Yolk androgens and begging behaviour in black-headed gull chicks: an experimental field study

Black-headed gulls, Larus ridibundus, produce clutches of three eggs, which contain high levels of maternal androgens in the yolk. These levels increase with laying order and the eggs hatch asynchronously. Experiments have supported the hypothesis that this within-clutch variation in maternal androgens mitigates the disadvantage of last-hatched chicks in sibling competition, by enhancing embryonic development and early posthatching growth. We hypothesized that these effects come about by the stimulating effects of maternal androgens on begging behaviour and competitive ability. In the field, we injected first-laid eggs of a clutch (which have a low androgen level) with either an androgen solution (T eggs) or vehicle (Oil eggs). We then created pairs of chicks hatched from Oil and T eggs, matched for egg mass and hatching date. Parent–chick interactions were recorded from observation hides. Chicks from T eggs hatched almost half a day sooner than those from Oil eggs. Furthermore, chicks from T eggs were more active during the first week after hatching, were more often the first to react to the approaching parent, begged more frequently, and obtained the larger share of food. We conclude that the enhancing effect of yolk androgens on growth in this species arises at least partly through androgen-mediated effects on the chicks’ behaviour.

An experimental study on the causes of sex-biased mortality in the black-headed gull − the possible role of testosterone

1. During early development male offspring in avian species often suffer from enhanced mortality compared to female offspring. This has been attributed to different nutritional requirements, as sex-biased mortality has been reported particularly in sexually size-dimorphic species. However, other traits of the male phenotype, such as the embryonic hormone profile, have been suggested to contribute to this male disadvantage. In particular the negative effects of sex steroids on immune function may be causally involved. 2. We investigated the role of testosterone in the expression of male phenotype disadvantage through an experimental reduction of the availability of testosterone receptors by in-ovo injection of an anti-androgen (Flutamide©). Experimental nests contained a male and a female chick hatching from control treated eggs and a male and a female chick hatching from Flutamide treated eggs. 3. Male-biased mortality occurred in control chicks at a stage where the sexes did not yet differ in their growth pattern, suggesting that sex-specific nutritional requirements are not necessary for male-biased mortality to occur. Control males and control females did not differ in their cell-mediated immunity (CMI). This renders it unlikely that the observed skewed mortality was due to sex-specific differences in the CMI. 4. Treatment of the eggs with flutamide antagonistically affected males and females. Flutamide treatment positively affected male development in particular through an enhanced growth rate, indicating that testosterone is involved in the expression of the male phenotype disadvantage. Female chicks hatching from Flutamide treated eggs were disadvantaged in growth and CMI. The possible pathways of this sex-specific effect of Flutamide are discussed with regard to differential consequences of blocking the beneficial effects of maternal androgens and the interference with the process of sexual differentiation in males.

Maternal androgens in egg yolks: relation with sex, incubation time and embryonic growth

Hormones of maternal origin are known to be transferred to the egg yolks of oviparous species. Several studies have shown that within and between clutch variation of maternal androgens may be adaptively tuned. Moreover, it has recently been hypothesized that sex steroids of maternal origin may play a role in adaptive sex ratio manipulation. For sex determination the eggs have to be incubated to allow the germinal disc to grow and thus extract sufficient DNA. This means that yolk hormone levels are determined after a number of days of incubation and this may hamper interpretation of the data. If yolk utilization or embryonic hormone production are influenced by the sex of the embryo, differences in hormone content at a certain stage of incubation do not reflect the mother's initial allocation. In this experiment we show that testosterone levels in chicken eggs do not change with incubation period. A4 levels decrease between 3 and 5 days of incubation, which we cannot explain. Male eggs did not contain higher levels of testosterone or androstenedione than female eggs, in contrast to the data reported for another galliform species, the peacock. We conclude that it is unlikely that maternal androgens are a key factor in the avian sex determination mechanism.

Lesser double-collared sunbirds Nectarinia chalybea do not compensate for hatching asynchrony by adjusting egg mass or yolk androgens

Substantial amounts of maternal androgens are found in birds’ eggs and have been shown to benefit offspring development. Within-clutch patterns of increasing androgen concentrations over the laying sequence are often hypothesized to compensate for the negative effects of hatching asynchrony. However, detrimental effects to offspring fitness of exposure to high yolk androgen levels have also been demonstrated. This suggests that mothers should forego these costs to their offspring when the need for compensation for hatching asynchrony is low or when alternative compensatory strategies, e.g. in terms of increasing egg mass, are available. Here we show that in the south-temperate lesser double-collared sunbird Nectarinia chalybea, a species with hatching asynchrony but also with high survival of last-hatched chicks, mothers do not deposit resources differentially in terms of either yolk androgen concentration or egg mass across the laying sequence. We discuss to what extend this challenges the original explanation of within-clutch variation in these egg parameters and offer some explanation for their between-clutch variation which was related to female body mass.

Balancing between costs and benefits of maternal hormone deposition in avian eggs

Avian eggs contain substantial amounts of maternal androgens, and several studies have indicated that these are beneficial for the chick. Nevertheless, there is a large and systematic variation in maternal hormone concentrations both within and between clutches. If maternal androgens also involve costs, this might explain why not all mothers put high levels of androgens in their clutches. However, the simultaneous occurrence of both benefits and costs has not yet been convincingly demonstrated. We show experimentally that yolk androgens suppress immune function and simultaneously stimulate growth in black-headed gull chicks. Thus, mothers face a trade-off between these costs and benefits and may tune hormone deposition to prevailing conditions that influence chick survival.

Consistent variation in yolk androgens in the Australian Brush-turkey, a species without sibling competition or parental care

Maternal hormones are an excellent pathway for the mother to influence offspring development, and birds provide exceptional opportunities to study these hormone-mediated maternal effects. Two dominant hypotheses about the function of yolk androgens in avian eggs concern maternal manipulation of sibling competition and post hatching paternal care. In megapodes, however, neither sibling competition nor post hatching parental care exists. Eggs are incubated by external heat sources, and chicks dig themselves out of their underground nest and live independently of their parents and their siblings. In this first study on egg androgens of such a megapode, the Australian Brush-turkey Alectura lathami, we found nevertheless substantial amounts of maternal androgens. Since size of the incubation mound, incubation temperature, egg size and laying date greatly vary in this species, we analysed variation in testosterone (T), androstenedione (A4) and dihydrotestosterone (DHT) in relation to these factors. T concentrations were significantly higher in eggs from bigger mounds and laid at greater depth, which may compensate via anabolic effects for the longer duration and higher energetic requirements of chicks when digging themselves out. T concentrations were higher in smaller eggs, and both yolk A4 and T concentrations increased with laying date, perhaps as a compensatory measure, while DHT concentrations only varied across different mounds. These results indicate that maternal androgens may influence offspring development outside the contexts of sibling competition or parental care.

Steroids for free? No metabolic costs of elevated maternal androgen levels in the black-headed gull

Within- and between-clutch variation in yolk titres of hormones of maternal origin has been found in many avian species. So far, experiments have revealed mainly beneficial effects of maternal androgens. This would also apply to black-headed gulls (Larus ridibundus). Previous experiments have shown that chicks benefit from these higher levels since their competitive abilities are improved and growth and survival probabilities thus enhanced. However, not all females show the same increase in yolk hormones from first to last egg or invest equally high amounts of androgens in their clutches. Possibly, there is a trade-off between the beneficial effects of high androgen levels and potential costs, such as increased metabolic rates. We studied possible metabolic costs of experimentally elevated yolk androgen levels for chicks of several age classes, starting three days prior to hatching until fledging at an age of approximately 30 days. Daily energy expenditure in the field, measured using the doubly labelled water technique, did not differ between treatments or between sexes. Oxygen consumption measured in birds at rest in the lab (RMR) did not vary between chicks hatched from androgen-injected (T) or oil-injected (Oil) control eggs at any age in thermo-neutral or below thermo-neutral conditions. Males showed a lower RMR than females towards the fledging age. We conclude that it is unlikely that the costs of high maternal androgen levels can be found in higher energy expenditure in the chick.

Maternal yolk androgens in European starlings: affected by social environment or individual traits of the mother?

Social competition among female birds has been shown to positively affect yolk androgen levels, perhaps providing a mechanism to communicate environmental conditions to offspring. Whether this relationship is due to social density or to differences among mothers that breed in different social situations is unclear. We manipulated breeding density to test these alternative explanations. Yolk androgens were measured in clutches of European starlings, Sturnus vulgaris, breeding in consecutive years in outdoor aviaries of different sizes and with varying numbers of breeding pairs. Testosterone (T) levels increased significantly with increasing density. Dihydrotestosterone (DHT) levels increased with the number of nest boxes available. The number of nest boxes monopolized by a breeding male negatively affected androstenedione (A4) levels, positively affected DHT levels and did not affect T levels. Other factors related to social interactions or competition among females (including polygyny) did not influence yolk hormone levels. Within-female yolk androgen levels were highly consistent over two consecutive years even though females were breeding in opposite breeding densities during each year, suggesting that individual characteristics are important determinants of variation in maternal androgen allocation in addition to potential effects of environmental conditions. This within-female consistency of yolk androgen levels across years has important implications.