Differential effects of testosterone metabolites oestradiol and dihydrotestosterone on oxidative stress and carotenoid-dependent colour expression in a bird

Despite extensive research, the potential costs that keep secondary sexual traits honest and evolutionary stable remain somewhat elusive. Many carotenoid-based signals are regulated by testosterone (T), which has been suggested to impose a cost to the signaller by suppression of the immune system or an increase in oxidative stress. Results are, however, inconsistent, which may be due to the fact that T can be metabolised to both 5α-dihydrotestosterone (DHT, a potent androgen) and oestradiol (E2, a potent oestrogen). To evaluate for the first time the independent effect of these testosterone metabolites on oxidative status, circulating carotenoids and a carotenoid-dependent sexual signal, we administered DHT and E2 to captive non-breeding adult kestrels Falco tinnunculus of both sexes. E2 increased oxidative damage and downregulated the antioxidant barrier without affecting colouration or circulating carotenoids. In contrast, DHT did not affect oxidative status, but increased skin redness, again without affecting circulating carotenoids. No sex-specific effects were found. These results suggest that the pro-oxidant activity of T could be induced indirectly by its metabolite, E2, whereas the other metabolite, DHT, stimulates signal expression. Finally, the study shows that changes in oxidative damage or antioxidant status of plasma were not correlated with either skin redness or circulating carotenoids.     

Erratum to: Beak colour reflects circulating carotenoid and vitamin A levels in spotless starlings (Sturnus unicolor)

Many colourful sexually selected signals in animals are carotenoid-dependent and, because carotenoids function as antiradicals and immunostimulating molecules, carotenoid-dependent signals may honestly reflect the health state of individuals. Some others nutrients like vitamin A may also enhance health and colouration, but these have rarely been tested alongside carotenoids in colourful birds. Here, we examined whether beak colour of the spotless starling (Sturnus unicolor) reflected circulating levels of carotenoids and/or vitamin A (retinol). Spotless starlings are polygynous, sexually dimorphic birds (i.e. length of chest feathers). The tip of the beaks of male and female spotless starlings is more intensely coloured at the beginning of the breeding season and becomes dull after mating, which may suggest a sexual function. We found that females have a more intensely coloured beak and higher plasma carotenoid concentration than males during mating, and, despite the finding that carotenoid and vitamin A levels were not significantly related; colour intensity was positively correlated with plasma concentration of carotenoids and vitamin A in both sexes. However, adult beak coloration was not associated with carotenoid and vitamin A concentrations after nestlings were hatched. Therefore, beak colouration of spotless starlings provides information about circulating levels of carotenoids and vitamins during the mating season and may potentially function as a reliable signal of physiological status in the context of sexual selection.     

Carotenoid-derived ornaments reflect parental quality in male and female yellow-eyed penguins (<Emphasis Type="Italic">Megadyptes antipodes</Emphasis>)

The handicap principle suggests that ornamental traits that function as honest signals in mate selection must be costly to be effective. We evaluated in the sexually monochromatic yellow-eyed penguin (Megadyptes antipodes) whether the carotenoid-derived plumage and eye coloration predicts parental quality and whether males and females within pairs mate assortatively in relation to these carotenoid-derived ornaments. In addition, we investigated whether age or body condition was related to the coloration of the ornamental traits. In yellow-eyed penguins, parental quality of males and females was predicted by eye and head plumage coloration. Even when we controlled for gender- and age-specific differences, eye and head plumage coloration reflected honestly parental quality. Males and females mated assortatively in relation to these ornamental traits. While age influenced coloration of both the eye and head plumage, body condition was related only to the saturation of plumage coloration. These results provide evidence that the carotenoid-derived ornaments in yellow-eyed penguins reflect the parental abilities of birds and, therefore, may be costly signals. Potentially, female and male yellow-eyed penguins could use eye and plumage coloration as an indirect cue in assessing age and quality of individual birds during mate choice. This is only the second study to examine plumage coloration in relation to sexual selection in penguins, while conspicuous ornamental traits in other species of penguin beg the question whether they also play a role in sexual selection.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by C.R. Brown     

Male mate choice favors more colorful females in the gift-giving cabbage butterfly

While the phenomenon of male mate choice has attracted considerable attention in the last two decades, whether this sexual selection mechanism could drive the evolution of female ornaments remains poorly understood. Here, we used experimental manipulation of female wing coloration to investigate male mate choice in Pieris rapae, a gift-giving butterfly. Further, we tested whether males’ nutritional status influenced their mating preferences by subjecting larvae to short periods of starvation. We found that males showed significantly more mating approaches toward control females with more colorful wings (higher pteridine content), and that this preference was strongest in low-nutrition males. Additionally, a study of field-collected females revealed that pteridine-based wing coloration was positively correlated with female egg load, which suggests such ornaments may signal female quality. Pteridine-based ornaments are widespread in nature, however their potential as honest signals in male mate choice remains largely unexplored. This work furthers our understanding of how male mate choice and female ornamentation may evolve in species whose mating systems include nutritional nuptial gifts.     

Beak colour reflects circulating carotenoid and vitamin A levels in spotless starlings (Sturnus unicolor)

Many colourful sexually selected signals in animals are carotenoid-dependent and, because carotenoids function as antiradicals and immunostimulating molecules, carotenoid-dependent signals may honestly reflect the health state of individuals. Some others nutrients like vitamin A may also enhance health and colouration, but these have rarely been tested alongside carotenoids in colourful birds. Here, we examined whether beak colour of the spotless starling (Sturnus unicolor) reflected circulating levels of carotenoids and/or vitamin A (retinol). Spotless starlings are polygynous, sexually dimorphic birds (i.e. length of chest feathers). The tip of the beaks of male and female spotless starlings is more intensely coloured at the beginning of the breeding season and becomes dull after mating, which may suggest a sexual function. We found that females have a more intensely coloured beak and higher plasma carotenoid concentration than males during mating, and, despite the finding that carotenoid and vitamin A levels were not significantly related; colour intensity was positively correlated with plasma concentration of carotenoids and vitamin A in both sexes. However, adult beak coloration was not associated with carotenoid and vitamin A concentrations after nestlings were hatched. Therefore, beak colouration of spotless starlings provides information about circulating levels of carotenoids and vitamins during the mating season and may potentially function as a reliable signal of physiological status in the context of sexual selection.     

Multiple color signals may reveal multiple messages in male Schreiber’s green lizards, <Emphasis Type="Italic">Lacerta schreiberi</Emphasis>

Multiple traits may either signal different characteristics of a male or be redundant. These multiple signals may convey different messages if they are intended for different receivers (e.g., male or females) that have different interests. We examined the functions of multiple colorful visual traits of male Schreiber’s green lizard (Lacerta schreiberi). Results showed that interindividual variation in the characteristics of coloration of males can be related to variation in morphology, health state, dominance status, and pairing status, but that different relationships were found for each color signal. For example, dominant males had brighter “blue” throat and with higher values of ultraviolet (UV) and bluish coloration and darker and greenish dorsal coloration than subordinate males. Health state was also reflected in coloration; males with a higher immune response had “blue” throats with lower amounts of UV coloration, but had “yellow” chests with higher amounts of UV coloration. Males found guarding females also differed in coloration from males found alone. These data suggest that characteristics of coloration of the different multiple signals may reveal different messages for different receivers, either male or female conspecifics. The development of the different signals, based on different morphological and physiological mechanisms and trade-offs, may allow signal reliability of multiple colorful traits in different social contexts.     

Chemical ornaments of male lizards <Emphasis Type="Italic">Psammodromus algirus</Emphasis> may reveal their parasite load and health state to females

Evidence for parasite-mediated sexual selection has been found in many species that use visual ornaments to attract females. However, in many animals, variation in female responses to scents of parasitized males suggests that parasitic infections might also affect information conveyed by pheromones (i.e., chemical ornaments). Thus, pheromones might also function in parasite-mediated sexual selection. We show here that female lizards Psammodromus algirus responded differently to femoral gland secretions of males according to the parasite load and health of these males. Scents of healthier males elicited more tongue flicks (a chemosensory behavior) by females, suggesting that these scents were more attractive. Chemical analyses showed that parasite load and the T-cell-mediated immune response were related to the variability in the proportions of some lipids in secretions of males. Further trials testing the chemosensory responses of females to chemical standards indicated that females actually discriminated the chemicals related to males’ health from other chemicals found in secretions. We suggest that these chemical ornaments may provide reliable information on the health and degree of parasitic infection of a male.     

Antioxidant status, flight performance and sexual signalling in wild-type parrots

During mate choice, individuals are predicted to assess traits that honestly signal the quality of potential partners. Locomotor capacity may be such a trait, potentially signalling condition and ability to resist oxidative damage. In this study, we experimentally manipulated nutritional status: Male wild-type budgerigars, imported from Australia, were provided with either an enhanced (EQ) or reduced quality (RQ) diet varying in vitamin (particularly retinol and α-tocopherol) and mineral levels. Then, we assessed whether this influenced locomotor capacity, i.e. escape flight performance, and sexual attractiveness in male budgerigars Melopsittacus undulatus. Males in the EQ group showed significantly greater total antioxidant capacity and higher blood plasma concentrations of the dietary antioxidants retinol and α-tocopherol, but not carotenoids, than the RQ group. Over 8 weeks of flight training, males on the EQ diet showed significantly greater improvement on the most strenuous flight test than RQ males. In mate choice trials, females preferred EQ over RQ males. EQ males that were relatively fast in escape flight trials were more strongly preferred in the mate preference arena than their RQ competitors. Interestingly, males with high plasma carotenoid levels flew slower and were less attractive than males with low carotenoid levels. This might indicate that carotenoids are not effective antioxidants in birds. Overall, our results show that dietary-derived antioxidants can influence sexual attractiveness and other fitness-related traits through multiple pathways. Locomotor capacity appears to be an honest signal of male condition in birds.     

Is male rhesus macaque red color ornamentation attractive to females?

Male sexually selected traits can evolve through different mechanisms: conspicuous and colorful ornaments usually evolve through intersexual selection, while weapons usually evolve through intra-sexual selection. Male ornaments are rare among mammals in comparison to birds, leading to the notion that female mate choice generally plays little role in trait evolution in this taxon. Supporting this view, when ornaments are present in mammals, they typically indicate social status and are products of male-male competition. This general mammalian pattern, however, may not apply to rhesus macaques (Macaca mulatta). Males of this species display conspicuous skin coloration, but this expression is not correlated to dominance rank and is therefore unlikely to have evolved due to male-male competition. Here, we investigate whether male color expression influences female proceptivity toward males in the Cayo Santiago free-ranging rhesus macaque population. We collected face images of 24 adult males varying in dominance rank and age at the peak of the mating season and modeled these to rhesus macaque visual perception. We also recorded female sociosexual behaviors toward these males. Results show that dark red males received more sexual solicitations, by more females, than pale pink ones. Together with previous results, our study suggests that male color ornaments are more likely to be a product of inter- rather than intra-sexual selection. This may especially be the case in rhesus macaques due to the particular characteristics of male-male competition in this species.     

Female sensory bias may allow honest chemical signaling by male Iberian rock lizards

Some mate choice theories propose that only male signals that are honest and condition-dependent can be stable, while another hypothesis states that males evolve signals that exploit the sensory system of females. However, sensory traps might evolve into honest signals if they are differentially costly for males. We tested whether a pre-existing sensory bias for food chemicals explained chemosensory preferences of female Iberian rock lizards for male scents. We manipulated hunger levels of females and found that food-deprived females had increased chemosensory responses to chemical stimuli from both invertebrate prey and femoral secretions of males, but not to control water. Further tests suggested that cholesta-5,7-dien-3-ol (provitamin D₃), a lipid found in both prey and males’ scent, may be one of the chemicals eliciting these responses. Moreover, hungry females spent more time on scent marks of males that had experimentally increased cholesta-5,7-dien-3-ol than on scent marks of males alone, whereas for control females this effect was not significant. We suggest that preexisting sensory bias for essential nutrients (i.e., provitamin D) may be the origin of similar female responses to male chemicals. However, previous studies have suggested that the allocation of these chemicals to ornaments is costly and only high quality males can afford it. Therefore, preexisting sensory bias for essential nutrients may further allow the evolution and maintenance of honest sexual displays.     

Carotenoid-based ornamentation as a dynamic but consistent individual trait

Carotenoid-based ornaments act as signals of quality in many animal species. In contrast to feathers, which are relatively stable structures, carotenoid-pigmented integuments (e.g. bills, lores, tarsi) can change colour rapidly and may better reflect changes in physiological condition. I studied the seasonal variations in plasma carotenoids in red-legged partridges (Alectoris rufa) kept on a constant diet and free of intestinal parasites. Furthermore, I analyzed whether seasonal changes in circulating carotenoids were mirrored by the carotenoid-based coloration of eye rings and bill of this species. Plasma carotenoids showed seasonal variation, with higher levels coinciding with the end of the mating and the start of the laying season. Eye ring pigmentation was related to plasma carotenoid levels, and changes in bill hue (but not changes in UV or red bill chroma) mirrored the variation in plasma carotenoids during the breeding season. Despite the seasonal variation, individual differences in eye ring pigmentation and bill hue, UV and red chroma were consistent throughout the breeding season. Similarly, individual differences in eye ring pigmentation and bill hue and red chroma remained consistent between consecutive years. These results suggest that carotenoid based integumentary colorations act as dynamic traits that accurately reflect the carotenoid-status of individuals, thus reliably indicating consistent differences in individual quality. Furthermore, variability in signal expression appears to have a relevant genetic/phenotypic basis independently of environmental conditions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Patterns of egg yolk antioxidant co-variation in an avian brood parasite–host system

Despite their importance to offspring fitness outcomes, there has been little previous work on egg-mediated maternal effects in avian brood parasites which lay their eggs in the nests of other species. Here, we examine patterns of egg yolk antioxidant deposition in an avian host (red-winged blackbird, Agelaius phoeniceus) and their brood parasite (brown-headed cowbird, Molothrus ater). Cowbird nestlings compete directly for food with host blackbird chicks and succeed, despite hatching from a smaller egg, by begging more intensely and growing as or more rapidly than their hosts. Dietary-derived antioxidants, such as carotenoids and vitamins E and A, deposited in egg yolk, may enhance growth rate and reduce oxidative stress, and therefore provide a potential mechanism by which female cowbirds could enhance the competitiveness of their young in host nests. However, provision of antioxidants to eggs is costly so we predicted that female cowbirds should adjust the amount of antioxidants in response to variation in host quality. We found that whilst red-winged blackbird clutches parasitised by cowbirds did not differ in the pattern of antioxidant deposition compared to non-parasitised clutches, levels of vitamin E deposited in cowbird eggs were closely matched to that of the individual host clutch in which cowbirds laid their egg. This supports the prediction that female cowbirds adjust the antioxidant concentration of their eggs to increase the likely competitiveness of their offspring in the host nest into which they are laid and may help explain the success of cowbirds in competing with larger host young.     

Carotenoid-based colouration and ultraviolet reflectance of the sexual ornaments of grouse

Among the most familiar sexual signals are red, yellow, and orange sexual traits pigmented by carotenoids. Many birds can detect near-ultraviolet (UV) light, and UV signals can play key roles in mate choice. Grouse (Tetraonidae) exhibit bright carotenoid-dependent sexual ornaments, their supra-orbital combs, which to humans appear orange-red. Combs also reflect in the UV, which is not visible to humans but is likely to be visible to grouse. In male red grouse Lagopus lagopus scoticus, we show that comb UV reflectance decreases with increasing comb size and redness. By removing the epidermis of combs, where carotenoid pigments are, we show that the UV reflectance is a property of the dermis, underneath the red pigmented epidermis. Carotenoid pigmentation of combs acted as a mask to reduce reflectance by the dermis in the range 400–550 nm and in the UV, 300–400 nm. Patagium skin (non-ornamental skin under the wing) also reflects in the UV, but epidermis removal on this bare part tended to reduce UV reflectance, whereas removal of the red epidermis of combs increased UV reflectance. Males in better condition (greater body mass relative to size) had bigger and redder combs, but with less UV. Thus, carotenoid pigments of grouse combs are deposited on a white background with significant UV reflectance, which can influence how the signal is perceived by conspecifics. Carotenoid-based traits exhibit UV reflectance in a number of species, but how UV reflectance and carotenoid pigmentation influence colour remains little known for integumentary ornaments compared to plumage traits. UV vision is not uncommon in birds and other animals, so future studies should investigate how UV reflectance influences the perception of carotenoid-based signals of quality.     

A simple and inexpensive chemical test for behavioral ecologists to determine the presence of carotenoid pigments in animal tissues

Animals use several different types of pigments to acquire their colorful ornaments. Knowing the types of pigments that generate animal colors often provides valuable information about the costs of developing bright coloration as well as the benefits of using these signals in social or sexual contexts. It is often assumed that red, orange, and yellow colors in animals are derived from carotenoid pigments, when in fact there are other pigments that confer similar colors on animals. These include the pteridine pigments in a wide range of organisms, hemoglobin in blood-filled sinuses, the psittacofulvins of parrot feathers, and the phaeomelanin pigments in rufous or yellow feathers and fur. In this paper, we describe a quick and easy, two-step chemical method for field biologists to determine if their study species uses carotenoid pigments as integumentary colorants. This laboratory procedure first employs a thermochemical extraction technique, in which acidified pyridine is used under high temperature to free carotenoid pigments from tissue to produce a colorful, pigmented solution. Red, orange, or yellow tissues containing pteridines, hemoglobin, or eumelanins do not release colored pigments into heated pyridine. However, psittacofulvins, and occasionally phaeomelanins, will also solubilize using this method. Thus, a follow-up test is needed, using solvent transfer, to confirm the presence of carotenoids in animal tissues. The use of absorbance spectrophotometry on the colorful solution may also provide information about the predominant carotenoids that bestow color on your study animal.     

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.     

Pigment allocation to eggs decreases plasma antioxidants in a songbird

Although evidence is accumulating on the adaptive function of female ornamentation, very little is known about maternal allocation decisions involving sexual signaling and other reproductive functions. Blue egg coloration has been suggested as a sexually selected signal of female quality to males, and some recent studies are in accordance with this hypothesis. Blue eggshell coloration results from the deposition of biliverdin pigment by laying females, which is a potent antioxidant. Thus, egg pigmentation should be costly in terms of antioxidants, an assumption of the signaling hypothesis that has not been tested yet. We induced increased reproductive effort in a set of female pied flycatchers Ficedula hypoleuca through nest removal and measured egg pigmentation and plasma antioxidant levels in relation with a control group. Experimental females showed a negative association between egg color and plasma antioxidant levels, while there was no relationship for control birds. This supports that egg pigmentation is costly in terms of general antioxidant defenses and suggests a tradeoff between the allocations to both traits. Furthermore, experimental females with more colorful eggs raised more fledglings, especially when breeding early. Controls did not show a relationship between egg color and reproductive success. Females laying more colorful eggs could have shifted their allocation decisions towards current reproduction, at the expense of their own antioxidant defenses. Our results highlight that blue egg coloration is a life-history trait, subject to tradeoffs with other attributes, and seems to be especially informative under harsh breeding conditions.     

Vitamin E supplementation enhances growth and condition of nestling barn swallows (Hirundo rustica)

To optimize fitness, organisms may have to trade the number and quality of individual offspring against their own condition and survival. Limiting micronutrients such as antioxidants may be crucial to this trade-off. We investigated whether vitamin E, a major antioxidant in the diet of vertebrates, is limiting to barn swallow (Hirundo rustica) nestlings. We manipulated brood size to alter the intensity of sib–sib competition and supplemented nestlings with two different physiological doses of vitamin E while establishing a control group. Treatment effects were measured on body mass and size, feather growth, T cell-mediated immune response and hematocrit. Supplementation with vitamin E at intermediate physiological doses improved nestling mass and condition and feather growth, whereas higher physiological doses did not enhance offspring quality compared to a control treatment. The positive effects of vitamin E on body mass and condition were only detectable from days 6 to 10 when maximum growth rate is attained. Experimental enlargement of broods reduced body mass and size and T cell-mediated immune response only during the late nestling period. The effect of vitamin E supplementation did not depend on brood size manipulation, as revealed by the nonsignificant statistical interaction. This result contradicts the hypothesis that availability of vitamin E depends on intrabrood competition and instead suggests that it depends on concentration of vitamin E in the insect prey of swallows. Thus, antioxidants may be available in limited amounts to barn swallow nestlings and such limitation affects growth. In addition, present results confirm that barn swallow parents trade progeny number against growth and immunity of individual offspring.     

Feather coloration in museum specimens is related to feather corticosterone

Colorful ornaments in birds are often sexually selected signals of quality, and variation in ornament expression may be mediated by physiological stress through the secretion of corticosterone. However, testing for links between ornamentation and corticosterone often requires sampling live animals, and such physiological measures may not be matched in the time span in which they were sampled (e.g., very dynamic plasma corticosterone vs. plumage coloration, which is relatively static). Here, we use museum specimens to test for a link between the color of a sexual ornament and feather corticosterone at the time of ornament formation. In red-winged blackbirds, Agelaius phoeniceus, carotenoid-based epaulets appear to be important in male–male social interactions, territory maintenance, and female choice. We measured reflectance spectra of adult male epaulets and plucked adjacent feathers for corticosterone analysis via radioimmunoassay. We controlled for differences in the number of mates, specimen age, and geography by selecting only males with one mate and only birds collected in Florida during a 3-year period. Epaulet hue and red chroma did not vary with feather corticosterone, but males whose epaulets scored high for mean brightness and red brightness had significantly lower corticosterone than males with low brightness scores. This correlation with brightness but not hue or chroma is consistent with an effect of corticosterone (CORT) on feather microstructure, with elevated CORT leading to lower reflectance of white light from the keratin matrix surrounding the carotenoid pigments.     

Responses of female lizards,<Emphasis Type="Italic"> Lacerta monticola</Emphasis>, to males' chemical cues reflect their mating preference for older males

Female mate choice based on visual traits appears to be rare in lizards. Field observations suggest that females of the lizard Lacerta monticola preferred to mate with larger/older males. Although older males are usually green and larger, and younger males brown and smaller, there is some overlap in size and coloration between age classes. Thus, visual cues may not always be reliable indicators of a male's age. We hypothesized that female mate-choice preferences may be based on males' pheromones, which might transmit information about characteristics such as age. In a laboratory experiment, we analyzed the effect of age of males on attractiveness of their scents to females. When we offered scents of two males of different age, females associated preferentially with scents of older males. This suggested that females were able to assess the age of males by chemical signals alone, and that females preferred to be in areas scent-marked by older males. Thus, females may increase their opportunities to mate with males of high quality, or may avoid harassment by sneaking young males. This result agreed with field observations on females mating with old males, and rejection of advances by young males. Our results also suggested that female preference for older males may depend on their own body size. Large females showed a strong preference for older males, whereas smaller females were not so selective. This, together with males' preference for large females, might lead to size-assortative matings. We suggest that the quality and/or quantity of male pheromones could communicate to the female heritable male genetic quality (i.e. age) and thereby serve as the basis of adaptive female choice in lizards.Communicated by W.E. Cooper