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Independent and combined effects of egg pro- and anti-oxidants on gull chick phenotype
Abstract
Oviparous mothers transfer to the eggs components that have both independent and combined effects on offspring phenotype. Functional interactions between egg components, such as antioxidant and hormones, lead to expect that a change in the concentration of one component has effects on offspring traits that depend on the concentration of other interacting components. However, the combined effects of variation in different egg components are virtually unknown. Bird eggs contain vitamin E, a major antioxidant, and also maternal corticosterone. The independent consequences of variation in the egg concentrations of these compounds for offspring phenotype are largely unknown and no study has investigated their combined effects. We manipulated the concentration of vitamin E and corticosterone in the eggs of the yellow-legged gull (Larus michahellis) by administering a physiological (2 standard deviations) dose both independently and in combination. We tested for an effect on chick post-natal growth, plasma antioxidant capacity (TAC) and oxidative compounds (TOS). Separate administration of vitamin E or corticosterone caused a reduction in body mass relative to controls, whereas the combined administration of the two compounds reversed their negative effects. These results suggest thatmaternal egg components, such as antioxidants and steroid hormones, interact and mothers must balance their concentrations in order to achieve optimal offspring phenotype.The functional relationship between vitamin E and corticosterone is corroborated by the observation of positive covariation between these compounds.
... This approach has been pivotal for advancing our knowledge of how mothers affect offspring phenotype. However, maternal effects are by nature multivariate (reviewed by [33,34]), with different groups of yolk components influencing similar nestling traits, i.e., by having interactive effects on offspring phenotype [35][36][37]. For instance, hatchling mass was reduced and oxidative stress increased in Japanese quails (Coturnix japonica) hatching from eggs injected with either testosterone or carotenoids [35]. ...
... But when both components were administered together, neither hatchling mass nor oxidative stress were affected. Similar compensatory effects were found in yellow-legged gulls (Larus michahellis) after eggs were simultaneously injected with corticosterone and vitamin E [36,37]. Such studies suggest that the consequences of maternal effects on offspring phenotype known from studies that focus on single components might actually be absent, weakened or potentiated if the presence and actions of other components are also considered. ...
Background
In egg-laying animals, mothers can influence the developmental environment and thus the phenotype of their offspring by secreting various substances into the egg yolk. In birds, recent studies have demonstrated that different yolk substances can interactively affect offspring phenotype, but the implications of such effects for offspring fitness and phenotype in natural populations have remained unclear. We measured natural variation in the content of 31 yolk components known to shape offspring phenotypes including steroid hormones, antioxidants and fatty acids in eggs of free-living great tits (Parus major) during two breeding seasons. We tested for relationships between yolk component groupings and offspring fitness and phenotypes.
Results
Variation in hatchling and fledgling numbers was primarily explained by yolk fatty acids (including saturated, mono- and polyunsaturated fatty acids) - but not by androgen hormones and carotenoids, components previously considered to be major determinants of offspring phenotype. Fatty acids were also better predictors of variation in nestling oxidative status and size than androgens and carotenoids.
Conclusions
Our results suggest that fatty acids are important yolk substances that contribute to shaping offspring fitness and phenotype in free-living populations. Since polyunsaturated fatty acids cannot be produced de novo by the mother, but have to be obtained from the diet, these findings highlight potential mechanisms (e.g., weather, habitat quality, foraging ability) through which environmental variation may shape maternal effects and consequences for offspring. Our study represents an important first step towards unraveling interactive effects of multiple yolk substances on offspring fitness and phenotypes in free-living populations. It provides the basis for future experiments that will establish the pathways by which yolk components, singly and/or interactively, mediate maternal effects in natural populations.
... lower hatching mass) but when eggs were treated simultaneously with both carotenoids and testosterone, the detrimental effects were mitigated (Giraudeau et al., 2017). Possenti et al. (2018) manipulated both an anti-oxidant (vitamin E) and a putative pro-oxidant (corticosterone) and found that administration of vitamin E or corticosterone alone caused a reduction in body mass relative to controls, whereas the combined treatment reversed the negative effects. ...
... In summary, three studies (this study; Giraudeau et al., 2017;Possenti et al., 2018) have failed to provide compelling evidence for strong, or long-term, synergistic effects of multiple, maternally derived egg components, although the latter two studies provide some evidence for short-term antagonistic effects. Although the task of fully investigating the concept of the 'multivariate egg' (sensu Postma et al. 2014;Williams and Groothuis, 2015) is clearly challenging, these studies highlight the importance of being able to predict a priori the direction and magnitude of effects of single egg components, to enable clear predictions about multivariate effects (see Table 1). ...
Eggs are 'multivariate' in that they contain multiple maternally derived egg components (e.g. hormones, antibodies, mRNA, antioxidants) which are thought to influence offspring phenotype. However, most studies have focused on single egg components and on short-term effects. Here, we simultaneously manipulated two egg components, maternally derived antibodies (MAb) and yolk testosterone, to assess potential synergistic or antagonistic effects on zebra finch offspring phenotype from hatching to sexual maturity. We found no evidence for short- or long-term effects of either MAb or yolk testosterone alone, or their interaction, on hatching mass, size at fledging (tarsus length), body mass at sexual maturity (day 82), chick survival, humoral immune function or any measured female reproductive trait at sexual maturity. There was a positive effect of yolk testosterone, but not MAb, on offspring phytohaemagglutinin (PHA) response at 26 days of age but at 82 days of age, MAb, but not yolk testosterone, had a positive effect on PHA response. There was also a MAb×sex interaction on 30 day chick mass, and a positive effect of yolk testosterone on male courtship behaviour at sexual maturity. However, we found no evidence for synergy, i.e. where offspring treated with both MAb and yolk testosterone had higher trait values than offspring treated with either MAb or yolk testosterone alone for any measured trait. Similarly, evidence for antagonistic (compensatory) effects, where offspring treated with both MAb and yolk testosterone had intermediate trait values compared with offspring treated with either MAb or yolk testosterone alone, was equivocal.
... Androgen hormones such as testosterone have been suggested to be the main mediator of maternal effects in poultry species 19 ; however, other egg components have also been investigated (glucocorticoid hormones 20 , thyroid hormones 21,22 , antioxidants 23 and immunoglobulins 24 ). ...
Maternal effects have been reported to alter offspring phenotype in laying hens. In this study, we investigated the effects of maternal environment and maternal age on egg traits and offspring development and behaviour. For this, we ran two experiments. First (E1), commercial hybrid hens were reared either in aviary or barren brooding cages, then housed in aviary, conventional cages or furnished (enriched) cages, thus forming different maternal housing treatments. Hens from each treatment were inseminated at three ages, and measures of egg composition, yolk testosterone concentration and offspring’s development, anxiety and fearfulness were assessed. In experiment 2 (E2), maternal age effects on offspring's growth and behaviour were further investigated using fertile eggs from commercial breeder flocks at three ages. Results from E1 showed that maternal age affected the majority of measures including egg composition, yolk testosterone, offspring growth, anxiety and fearfulness. Maternal rearing and housing affected fewer measures but included egg characteristics, offspring weight and behaviour. Effects of maternal age were not replicated in E2, possibly due to higher tolerance to maternal effects in commercial breeders. Overall, our research confirms that maternal environment and maternal age affect the offspring of laying hens in a variety of ways. These effects may be mediated by the natural decrease in egg yolk testosterone concentration over time.
... This diversity of egg components that can putatively modulate the phenotype of the developing organism (in addition to their interactions) has led to the concept of the 'multivariate egg' (Postma et al., 2014;Williams and Groothuis, 2015). Because virtually all components of the egg may have their own effects, and potentially act in synergistic or antagonistic ways, this requires integration of multiple interactive factors when studying maternal effects on offspring phenotype (Possenti et al., 2018;Torres et al., 2019). For example, manipulation of yolk-deposited carotenoids and testosterone showed that their independent deleterious effects on chick oxidative stress while triggering a higher growth rate were buffered when both factors were increased simultaneously (Giraudeau et al., 2017). ...
Avian eggs contain a large number of molecules deposited by the mother that provide the embryo with energy but also potentially influence its development via the effects of maternally-derived hormones and antibodies: the avian egg is “multivariate”. Multivariate effects on offspring phenotype were evaluated in a sister study on captive zebra finches, by simultaneously manipulating maternally derived antibodies after LPS-treatment of mothers and egg-treatment using yolk testosterone injection. LPS-treatment had a positive effect on body mass growth at 30 days after hatching and immune response at sexual maturity, while egg-testosterone positively influenced immune response at fledging and courtship behaviour in sexually mature male offspring. Maternal effects are known to modulate offspring telomere length. Still, the multivariate effects of egg-derived maternal components on offspring telomere dynamics from hatching to sexual maturity are undefined. Here, using the data of the sister study completed with telomere measurements, we tested a) the effects of LPS and T treatments on telomere length (TL) from hatching to sexual maturity (day 82), b) how LPS treatment modulated telomere length over reproduction in adult females, and c) the relationship between maternal and offspring TL. We predicted, a) TL would be shorter in LPS fledglings (as a cost of faster growth), and b) TL would be longer in sexually mature adults from T-treated (as a proxy of individual quality). In adult females, there was an overall negative relationship between laying and rearing investments and telomere length, this relationship being weaker in LPS treated females. In chicks, there was an overall negative effect of LPS treatment on telomere length measured at fledging and sexual maturity (day 25 – 82). In addition, at fledging, there was a sex x LPS x T-treatment interaction, suggesting the existence of antagonistic effects of our treatments. Our data partially support the hypothesis of telomeres are proxies of individual quality and that individual differences in telomere length are set-up very early in life.
... Cottrell and Seckl (15) proposed two major hypotheses to explain the association between maternal stress and postnatal effects on offspring: fetal malnutrition and overexposure to glucocorticoid hormones. More recently, studies in avian species have shown that maternal stress can also be linked to the increase in other biological components in the egg, such as androgens (16), thyroid hormones (17), antioxidants (18), and immunoglobulins (19). Nevertheless, although glucocorticoid hormones are not a synonym for "stress" (20), corticosterone remains as one of the most analyzed mediators of maternal stress in the literature due to their pleiotropic role in regulating physiological responses to the environment and in the development and maturation of vital organs [reviewed in (6,16,21)]. ...
Maternal stress can affect the offspring of birds, possibly due to hormone deposition in the egg. Additionally, phenotypic diversity resulting from domestication and selection for productivity has created a variety of poultry lines that may cope with stress differently. In this study, we investigated the effects of maternal stress on the behavior of different strains of laying hens and the role of corticosterone as its mediator. For this, fertilized eggs of five genetic lines—two brown (Brown 1 and 2), two white (White 1 and 2), and one pure line White Leghorn—were reared identically as four flocks of 27 birds (24F: 3M) per strain. Each strain was equally separated into two groups: Maternal Stress (“MS”), where hens were subjected to a series of daily acute psychological stressors for 8 days before egg collection, and “Control,” which received routine husbandry. Fertile eggs from both treatments were collected at three different ages forming different offspring groups that were treated as replicates; additional eggs from Control were injected either with corticosterone diluted in a vehicle solution (“CORT”) or just “Vehicle.” Eggs from each replicate were incubated and hatched, and offspring (N = 1,919) were brooded under identical conditions. To measure the effects of maternal stress on anxiety and fear-like behavior, offspring were subjected to a social isolation test (SI) between 5 and 10 days of age and a tonic immobility test (TI) at 9 weeks of age. Compared to Control, MS decreased the number of distress vocalizations emitted by White 2 in SI. No effects of MS were observed in TI, and no effects of CORT were observed in any tests. Overall, brown lines vocalized more in SI and remained in TI for a longer duration than white strains, suggesting genetic differences in fear behavior. Females vocalized more than males in TI and showed a trend toward significance for the same trait in SI. Overall, results suggest that the effects of maternal stress on fearfulness are not directly mediated by corticosterone. Moreover, it highlights behavioral differences across various strains of laying hens, suggesting that fear responses are highly dependent on genotype.
... The effects of maternal stress on egg composition have been extensively studied in birds (e.g., hormones (Carter et al., 2018), antioxidants (Possenti et al., 2018), and immunoglobulins (Roth et al., 2018)). Gonadal steroid hormones, and particularly androgens, are known to be important mediators of maternal effects. ...
Early-life exposure to stressors can shape the phenotype of the offspring resulting in changes that may affect their prehatch and posthatch development. This can be modeled indirectly through maternal exposure to stressors (natural model) or by offspring exposure to stress hormones (pharmacological model). In this study, both models were used to investigate the effects of genetic line on hatchability, late embryonic mortality, sex ratio, and body weight until 17 wk of age. To form the parent stock, fertilized eggs of 4 commercial genetic lines — two brown (brown 1 and 2), two white (white 1 and 2), and a pure line White Leghorn — were incubated, hatched, and housed identically in 4 flocks of 27 birds (24 females and 3 males) per strain. Each strain was equally separated into 2 groups: “maternal stress,” where hens were subjected to a series of acute psychological stressors (e.g., physical restraint, transportation) for 8 D before egg collection, and “control,” where hens received routine husbandry. At 3 maternal ages, fertile eggs from both treatments were collected, and additional eggs from the control group were injected with corticosterone (10 ng/mL egg content) (“CORT”). A “vehicle” treatment was included to account for effects of egg manipulation. Each maternal age comprised a replicate over time. Eggs were incubated and hatched, and the offspring (N = 1,919) were brooded until 17 wk under identical conditions. The results show that prenatal stress interacted with strain to decrease embryonic survival and growth. Among all strains, brown 2 was consistently the most affected line in both prehatch and posthatch development. Our study shows that embryonic survival and offspring growth are mostly affected by the pharmacological model and that strain differences may increase susceptibility to prenatal stress. Moreover, it suggests that the natural stressor model may be useful for quantifying the response of the mother to stressors, whereas the pharmacological model may be useful for quantifying the response of the embryo to increased levels of corticosterone.
... While the importance of oxidative status particularly for the hatchlings is well established, surprisingly few studies have focused on the changes in physiological variables that reflect oxidative status, including total antioxidant capacity and concentration of pro-oxidant molecules, during the early post-natal stages (e.g. Parolini et al. 2017a,b;Possenti et al. 2018). ...
The perinatal period is critical to survival and performance of many organisms. In birds, rapid post-natal growth and sudden exposure to aerial oxygen around hatching markedly affect the chick redox status, with potentially negative consequences on physiology mediated by oxidative stress. In addition, telomere length undergoes reduction during birds’ early life, partly depending on oxidative status. However, relatively few studies have focused specifically on the changes in oxidative status and telomere length that occur immediately after hatching. In the present study of the yellow-legged gull Larus michahellis, we found that chicks undergo a marked increase in plasma total antioxidant capacity and a marked decrease in the concentration of pro-oxidant molecules during the first days after hatching. In addition telomere length in erythrocytes decreased by one standard deviation over the 4 days post-hatching. Body mass and tarsus length covaried with total antioxidant capacity and concentration of pro-oxidants in a complex way, that partly depended on sex and laying order, suggesting that oxidative status can affect growth. Moreover, telomere length positively covaried with the concentration of pro-oxidant molecules, possibly because retention of high concentrations of pro-oxidant molecules results from mechanisms of prevention of their negative effects, including reduction in telomere length. Thus, this study shows that chicks undergo marked variation in oxidative status, which predicts growth and subsequent telomere length, prompting for more studies of the perinatal changes in the critical post-hatching stages.
Urbanization can cause innumerable abiotic and biotic changes that have the potential to influence the ecology, behavior, and physiology of native resident organisms. Relative to their rural conspecifics, urban Side-blotched Lizard (Uta stansburiana) populations in southern Utah have lower survival prospects and maximize reproductive investment via producing larger eggs and larger clutch sizes. While egg size is an important predictor of offspring quality, physiological factors within the egg yolk are reflective of the maternal environment and can alter offspring traits, especially during energetically costly processes, such as reproduction or immunity. Therefore, maternal effects may represent an adaptive mechanism by which urban-dwelling species can persist within a variable landscape. In this study, we assess urban and rural differences in egg yolk bacterial killing ability (BKA), corticosterone (CORT), oxidative status (d-ROMs), and energy metabolites (free glycerol and triglycerides), and their association with female immune status and egg quality. Within a laboratory setting, we immune challenged urban lizards via lipopolysaccharide injection (LPS) to test whether physiological changes associated with immune system activity impacted egg yolk investment. We found urban females had higher mite loads than rural females, however mite burden was related to yolk BKA in rural eggs, but not urban eggs. While yolk BKA differed between urban and rural sites, egg mass and egg viability (fertilized vs. unfertilized) were strong predictors of yolk physiology and may imply tradeoffs exist between maintenance and reproduction. LPS treatment caused a decrease in egg yolk d-ROMs relative to the control treatments, supporting results from previous research. Finally, urban lizards laid a higher proportion of unfertilized eggs, which differed in egg yolk BKA, CORT, and triglycerides in comparison to fertilized eggs. Because rural lizards laid only viable eggs during this study, these results suggest that reduced egg viability is a potential cost of living in an urban environment. Furthermore, these results help us better understand potential downstream impacts of urbanization on offspring survival, fitness, and overall population health.
Recent studies have shown that the egg yolk maternal components, which are a mixture of substances that can affect the developing embryo, do not act separately but are interconnected and co-adapted. Surprisingly, no study to date has focused on the associations between maternally derived albumen steroids and albumen and eggshell compounds with pleiotropic effects. Eggshell pigment protoporphyrin (PROTO IX) should provide primary antimicrobial protection for eggs, but as a proven pro-oxidant, it may compromise female fitness. Abundant albumen proteins ovotransferrin (OVOTR) and lysozyme (LSM) have been shown to have antimicrobial, antioxidant, immunoregulatory and growth-regulatory roles. To investigate associations between albumen steroids and OVOTR, LSM and eggshell cuticle PROTO IX, we used chicken eggs with differently pigmented eggshells. We found that albumen steroid hormones were strongly intercorrelated. In addition, we revealed that albumen LSM and testosterone (T) were positively associated, while a negative association was found between albumen LSM and pregnenolone (P5). Eggshell cuticle PROTO IX was negatively associated with the concentration of albumen 17α-hydroxypregnenolone (17-OHP5). Finally, of all the hormones tested, only the concentration of albumen 17-OHP5 correlated negatively with egg volume and varied with eggshell colour and chicken breed. Although experimental evidence for the effect of maternal albumen steroids on avian developing embryo is still scarce, our study is the first to highlight co-variation and potential co-adjustment of maternally derived albumen steroids, proteins and eggshell cuticle pigment suggesting similar allocation mechanisms known for yolk maternal compounds with the potential to influence the avian embryo and offspring phenotype.
Maternal effects have been reported to alter offspring phenotype in laying hens. In this study, we investigated the effects of maternal environment and maternal age on egg traits and offspring development and behaviour. For this, we ran two experiments. First (E1), commercial hybrid hens were reared either in aviary or barren brooding cages, then housed in aviary, conventional cages or furnished (enriched) cages, thus forming different maternal housing treatments. Hens from each treatment were inseminated at three ages, and measures of egg composition, yolk testosterone concentration and offspring’s development, anxiety and fearfulness were assessed. In experiment 2 (E2), maternal age effects on offspring's growth and behaviour were further investigated using fertile eggs from commercial breeder flocks at three different ages. Results from E1 showed that Old hens laid heavier eggs with less yolk testosterone and produced offspring with fewer indicators of anxiety and fearfulness. Maternal rearing and housing affected egg traits, offspring weight and behaviour, but not in a consistent way. Effects of maternal age were not replicated in E2, possibly due to differences in management or higher tolerance to maternal effects in commercial breeders. Overall, our research confirms that maternal age and maternal environment affects egg composition, with maternal age specifically affecting yolk testosterone concentration, which may mediate physical and behavioural effects in offspring.
Maternally-derived hormones induce variation in offspring phenotype, with consequences that can carry-over into post-natal life and even into adulthood. In birds, maternal egg corticosterone (CORT) is known to exert contrasting effects on offspring morphology, physiology and behaviour after hatching. However, information on the effects of CORT exposure on pre-hatching embryonic development is limited. We experimentally increased yolk CORT levels in yellow-legged gull (Larus michahellis) eggs, and assessed the effects on embryo pre-hatching development and oxidative status of brain and liver. CORT-supplemented embryos reached a larger skeletal size and liver mass compared to controls. Embryos from CORT-injected last-laid eggs showed decreased activity of the hepatic antioxidant enzymes superoxide dismutase and catalase, while intermediate-laid eggs showed an increased levels of lipid peroxidation. However, elevated yolk CORT did not affect oxidative stress endpoints in the brain. Our results indicate that elevated yolk CORT levels affect prenatal embryo development by promoting skeletal growth, and induce laying sequence- and organ-specific oxidative imbalance, with potential adverse consequences during postnatal life especially for late-hatched offspring.
Stressors experienced by layer breeders during egg production can lead to changes in the egg hormone content, potentially impacting their offspring, the commercial layers. Genetic differences might also affect the offspring's susceptibility to maternal experiences. In this study, we tested if maternal stress affects measures of stress and fear in five strains of layer breeders: commercial brown 1 & 2, commercial white 1 & 2 and a pure line White Leghorn. Each strain was equally separated into two groups: "Maternal Stress" (MS), where hens were subjected to a series of 8 consecutive days of acute psychological stressors, and "Control," which received routine husbandry. Additional eggs from Control were injected either with corticosterone diluted in a vehicle solution ("CORT") or just "Vehicle." Stress- and fear-responses of the offspring were measured in a plasma corticosterone test and a combined human approach and novel object test. While the stress treatments did not affect the measured endpoints in the offspring, significant strain differences were found. The offspring of the white strains showed a higher physiological response compared to brown strains and the White 2 offspring was the least fearful strain in the human approach test. Our study found that neither the acute psychological stressors experienced by layer breeders nor the egg injections of corticosterone affected the parameters tested in their offspring. Post hoc power analyses suggest that the lack of treatment effects might be due to a small sample size (type II error). Although studies on larger flocks of layers are still needed, our results provide an initial understanding of an important subject, as in poultry production, layer breeders are often subjected to short-term stressors. In addition, our results suggest the dissociation between the physiological and behavioural parameters of stress response in laying hens, showing that increased concentrations of plasma corticosterone in response to stress might not be directly associated with high levels of fear.
Variation in the concentration of antioxidants and hormones of maternal origin in the eggs of birds can have a profound influence on offspring phenotype both prenatally and postnatally. Egg maternal substances can have interacting effects, but experimental studies of the consequences of the combined variation in the egg concentration of such molecules are extremely rare, particularly as far as prenatal stages are concerned. We manipulated the yolk concentration of vitamin E and corticosterone, which are, respectively, the main antioxidant and the main glucocorticoid hormone in bird eggs, both independently and simultaneously, and we tested their separate and combined effects on growth and oxidative status in the liver and in the brain of yellow-legged gull (Larus michahellis) embryos. Egg supplementation of relatively large physiological doses of corticosterone depressed embryo growth (total body mass, tarsus length and liver mass), whereas administration of vitamin E in association with corticosterone restored normal growth. Vitamin E did not affect embryo growth when administered alone. We further analysed the independent and combined effects of vitamin E and corticosterone on liver and brain total antioxidant capacity, the concentration of reactive oxygen molecules and lipid peroxidation. Vitamin E significantly reduced liver total antioxidant capacity, while corticosterone depressed brain lipid peroxidation. Prenatal exposure to vitamin E and corticosterone appears to have antagonistic effects on body growth, although vitamin E is not limiting in yellow-legged gull eggs. In combination with the results of previous experiments on the same species applying smaller experimental doses or focusing on the postnatal rather than prenatal life stages, our findings indicate that the effects of a physiological increase in the egg concentration of these substances can be life stage and dose specific, implying that generalizing prenatal effects of egg compounds may not be feasible.
Oxidative stress experienced during early development can negatively affect diverse life-history traits, and organisms have evolved complex defence systems against its detrimental effects. Bird eggs contain maternally derived exogenous antioxidants that play a major role in embryo protection from oxidative damage, including the negative effects on telomere dynamics. In this study on the yellow-legged gull (Larus michahellis), we manipulated the concentration of vitamin E (VE) in the egg yolk and analysed the consequences on oxidative status markers and telomere length in the hatchlings. This study provides the first experimental evidence that, contrary to the expectation, a physiological increase in yolk VE concentration boosted total antioxidant capacity and reduced the concentration of pro-oxidant molecules in the plasma, but did not reduce telomere attrition or ameliorate oxidative damage to proteins and lipids in the early postnatal period.
Organisms have evolved complex defense systems against oxidative stress. Bird eggs contain maternally derived antioxidants that protect embryos from oxidative damage. The antioxidant system components are thought to be integrated, but few studies have analyzed the covariation between antioxidant concentrations, embryo 'oxidative status' and morphology. In addition, no study has tested the effects of experimental change in yolk antioxidant concentration on other antioxidants, on their reciprocal relationships and on their relationships with embryo oxidative status or growth, which are expected if antioxidants defenses are integrated. In yellow-legged gull (Larus michahellis) embryos, we analyzed the covariation between several antioxidants, markers of 'oxidative status' [total antioxidant capacity (TAC), concentration of pro-oxidants (TOS), lipid peroxidation (LPO) and protein carbonylation (PC)] in the yolk, liver and brain, and morphology. Yolk and liver antioxidant concentrations were positively correlated reciprocally and with embryo size, and positively predicted TAC but not oxidative status. TOS and LPO were positively correlated in the liver, while TAC and LPO were negatively correlated in the brain. Weak relationships existed between antioxidants and TOS, PC and LPO. The effects of antioxidants on oxidative status and morphology were non-synergistic. An experimental physiological increase in yolk Vitamin E had very weak effects on the relationships between other antioxidants or oxidative status and Vitamin E concentration, the concentration of other antioxidants or oxidative status; the covariation between other antioxidants and oxidative status, and relationships between morphology or oxidative status and other antioxidants, challenging the common wisdom of strong functional relationships among antioxidants, at least for embryos in the wild.
Conditions experienced during prenatal development can have long-lasting organizational effects on offspring. Maternal carotenoids deposited in the eggs of birds and other oviparous species play an important role during fast embryonic growth and chick development through their antioxidant properties. However, the long-term consequences of variation in maternal carotenoid transfer for the offspring have seldom been considered. Since plasma carotenoid levels at adulthood are known to influence testis size and yolk carotenoid levels influence the ability to extract carotenoids later in life, we hypothesized that maternally transmitted carotenoids might influence gonad size at adulthood. Here, we showed that male Japanese quail (Coturnix japonica) originating from a carotenoid-enriched egg had smaller testes than control individuals at adulthood. This result shows that yolk carotenoids have long-term organizational effects. In addition, given that carotenoid intake at sexual maturity increases sperm quality and that a decreased testis size is associated with a lower sperm production, we propose that carotenoid exposure during embryo development might influence a trade-off between ejaculate size and sperm quality.
We discuss the long-term effects of stress exposure in pre- and early postnal life. We present an evolutionary framework within which such effects can be viewed, and describe how the outcomes might vary with species life histories. We focus on stressors that induce increases in glucocorticoid hormones and discuss the advantages of an experimental approach. We describe a number of studies demonstrating how exposure to these hormones in early life can influence stress responsiveness and have substantial long-term, negative consequences for adult longevity. We also describe how early life exposure to mild levels of stressors can have beneficial effects on resilience to stress in later life, and discuss how the balance of costs and benefits is likely dependent on the nature of the adult environment.
1.Parental phenotype and environment have cascading effects on offspring performance. Such ‘maternal’ effects are often mediated by the size and quality of the eggs in terms of maternal endogenous substances or exogenous components.2.Maternal effects can function to maximize parental fitness via adaptive allocation of components among sibling eggs. In birds, the often documented variation in concentration of egg dietary antioxidants or other egg compounds with laying order may thus reflect adaptive parental favouritism or maternal physiological constraints.3.Vitamins are found in the yolk at high concentrations and accomplish important physiological functions. Tocopherols (‘vitamin E’) have major roles in antioxidant protection, but the consequence of variation in their concentration for chick phenotype has never been experimentally investigated under a natural selection regime.4.In the yellow-legged gull (Larus michahellis) free-ranging population we studied, vitamin E concentration and egg size decline from the first to the third (last) egg. Chicks from third eggs are smaller and have lower viability compared to those from sibling eggs. Here, we tested the effect of injecting a physiological dose of vitamin E in the egg yolk on chick body size at hatching by comparison with chicks from sham-injected eggs.5.As predicted, chicks from vitamin E injected third eggs were larger than chicks from control third eggs, whereas vitamin E had no effect on chicks from the other eggs. Thus, vitamin E injection disrupted the brood size hierarchy according to laying order normally found in this species.6.In species that adopt a brood reduction strategy, differential allocation of vitamin E to the eggs can contribute to establish a hierarchy of reproductive value among progeny members functioning to maximize parental fitness. The present results therefore have general implications for the study of the evolution of maternal effects mediated by transfer of potentially limiting maternal antioxidants to the eggs.This article is protected by copyright. All rights reserved.
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.
Empirical evidence is growing that the offspring sex ratio in birds can be biased in relation to the body condition of parents
during breeding. The sex ratio bias may come about because (1) the actual production of the two sexes may be skewed and/or
(2) there may be a sex bias in early nestling mortality contingent on parental condition. By manipulating parental condition
and giving them a control brood to rear, thereby eliminating effects operating via the eggs, we examined the extent to which
parental condition influences the post-hatching survival of male and female lesser black-backed gulls, Larus fuscus. We found that the pre-fledging survival of male chicks was strongly reduced in all-male broods reared by parents in poor
condition. Pre-fledging survival of female chicks was, however, unaffected by parental condition or brood sex composition.
Thus, independently of any production biases, sex differences in nestling mortality alone can bias the offspring sex ratio
at fledging in relation to the prevailing rearing conditions. In other studies on gulls we have, however, also shown that
females in poor condition at laying preferentially produce female eggs. Clearly a bias in fledging sex ratio can occur within
the same species due to a combination of differential production and differential post-laying mortality; the latter can involve
a differential effect of poor egg quality on male and female offspring, differential effects of brood sex composition on their
survival and a difference in the capacity of parents to rear males and females. All of these processes need to be taken into
account in attempting to understand offspring sex ratios.
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 days6 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.
Early embryonic exposure to maternal glucocorticoids can broadly impact physiology and behaviour across phylogenetically diverse taxa. The transfer of maternal glucocorticoids to offspring may be an inevitable cost associated with poor environmental conditions, or serve as a maternal effect that alters offspring phenotype in preparation for a stressful environment. Regardless, maternal glucocorticoids are likely to have both costs and benefits that are paid and collected over different developmental time periods. We manipulated yolk corticosterone (cort) in domestic chickens (Gallus domesticus) to examine the potential impacts of embryonic exposure to maternal stress on the juvenile stress response and cellular ageing. Here, we report that juveniles exposed to experimentally increased cort in ovo had a protracted decline in cort during the recovery phase of the stress response. All birds, regardless of treatment group, shifted to oxidative stress during an acute stress response. In addition, embryonic exposure to cort resulted in higher levels of reactive oxygen metabolites and an over-representation of short telomeres compared with the control birds. In many species, individuals with higher levels of oxidative stress and shorter telomeres have the poorest survival prospects. Given this, long-term costs of glucocorticoid-induced phenotypes may include accelerated ageing and increased mortality.
ABSTRACT:
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.
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.
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.
Conditions experienced during early life can influence the development of an organism and several physiological traits, even in adulthood. An important factor is the level of oxidative stress experienced during early life. In birds, extra-genomic egg substances, such as the testosterone hormone, may exert a widespread influence over the offspring phenotype. Interestingly, testosterone can also upregulate the bioavailability of certain antioxidants but simultaneously increases the susceptibility to oxidative stress in adulthood. However, little is known about the effects of maternally derived yolk testosterone on oxidative stress in developing birds. Here, we investigated the role of yolk testosterone on oxidative stress of yellow-legged gull chicks during their early development by experimentally increasing yolk testosterone levels. Levels of antioxidants, reactive oxygen species and lipid oxidative damage were determined in plasma during nestlings' growth. Our results revealed that, contrary to control chicks, birds hatched from testosterone-treated eggs did not show an increase in the levels of oxidative damage during postnatal development. Moreover, the same birds showed a transient increase in plasma antioxidant levels. Our results suggest that yolk testosterone may shape the oxidative stress-resistance phenotype of the chicks during early development owing to an increase in antioxidant defences and repair processes.
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.
The costs of coping with stressful situations are traded-off against other functions such as immune responses. This trade-off may explain why corticosterone secretion reduces immune reactions. Corticosterone differentially affects various immunity components. However, which component is suppressed varies between studies. It remains unclear whether the trade-off in energy, nutrition, autoimmunity or oxidative stress accounts for differential immunosuppression. In this study, we investigated whether corticosterone differentially affects the constitutive innate and humoral acquired immunity. We used barn owl nestlings, implanting 50% with a corticosterone-releasing pellet and the other 50% with a placebo pellet. To measure the effect on humoral immunity we vaccinated 50% of the corticosterone-nestlings and 50% of the placebo-nestlings with the antigens 'Tetravac' and the other 50% were injected with PBS. To assess the costs of elevated corticosterone, we measured body mass and resistance to oxidative stress. Administration of corticosterone increased corticosterone levels whereas vaccination induced the production of antibodies. Corticosterone reduced the production of antibodies, but it did not significantly affect the constitutive innate immunity. Corticosterone reduced body growth and resistance to oxidative stress. Under stressful conditions barn owl nestlings seem to keep the constitutive innate immunity, whereas elevated corticosterone levels negatively affected inducible immune responses. We found evidence that mounting a humoral immune reaction is not costly in terms of growth, but reduces the resistance to oxidative stress independently of corticosterone administration. We suggest that humoral immunity is suppressed because the risk of immunopathologies may be disproportionately high when mounting an antibody response under stressful situations.
The present study had been conducted to explore the effect of corticosterone (CORT) on the induction of lipid peroxidation in skeletal muscle. The experimental chickens were subjected to 1 single s.c. injection of CORT (4 mg/kg of BW). Blood samples were obtained at the beginning and end of a 3-h experimental period. Muscle samples were obtained from musculus pectoralis major and musculus biceps femoris (BF) before and immediately after slaughter, and at 24- and 48-h time points postmortem. The result showed that plasma level of TBA reacting substances (TBARS) was significantly increased, whereas the activity of super-oxide dismutase was decreased after CORT injection. Plasma level of total antioxidant power, as reflected by the ferric reducing-antioxidant power, was increased by CORT administration. The preslaughter level of TBARS could be increased by CORT administration in pectoralis major. In contrast, although the preslaughter level of TBARS was not significantly changed by CORT administration in BF, the augmented TBARS level was detected at 48 h postmortem in BF of CORT chickens. During the period of storage, the concentrations of TBARS increased, whereas ferric reducing-antioxidant power level decreased over time regardless of treatment. In conclusion, the data suggest that elevated CORT due to preslaughter stress would affect the redox balance in skeletal muscle. The result suggested that the oxidative stability during storage of meat is associated with the alteration in muscle physiology induced by CORT administration.
Oxidation of low density lipoprotein (LDL) may be involved in the development of atherosclerosis. It has recently been shown
that α-tocopherol (α-TOH) can act either as an antioxidant or prooxidant for isolated low density lipoprotein (LDL). In the
absence of an effective co-antioxidant, α-TOH is a prooxidant and this activity is evidently due to reaction of the α-tocopheroxyl
radical (α-TO•) with the LDL's polyunsaturated lipids (Bowry, V. B., and Stocker, R.(1993) J. Am. Chem. Soc. 115, 6029-6045). Herein we examined the effectiveness of selected natural and synthetic radical scavengers as co-antioxidants for inhibiting peroxyl radical-induced peroxidation in LDL that is devoid of ubiquinol-10 (an effective endogenous co-antioxidant)
but still contains most of its natural complement of α-TOH. Various quinols, catechols, and aminophenols, as well as ascorbate,
6-palmityl ascorbate, and bilirubin, were very effective co-antioxidants under our test conditions, whereas ordinary phenolic
antioxidants, including short-tailed α-TOH homologues, were less effective. Reduced glutathione, urate, and Probucol were
ineffective. These findings confirm that the prooxidant activity of α-TOH in LDL relies heavily on the segregation of water-insoluble
radicals (particularly α-TO•) into individual LDL particles, since it was those compounds that are expected to either irreversibly
reduce α-TO• or accelerate the diffusion of radicals between particles which most effectively inhibited the tocopherol-mediated phase of peroxidation. Theoretical and practical implications
of these findings are discussed, as is their relevance to the “LDL oxidation” hypothesis of atherogenesis.
The relationship between the dietary level of vitamin E (VE) and the immune response of broilers was studied in three experiments. Immunity was assessed as antibody production to infectious bronchitis virus (IBV), SRBC, and Brucella abortus (BA) antigens, mitogenic response to phytohemagglutinin A (PHA) and concanavalin A (Con A), cutaneous basophil hypersensitivity (CBH) to PHA, and lipopolysaccharide induction of acute-phase proteins (APP) and heterophilia. A range of VE (0, 10, 17.5, 25, 37.5, 50, 100, and 200 IU/kg) levels were supplemented to a basal diet (corn-soy) containing 10.2 IU of VE/kg. We found a dose-dependent increase in antibody production in response to attenuated IBV between 0 and 25 IU/kg of supplemented VE and no further increase at higher levels. Antibody levels to SRBC were higher in birds supplemented with 50 IU of VE/kg compared to those supplemented with 0 or 200 IU/kg of VE. Antibody production in response to BA antigens was not influenced by VE. Mitogenic responses were suppressed by supplemented VE in Experiment 1 for PHA (25 IU/kg diet) and Con A (25 and 50 IU/kg diets). CBH and APP levels were not affected by VE. Heterophilia was lowest at 50 IU/kg 6 h after lipopolysaccharide injection (Experiment 1). Our study showed that moderate (25 to 50 IU/kg) levels of VE supplementation were most immunomodulatory and that high levels were less effective.
Egg quality is a phenotype of, and can profoundly influence fitness in, both mother and offspring. However, the physiological mechanisms that underlie this maternal effect are poorly understood. Carotenoids are hypothesized to enhance antioxidant activity and immune function, and are responsible for the pigmentation of egg yolk. The proximate basis and consequences of this maternal investment, however, have not previously been studied in wild birds. In this supplemental feeding study of lesser black-backed gulls, Larus fuscus, carotenoid-fed females are shown to have increased integument pigmentation, higher plasma concentrations of carotenoids and antioxidant activity, and lower plasma concentrations of immunoglobulins (Igs) in comparison with controls. In turn, carotenoid-fed females produced eggs containing high carotenoid but low Ig concentrations (i.e. passive immunity), whereas control females produced eggs containing low carotenoid but high Ig concentrations. Within-clutch patterns of these resources varied over the laying sequence in a similar manner in both carotenoid-fed and control nests. Our results suggest that carotenoids could be one resource responsible for egg quality maternal effects in birds. We discuss the possible implications of carotenoid-mediated effects on phenotype for fitness in mothers and their offspring.
Carotenoids provide pigmentation to avian species, and also have immunomodulatory potential, although experimental results are often inconsistent. Therefore, dietary carotenoid deposition into immune tissue of growing chicks was examined in relation to their maternal carotenoid status (i.e., yolk carotenoid level). Single-comb white leghorn chicks were hatched from carotenoid-replete (C+) or carotenoid-deplete (C-) eggs. For 4 wk posthatch, chicks were fed diets whose carotenoid level ranged from 0 to 38 mg total carotenoid/kg. Carotenoid additions consisted of lutein + canthaxanthin at a ratio of 4:1. After 4 wk, the carotenoid concentration of thymus, bursa, liver, plasma and shank epithelium was measured by HPLC. Egg yolk-derived carotenoids were detectable in chicks fed 0 dietary carotenoids for 4 wk. Chicks hatched from C+ eggs had significantly greater tissue lutein, zeaxanthin and/or canthaxanthin for all tissues (P < 0.05), compared to chicks hatched from C- eggs. Only bursa carotenoids were not dependent on chick diet (P = 0.24); for all other tissues, C+ chicks incorporated dietary carotenoids in a dose-dependent manner (P < 0.01), whereas C- chicks never achieved the same level of carotenoid incorporation. This study demonstrated the importance of maternal carotenoid status on incorporation of yolk- and diet-derived tissue carotenoids in an avian model, and may explain some variability in carotenoid-based research, given that maternal carotenoid status is rarely controlled.
The effects of maternal vitamin E supplementation on the antioxidant status of chicks were investigated. Female breeder chicks were fed corn-soybean growing diets without supplemental vitamin E for a 17-wk developmental period. After 17 wk, the birds were randomly assigned to 5 treatments and fed corn-soybean diets supplemented with 0, 40, 80, 120, and 160 mg/kg vitamin E (all-rac-alpha-tocopherol acetate), respectively. Blood samples were collected and pullets were artificially inseminated at 35 wk of age. Eggs laid beginning on d 2 after insemination were placed in an incubator. At the time of hatching, 12 chicks from each treatment were randomly sampled and killed. Livers and brains of chicks were collected for the subsequent evaluation of antioxidant status. Plasma vitamin E concentrations increased linearly (P < 0.001; r = 0.997) with the increase in supplemental vitamin E, but those in egg yolk reached a plateau at 120 mg/kg supplemental vitamin E. The malondialdehyde (MDA) concentration, an indicator of lipid peroxidation, of chick brain decreased linearly (P < 0.01; r = -0.909) with the increase in supplemental vitamin E. Pullets given 160 mg/kg supplemental vitamin E had lower plasma MDA concentrations than those given 0 mg/kg (P < 0.05). Similar results were found for the reactive oxygen species levels, an indicator of oxidative stress, of chick brain and liver. For antioxidant enzymes, chicks of pullets given 120 mg/kg supplemental vitamin E had higher (P < 0.05) activities of liver catalase than those given 0-80 mg/kg. Chicks of pullets given 160 mg/kg supplemental vitamin E had higher (P < 0.05) activities of brain superoxide dismutase than those given 0-40 mg/kg. These results indicated that maternal supplementation with high levels of vitamin E (120-160 mg/kg) enhances antioxidant capability and depresses oxidative stress in chicks.
Behavioral lateralization is widespread across vertebrates. The development of lateralization is affected by both genetic and environmental factors. In birds, maternal substances in the egg can affect offspring lateralization via activational and/or organizational effects. Corticosterone affects the development of brain asymmetry, suggesting that variation in yolk corticosterone concentration may also influence post-natal behavioral lateralization, a hypothesis that has never been tested so far. In the yellow-legged gull (Larus michahellis), we increased yolk corticosterone concentration within physiological limits and analyzed the direction of lateralization of hatchlings in reverting from supine to prone position ('RTP' response) and in pecking at dummy parental bills to solicit food provisioning ('begging' response). We found that corticosterone treatment negatively affects the frequency of begging and it may cause a slight leftward lateralization. However, the direction of lateralization of the RTP response was not affected by corticosterone administration. Thus, our study shows a maternal effect mediated by corticosterone on a behavioral trait involved in parent-offspring communication during food provisioning events. The findings on lateralization are not conclusive due to the weak effect size but provide information for further ecological and evolutionary studies, investigating mechanisms underlying the development of lateralization.
Mothers can shape the developmental trajectory of their offspring through the transmission of resources such as hormones, antioxidants or immunoglobulins. Over the last two decades, an abundant literature on maternal effects in birds has shown that several of these compounds (i.e. androgens, glucocorticoids and antioxidants) often influence the same offspring phenotypic traits (i.e. growth, immunity or oxidative stress levels), making interaction effects between egg components a likely scenario. However, the potential interactive effects of maternally transmitted compounds on offspring development and potential co-adjustment of these compounds within an egg are still poorly understood. Here, we report the results of an interspecific comparative analysis on birds' egg yolk composition (i.e. androgens and antioxidants) where we found that yolk carotenoid and vitamin E concentrations are positively associated, supporting the hypothesis that these two antioxidants act in synergy. The concentrations of vitamin E also increased with increasing concentrations of testosterone. This last result confirms the emerging idea that androgens and antioxidants are co-adjusted within eggs and that maternally transmitted antioxidants might limit the potential direct and indirect effects of prenatal exposure to high testosterone levels on oxidative stress.
The function of carotenoids in the barn swallow (Hirundo rustica) A signal will evolve and spread only if costly, since its costs will prevent cheating, thereby making the signal reliable. In this thesis I investigate carotenoid-based signals that affect fitness in the barn swallow. Carotenoids are coloured molecules synthesised by photosynthetic organisms, and animals therefore have to ingest carotenoids with their diet. Apart from being important in reproduction, they act as free radical scavengers and have several physiological functions related to immune defence. Rarity of carotenoids in the environment leads to a trade-off between carotenoid use in reproduction, allocation to signal coloration and maintenance of health. Simply stated, only high quality individuals are hypothesised to acquire a sufficient amount of carotenoids to invest in coloured signals while maintaining prime health status. Firstly, I focused on carotenoid availability in the diet of barn swallows, to investigate whether they are a limited resource. My results indicate that while carotenoids are present in their diet, the carotenoid content of prey that barn swallows obtain are on average lower than that found in randomly collected samples within their foraging range. Greater variance in carotenoids in prey among than within nests indicates that individuals vary in their ability to obtain carotenoid-rich food. Secondly, I focused on the arrival date of barn swallow at their breeding grounds. Arrival date has previously been shown to affect fitness in this species. Early arrival after migration is a costly activity and the high metabolism associated with migration is likely to increase oxidative stress due to free radical production. Concentration and variation in carotenoids in blood of recently arrived adult males suggest a role of carotenoids in determining arrival date. If low quality individuals suffer more from peroxidation due to higher production of free radicals than high quality individuals, they cannot advance their arrival, because of greater need for carotenoids and other antioxidants. In addition, I analysed amount and variation of carotenoids in blood of adult females during reproduction. Carotenoids are invested both in eggs and maintenance of female health during reproduction. I report a decrease in carotenoid concentration in blood of females during reproduction and a significant difference between sexes during reproduction. During laying, females had a significant lower concentration of circulating carotenoids than males indicating difference in use of carotenoids between the sexes. Finally I investigated whether sexual dichromatism exists in colour of carotenoid-based red facial feathers of adult barn swallows. Males developed a more saturated colour, with a higher absorbance in the lutein band, and a hue more shifted to the red than females. Pigments also covered a relatively greater part of each feather in males than in females. My results show that males with a higher absorbance in the lutein band have a larger probability of obtaining a mate, indicating that the carotenoid-based red facial coloration could have evolved through female choice for more conspicuous male coloration. Together, these results suggest that carotenoids play a central role in the evolution of reliable signals in the barn swallow.
This chapter reviews maternal hormones in avian eggs, their effects, their regulating mechanisms, and their potential Darwinian function, i.e., the way in which they maximize survival and reproductive success. An avian egg contains all the substances needed to produce a fully functioning organism. In addition to basic resources (proteins, lipids, water, and minerals), eggs contain a range of other substances, such as vitamins, antibodies, and enzymes. These appear to fulfill important functions, acting as regulators of embryonic development and protecting the embryo against pathogens, or may represent metabolic waste products. The eggs also contain substantial levels of maternal hormones. Their levels vary in relation to a range of factors such as laying order, food availability, breeding density, and male quality. Yolk hormones subsequently influence offspring growth, behavior, physiology, immune system, and ultimately survival and reproduction. Separate control over hormone levels in circulation and yolk allows independent effects on offspring and mothers, while regulation of the response to maternal hormones allows offspring to fine-tune the effects on different traits.
Nutrient administration in-ovo could be considered as an alternative method to improve hatchability and duckling weights followed by better economic performance. On the 12th day of incubation, fertile duck eggs (n= 500) were distributed into 5 groups: uninjected control; 0.1 ml corn oil; 0.1 ml corn oil plus 10 mg vitamin E; 0.1 ml saline; and 0.1 ml saline plus 3 mg ascorbic acid. In-ovo injection of vitamin E but not ascorbic acid resulted in higher hatchability percentage compared to the uninjected control; however this was not statistically confirmed. In-ovo injection of either vitamin E or ascorbic acid resulted in significantly (P < 0.05) higher body weights at hatch, final body weights, and feed intakes than the uninjected control group. However, both male and female ducklings hatched from eggs injected with either vitamin E or ascorbic acid had better feed conversion during the starting period only. The carcass characteristics and relative lymphoid organ weights did not differ (P < 0.05) between the in ovo injected groups and the uninjected controls. In-ovo injection of either vitamin E or ascorbic acid resulted in significant increase (P < 0.05) in the geometric mean of the 1st antibodies titers of males and the 1st and 2nd antibodies titers of females compared to the uninjected control. It could be concluded that in-ovo injection of vitamin E and ascorbic acid improve the embryonic and post-hatch growth of Muscovy ducks.
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.
In a 2-yr study of the survival of Glaucous-winged Gull (Larus glaucescens) chicks, pecking of trespassing chicks by neighboring adults was the major cause of chick mortality. In years of both low and high food availability chick survival was strongly correlated with growth rates. Chicks that grew slowly were more likely to be killed by neighbors than fast-growing chicks. In the year of low food availability, among slow-growing chicks, those hatched early in the season on large territories had better survival rates than chicks hatched late in the season on small territories. In the year of high food availability, timing of breeding and territory size had little bearing on chick survival. A model of chick survival in relation to timing of breeding predicts that when neighbor interference is the major cause of chick mortality, chicks hatched early in the season will have the highest probability of surviving. When predators are the major cause of chick mortality, chicks hatching in the middle of the breeding season will have highest survival. A second model relates chick survival to territory size. Chicks raised on large territories will most likely avoid neighbor interference, while predation will select for either small or large territory size, depending upon the effectiveness of group mobbing against the predator.
Most organisms, including ourselves, are exposed to environmental stressors at various points during life, and responses to such stressors have been optimised by evolution to give the best fitness outcomes. It is expected that environmental change will substantially increase long-term stress exposure in many animal groups in the coming decades. A major challenge for biologists is to understand and predict how this will influence individuals, populations and ecosystems, and over what time scale such effects will occur. This requires a multi-disciplinary approach, combining studies of mechanisms with studies of fitness consequences for individuals and their descendants. In this review, I discuss the positive and negative fitness consequences of responses to stressful environments, particularly during early life, and with an emphasis on studies in birds. As many of the mechanisms underlying stress responses are highly conserved across the vertebrate groups, the findings from these studies have general applicability when interpreted in a life history context. One important route that has recently been identified whereby chronic stress exposure can affect health and longevity over long time frames is via effects on telomere dynamics. Much of this work has so far been done on humans, and is correlational in nature, but studies on other taxa, and experimental work, are increasing. I summarise the relevant aspects of vertebrate telomere biology and critically appraise our current knowledge with a view to pointing out important future research directions for our understanding of how stress exposure influences life histories.
Oxidation of human low-density lipoprotein (LDL) is implicated as an initiator of athcrosclerosis. Alpha-tocopherol (alpha-TocH) may thus inhibit atherosclerosis because it is the major and most active chain-breaking antioxidant in extracted LDL lipid. Our studies show, however, that alpha-TocH can bc a strong prooxidant for the LDL itself, i.e., an aqueous dispersion of lipid-bearing particles. Thus, a steady flux (R(g)) of alkylperoxyl radicals (ROO.) generated from a water-soluble azo initiator induced lipid peroxidation in LDL which was faster in the presence of alpha-TocH than in its absence (for R(g) < 2 nM s-1), insensitive to R(g) and [O2], and inhibited by vitamin C, ubiquinol-10 (normally present in fresh LDL), and small phenolic antioxidants but not inhibited by the aqueous radical scavenger uric acid. Furthermore, LDL peroxidation induced by a water- or lipid-soluble azo initiator or by transition metals in Ham's F-10 cell culture medium was accelerated by increasing the concentration of alpha-TocH in LDL. We propose that LDL peroxidation is initiated by the reaction of ROO. with alpha-TocH and that the inability of the alpha-Toc. formed in this reaction to escape from an LDL particle then forces alpha-Toc. to propagate a radical chain via its reaction with PUFA lipid (LH) within the particle (alpha-Toc. + LH + O2 --> alpha-TocH + LOO.). Termination of a radical chain occurs when a peroxidizing LDL particle captures a second radical from the aqueous medium (ROO. + alpha-Toc. --> nonradical products). Steady-state kinetic analysis of this mechanism yields a theoretical model for tocopherol-mediated peroxidation (TMP) in lipid dispersions which fully explains our findings for LDL. We conclude that peroxidation of LDL lipid can (only) be prevented by agents which eliminate the alpha-Toc. radical: vitamin C and LDL-associated ubiquinol-10 do so by ''exporting the radical'' into the aqueous medium, whereas small phenolic antioxidants (e.g., butylated hydroxytoluene) accelerate the transfer of radicals between particles. The theoretical and practical implications of TMP in LDL, dispersions, and bulk lipids are discussed.
Avian eggs are rich in carotenoids, which derive from maternal diet where they may be available in limiting amounts. Egg carotenoids may accomplish major roles in antioxidant protection or modulate physiological functions and growth, interfering with offspring redox status, potentially in a sex-dependent way. In this study of maternal effects in relation to sex and laying order of yellow-legged gull (Larus michahellis) chicks, we analyzed the consequences of increased yolk lutein concentration on plasma antioxidant capacity (AOC) and an index of early oxidative damage (reactive oxygen metabolites, ROM), till 9 days after hatching. To this end, for the first time we directly manipulated yolk lutein, thus avoiding any effect on other components of egg quality due to maternal supplementation before laying. Lutein did not increase AOC but increased ROM in males and in first-laid chicks. Hence, lutein did not act as an antioxidant and determined increased early oxidative damage, possibly because of upregulation of immune or other physiological functions, but these effects were sex-related and apparent in first-laid chicks with larger yolk lutein supply. ROM positively covaried with AOC, suggesting a trade-off between AOC and oxidative damage. Moreover, lutein injection altered the covariation between body size or immunity and AOC or ROM. Carotenoids may thus not be major antioxidants in birds and rather affect redox status by increasing oxidative damage in a sex-dependent way and interfere with the resolution of growth trade-offs. In the absence of sex-related allocation, maternal decisions on egg carotenoid concentration may depend on the balance between divergent effects on either sex. Copyright 2011, Oxford University Press.
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.
We tested the hypothesis that exposure of chick embryos to corticosterone leads to increased fear, reduced competitive ability, reduced ability to cross a barrier and reduced growth in juvenile chicks. Behaviour was studied in birds subjected to three different egg injection treatments: a negative control (no treatment of eggs), a positive control (100 ml sesame oil vehicle) and a corticosterone treatment (0.6 mg corticosterone in 100 ml sesame oil). Eggs were injected prior to incubation and the behaviour of chicks was studied during the first 4 weeks of life. Corticosterone treatment increased fear in chicks, as indicated by greater avoidance of an observer in the home pen at 2 weeks of age (P < 0.0001), reduced ability to cross a wall to access feed at 2 weeks of age (P < 0.05) and reduced ability to compete for a wormlike object at 4 weeks of age (P < 0.01). Treatment with corticosterone also reduced body weight at 1 week of age (P < 0.003) and 4 weeks of age (P < 0.04), but not at hatch (P < 0.28). The sesame oil vehicle reduced fear (P < 0.0001), but had no other significant effects. These results indicate that embryonic exposure to corticosterone leads to behavioural and growth deficits in chicks.
Experimental models and observational studies suggest that vitamin E supplementation may prevent cardiovascular disease and cancer. However, several trials of high-dosage vitamin E supplementation showed non-statistically significant increases in total mortality.
To perform a meta-analysis of the dose-response relationship between vitamin E supplementation and total mortality by using data from randomized, controlled trials.
135,967 participants in 19 clinical trials. Of these trials, 9 tested vitamin E alone and 10 tested vitamin E combined with other vitamins or minerals. The dosages of vitamin E ranged from 16.5 to 2000 IU/d (median, 400 IU/d).
PubMed search from 1966 through August 2004, complemented by a search of the Cochrane Clinical Trials Database and review of citations of published reviews and meta-analyses. No language restrictions were applied.
3 investigators independently abstracted study reports. The investigators of the original publications were contacted if required information was not available.
9 of 11 trials testing high-dosage vitamin E (> or =400 IU/d) showed increased risk (risk difference > 0) for all-cause mortality in comparisons of vitamin E versus control. The pooled all-cause mortality risk difference in high-dosage vitamin E trials was 39 per 10,000 persons (95% CI, 3 to 74 per 10,000 persons; P = 0.035). For low-dosage vitamin E trials, the risk difference was -16 per 10,000 persons (CI, -41 to 10 per 10,000 persons; P > 0.2). A dose-response analysis showed a statistically significant relationship between vitamin E dosage and all-cause mortality, with increased risk of dosages greater than 150 IU/d.
High-dosage (> or =400 IU/d) trials were often small and were performed in patients with chronic diseases. The generalizability of the findings to healthy adults is uncertain. Precise estimation of the threshold at which risk increases is difficult.
High-dosage (> or =400 IU/d) vitamin E supplements may increase all-cause mortality and should be avoided.
Prolonged high secretion of glucocorticoids normally reflects a state of chronic stress, which has been associated with an increase in disease susceptibility and reduction in Darwinian fitness. Here, we hypothesize that an increase in oxidative stress accounts for the detrimental effects of prolonged high secretion of glucocorticoids. We performed a meta-analysis on studies where physiological stress was induced by administration of glucocorticoids to evaluate the magnitude of their effects on oxidative stress. Glucocorticoids have a significant effect on oxidative stress (Pearson r = 0.552), although this effect depends on the duration of treatment, and is larger in long-term experiments. Importantly, there was a significant effect on tissue, with brain and heart being the most and the least susceptible to GC-induced oxidative stress, respectively. Furthermore, effect size was larger (1) in studies using both sexes compared to males only, (2) when corticosterone rather than dexamethasone was administered and (3) in juveniles than in adults. These effects were not confounded by species, biochemical biomarker, or whether wild or laboratory animals were studied. In conclusion, our meta-analysis suggests that GC-induced oxidative stress could be a further mechanism underlying increases in disease susceptibility and decreases in Darwinian fitness observed under chronic stress.
The question of why maternal stress influences offspring phenotype is of significant interest to evolutionary physiologists. Although embryonic exposure to maternally derived glucocorticoids (i.e., corticosterone) generally reduces offspring quality, effects may adaptively match maternal quality with offspring demand. We present results from an interannual field experiment in European starlings (Sturnus vulgaris) designed explicitly to examine the fitness consequences of exposing offspring to maternally derived stress hormones. We combined a manipulation of yolk corticosterone (yolk injections) with a manipulation of maternal chick-rearing ability (feather clipping of mothers) to quantify the adaptive value of corticosterone-induced offspring phenotypes in relation to maternal quality. We then examined how corticosterone-induced "matching" within this current reproductive attempt affected future fecundity and maternal survival. First, our results provide support that low-quality mothers transferring elevated corticosterone to eggs invest in daughters as predicted by sex allocation theory. Second, corticosterone-mediated sex-biased investment resulted in rapid male-biased mortality resulting in brood reduction, which provided a better match between maternal quality and brood demand. Third, corticosterone-mediated matching reduced investment in current reproduction for low-quality mothers, resulting in fitness gains through increased survival and future fecundity. Results indicate that the transfer of stress hormones to eggs by low-quality mothers can be adaptive since corticosterone-mediated sex-biased investment matches the quality of a mother to offspring demand, ultimately increasing maternal fitness. Our results also indicate that the branding of the proximate effects of maternal glucocorticoids on offspring as negative ignores the possibility that short-term phenotypic changes may actually increase maternal fitness.
Egg quality may mediate maternal allocation strategies according to progeny sex. In vertebrates, carotenoids have important physiological roles during embryonic and post-natal life, but the consequences of variation in yolk carotenoids for offspring phenotype in oviparous species are largely unknown. In yellow-legged gulls, yolk carotenoids did not vary with embryo sex in combination with egg laying date, order and mass. Yolk lutein supplementation enhanced the growth of sons from first eggs but depressed that of sons from last eggs, enhanced survival of daughters late in the season, and promoted immunity of male chicks and chicks from small eggs. Lack of variation in egg carotenoids in relation to sex and egg features, and the contrasting effects of lutein on sons and daughters, do not support the hypothesis of optimal sex-related egg carotenoid allocation. Carotenoids transferred to the eggs may rather result from a trade-off between opposing effects on sons or daughters.
The eggs of the canary (Serinus canaria) contain variable doses of maternal testosterone. The reported experiments investigated whether testosterone influences nestling growth and how this interacts with differences of the growth of nest mates that are caused by asynchronous hatching. Injections of testosterone into the yolk of unincubated eggs enhanced the growth after hatching compared to nestlings that had hatched simultaneously from control eggs. These differences were established within 22 hr of hatching. Exogenous testosterone promoted growth in both sexes and there was no sexual difference in the growth of control birds. Testoster-one-treated chicks also begged more often for food. Previous studies have shown that the content of maternal testosterone increases in each subsequently laid egg in a clutch. Consistent with the results obtained by testosterone injections nestlings that hatched from eggs with higher concentrations of maternal testosterone grew faster compared to chicks that hatched synchronously from eggs with lower testosterone concentrations. However, more testosterone did not compensate for reduced growth that was caused by later hatching due to asynchronous incubation of clutches. This direct effect of maternal testosterone on growth in combination with a flexible onset of incubation allows to selectively enhance the growth and fitness of individual offspring of a brood.
The yolk of bird eggs contains maternal carotenoids that may act as antioxidants thus influencing offspring performance and survival. However, to our knowledge, this hypothesis has not been subjected to experimental tests and the function of transmission of carotenoids to the egg is largely unknown. We directly manipulated the concentration of the main carotenoid (lutein) in the eggs of barn swallows (Hirundo rustica) and analysed the effect of experimental manipulation on growth of nestlings and two fundamental components of their acquired immunity. Nestlings hatched from lutein-inoculated eggs had larger T-cell-mediated immune response compared with those of two control groups. T-cell-mediated immune response predicted nestling survival until fledging. However, lutein inoculation did not affect antibody response to an immunogen, body mass, tarsus length or plumage development. Nestling body mass and plumage development declined with egg laying order, but the effects of lutein inoculation were independent of egg laying order for all traits. Our results show that maternal yolk carotenoids can have a major effect in promoting a fundamental component of immunity that predicts offspring survival and suggests that adaptive early maternal effects can be mediated by transmission of antioxidants to eggs.
Many environmental perturbations may elevate plasma corticosterone in laying birds, including disease, poor body condition, high predator density, anthropogenic disturbance, and/or food scarcity. When adverse conditions are not dire enough to dictate foregoing reproduction, maternal corticosterone in egg yolk may phenotypically engineer offspring so as to maximize success under the constraints of the local environment. We tested the hypotheses that corticosterone in avian egg yolk should correlate with corticosterone in maternal circulation at the time of laying, and that high corticosterone in yolk should then influence offspring development and adult phenotype. We implanted female Japanese quail (Coturnix coturnix japonica) with corticosterone-filled or empty implants and measured concentrations of corticosterone in the yolk of their eggs. Then we incubated the eggs and raised the chicks to test for effects on growth and hypothalamo-pituitary-adrenal response to capture and restraint in adult offspring. We found that corticosterone implants significantly increased corticosterone in yolk. Furthermore, chicks of corticosterone-implanted mothers grew more slowly than controls and showed higher activity of the hypothalamo-adrenal axis in response to capture and restraint as adults. These results suggest that stress experienced by a laying bird has significant effects on offspring development and adult phenotype, possibly mediated by the transfer of maternal corticosterone to yolk.
By injecting a single 60 microg dose of corticosterone into the eggs of domestic chicks on day 18 of incubation, we have shown that elevated levels of this hormone affect the development of asymmetry in the visual projections from the thalamus to the Wulst regions in the left and right hemispheres of the forebrain. In vehicle-treated (control) embryos this visual pathway develops asymmetry in response to light stimulation during the final stages of incubation, when the embryo is oriented so that its left eye is occluded by its body and its right eye can be stimulated by light entering through the egg shell. Pre-hatching exposure to light leads to more projections from the left side of the thalamus to the right Wulst than from the right side of the thalamus to the left Wulst, as confirmed here by injection of the tracers Fluorogold and Rhodamine into the left and right Wulst followed by counting the number of labelled cell bodies in the thalamus (asymmetry greater in males than females). The chicks injected with corticosterone pre-hatching did not develop any group bias for asymmetry in response to light exposure before hatching. They were random with respect to presence/absence of lateralization and, when present, the lateralization was not as strong as in the controls and its direction was random. The corticosterone-treated group had fewer projections from the left side of the thalamus to the right Wulst than did the controls. The results are considered with respect to maternal deposits of the hormone in the yolk and pre-hatching stress of the embryo.
Eggs of vertebrates contain steroid hormones of maternal origin that may influence offspring performance. Recently, it has been shown that glucocorticoids, which are the main hormones mediating the stress response in vertebrates, are transmitted from the mother to the egg in birds. In addition, mothers with experimentally elevated corticosterone levels lay eggs with larger concentrations of the hormone, which produce slow growing offspring with high activity of the hypothalamo-adrenal axis under acute stress. However, the effects and function of transfer of maternal corticosterone to the eggs are largely unknown. In the present study, we injected corticosterone in freshly laid eggs of yellow-legged gulls (Larus michahellis), thus increasing the concentration of the hormone within its natural range of variation, and analyzed the effect of manipulation on behavioral, morphological, and immune traits of the offspring in the wild. Eggs injected with corticosterone had similar hatching success to controls, but hatched later. Mass loss during incubation was greater for corticosterone-treated eggs, except for the last laid ones. Corticosterone injection reduced rate and loudness of late embryonic vocalizations and the intensity of chick begging display. Tonic immobility response, reflecting innate fearfulness, was unaffected by hormone treatment. Elevated egg corticosterone concentrations depressed T-cell-mediated immunity but had no detectable effects on humoral immune response to a novel antigen, viability at day 10, or growth. Present results suggest that egg corticosterone can affect the behavior and immunity of offspring in birds and disclose a mechanism mediating early maternal effects whereby stress experienced by females may negatively translate to offspring phenotypic quality.
Avian eggs contain substantial amounts of maternal hormones and so provide an excellent model to study hormone-mediated maternal effects. We review this new and rapidly evolving field, taking an ecological and evolutionary approach and focusing on effects and function of maternal androgens in offspring development. Manipulation of yolk levels of androgens within the physiological range indicates that maternal androgens affect behaviour, growth, morphology, immune function and survival of the offspring, in some cases even long after fledging. Descriptive and experimental studies show systematic variation in maternal androgen deposition both within and among clutches, as well as in relation to the sex of the embryo. We discuss the potential adaptive value of maternal androgen transfer at all these three levels. We conclude that maternal androgen deposition in avian eggs provides a flexible mechanism of non-genetic inheritance, by which the mother can favour some offspring over others, and adjust their developmental trajectories to prevailing environmental conditions, producing different phenotypes. However, the literature is less consistent than often assumed and at all three levels, the functional explanations need further experimental testing. The field would greatly benefit from an analysis of the underlying physiological mechanisms.
To develop a new, colorimetric and automated method for measuring total oxidation status (TOS).
The assay is based on the oxidation of ferrous ion to ferric ion in the presence of various oxidant species in acidic medium and the measurement of the ferric ion by xylenol orange. The oxidation reaction of the assay was enhanced and precipitation of proteins was prevented. In addition, autoxidation of ferrous ion present in the reagent was prevented during storage. The method was applied to an automated analyzer, which was calibrated with hydrogen peroxide and the analytical performance characteristics of the assay were determined.
There were important correlations with hydrogen peroxide, tert-butyl hydroperoxide and cumene hydroperoxide solutions (r=0.99, P<0.001 for all). In addition, the new assay presented a typical sigmoidal reaction pattern in copper-induced lipoprotein autoxidation. The novel assay is linear up to 200 micromol H2O2 Equiv./L and its precision value is lower than 3%. The lower detection limit is 1.13 micromol H2O2 Equiv./L. The reagents are stable for at least 6 months on the automated analyzer. Serum TOS level was significantly higher in patients with osteoarthritis (21.23+/-3.11 micromol H2O2 Equiv./L) than in healthy subjects (14.19+/-3.16 micromol H2O2 Equiv./L, P<0.001) and the results showed a significant negative correlation with total antioxidant capacity (TAC) (r=-0.66 P<0.01).
This easy, stable, reliable, sensitive, inexpensive and fully automated method that is described can be used to measure total oxidant status.