Kevin J. McGraw

Carotenoids, Immunocompetence, and the Information Content of Sexual Colors: An Experimental Test

Many male birds use carotenoid pigments to acquire brilliant colors that advertise their health and condition to prospective mates. The direct means by which the most colorful males achieve superior health has been debated, however. One hypothesis, based on studies of carotenoids as antioxidants in humans and other animals, is that carotenoids directly boost the immune system of colorful birds. We studied the relationship between carotenoid pigments, immune function, and sexual coloration in zebra finches (Taeniopygia guttata), a species in which males incorporate carotenoid pigments into their beak to attract mates. We tested the hypotheses that increased dietary carotenoid intake enhances immunocompetence in male zebra finches and that levels of carotenoids circulating in blood, which also determine beak coloration, directly predict the immune response of individuals. We experimentally supplemented captive finches with two common dietary carotenoid pigments (lutein and zeaxanthin) and measured cell‐mediated and humoral immunity a month later. Supplemented males showed elevated blood‐carotenoid levels, brighter beak coloration, and increased cell‐mediated and humoral immune responses than did controls. Cell‐mediated responses were predicted directly by changes in beak color and plasma carotenoid concentration of individual birds. These experimental findings suggest that carotenoid‐based color signals in birds may directly signal male health via the immunostimulatory action of ingested and circulated carotenoid pigments.

Differential effects of endoparasitism on the expression of carotenoid- and melanin-based ornamental coloration

The striking diversity of sexual dimorphisms in nature begs the question: Why are there so many signal types? One possibility is that ornamental traits convey different sets of information about the quality of the sender to the receiver. The colourful, pigmented feathers of male birds seem to meet the predictions of this hypothesis. Evidence suggests that carotenoid pigmentation reflects the nutritional condition of males during moult, whereas in many instances melanin pigmentation is a reliable indicator of social status. However, as of yet there have been no experimental tests to determine how these two ornament types respond to the same form of environmental stress. In this study, we tested the effect of endoparasitic infection by intestinal coccidians (Isospora sp.) on the expression of both carotenoid– and melanin–based ornamental coloration in captive male American goldfinches (Carduelis tristis). We found that the carotenoid–based plumage and bill coloration of parasitized males was less saturated than that developed by unparasitized males, but that the brightness and size of melanin–based black caps did not differ between the groups. These findings provide the most robust empirical support to date for the notion that carotenoid and melanin ornaments reveal different information to conspecifics.

Maternally derived carotenoid pigments affect offspring survival, sex ratio, and sexual attractiveness in a colorful songbird

In egg-laying animals, mothers can influence the development of their offspring via the suite of biochemicals they incorporate into the nourishing yolk (e.g. lipids, hormones). However, the long-lasting fitness consequences of this early nutritional environment have often proved elusive. Here, we show that the colorful carotenoid pigments that female zebra finches (Taeniopygia guttata) deposit into egg yolks influence embryonic and nestling survival, the sex ratio of fledged offspring, and the eventual ornamental coloration displayed by their offspring as adults. Mothers experimentally supplemented with dietary carotenoids prior to egg-laying incorporated more carotenoids into eggs, which, due to the antioxidant activity of carotenoids, rendered their embryos less susceptible to free-radical attack during development. These eggs were subsequently more likely to hatch, fledge offspring, produce more sons than daughters, and produce sons who exhibited more brightly colored carotenoid-based beak pigmentation. Provisioned mothers also acquired more colorful beaks, which directly predicted levels of carotenoids found in eggs, thus indicating that these pigments may function not only as physiological ‘damage-protectants’ in adults and offspring but also as morphological signals of maternal reproductive capabilities.

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.

Testosterone upregulates lipoprotein status to control sexual attractiveness in a colorful songbird

A salient feature of many secondary sexual characteristics in animals is that their expression is controlled by sex-steroid hormones. However, for only a few types of ornaments do we know the precise molecular mechanism by which androgens like testosterone (T) enhance trait production. We studied the red carotenoid-based beak of male zebra finches (Taeniopygia guttata), which serves as a sexually selected trait and is thought to be T dependent. In previous research, we demonstrated that the extent to which male finches produce lipoproteins in the bloodstream, which bind carotenoid pigments acquired from the diet and transport them to peripheral tissues, regulates the accumulation of carotenoids in the body and beak pigmentation. Here we show that T acts to upregulate lipoprotein production and allows male zebra finches to display flashy, sexually attractive coloration. Levels of circulating T in blood positively and significantly predicted lipoprotein profile (as measured by cholesterol levels), blood carotenoid concentration, and beak color. Exogenous T administration elevated cholesterol and carotenoid status as well as beak redness. Last, experimental inhibition of T (using the anti-androgen flutamide) downregulated lipoprotein production and carotenoid circulation and faded the beak. This androgen- and lipoprotein-mediated system represents one of the more detailed physiological mechanisms underlying the development of a sex-steroid-dependent trait in animals.

Social environment during molt and the expression of melanin-based plumage pigmentation in male house sparrows (Passer domesticus)

Evolutionary biologists have shown much recent interest in the costliness and signal content of colorful plumage displays in birds. Although many studies suggest that both carotenoid- and structurally-based plumage colors are condition-dependent indicators of health and nutritional state at the time ornamental feathers are grown, there is little experimental evidence supporting the idea that melanin pigmentation is a reliable signal of condition during molt. Instead, melanin-based ornamental coloration often reveals the competitive ability and dominance of individuals throughout the year. However, this work does not indicate which proximate environmental factors shape the expression of melanin pigmentation at the time of feather growth. Because of the link between melanin coloration and the social environment, it is possible that the development of brightly colored plumage may be associated with aggressive social interactions during feather molt. Here, we show that melanin-based ornamental coloration in male house sparrows (Passer domesticus) is correlated with the degree to which individuals interact aggressively with conspecifics during molt. Males that were dominant (beta, but not alpha) within captive social groups during molt grew larger badges than subordinates. Groups of males that had higher rates of aggression during molt grew larger badges than less aggressive triads. To our knowledge, this is the first demonstration that melanin pigmentation and plumage-based status badges are related to the competitive history of individuals during feather development. By coupling badge size directly with aggressive experiences during molt, birds can use their status signal to honestly indicate their likelihood of winning agonistic encounters throughout the year.

An update on the honesty of melanin-based color signals in birds

The control mechanisms and information content of melanin‐based color signals in birds have generated much recent interest and controversy among evolutionary biologists. Initial experimental studies on this topic manipulated coarse metrics of an individual’s condition (i.e. food intake, disease state) and failed to detect significant condition‐dependence of melanin ornament expression. However, three new lines of research appear profitable and target specific factors associated with the production of melanin pigments. These include the role of (i) metals, (ii) amino acids, and (iii) testosterone and social interactions in shaping the extent and intensity of melanin‐colored plumage patches. Here, I review recent studies of and evidence for these honesty‐reinforcing mechanisms.

DIET, PLASMA CAROTENOIDS, AND SEXUAL COLORATION IN THE ZEBRA FINCH (TAENIOPYGIA GUTTATA)

Abstract Carotenoid-based colors serve important sexual-signaling functions in many animals, but the proximate factor(s) underlying their expression has sparked controversy. In particular, the relative contributions of dietary and physiological mechanisms have been questioned of late. However, no studies have concurrently quantified levels of food intake or pigment processing in any species to examine the comparative effects of pigment acquisition and use on integumetary coloration. Here, we studied within- and between-sex patterns of food intake and plasma pigment circulation in the Zebra Finch (Taeniopygia guttata) to assess how sexually dichromatic, carotenoid-based bill pigmentation serves as an indicator of pigment access in the diet and carotenoid transport through the bloodstream. First, in a food-choice study, we found that males and females did not consume different types or amounts of food, despite dramatic sex differences in bill coloration. Similarly, variability in carotenoid-based bill pigmentation within each sex was uncoupled from levels of food consumption. Next, we used high-performance liquid-chromatography (HPLC) to quantify the types and amounts of carotenoids circulating through blood. Male and female Zebra Finches circulated the same four major carotenoid pigments in blood plasma (lutein, zeaxanthin, anhydrolutein, and β-cryptoxanthin), but males circulated a significantly higher concentration of plasma carotenoids than did females. Within both sexes, individuals that circulated more carotenoid pigments displayed more brightly colored bills. In sum, these results suggest that physiological factors such as pigment transport may play a more important role in shaping variability in carotenoid-based bill coloration in this species than does diet. Future studies should be aimed at identifying the proximate determinants of plasma carotenoid circulation in these birds as well as how circulated pigments are used to produce maximum color displays.

Interspecific associations between circulating antioxidant levels and life-history variation in birds

Antioxidants play an important role in protecting tissues against aging‐associated oxidative damage and are thus prime candidates for relating physiological mechanisms to variation in life histories. We measured total antioxidant capacity, antioxidant response to stress, and levels of uric acid, vitamin E, and four carotenoids in 95 avian species, mostly passerines from Michigan or Panama. We compared antioxidant measures to seven variables related to life histories (clutch size, survival rate, incubation period, nestling period, basal metabolic rate, body mass, and whether the species lived in a tropical or temperate climate). Life‐history‐related traits varied over at least three statistically independent axes. Higher antioxidant levels were generally characteristic of more rapid development, lower survival rate, smaller body size, larger clutch size, and higher mass‐adjusted metabolic rate, but the relationships of particular antioxidants with individual life‐history traits showed considerable complexity. Antioxidant–life history associations differed between tropical and temperate species and varied with respect to taxonomic sampling. Vitamin E showed few relationships with life‐history traits. Overall, our results partly support the hypothesis that antioxidant levels evolve to mirror free radical production. Clearly, however, the complex patterns of physiological diversification observed here result from the interplay of many factors, likely including not just investment in somatic maintenance but also phylogenetic constraint, diet, and other aspects of ecology.

The physiological costs of being colourful: nutritional control of carotenoid utilization in the American goldfinch, Carduelis tristis

Many sexually selected traits are thought to be costly to produce, which ensures that they communicate information honestly to conspecifics. Carotenoid pigmentation is classically considered as a costly sexual signal in many fish and birds. It is often argued that carotenoid colours are ‘condition dependent’, with an individual’s nutritional or health state directly determining trait expression. However, few studies have investigated precisely how a compromised nutritional state affects an animal’s capacity to develop these sexually attractive colours. Here, we studied the effect of food restriction on the ability of male American goldfinches to physiologically process carotenoids during the period of feather growth. We used highperformance liquid-chromatography to determine the types and amounts of carotenoids circulating in blood during moult as well as in newly grown colourful feathers. We show that nutritional deprivation affects the degree to which male goldfinches transport carotenoids through the bloodstream. Foodrestricted males circulated significantly less blood carotenoids than controls. They also incorporated less carotenoids into feathers and grew less colourful plumage, but the decrease in plumage carotenoids did not significantly exceed the depressed amounts already present in blood. These results suggest that the means by which these yellow-coloured passerines either extract carotenoids from food (e.g. via lipoidal micelles) or transport them through blood (e.g. via lipoproteins) are more sensitive to changes in nutritional/energy state than are the mechanisms for metabolizing dietary pigments or depositing metabolites into feathers.