Food availability affects the maternal transfer of androgens and antibodies into eggs of a colonial seabird

Mothers can improve the quality of their offspring by increasing the level of certain components in their eggs. To examine whether or not mothers increase deposition of such components in eggs as a function of food availability, we food-supplemented black-legged kittiwake females (Rissa tridactyla) before and during egg laying and compared deposition of androgens and antibodies into eggs of first and experimentally induced replacement clutches. Food-supplemented females transferred lower amounts of androgens and antibodies into eggs of induced replacement clutches than did non-food-supplemented mothers, whereas first clutches presented no differences between treatments. Our results suggest that when females are in lower condition, they transfer more androgens and antibodies into eggs to facilitate chick development despite potential long-term costs for juveniles. Females in prime condition may avoid these potential long-term costs because they can provide their chicks with more and higher quality resources.     

Prenatal androgen exposure modulates cellular and humoral immune function of black-headed gull chicks

Avian eggs contain considerable amounts of maternal yolk androgens, which have been shown to beneficially influence the physiology and behaviour of the chick. As androgens may suppress immune functions, they may also entail costs for the chick. This is particularly relevant for colonial species, such as the black-headed gull (Larus ridibundus), in which the aggregation of large numbers of birds during the breeding season enhances the risk of infectious diseases for the hatching chick.To test the effect of maternal yolk androgens on the chick's immune function, we experimentally manipulated, in a field study, yolk androgen levels within the physiological range by in ovo injection of either androgens (testosterone and androstenedione) or sesame oil (control) into freshly laid eggs. We determined cell-mediated immunity (CMI) and humoral immunity of the chicks at the beginning of the nestling period to evaluate early modulatory effects of yolk androgens on immune function.Embryonic exposure to elevated levels of androgens negatively affected both CMI and humoral immunity in nestling gull chicks. Consequently, maternal yolk androgens not only entail benefits of enhanced competitiveness and growth as previously shown, but also costs in terms of immunosuppression. The outcome of embryonic yolk androgen exposure thus likely depends on the post-hatching circumstances for the developing offspring such as parasite exposure and degree of sibling competition.     

Maternal androgens in the pied flycatcher: timing of breeding and within-female consistency

Maternal hormones can have substantial phenotypic effects in the progeny of many vertebrates. It has been proposed that mothers adaptively adjust hormone levels experienced by particular young to optimize their reproductive output. In birds, systematic variation in egg hormone levels has been related to different female reproductive strategies. Because in many bird species prospects of the offspring change seasonally and with brood number, strategic adjustment of yolk androgen levels would be expected. To test this idea, we induced pied flycatcher (Ficedula hypoleuca) females to nest twice during the same season by removing their first clutches shortly after clutch completion. We collected eggs of first and replacement clutches to measure yolk concentrations of androstenedione (A4) and testosterone (T) and captured the females that laid these clutches for phenotypic measurements. Although average egg androgen levels were remarkably consistent within females, hormone patterns differed considerably between first and replacement clutches. Eggs of replacement clutches were heavier with larger yolks compared to first clutches, but they contained on average lower levels of androgens. Within clutches, androgen concentration increased over the laying sequence in the first clutch, but decreased or remained more constant over the laying sequence in the replacement clutch. Mean yolk T, but not A4 levels, were negatively associated with laying date for both breeding attempts. Moreover, females in good body condition produced eggs containing lower levels of androgens than females in poor condition. Our results are consistent with the idea that differences in yolk androgen levels may be one mechanism underlying seasonal variation in reproductive success and it is possible that changes in egg androgen patterns may reflect a change in female reproductive strategy. High within-female consistency also highlights the possibility that there may be some underlying genetic variation in yolk androgen levels.     

Maternal condition, yolk androgens and offspring performance: a supplemental feeding experiment in the lesser black-backed gull (Larus fuscus)

It has been proposed that the maternal androgens in avian egg yolk enhance offspring fitness by accelerating growth and improving competitive ability. Because egg quality is strongly influenced by maternal condition, we predicted that females in good condition would produce high-quality eggs with relatively high androgen content. We experimentally enhanced maternal condition by supplementary feeding lesser black-backed gulls (Larus fuscus) during egg formation and compared the concentrations of androstenedione (A4), 5alpha-dihydrotestosterone (DHT) and testosterone (T) in their eggs with those in eggs laid by control females. We also measured circulating levels of T in females immediately after laying. Egg androgens could affect offspring performance directly through chick development and/or indirectly through changes in the competitive ability of a chick relative to its siblings. To avoid confounding these two routes, and to separate effects operating through the egg itself with those operating through experimental changes in parental chick rearing capacity, we fostered eggs from both maternal treatment groups singly into the nests of unmanipulated parents. Contrary to expectation, mothers with experimentally enhanced body condition laid eggs with lower levels of androgens, while exhibiting higher circulating T concentrations post-laying. Despite these lower levels of egg androgen, offspring hatched from eggs laid by mothers in good condition did not show reduced growth or survival when reared in the absence of sibling competition. Our results demonstrate that yolk androgen concentrations vary with the body condition of the female at the time of egg formation and that females in good condition reduced the yolk androgen content of their eggs without altering offspring performance.     

Subtle manipulation of egg sex ratio in birds

Mothers are predicted to overproduce male or female eggs when the relative fitness gains from one sex are higher and outweigh the costs of manipulation. However, in birds such biases are often difficult to distinguish from differential embryo or chick mortality. Using a molecular technique to identify the sex of early embryos, we aim to determine the effect of maternal nutrition on zebra finch (Taeniopygia guttata) egg sex ratios after 2 days of incubation, which is as close to conception as is currently possible. We found no overall bias in the sex ratio of eggs laid and sex did not differ with relative laying order under any diet regime. However, mothers on a low-quality diet did produce a female bias in small clutches and a slight male bias in large clutches. On a high-quality diet, mothers produced a male bias in small clutches and a female bias in large clutches. Those on a standard diet produced a roughly even sex ratio, irrespective of clutch size. These observed biases in egg sex are partly in line with predictions that, in this species, daughters suffer disproportionately from poor rearing conditions. Thus, when relatively malnourished, mothers should only rear daughters in small broods and vice versa. Sex-ratio patterns in this species therefore appear to be subtle.     

Maternal androgens in black-headed gull (Larus ridibundus) eggs: consequences for chick development

We tested the hypothesis that mother birds counterbalance the negative effects of hatching asynchrony for later-hatched chicks by increasing the yolk androgen concentrations in consecutive eggs of their clutch. In doing so, they may adaptively tune each offspring's competitive ability and, thus, growth and survival. However, evidence in support of this hypothesis is contradictory. The yolk concentrations of maternal androgens in the eggs of black-headed gulls increase significantly with the laying order of the eggs in a clutch. We experimentally tested the functional consequences of this increase on chick development under natural conditions by injecting eggs with either an oil or androgen solution. We created experimental clutches in which androgen levels either stayed constant or increased with laying order while controlling for differences in egg quality by using only first-laid eggs. We then compared development, growth and survival between these broods. Androgen treatment enhanced embryonic development because androgen-treated eggs hatched half a day earlier than controls, while their size at hatching was similar to oil-treated controls. Androgen treatment did not increase chick survival, but it enhanced growth. Androgen-treated, third-hatched chicks had a higher body mass and longer legs than third-hatched chicks that hatched from oil-treated eggs. At the same time, growth of first chicks (which were all oil treated) was reduced by the presence of two androgen-treated siblings, suggesting that yolk androgens enhance the competitive ability of later-hatched chicks. Our results support the hypothesis that transfer of different amounts of androgens to the eggs of a clutch is a mechanism by which mothers maximize their reproductive output.     

Hatching asynchrony and maternal androgens in egg yolks of House Wrens

Synchronously and asynchronously hatching clutches of House Wrens Troglodytes aedon usually do not differ in reproductive success. Thus late-hatching nestlings in asynchronously hatching clutches somehow overcome any age- and size-related disadvantages of hatching after their nest-mates. One possible way for them to do this is for female House Wrens to add maternal androgens to the yolk of late-hatching eggs. We tested this hypothesis in a wild population of House Wrens that produces both asynchronously and synchronously hatching clutches. Yolks of eggs from both types of clutches were biopsied and the eggs returned to their nests to hatch. Radioimmunoassays revealed that total androgen levels in the yolk varied within and among clutches. However, total androgen levels in yolks did not vary predictably with egg position in either synchronously or asynchronously hatching clutches. Thus, deposition of androgens in yolk did not follow the expected pattern based on the potential for sibling competition in House Wrens.     

Avian mothers create different phenotypes by hormone deposition in their eggs

An increasing number of studies in a variety of taxa demonstrate the role of maternal sex steroids on offspring development. In avian species, mothers deposit substantial amounts of androgens in their eggs, and experimental evidence indicates that these maternal androgens influence the chick's early development. Despite the well-known organizing role of sex steroids on brain and behaviour, studies on avian maternal egg hormones almost exclusively focus on the chick phase. Here, we show experimentally that in Black-headed gulls maternal androgens in the egg enhance the development of the nuptial plumage and the frequency of aggressive and sexual displays almost 1 year after hatching. We conclude that maternal sex steroids may be a key factor for the determination of subtle but important individual differences within the same sex and species, which may have important consequences for Darwinian fitness and evolutionary processes.     

Interfemale variation in egg yolk androgen allocation in the European starling: do high-quality females invest more?

Androgenic hormones occur naturally in bird egg yolk and are known to enhance growth in canary and gull chicks. Gil et al. (1999, Science, 286, 126-128) have recently proposed that female allocation of androgens to eggs represents a form of costly maternal investment. This hypothesis predicts that females of high quality or high reproductive potential should invest more yolk androgens in their eggs than females of low quality or low reproductive potential. We tested these predictions by examining interfemale variation in allocation of the androgenic steroids testosterone and androstenedione to eggs in a wild population of European starlings, Sturnus vulgaris. We collected 30 full clutches of eggs and captured 22 of the females that laid these clutches for phenotypic measurements. In agreement with the hypothesis, we found that there was significant interfemale variation in yolk androgen concentrations. Furthermore, older females deposited more androstenedione and testosterone in eggs than 1-year-old females, and females laying early or large clutches deposited more testosterone in eggs than females laying late or small clutches. However, females in good body condition did not deposit more androgens in eggs than females in poor body condition. Large-yolked eggs had a higher total yolk androgen content than small-yolked eggs, but larger eggs did not have higher concentrations of yolk androgens. The results in general support the investment hypothesis of yolk androgen allocation. However, further data are needed regarding the costs of yolk androgen allocation and the effects of yolk androgens on offspring to substantiate the assumptions of the hypothesis. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Do mothers regulate facultative and obligate siblicide by differentially provisioning eggs with hormones?

We examined androgens in clutches of two booby species that differ in their sibling conflict. Blue-footed booby Sula nebouxii chicks show an aggression-submission relationship, aggression is normally moderate and siblicide is facultative. Brown booby Sula leucogaster chicks show an aggression-aggression relationship, aggression of both chicks can be relentless and siblicide is obligate. The parental favoritism hypothesis predicts that egg mass, yolk mass and yolk androgens should decline with laying order less in the blue-footed booby than in the brown booby, to promote the survival of the former's junior chick. The eggs of the blue-footed booby had higher yolk concentrations of 5α-dihydrotestosterone (DHT) and lower concentrations of testosterone (T); both species had similarly high yolk concentrations of androstenedione (A). Intra-clutch variation in yolk DHT, T and A failed to support our predictions. In both species, first and second eggs showed similar concentrations of all three hormones and were of similar size, although in the blue-footed booby (only) yolk masses declined with laying order. Brown booby mothers make junior chicks vulnerable to siblicide by hatching them 5 d after their broodmates, but not by differential allocation of egg androgens or nutrients. Blue-footed booby mothers appear to prepare junior chicks for thriving in a subordinate non-provocative role by hatching them 4 d after their broodmates and providing them with 10% less yolk. To orchestrate agonism within the brood, these boobies may rely more on hatch intervals and yolk provision than on androgens.     

Multiple Paternity Benefits Female Marbled Salamanders by Increasing Survival of Progeny to Metamorphosis

Multiple paternity occurs in most species and animal groups that have been studied. Because mating involves fitness costs to individual females, theory predicts that polyandrous females gain greater fitness benefits than costs, allowing the behavior to be maintained. Genetic, rather than material, benefits often occur in species where males provide females with little more than sperm and seminal fluid. We compared fitness correlates of single‐ and double‐sire clutches from female marbled salamanders (Ambystoma opacum) at the egg, hatchling, and metamorph stages of offspring development. Because clutches were collected from experimental breeding groups, strict paternity exclusion of offspring using microsatellite data allowed us to categorize each clutch as having either one or two fathers. Early offspring viability and size of hatchlings were not different between single‐ and multiple‐paternity clutches. Larvae from the two clutch types were allowed to develop together in field enclosures until metamorphosis. Although there was no difference in size at metamorphosis, survival to metamorphosis was significantly higher in multiple‐paternity clutches (44% vs. 40%) suggesting a benefit for females. The results were consistent with genetic benefits, although maternal effects could not be ruled out. The data did not support predictions of the genetic bet‐hedging and good sperm hypotheses for genetic benefits of polyandry.     

Elevated yolk androgen levels benefit offspring development in a between-clutch context

The field of androgen deposition in avian eggs and its consequencesfor offspring development has received a lot of attention inrecent research. However, although variation within clutchesin yolk androgens is relatively well understood, the adaptivesignificance of patterns of variation between clutches remainsrather unclear. Furthermore, it has been hypothesized that yolkandrogens act as a means of an adaptive maternal effect to adjustoffspring to a given posthatching environment. Thus, the consequencesof maternal yolk androgens for offspring development are likelyto depend on the specific environment of a given brood. We experimentallymanipulated yolk androgen concentrations in spotless starlingeggs, using a between-brood design in which full broods weremanipulated applying either an androgen or sham treatment, inorder to test the effects of between-brood variation in yolkandrogen levels. We also included in the analysis several femalecharacters that have been shown to affect androgen deposition.Androgen-chicks tended to gain more mass, a similar effect tothat shown in previous studies where direct competition betweenchicks belonging to different treatments was allowed, but didnot gain a survival benefit. Androgen-chicks had wider beakflanges, an effect that has not been described previously andcould play an important role in food acquisition. In addition,androgen-chicks had higher endogenous plasma levels of androgens,which could induce higher begging intensity. We hypothesizethat these effects are an important mechanistic link for ourunderstanding of how yolk androgens exert their effects on offspringdevelopment after hatching. Contrary to our predictions, wefound no evidence that the observed effects depended on theenvironment under which it was investigated. Because high yolkandrogen levels seem to be beneficial for nestlings in thisspecies, we hypothesize that yolk hormone deposition could becostly for females or alternatively that potential negativeeffects later in adulthood may constrain maternal hormone allocation.     

The multivariate egg: quantifying within- and among-clutch correlations between maternally derived yolk immunoglobulins and yolk androgens using multivariate mixed models

Egg components are important mediators of prenatal maternal effects in birds and other oviparous species. Because different egg components can have opposite effects on offspring phenotype, selection is expected to favour their mutual adjustment, resulting in a significant covariation between egg components within and/or among clutches. Here we tested for such correlations between maternally derived yolk immunoglobulins and yolk androgens in great tit (Parus major) eggs using a multivariate mixed-model approach. We found no association between yolk immunoglobulins and yolk androgens within clutches, indicating that within clutches the two egg components are deposited independently. Across clutches, however, there was a significant negative relationship between yolk immunoglobulins and yolk androgens, suggesting that selection has co-adjusted their deposition. Furthermore, an experimental manipulation of ectoparasite load affected patterns of covariance among egg components. Yolk immunoglobulins are known to play an important role in nestling immune defence shortly after hatching, whereas yolk androgens, although having growth-enhancing effects under many environmental conditions, can be immunosuppressive. We therefore speculate that variation in the risk of parasitism may play an important role in shaping optimal egg composition and may lead to the observed pattern of yolk immunoglobulin and yolk androgen deposition across clutches. More generally, our case study exemplifies how multivariate mixed-model methodology presents a flexible tool to not only quantify, but also test patterns of (co)variation across different organisational levels and environments, allowing for powerful hypothesis testing in ecophysiology.     

Maternal effects mediated by egg quality in the Yellow-legged Gull Larus michahellis in relation to laying order and embryo sex

Background

Maternal effects mediated by egg size and quality may profoundly affect offspring development and performance, and mothers may adjust egg traits according to environmental or social influences. In avian species, context-dependency of maternal effects may result in variation in egg composition, as well as in differential patterns of covariation among selected egg components, according to, for example, position in the laying sequence or offspring sex. We investigated variation in major classes of egg yolk components (carotenoids, vitamins and steroid hormones) in relation to egg size, position in the laying sequence and embryo sex in clutches of the Yellow-legged Gull (Larus michahellis). We also investigated their covariation, to highlight mutual adjustments, maternal constraints or trade-offs in egg allocation.

Results

Laying sequence-specific patterns of allocation emerged: concentration of carotenoids and vitamin E decreased, while concentrations of androgens increased. Vitamin A, estradiol and corticosterone did not show any change. There was no evidence of sex-specific allocation or covariation of yolk components. Concentrations of carotenoids and vitamins were positively correlated. Egg mass decreased along the laying sequence, and this decrease was negatively correlated with the mean concentrations of carotenoids in clutches, suggesting that nutritionally constrained females lay low quality clutches in terms of carotenoid content. Finally, clutches with smaller decline in antioxidants between first- and last-laid eggs had a larger increase in yolk corticosterone, suggesting that a smaller antioxidant depletion along the laying sequence may entail a cost for laying females in terms of increased stress levels.

Conclusions

Since some of the analyzed yolk components (e.g. testosterone and lutein) are known to exert sex-specific phenotypic effects on the progeny in this species, the lack of sex-specific egg allocation by mothers may either result from trade-offs between contrasting effects of different egg components on male and female offspring, or indicate that sex-specific traits are controlled primarily by mechanisms of sexual differentiation, including endogenous hormone production or metabolism of exogenous antioxidants, during embryonic development.     

Functional Value of Matriphagy in the Spider Amaurobius ferox

The adaptive value of matriphagy, the consumption of the mother by her offspring, in the sub-social spider Amaurobius ferox (Araneae, Amaurobiidae) was experimentally evaluated in terms of the benefits to the offspring and the costs to the mother. Matriphagy resulted in a 2.5-fold weight gain in the offspring over their initial weight, advancement of their moulting time, and larger body mass at dispersal in comparison with clutches deprived of matriphagy, but otherwise well provisioned with prey. Matriphagous offspring were also more successful at capturing large prey items, had a more extended social period, and a higher survival rate at dispersal. Mothers separated from their broods just prior to matriphagy were able to produce second clutches, effectively producing 33% more viable offspring (compared with the first clutches). However, the estimated reproductive outputs of the alternative maternal strategies (being devoted to the first clutch vs. deserting the first and producing a second one) suggest that mothers of A. ferox that are cannibalized by their broods enjoy greater reproductive success than those that escape cannibalism and produce second clutches.     

Brief communication: Fecal androgen excretion and fetal sex effects during gestation in wild Assamese macaques (Macaca assamensis)

In placental mammals, pregnancy usually is associated with an increase in maternal androgens, which may significantly impact fetal growth and differentiation, and affect postnatal development and behavior. Owing to their slow life histories and challenging social conditions, determination of maternal androgens and potential interference effects of fetal androgen production are of particular interest in wild primates. However, androgen production has been rarely investigated in wild female primates, and studies on maternal androgens during gestation in particular often do not span the entire pregnancy. Here, we characterize fecal androgen production throughout gestation in wild Assamese macaques (Macaca assamensis) using noninvasive hormone analysis and, furthermore, examine fetal sex effects on maternal androgen excretion. A total of 207 fecal samples were analyzed from seven females for concentrations of immunoreactive epiandrosterone (iEA). Fecal iEA concentrations, as predicted based on cercopithecine blood-serum patterns, increased during early gestation and were significantly higher during the first trimester compared with preconception concentrations and those recorded during later stages of gestation. Further, during the third trimester, male-carrying mothers showed significantly higher iEA concentrations compared with female-carrying mothers. This first characterization of fecal androgen excretion during gestation in Assamese macaques indicates both a maternal and fetal effect on androgen production. Although our sample size is small, our results, nevertheless, provide the basis for assessing potential influences of maternal androgens on postnatal offspring development and behavior.     

Within-clutch patterns of yolk testosterone vary with the onset of incubation in black-headed gulls

Hatching asynchrony in birds produces an age and size hierarchyamong siblings. Later-hatching chicks have a competitive disadvantage,and brood reduction may occur when food availability is insufficientto raise all chicks. When early-hatched chicks fail to surviveor if the circumstances allow raising all chicks, mothers shouldreverse the disadvantage to later-hatched chicks. Increasingdeposition of maternal androgens with the laying sequence hasbeen suggested to compensate for detrimental effects of hatchingasynchrony, allowing a more precise adjustment of the survivalprobabilities of each chick. Here, we show for black-headedgulls that the increase in yolk testosterone with each successiveegg is greater when the mother incubates longer before clutchcompletion, which is the major determinant of the degree ofhatching asynchrony. This finding supports the idea that yolktestosterone has a compensatory function in the context of hatchingasynchrony. Our data further show that if the time needed tocomplete a clutch is lengthened, the developmental differencesdue to incubation between the first- and the last-laid eggsincrease. In addition, the onset of incubation before clutchcompletion occurs sooner as the breeding season progresses.Both long inter-egg intervals and the seasonal shift in incubationbehavior enhance the necessity of compensation for later-hatchingchicks. Indeed, yolk levels of testosterone increased more steeplyover the laying order, if the duration of the egg-laying periodwas extended and in later-laid clutches. We suggest that prolactinplays a key role in the adjustment of testosterone allocationto the incubation pattern.     

Maternal allocation in eggs when counting on helpers in a cooperatively breeding bird

For cooperatively breeding birds, it has been proposed that breeders should reduce their investment in eggs when they count on helpers, because this can be compensated for by helpers provisioning of nestlings. Data from some species have supported this prediction, but this is not the case in others. It has also been proposed that mothers should not reduce but rather increase investment if the presence of helpers enhances the reproductive value of offspring, a pattern that might also influence egg production as long as helpers are predictable for laying females. Here, we studied maternal expenditure in eggs and clutches in the Iberian magpie, to see whether mothers reduce their expenditure at the egg stage in the presence of helpers. Our results show that investment in clutches varied depending on the year, date in the season and age of the mother, but there were no reductions in maternal expenditure per individual egg when they counted on helpers. On the contrary, a pattern emerged in the opposite direction of more investment in eggs associated with the future presence of helpers at the nestling stage. Our data suggest that the predictability of helpers, along with the type of benefits accrued from the contribution of helpers, may be crucial to understanding the reaction of mothers at egg production.     

Do Genetic Diversity Effects Drive the Benefits Associated with Multiple Mating? A Test in a Marine Invertebrate

Background

Mothers that mate with multiple males often produce higher quality offspring than mothers that mate with a single male. By engaging in polyandry, mothers may increase their chances of mating with a compatible male or promote sperm competition - both of which act to increase maternal fitness via the biasing of the paternity of offspring. Surprisingly, mating with multiple males, can carry benefits without biasing paternity and may be due simply to differences in genetic diversity between monandrous and polyandrous clutches but role of genetic diversity effects in driving the benefits of polyandry remains poorly tested. Disentangling indirect, genetic benefits from genetic diversity effects is challenging but crucial if we are to understand the selection pressures acting to promote polyandry.

Methodology/Principal Findings

Here, we examine the post-fertilisation benefits of accessing the sperm of multiple males in an externally fertilising polychaete worm. Accessing the sperm of multiple males increases offspring performance but this benefit was driven entirely by genetic diversity effects and not by the biasing of paternity at fertilisation.

Conclusions/Significance

Previous studies on polyandry should be interpreted cautiously as genetic diversity effects alone can explain the benefits of polyandry yet these diversity effects may be difficult to disentangle from other mechanisms. We suggest that future studies use a modified experimental design in order to discriminate between genetic diversity effects and indirect, genetic benefits.     

Maternal allocation of androgens and antagonistic effects of yolk androgens on sons and daughters

Mothers can influence the phenotype of their offspring by adjustingthe quality of their eggs in relation to sex and reproductivevalue of the progeny. Maternal androgens in the eggs of vertebratesmay mediate such adaptive early maternal effects. However, theevolution of early maternal effects mediated by egg androgensmay be constrained by the inability of mothers to differentiallyallocate androgens to eggs with a male or a female if androgenshave different effects on sons and daughters. In this study,we increased the concentration of androgens in the eggs of barnswallows (Hirundo rustica) within the physiological range ofvariation and analyzed the effect on nestling growth and beggingbehavior. Egg androgens increased body size and mass of sonsbut reduced these characters in daughters when compared to twocontrol groups in a repeated-measures analysis of variance ofdata collected at different ages. However, the differentialeffect of androgen on the two sexes was no longer significantwhen the analysis was restricted to the age of 12 days, whenfinal body size is attained. In a second experiment, we testedwhether mothers differentially allocated androgens to eggs withsons rather than daughters while manipulating a paternal secondarysexual character. Androgen concentration did not vary in relationto paternal ornamentation or embryo sex. Hence, antagonisticeffects of egg androgens on sons and daughters may exist inthe very early posthatching life and may constrain the evolutionof adaptive maternal effects because mothers do not differentiallyallocate androgens in relation to embryo sex.