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.     

Biliverdin-based egg coloration is enhanced by carotenoid supplementation

Avian eggshell color is remarkably variable among and within species and its possible function has long been controversial. Female birds use biliverdin pigment to color their eggs blue and green. Although evidence is accumulating that blue-green eggshell coloration is costly to produce, the exact mechanisms underlying its expression are little studied. Biliverdin pigment is involved in important maintenance functions such as immune and antioxidant defenses and shows similar properties to carotenoid pigments. Carotenoids play a role as immunoenhancers and have the potential to ameliorate the impact of oxidative processes, although their antioxidant function could be minor for some bird species. Important life-history components can be limited by carotenoid availability, including sexual displays and fecundity. Here we explored if biliverdin-based eggshell pigmentation was affected by carotenoid availability in a seabird, by performing a carotenoid supplementation experiment in female blue-footed boobies (Sula nebouxii) after laying their first egg. In this species, blue eggshell color is associated with incubation patterns, and carotenoid availability affects the crucial trade-off between female foot ornamentation and fecundity. We found that under natural conditions, there was a decline in eggshell color with laying order, suggesting pigment limitation for females. However, carotenoid-supplemented females had enhanced second egg coloration compared to controls, and the color increase was independent of their previous pigment allocation to the first egg. Our results suggest that biliverdin-based eggshell coloration is costly to produce and can be alleviated by current carotenoid availability.     

Egg coloration is correlated with female condition in eastern bluebirds (Sialia sialis)

Egg coloration has been hypothesized to reflect female condition. Because of the proposed physiological costs associated with deposition of biliverdin pigments and because of their conspicuousness, eggs with blue-green coloration may reliably convey information about female or brood quality. We tested the hypothesis that expression of blue-green coloration of eastern bluebird (Sialia sialis) eggs positively correlates to female condition. First, we documented the incidence of egg color polymorphism within the population. We observed that 98% of females laid blue-green eggs while less than 2% laid white eggs and less than 1% laid pink eggs. In a subset of clutches, we used full spectrum reflectance spectrometry (300–700 nm) to compare eggshell coloration to measures of female condition. We found that the color of eggs within clutches was more similar than the color of eggs from different clutches, and that the blue-green eggs have spectral peaks that are consistent with the characteristic absorbance spectra of biliverdin pigmentation. Females in better body condition and older females laid more colorful eggs. Moreover, individual females laid more colorful eggs later in the laying sequence. Overall, these data indicate that egg coloration covaries with female condition, suggesting that egg coloration could function as a reliable signal of female quality or that egg coloration may allow females to recognize eggs laid by conspecific brood parasites.     

Are blue eggs a sexually selected signal of female collared flycatchers? A cross-fostering experiment

Impressive variation in egg colouration among birds has puzzled evolutionary biologists for a long time. The most frequently studied selective forces moulding egg colouration—predation and brood parasitism—have either received little empirical support or may play a role in only a minority of species. A novel hypothesis has suggested that egg colour may be significantly influenced by sexual selection. Females may deposit a blue-green pigment biliverdin into eggshells instead of using it for themselves as a powerful antioxidant. By this handicap, females may signal their quality to males, which are then hypothesized to increase their paternal effort. We experimentally tested the hypothesis in the collared flycatcher (Ficedula albicollis), a species laying blue-green eggs. We cross-fostered clutches between nests to disentangle effects of female/territory quality and egg colour on paternal effort and nestling quality. The results supported two assumptions of sexual signalling through egg colour hypothesis: Blue pigment seems to be a limited resource for females, and female quality is positively correlated with the intensity of the blue-green colour. However, we did not find support for the main prediction of the hypothesis, as male parental effort parameters (feeding frequencies to nestlings and intensity of nest defence) were unrelated to egg colour. We discuss possible reasons for the discrepancy between our results and previous correlative analyses that supported the hypothesis that blue egg colour may be a postmating, sexually selected signal in females.     

Maternal investment in eggs is affected by male feet colour and breeding conditions in the blue-footed booby, <Emphasis Type="Italic">Sula nebouxii</Emphasis>

Females are expected to vary investment in offspring according to variables that may influence the offspring fitness in a way that optimises her inclusive fitness for a particular context. Thus, when sexual ornaments signal the quality of the male, females might invest in reproduction as a function of the attractiveness of their mate. We tested whether breeding conditions and male feet colour influence reproductive decisions of blue-footed booby females. In the blue-footed booby, male feet colour is a dynamic condition-dependent sexually selected trait that is related to paternal effort. During two consecutive years, an El Niño year (poor breeding conditions) and a year with good breeding conditions, we experimentally reduced male attractiveness by modifying their feet colour after the first egg was laid and recorded female investment in the second egg. We found that, relative to the first egg in the clutch, females laid heavier second eggs during the poor year than during the good year. Females paired with males with duller feet colour reduced second-egg mass and volume and delayed the laying of the second egg, independently of the year. Absolute yolk androstenedione (A4) concentration (but not testosterone, T) in second eggs was higher during a poor year than during a good year. Only during a year with poor breeding conditions, females paired with experimental males decreased the relative A4 concentration (but not T) in the second egg compared to control females. Thus, blue-footed booby females probably favour brood reduction by decreasing egg quality and increasing size asymmetry between chicks when the breeding and the mate conditions are poor.     

Sexual selection based on egg colour: physiological models and egg discrimination experiments in a cavity-nesting bird

It has been proposed that the blue-green bird egg colourations of many avian species may constitute a sexually selected female signal that males can use to modulate their parental investment. A fundamental untested assumption for the validation of this hypothesis is that males can accurately assess differences in the colour of eggs. A recent review suggests that this could be particularly problematic when egg clutches were located within a dimly lit nest cavity, due to limitations of the visual system in low light conditions. Here, we first used a photoreceptor noise-limited model of colour discrimination ability that accounts for visual performance under low light conditions to study whether a typical cavity-nesting passerine, the spotless starling Sturnus unicolor, can discriminate their eggs under the ambient illumination in their nest-holes. Secondly, we tested the validity of model predictions with behavioural data collected in two egg discrimination experiments performed in this species. Estimated egg detectability depended entirely on model assumptions about visual limitations linked to light intensity. Starlings would not be able to discriminate egg differences in their nests if the model was based on the assumption that light intensity limited detectability, whereas they could potentially perceive as different many possible pairwise clutch comparisons if the model assumption was that light intensity did not limit detectability. Results of behavioural experiments fitted the prediction of the visual model where light intensity did not limit detectability. Our results suggest that photoreceptor noise-limited colour models based on stimulation of single photoreceptors cannot, at present, be used to predict egg discrimination ability in spotless starlings under low light conditions. Future studies aiming to test egg discrimination constraints in the frame of the sexual selection hypothesis should therefore combine both modelling and behavioural experiments to determine which are the components of the models that produce the mismatch with the behavioural conditions.     

Intra-specific interactions influence egg composition in the lesser black-backed gull (Larus fuscus)

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.Communicated by J. Graves     

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.     

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.     

Are yolk androgens and carotenoids in barn swallow eggs related to parental quality?

Recently, evidence is mounting that females can adaptively engineer the quality of their offspring via the deposition of yolk compounds, including carotenoids and androgens. In this study, we simultaneously consider how both carotenoids and androgens in egg yolk relate to parental quality in barn swallows (Hirundo rustica erythrogaster). First, we found no relationship between concentrations or amounts of yolk androgens and carotenoids. Yolk carotenoids decreased with laying order, whereas we found no relationship between yolk androgens and laying order. Second, we tested the Investment Hypothesis, which predicts that high-quality females or females paired to high quality mates, allocate differentially more of these yolk compounds to their offspring. For carotenoids, we mostly found evidence to counter predictions of the Investment hypothesis: (1) Carotenoid concentrations varied among females, (2) heavier eggs contained lower carotenoid concentrations, although heavier yolks contained greater amounts of carotenoids, (3) eggs of earlier-laying females had lower concentrations in their eggs, and (4) yolk carotenoids were not correlated with clutch size or male plumage ornamentation. For androgens, we found weak support for the Investment Hypothesis: (1) Yolk androgens varied among females, (2) heavier eggs and yolks contained greater amounts, although not concentrations of androgens, (3) females paired to more colorful males laid eggs with greater concentrations of androgens, and (4) no effects of laying date or morphological correlates of female quality on androgen concentrations in egg yolks. Overall, these findings suggest that each yolk compound may have different functions and therefore may be regulated by different mechanisms.     

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.     

Carotenoids in the egg yolks of collared flycatchers (<Emphasis Type="Italic">Ficedula albicollis</Emphasis>) in relation to parental quality,environmental factors and laying order

Birds may influence the fitness of their offspring by transmission of different amounts of carotenoids to their eggs. Carotenoids play crucial roles in antioxidant protection and immune defence mechanisms, but they may be available to females in limiting amounts. Therefore, their allocation to the eggs may be influenced by the female’s condition, age and environmental circumstances. Furthermore, the quality of the male parent, which affects the reproductive value of the offspring, may also influence this investment. In this correlational study, we investigated proximate and ultimate factors that may lead to variation in yolk lutein, zeaxanthin and β-carotene concentrations among and within clutches of a wild passerine, the collared flycatcher (Ficedula albicollis). We found that carotenoid concentration was positively associated with caterpillar supply at the time of egg formation, which suggests a proximate constraint of carotenoid availability on yolk composition. Neither female condition, body size, age, nor male plumage ornamentation, age and body size correlated with carotenoid deposition. Yolk β-carotene concentration was found to be positively linked to yolk testosterone concentration. We suggest that females allocated more β-carotene to their eggs to mitigate the potentially detrimental effects of elevated steroid concentration. We found that concentration of β-carotene increased with laying order. The possible function of this pattern may be to enhance the resistance to oxidative stress and pathogens of the disadvantaged last-hatching nestling, suggesting that collared flycatchers pursue a compensatory, “brood survival” strategy.     

Female house martins (Delichon urbica) reduce egg androgen deposition in response to a challenge of their immune system

Female birds deposit in the yolks of eggs substantial amounts of androgens, such as testosterone and androstenedione. These androgens have been shown to speed up nestling development, induce a fast development of ornaments and increase dominance in adults. Experiments in several species have reported that females invest greater amounts of androgens in the eggs fathered by attractive males, suggesting that yolk androgen is a costly investment for either the offspring or the mother. There is some evidence that nestling immunocompetence may be partially suppressed by high levels of yolk androgens, but it is not known whether this is also the case for females. We tested this hypothesis in the house martin by inducing an immune challenge through an injection of sheep red blood cells, a standard challenge of the humoral immune system. Experimental birds laid eggs with lower amounts of yolk androstenedione than controls, and there was a similar non-significant trend for testosterone. Furthermore, the probability of laying a replacement clutch was higher for birds that had laid a first clutch with relatively high levels of yolk testosterone. These results suggest that yolk androgen deposition is limited by immune costs in the female, and that only females in good condition may afford to invest high levels of androgen in eggs in this species.     

Detecting pigments from colourful eggshells of extinct birds

The known chemical basis of diverse avian eggshell coloration is generated by the same two classes of tetrapyrrole pigments in most living birds. We aimed to extend the evolutionary scope of these patterns by detecting pigments from extinct birds’ eggs. In our samples biliverdin was successfully extracted from subfossil shell fragments of the blue-green egg-laying upland moa Megalapteryx didinus, while protoporphyrin was extracted from the beige eggs of two other extinct moa species. Our data on pigment detection from eggshells of other extant paleognath birds, together with published information on other modern lineages, confirm tetrapyrroles as ubiquitous and conserved pigments contributing to diverse eggshell colours throughout avian evolution.     

Male sexual attractiveness affects the investment of maternal resources into the eggs in peafowl (<Emphasis Type="Italic">Pavo cristatus</Emphasis>)

According to the differential investment hypothesis, females paired with attractive mates are expected to invest more in the current reproduction relative to females paired with unattractive males. We experimentally tested this hypothesis in the peafowl (Pavo cristatus) by providing females with males that differed in sexual attractiveness. In agreement with the differential allocation hypothesis, females paired with more ornamented males laid larger eggs, and deposited higher amounts of testosterone into the egg yolk, independently of the sex of the embryo. These results show that the association between paternal phenotype and offspring quality could arise via a differential maternal investment. They also suggest that, if ornamented males do transmit good genes to the progeny, the maternal differential investment can amplify the effect of such good genes on the offspring fitness.     

Environmental conditions influence egg color of reed warblers <Emphasis Type="Italic">Acrocephalus scirpaceus</Emphasis> and their parasite,the common cuckoo <Emphasis Type="Italic">Cuculus canorus</Emphasis>

The outer layer of the eggshell in birds is in many cases covered by pigments that are assumed to be genetically determined traits with a negligible environmental component. To test the hypothesis that spring environmental conditions (i.e., temperature and rainfall) may affect bird egg pigmentation, we measured by spectrophotometry and photography egg coloration and spottiness on reed warbler (Acrocephalus scirpaceus L.) clutches parasitized by the common cuckoo (Cuculus canorus L.) collected over a period of 24 years and preserved in the Zoological Museum, Copenhagen, Denmark. In addition, we investigated whether spring environmental conditions may influence the coevolutionary relationship between the cuckoo and its host via changes in cuckoo–host egg matching. Generalized mixed models revealed that reed warbler eggs were more brilliant in those springs with a higher rainfall and tended to be bluer and greener in springs with a lower relative temperature. On the other hand, cuckoo eggs were bluer and greener in springs with a higher rainfall. Cuckoo–host egg matching in blue-greenness and spottiness was better in springs with a higher rainfall. These results provide support for the existence of an environmental component on bird egg coloration and suggest that environmental factors may potentially affect the outcome of important features of the arms race between cuckoos and reed warblers.     

Effects of elevated yolk testosterone levels on survival, growth and immunity of male and female yellow-legged gull chicks

Androgen hormones of maternal origin contained in the eggs of avian species are considered to have positive effects on offspring characteristics and performance. However, negative consequences have also been reported, suggesting that mothers may experience a trade-off between beneficial and detrimental effects of egg androgens to offspring fitness. We studied the effects of elevated yolk testosterone (T) concentration on survival, development and phenotype of male and female yellow-legged gull (Larus michahellis) chicks by injecting egg yolks with physiological doses of the hormone. Elevated yolk T resulted in a male-biased post-hatching sex ratio, T-treated clutches producing a greater proportion of males compared to control ones at day 4 post-hatching, likely resulting from a reduction of female embryonic survival, whereas no effect of hormone treatment on hatching success or short-term chick survival was observed. In addition, T depressed post-hatching body mass in both sexes but had no effects on the intensity of the cell-mediated immune response or skeletal growth. No sex differences in egg characteristics or chick phenotype were detected. Time to hatching was not affected by T, but females originating from first laid eggs hatched earlier than males of the same laying order, independently of hormone treatment. However, the implications of sex differences in hatching times are unclear in the study species. Taken together, our results suggest that female yellow-legged gulls may be constrained in transferring androgens to their eggs by negative consequences on the viability of female offspring and growth of chicks of the two sexes.     

Long-lasting consequences of elevated yolk testosterone levels on female reproduction

Maternal yolk androgens in bird eggs represent an important pathway along which offspring phenotype is shaped. Most of the hormone-mediated maternal effects are highly important in the context of sibling competition. However, there is also increasing evidence for long-lasting effects far beyond the nestling period, and these effects may have important consequences on the reproductive success of the offspring. Here, we investigated the effects of experimentally elevated yolk testosterone concentrations on growth and reproduction in female canaries. Elevated yolk testosterone concentrations enhanced the post-natal growth rate, but not the asymptotic mass, and reduced the survival probability. The latter may be a consequence of the higher growth rate, which may have rendered females hatching from testosterone-treated eggs (T-females) more vulnerable to harsh environmental conditions. Adult T-females made a larger investment in their clutch by laying more but not heavier eggs than females hatching from control-treated eggs. Our results suggest that the observed long-lasting effect on clutch size relates to changes in the growth trajectory rather than being a direct consequence of testosterone, since studies manipulating early growth conditions obtained similar results. Clearly, further studies are now required in order to investigate the intriguing relationship between yolk testosterone, elevated growth rates, and clutch size.     

Eggshell spotting in brood parasitic shiny cowbirds (<Emphasis Type="Italic">Molothrus bonariensis</Emphasis>) is not linked to the female sex chromosome

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

Consequences of experimentally elevated yolk testosterone levels for intra- and inter-sexual selection in canaries

Hormones of maternal origin transferred to the eggs of oviparous species have been shown to significantly affect offspring development. Furthermore, there is now increasing evidence that these effects may last into adulthood. This underlines the persistence of yolk hormone-mediated maternal effects as well as their trans-generational potential as these changes may involve fitness-related traits such as mate choice behaviour, reproductive traits and longevity. Here, we tested the potential of yolk testosterone to affect sexual selection by experimentally increasing the yolk testosterone levels via egg injections. We focused on two central axes of sexual selection, male–male competition for access to a female (intra-sexual selection) and female mate choice behaviour (inter-sexual selection), using canaries (Serinus canaria) as a model species. Neither male agonistic behaviour nor access to the opposite sex, as measured in staged male–male encounters in the presence of a female, were affected by experimentally elevating yolk testosterone levels. We did not find any evidence for effects on female mate choice behaviour either, given the lack of significant effects on mate choice activity, consistency in female mate choice or choosiness. In conclusion, our results indicate that the consequences of yolk testosterone for sexual selection through changes in behavioural traits, which are expressed during pair formation or male–male competition, are probably limited.