolvable means to gain red coloration using carotenoids.     

Lutein-based plumage coloration in songbirds is a consequence of selective pigment incorporation into feathers

Many birds obtain colorful carotenoid pigments from the diet and deposit them into growing tissues to develop extravagant red, orange or yellow sexual ornaments. In these instances, it is often unclear whether all dietary pigments are used as integumentary colorants or whether certain carotenoids are preferentially excluded or incorporated into tissues. We examined the carotenoid profiles of three New World passerines that display yellow plumage coloration—the yellow warbler (Dendroica petechia), common yellowthroat (Geothlypis trichas) and evening grosbeak (Coccothraustes vespertinus). Using high-performance liquid chromatography, we found that all species used only one carotenoid—lutein—to color their plumage yellow. Analyses of blood carotenoids (which document those pigments taken up from the diet) in two of the species, however, revealed the presence of two dietary xanthophylls—lutein and zeaxanthin—that commonly co-occur in plants and animals. These findings demonstrate post-absorptive selectivity of carotenoid deposition in bird feathers. To learn more about the site of pigment discrimination, we also analyzed the carotenoid composition of lipid fractions from the follicles of immature yellow-pigmented feathers in G. trichas and D. petechia and again detected both lutein and zeaxanthin. This suggests that selective lutein incorporation in feathers is under local control at the maturing feather follicle.     

Pterin-based pigmentation in animals

Pterins are one of the major sources of bright coloration in animals. They are produced endogenously, participate in vital physiological processes and serve a variety of signalling functions. Despite their ubiquity in nature, pterin-based pigmentation has received little attention when compared to other major pigment classes. Here, we summarize major aspects relating to pterin pigmentation in animals, from its long history of research to recent genomic studies on the molecular mechanisms underlying its evolution. We argue that pterins have intermediate characteristics (endogenously produced, typically bright) between two well-studied pigment types, melanins (endogenously produced, typically cryptic) and carotenoids (dietary uptake, typically bright), providing unique opportunities to address general questions about the biology of coloration, from the mechanisms that determine how different types of pigmentation evolve to discussions on honest signalling hypotheses. Crucial gaps persist in our knowledge on the molecular basis underlying the production and deposition of pterins. We thus highlight the need for functional studies on systems amenable for laboratory manipulation, but also on systems that exhibit natural variation in pterin pigmentation. The wealth of potential model species, coupled with recent technological and analytical advances, make this a promising time to advance research on pterin-based pigmentation in animals.     

Making extra room for carotenoids in plant cells: New opportunities for biofortification

Plant carotenoids are essential for photosynthesis and photoprotection and provide colors in the yellow to red range to non-photosynthetic organs such as petals and ripe fruits. They are also the precursors of biologically active molecules not only in plants (including hormones and retrograde signals) but also in animals (including retinoids such as vitamin A). A carotenoid-rich diet has been associated with improved health and cognitive capacity in humans, whereas the use of carotenoids as natural pigments is widespread in the agrofood and cosmetic industries. The nutritional and economic relevance of carotenoids has spurred a large number of biotechnological strategies to enrich plant tissues with carotenoids. Most of such approaches to alter carotenoid contents in plants have been focused on manipulating their biosynthesis or degradation, whereas improving carotenoid sink capacity in plant tissues has received much less attention. Our knowledge on the molecular mechanisms influencing carotenoid storage in plants has substantially grown in the last years, opening new opportunities for carotenoid biofortification. Here we will review these advances with a particular focus on those creating extra room for carotenoids in plant cells either by promoting the differentiation of carotenoid-sequestering structures within plastids or by transferring carotenoid production to the cytosol.     

CHROMOPLAST ULTRASTRUCTURE AS AFFECTED BY GENES CONTROLLING GRANA RETENTION AND CAROTENOIDS IN FRUITS OF CAPSICUM ANNUUM

Plastids in the fruits of isogenic lines of pepper (Capsicum annuum) were examined by electron microscopy with reference to four genotypes determining the carotenoid composition and the colors red, yellow, brown, and green of the ripe fruit. One gene pair (y⁺/y) influences carotenoid content and the other pair (cl⁺/cl) controls the chlorophyll. The retention of the grana and chlorophyll in the ripe fruits of the brown and green phenotypes is correlated with the cl cl genotype. The y⁺ gene increases the total carotenoids and promotes the formation of red pigments. Giant grana were found in the yellow and green phenotypes, but during ripening these disappeared in the yellow. Unusual dichotomous and concentric grana were observed in the green. Globule-associated carotenoids forming fibrillar crystalloids were present in all color types, although to a lesser degree in the yellow fruit. Membrane-associated carotenoids occurred only in the yellow and green phenotypes.     

Pteridine, not carotenoid, pigments underlie the female-specific orange ornament of striped plateau lizards (Sceloporus virgatus)

Indicator models of sexual selection suggest that signal honesty is maintained via costs of ornament expression. Carotenoid-based visual signals are a well-studied example, as carotenoids may be environmentally limited and impact signaler health. However, not all bright yellow, orange and red ornaments found in vertebrates are carotenoid-based; pteridine pigments may also produce these colors. We examine the contribution of carotenoid and pteridine pigments to the orange reproductive color of female striped plateau lizards (Sceloporus virgatus). This color ornament reliably indicates female mate quality, yet costs maintaining signal honesty are currently unknown. Dietary carotenoid manipulations did not affect orange color, and orange skin differed from surrounding white skin in drosopterin, not carotenoid, content. Further, orange color positively correlated with drosopterin, not carotenoid, concentration. Drosopterin-based female ornaments avoid the direct trade-offs of using carotenoids for ornament production vs egg production, thus may relax counter-selection against color ornament exaggeration in females. Direct experimentation is needed to determine the actual costs of pteridine-based ornaments. Like carotenoids, pteridines influence important biological processes, including immune and antioxidant function; predation and social costs may also be relevant.     

Melanin coloration in New World orioles II: ancestral state reconstruction reveals lability in the use of carotenoids and phaeomelanins

Although many animals use carotenoids to produce bright yellow, orange, and red colors, an increasing number of studies have found that other pigments, such as melanins, may also be used to produce bright colors. Yet, almost nothing is known about the evolutionary history of this colorful melanin use. We used reflectance spectrometry to determine whether colors in New World orioles were predominantly due to carotenoids, colorful melanins, or a mixture of both. We then used ancestral state reconstruction to infer the directionality of any pigment changes and to test for phylogenetic signal. We found that three oriole taxa likely switched from carotenoid- to melanin-based colors. Several other oriole taxa apparently gained localized melanin coloration, or had coloration that seemed to be produced by a mixture of carotenoids and melanins. We also found little phylogenetic signal on the use of carotenoids or melanins to produce color. However, all pigment changes occurred within one of three major clades of the oriole genus, suggesting there may be signal at deeper phylogenetic levels. These repeated independent switches between carotenoid and melanin colors are surprising in light of the important signaling role that color pigments (especially carotenoids) are thought to play across a wide range of taxa.     

Concerted Shifts in Absorption Maxima of Yellow and Red Plumage Carotenoids Support Specialized Tuning of Chromatic Signals to Different Visual Systems in Near-Passerine Birds

Recent evidence that absorption maxima (λR_min) expressed by colorful plumage pigments align to diagnostic cone sensitivities of affiliated visual systems suggests that birds employ specialized signals in relation to their color vision. However, these studies compared different pigments and clades for the violet (porphyrins in non-passerines) and ultraviolet (carotenoids in passerines) sensitive system, which confounds chemistry and phylogeny with tuning patterns. To test whether signal alignments to violet (VS) and ultraviolet (UVS) systems transcend confounding factors, parallel analyses were conducted for a diversity of near-passerines, a group in which plumage carotenoids occur in taxa with either visual system. Conventional and phylogenetically informed analyses confirmed earlier findings: short wavelength absorbing (yellow carotenoid) pigments aligned λR_min with the violet-sensitive (V) cone of VS species but with the short wavelength-sensitive (S) cone of UVS species, whereas long wavelength-absorbing (red carotenoid) pigments aligned only with the S cone of VS species. More extensive variation among VS yellow carotenoids produced λR_min alignments to cone sensitivities that differed at shorter (peaks) versus longer (overlaps) wavelengths. Ancestral trait reconstructions indicated that signals evolved to match pre-existing VS systems, but did not resolve scenarios for UVS systems. Regardless of historical details, alignments expressed a higher-level pattern in which λR_min values were blue-shifted for yellow and red carotenoids in VS compared to UVS species, a pattern opposite that expressed by receptor sensitivities between systems. Thus, generalized functional designs attributed to avian color vision allow for specialized visual communication through the development of chromatic signals suited to each perceptual system.     

On the ecological basis of interspecific homoplasy in carotenoid-bearing signals

Evidence that similar color patterns occur in unrelated animals with different habits undermines the traditional view that homoplasy evolves through shared ecological selection pressures. Carotenoid pigments responsible for many yellow to red signals exhibit two related properties that could link ecology with appearance by nontraditional means. Ecologic homoplasy could arise through ecophenotypy because all animals must obtain carotenoids through their diet. Such homoplasy also could be hidden from view because increased carotenoid levels are more strongly encoded by decreased reflectance over ultraviolet (UV) wavelengths invisible to humans. To explore these possibilities, I examined apparent matches or mismatches between color and ecology among insectivorous (low carotenoid diet) and frugivorous (high carotenoid diet) bird species in relation to the typical yellow and black plumage pattern of insectivorous, UV-sensitive titmice (Paridae). Diagnostic features of reflectance spectra indicated that all yellow plumages resulted from carotenoids, black plumages from melanins, and olive green plumages from codeposition of both pigments. However, reflectance by carotenoid-bearing plumages correlated with diet independent of plumage pattern; compared to the insectivores, frugivores had reduced amounts of UV reflectance, and to a lesser extent, "red shifts" in longer-wavelength reflectance. Furthermore, an asymptotic decrease in amount of UV with increased redness implied that plumage reflectance of insectivorous species differed more over UV wavelengths, whereas that of frugivorous species differed more over longer wavelengths. I verified that dietary links to plumage reflectance resulted from greater amounts of plumage carotenoids in frugivores, presumably due to their carotenoid-rich diets. All of these ecological associations transcended post-mortem or post-breeding color change, and phylogeny. Thus, predictable associations between avian-visible plumage reflectance, pigmentation, and diet across evolutionary scales may arise directly (diet per se) or indirectly (honest signaling of diet) by ecophenotypy, although various genetic factors also may play a role.     

Carotenoids, colour and conservation in an endangered passerine, the hihi or stitchbird (Notiomystis cincta)

Carotenoids are essential dietary components utilized not only in pigmentation but also as immuno-stimulants and antioxidants. Reduced availability can have consequences on individual health and survival, thus making carotenoids a good indicator of environmental stress. We compared carotenoid profiles and plumage colour characteristics of an endangered passerine species in New Zealand, between its remnant island source population and two reintroduced island populations. Circulating carotenoids were predominantly lutein (mean of 82.2%) and zeaxanthin (mean of 14.8%), and these were the major carotenoids present as yellow pigments in the males' plumage. There were clear differences in total carotenoid concentrations and plumage colour among the three populations. Circulating carotenoid concentration was significantly higher in one of the reintroduced populations, and the yellow plumage of males was significantly higher in both reintroduced populations in comparison with the remnant population (reflected as a significant increase in hue). Understanding how these differences arise may be of importance to this species given the health benefits carotenoids impart and our ability to select plant species containing these compounds or artificially supplement them.     

Carotenoid pigments and the selectivity of psittacofulvin-based coloration systems in parrots

Carotenoid pigments are commonly used as colorants of feathers and bare parts by birds. However, parrots (Aves: Psittaciformes) use a novel class of plumage pigments (called psittacofulvins) that, like carotenoids, are lipid-soluble and red, orange, or yellow in color. To begin to understand how and why parrots use these pigments and not carotenoids in their feathers, we must first describe the distribution of these two types of pigments in the diet, tissues, and fluids of these birds. Here, we studied the carotenoid content of blood in five species of parrots with red in their plumage to see if they show the physiological ability to accumulate carotenoids in the body. Although Scarlet (Ara macao) and Greenwing Macaws (Ara chloroptera) and Eclectus (Eclectus roratus), African Gray (Psittacus erithacus) and Blue-fronted Amazon (Amazona aestiva) Parrots all use psittacofulvins to color their feathers red, we found that they also circulated high concentrations of both dietary (lutein, zeaxanthin, beta-cryptoxanthin) and metabolically derived (anhydrolutein, dehydrolutein) carotenoids through blood at the time of feather growth, at levels comparable to those found in many other carotenoid-colored birds. These results suggest that parrots have the potential to use carotenoids for plumage pigmentation, but preferentially avoid depositing them in feathers, which is likely under the control of the maturing feather follicle. As there is no evidence of psittacofulvins in parrot blood at the tune of feather growth, we presume that these pigments are locally synthesized by growing feathers within the follicular tissue.