The price of defence: toxins, visual signals and oxidative state in an aposematic butterfly.

In a variety of aposematic species, the conspicuousness of an individual's warning signal and the quantity of its chemical defence are positively correlated. This apparent honest signalling is predicted by resource competition models which assume that the production and maintenance of aposematic defences compete for access to antioxidant molecules that have dual functions as pigments and in protecting against oxidative damage. To test for such trade-offs, we raised monarch butterflies (Danaus plexippus) on different species of their milkweed host plants (Apocynaceae) that vary in quantities of cardenolides to test whether (i) the sequestration of cardenolides as a secondary defence is associated with costs in the form of oxidative lipid damage and reduced antioxidant defences; and (ii) lower oxidative state is associated with a reduced capacity to produce aposematic displays. In male monarchs conspicuousness was explained by an interaction between oxidative damage and sequestration: males with high levels of oxidative damage became less conspicuous with increased sequestration of cardenolides, whereas those with low oxidative damage became more conspicuous with increased levels of cardenolides. There was no significant effect of oxidative damage or concentration of sequestered cardenolides on female conspicuousness. Our results demonstrate a physiological linkage between the production of coloration and oxidative state, and differential costs of sequestration and signalling in monarch butterflies.

Small increases in ambient temperature reduce offspring body mass in an equatorial mammal.

Human-induced climate change is leading to temperature rises, along with increases in the frequency and intensity of heatwaves. Many animals respond to high temperatures through behavioural thermoregulation, for example by resting in the shade, but this may impose opportunity costs by reducing foraging time (therefore energy supply), and so may be most effective when food is abundant. However, the heat dissipation limit (HDL) theory proposes that even when energy supply is plentiful, high temperatures can still have negative effects. This is because dissipating excess heat becomes harder, which limits processes that generate heat such as lactation. We tested predictions from HDL on a wild, equatorial population of banded mongooses (Mungos mungo). In support of the HDL theory, higher ambient temperatures led to lighter pups, and increasing food availability made little difference to pup weight under hotter conditions. This suggests that direct physiological constraints rather than opportunity costs of behavioural thermoregulation explain the negative impact of high temperatures on pup growth. Our results indicate that climate change may be particularly important for equatorial species, which often experience high temperatures year-round so cannot time reproduction to coincide with cooler conditions.

Consistent measures of oxidative balance predict survival but not reproduction in a long-distance migrant.

Physiological processes, including those that disrupt oxidative balance, have been proposed as key to understanding fundamental life-history trade-offs. Yet, examination of changes in oxidative balance within wild animals across time, space and major life-history challenges remains uncommon. For example, migration presents substantial physiological challenges for individuals, and data on migratory individuals would provide crucial context for exposing the importance of relationships between oxidative balance and fitness outcomes. Here we examined the consistency of commonly used measures of oxidative balance in longitudinally sampled free-living individuals of a long-lived, long-distance migrant, the Brent goose Branta bernicla hrota over periods of months to years. Although inter-individual and temporal variation in measures of oxidative balance were substantial, we found high consistency in measures of lipid peroxidation and circulating non-enzymatic antioxidants in longitudinally sampled individuals. This suggests the potential for the existence of individual oxidative phenotypes. Given intra-individual consistency, we then examined how these physiological measures relate to survival and reproductive success across all sampled individuals. Surprisingly, lower survival was predicted for individuals with lower levels of damage, with no measured physiological metric associated with reproductive success. Our results demonstrate that snapshot measurements of a consistent measure of oxidative balance can inform our understanding of differences in a key demographic trait. However, the positive relationship between oxidative damage and survival emphasises the need to investigate the relationships between the oxidative system and fitness outcomes in other species undergoing similar physiologically challenging life cycles. This would highlight the extent to which variation in such traits and resource allocation trade-offs is a result of adaptation to different life-history strategies.

No evidence of quantitative signal honesty across species of aposematic burnet moths (Lepidoptera: Zygaenidae).

Many defended species use conspicuous visual warning signals to deter potential predators from attacking. Traditional theory holds that these signals should converge on similar forms, yet variation in visual traits and the levels of defensive chemicals is common, both within and between species. It is currently unclear how the strength of signals and potency of defences might be related: conflicting theories suggest that aposematic signals should be quantitatively honest, or, in contrast, that investment in one component should be prioritized over the other, while empirical tests have yielded contrasting results. Here, we advance this debate by examining the relationship between defensive chemicals and signal properties in a family of aposematic Lepidoptera, accounting for phylogenetic relationships and quantifying coloration from the perspective of relevant predators. We test for correlations between toxin levels and measures of wing colour across 14 species of day-flying burnet and forester moths (Lepidoptera: Zygaenidae), protected by highly aversive cyanogenic glucosides, and find no clear evidence of quantitative signal honesty. Significant relationships between toxin levels and coloration vary between sexes and sampling years, and several trends run contrary to expectations for signal honesty. Although toxin concentration is positively correlated with increasing luminance contrast in forewing pattern in 1 year, higher toxin levels are also associated with paler and less chromatically salient markings, at least in females, in another year. Our study also serves to highlight important factors, including sex-specific trends and seasonal variation, that should be accounted for in future work on signal honesty in aposematic species.

Measures of oxidative state are primarily driven by extrinsic factors in a long-distance migrant.

Oxidative stress is a likely consequence of hard physical exertion and thus a potential mediator of life-history trade-offs in migratory animals. However, little is known about the relative importance of intrinsic and extrinsic stressors on the oxidative state of individuals in wild populations. We quantified the relationships between air temperature, sex, body condition and three markers of oxidative state (malondialdehyde, superoxide dismutase and total antioxidant capacity) across hundreds of individuals of a long-distance migrant (the brent goose Branta bernicla hrota) during wintering and spring staging. We found that air temperature and migratory stage were the strongest predictors of oxidative state. This emphasizes the importance of extrinsic factors in regulating the oxidative state of migrating birds, with differential effects across the migration. The significance of abiotic effects demonstrates an additional mechanism by which changing climates may affect migratory costs.

Effects of an early-life paraquat exposure on adult resistance to oxidative stress, plumage colour and sperm performance in a wild bird.

Early-life stressful conditions can shape individual phenotypes and ultimately influence fitness. Oxidative stress is a pervasive threat that affects many fitness-related traits and can modulate life-history trade-offs. Yet, the extent to which exposure to oxidative stress during early life can have long-lasting effects on key fitness-related traits remains to be elucidated, particularly in natural populations of vertebrates. Using a wild population of great tits Parus major, we experimentally dosed 11-day-old birds with paraquat, a pro-oxidant molecule, aiming at increasing oxidative stress. One year later, we recaptured 39 of them as adult recruiting breeders and quantified effects of the paraquat exposure on their resistance to oxidative stress, carotenoid-based plumage coloration and male sperm performance. Despite the absence of a short-term effect of paraquat on oxidative stress measured two days later, the pre-fledging exposure to paraquat induced a reduction in individual oxidative damage measured at adulthood. Paraquat-dosed individuals also had brighter plumage, but no effect was observed on male sperm performance. For the first time in a natural population of vertebrates, we experimentally show that an early-life acute exposure to a pro-oxidant has long-lasting effects on individual resistance to oxidative stress at adulthood. Our results are in line with the environmental matching and the hormesis hypotheses but may also reflect selective disappearance of individuals with lower resistance to oxidative stress.

Marker-dependent associations among oxidative stress, growth and survival during early life in a wild mammal.

Oxidative stress (OS) is hypothesized to be a key physiological mechanism mediating life-history trade-offs, but evidence from wild populations experiencing natural environmental variation is limited. We tested the hypotheses that increased early life growth rate increases OS, and that increased OS reduces first-winter survival, in wild Soay sheep (Ovis aries) lambs. We measured growth rate and first-winter survival for four consecutive cohorts, and measured two markers of oxidative damage (malondialdehyde (MDA), protein carbonyls (PC)) and two markers of antioxidant (AOX) protection (total AOX capacity (TAC), superoxide dismutase (SOD)) from blood samples. Faster lamb growth was weakly associated with increased MDA, but not associated with variation in the other three markers. Lambs with higher SOD activity were more likely to survive their first winter, as were male but not female lambs with lower PC concentrations. Survival did not vary with MDA or total TAC. Key predictions relating OS to growth and survival were therefore supported in some OS markers, but not others. This suggests that different markers capture different aspects of the complex relationships between individual oxidative state, physiology and fitness, and that overarching hypotheses relating OS to life-history variation cannot be supported or refuted by studying individual markers.

The oxidative costs of reproduction are group-size dependent in a wild cooperative breeder.

Life-history theory assumes that reproduction entails a cost, and research on cooperatively breeding societies suggests that the cooperative sharing of workloads can reduce this cost. However, the physiological mechanisms that underpin both the costs of reproduction and the benefits of cooperation remain poorly understood. It has been hypothesized that reproductive costs may arise in part from oxidative stress, as reproductive investment may elevate exposure to reactive oxygen species, compromising survival and future reproduction and accelerating senescence. However, experimental evidence of oxidative costs of reproduction in the wild remains scarce. Here, we use a clutch-removal experiment to investigate the oxidative costs of reproduction in a wild cooperatively breeding bird, the white-browed sparrow weaver, Plocepasser mahali. Our results reveal costs of reproduction that are dependent on group size: relative to individuals in groups whose eggs were experimentally removed, individuals in groups that raised offspring experienced an associated cost (elevated oxidative damage and reduced body mass), but only if they were in small groups containing fewer or no helpers. Furthermore, during nestling provisioning, individuals that provisioned at higher rates showed greater within-individual declines in body mass and antioxidant protection. Our results provide rare experimental evidence that reproduction can negatively impact both oxidative status and body mass in the wild, and suggest that these costs can be mitigated in cooperative societies by the presence of additional helpers. These findings have implications for our understanding of the energetic and oxidative costs of reproduction, and the benefits of cooperation in animal societies.

Telomere length in early life predicts lifespan.

The attrition of telomeres, the ends of eukaryote chromosomes, is thought to play an important role in cell deterioration with advancing age. The observed variation in telomere length among individuals of the same age is therefore thought to be related to variation in potential longevity. Studies of this relationship are hampered by the time scale over which individuals need to be followed, particularly in long-lived species where lifespan variation is greatest. So far, data are based either on simple comparisons of telomere length among different age classes or on individuals whose telomere length is measured at most twice and whose subsequent survival is monitored for only a short proportion of the typical lifespan. Both approaches are subject to bias. Key studies, in which telomere length is tracked from early in life, and actual lifespan recorded, have been lacking. We measured telomere length in zebra finches (n = 99) from the nestling stage and at various points thereafter, and recorded their natural lifespan (which varied from less than 1 to almost 9 y). We found telomere length at 25 d to be a very strong predictor of realized lifespan (P < 0.001); those individuals living longest had relatively long telomeres at all points at which they were measured. Reproduction increased adult telomere loss, but this effect appeared transient and did not influence survival. Our results provide the strongest evidence available of the relationship between telomere length and lifespan and emphasize the importance of understanding factors that determine early life telomere length.

Defence against the intergenerational cost of reproduction in males: oxidative shielding of the germline.

Reproduction is expected to carry an oxidative cost, yet in many species breeders appear to sustain lower levels of oxidative damage compared to non-breeders. This paradox may be explained by considering the intergenerational costs of reproduction. Specifically, a reduction in oxidative damage upon transitioning to a reproductive state may represent a pre-emptive shielding strategy to protect the next generation from intergenerational oxidative damage (IOD) - known as the oxidative shielding hypothesis. Males may be particularly likely to transmit IOD, because sperm are highly susceptible to oxidative damage. Yet, the possibility of male-mediated IOD remains largely uninvestigated. Here, we present a conceptual and methodological framework to assess intergenerational costs of reproduction and oxidative shielding of the germline in males. We discuss variance in reproductive costs and expected payoffs of oxidative shielding according to species' life histories, and the expected impact on offspring fitness. Oxidative shielding presents an opportunity to incorporate intergenerational effects into the advancing field of life-history evolution.

Fat provisioning in winter impairs egg production during the following spring: a landscape-scale study of blue tits.

1. Provisioning of garden birds is a growing phenomenon, particularly during winter, but there is little empirical evidence of its true ecological impacts. One possibility is that winter provisioning could enhance subsequent breeding performance, but this seems likely to depend on the types of nutrients provided. For example, whereas effects of macronutrients such as fat are unlikely to be carried over to influence breeding in small passerines, micronutrients such as dietary vitamin E (an antioxidant) may be stored or have lasting health benefits. 2. Here, we examine the carry-over effects of winter food supplements on egg production in wild populations of blue tits (Cyanistes caeruleus). Over three consecutive years, birds were provisioned with fat, fat plus vitamin E or remained unfed (controls). 3. The provision of fat in winter resulted in smaller relative yolk mass in larger eggs and reduced yolk carotenoid concentrations in early breeders. However, these effects were not seen in birds provisioned with fat plus vitamin E. Lay date, clutch size, egg mass and yolk vitamin E concentrations were not significantly affected by winter provisioning treatment. 4. Our results indicate that winter provisioning can have important downstream consequences, in particular affecting investment in egg production several weeks or months later. 5. Provisioning is widely applied to support garden bird populations and for the conservation management of endangered species. However, our results challenge the assumption that such practices are always beneficial at the population level and emphasize how the ecological impacts can depend on the specific nutritional profile of provisioned foods.

Nestling erythrocyte resistance to oxidative stress predicts fledging success but not local recruitment in a wild bird.

Stressful conditions experienced by individuals during their early development have long-term consequences on various life-history traits such as survival until first reproduction. Oxidative stress has been shown to affect various fitness-related traits and to influence key evolutionary trade-offs but whether an individual's ability to resist oxidative stress in early life affects its survival has rarely been tested. In the present study, we used four years of data obtained from a free-living great tit population (Parus major; n = 1658 offspring) to test whether pre-fledging resistance to oxidative stress, measured as erythrocyte resistance to oxidative stress and oxidative damage to lipids, predicted fledging success and local recruitment. Fledging success and local recruitment, both major correlates of survival, were primarily influenced by offspring body mass prior to fledging. We found that pre-fledging erythrocyte resistance to oxidative stress predicted fledging success, suggesting that individual resistance to oxidative stress is related to short-term survival. However, local recruitment was not influenced by pre-fledging erythrocyte resistance to oxidative stress or oxidative damage. Our results suggest that an individual ability to resist oxidative stress at the offspring stage predicts short-term survival but does not influence survival later in life.

Antioxidant availability trades off with warning signals and toxin sequestration in the large milkweed bug (Oncopeltus fasciatus).

In some aposematic species the conspicuousness of an individual's warning signal and the concentration of its chemical defense are positively correlated. Several mechanisms have been proposed to explain this phenomenon, including resource allocation trade-offs where the same limiting resource is needed to produce both the warning signal and chemical defense. Here, the large milkweed bug (Oncopeltus fasciatus: Heteroptera, Lygaeinae) was used to test whether allocation of antioxidants, that can impart color, trade against their availability to prevent self-damage caused by toxin sequestration. We investigated if (i) the sequestration of cardenolides is associated with costs in the form of changes in oxidative state; and (ii) oxidative state can affect the capacity of individuals to produce warning signals. We reared milkweed bugs on artificial diets with increasing quantities of cardenolides and examined how this affected signal quality (brightness and chroma) across different instars. We then related the expression of warning colors to the quantity of sequestered cardenolides and indicators of oxidative state-oxidative lipid damage (malondialdehyde), and two antioxidants: total superoxide dismutase and total glutathione. Bugs that sequestered more cardenolides had significantly lower levels of the antioxidant glutathione, and bugs with less total glutathione had less luminant orange warning signals and reduced chroma of their black patches compared to bugs with more glutathione. Bugs that sequestered more cardenolides also had reduced red-green chroma of their black patches that was unrelated to oxidative state. Our results give tentative support for a physiological cost of sequestration in milkweed bugs and a mechanistic link between antioxidant availability, sequestration, and warning signals.

Fine-scale thermal adaptation in a green turtle nesting population.

The effect of climate warming on the reproductive success of ectothermic animals is currently a subject of major conservation concern. However, for many threatened species, we still know surprisingly little about the extent of naturally occurring adaptive variation in heat-tolerance. Here, we show that the thermal tolerances of green turtle (Chelonia mydas) embryos in a single, island-breeding population have diverged in response to the contrasting incubation temperatures of nesting beaches just a few kilometres apart. In natural nests and in a common-garden rearing experiment, the offspring of females nesting on a naturally hot (black sand) beach survived better and grew larger at hot incubation temperatures compared with the offspring of females nesting on a cooler (pale sand) beach nearby. These differences were owing to shallower thermal reaction norms in the hot beach population, rather than shifts in thermal optima, and could not be explained by egg-mediated maternal effects. Our results suggest that marine turtle nesting behaviour can drive adaptive differentiation at remarkably fine spatial scales, and have important implications for how we define conservation units for protection. In particular, previous studies may have underestimated the extent of adaptive structuring in marine turtle populations that may significantly affect their capacity to respond to environmental change.

Environmental effects shape the maternal transfer of carotenoids and vitamin E to the yolk.

INTRODUCTION: Maternal effects occur when the phenotype of the offspring is influenced by the phenotype of the mother, which in turn depends on her heritable state as well as on influences from the current and past environmental conditions. All of these pathways may, therefore, form significant sources of variation in maternal effects. Here, we focused on the maternal transfer of carotenoids and vitamin E to the egg yolk, using canaries as a model species. Maternal yolk carotenoids and vitamin E are known to generate significant phenotypic variation in offspring, representing examples of maternal effects. We studied the intra-individual consistency in deposition patterns across two years and the mother-daughter resemblance across two generations in order to estimate the level of heritable variation. The effects of the current environmental conditions were studied via a food supplementation experiment, while the consequences of past environmental conditions were estimated on the basis of the early growth trajectories. RESULTS: There was a significant effect of the current environmental conditions on the yolk carotenoid and vitamin E deposition, but this effect varied between antioxidant components. The deposition of yolk carotenoids and vitamin E were linked to the process of yolk formation. Past environmental conditions did not contribute to the variation in yolk carotenoid and vitamin E levels nor did we find significant heritable variation. CONCLUSIONS: The transfer of carotenoids or vitamin E may be an example where current environmental variation is largely passed from the mother to the offspring, despite the numerous intermediate physiological steps that are involved. Differences in the effect of the environmental conditions as experienced by the mother during laying may be due to differences in availability as well as physiological processes such as competitive exclusion or selective absorption.

Methodological confounds of measuring urinary oxidative stress in wild animals.

Biomarkers of oxidative stress (OS) are useful in addressing a wide range of research questions, but thus far, they have had limited application to wild mammal populations due to a reliance on blood or tissue sampling. A shift toward non-invasive measurement of OS would allow field ecologists and conservationists to apply this method more readily. However, the impact of methodological confounds on urinary OS measurement under field conditions has never been explicitly investigated. We combined a cross-sectional analysis with a field experiment to assess the impact of four potential methodological confounds on OS measurements: (1) time of sampling, (2) environmental contamination from foliage; (3) delay between sample collection and flash-freezing in liquid nitrogen; and (4) sample storage of up to 15 months below -80°C. We measured DNA oxidative damage (8-hydroxy-2'-deoxyguanosine, 8-OHdG), lipid peroxidation (malondialdehyde, MDA), total antioxidant capacity (TAC), and uric acid (UA) in 167 urine samples collected from wild Zanzibar red colobus (Piliocolobus kirkii). We found that MDA was higher in samples collected in the morning than in the afternoon but there were no diurnal patterns in any of the other markers. Contamination of samples from foliage and length of time frozen at -80°C for up to 15 months did not affect OS marker concentrations. Freezing delay did not affect OS levels cross-sectionally, but OS values from individual samples showed only moderate-to-good consistency and substantial rank-order reversals when exposed to different freezing delays. We recommend that diurnal patterns of OS markers and the impact of storage time before and after freezing on OS marker concentrations be considered when designing sampling protocols. However, given the high stability we observed for four OS markers subject to a variety of putative methodological confounds, we suggest that urinary OS markers provide a valuable addition to the toolkit of field ecologists and conservationists within reasonable methodological constraints.

Untangling the oxidative cost of reproduction: An analysis in wild banded mongooses.

The cost of reproduction plays a central role in evolutionary theory, but the identity of the underlying mechanisms remains a puzzle. Oxidative stress has been hypothesized to be a proximate mechanism that may explain the cost of reproduction. We examine three pathways by which oxidative stress could shape reproduction. The "oxidative cost" hypothesis proposes that reproductive effort generates oxidative stress, while the "oxidative constraint" and "oxidative shielding" hypotheses suggest that mothers mitigate such costs through reducing reproductive effort or by pre-emptively decreasing damage levels, respectively. We tested these three mechanisms using data from a long-term food provisioning experiment on wild female banded mongooses (Mungos mungo). Our results show that maternal supplementation did not influence oxidative stress levels, or the production and survival of offspring. However, we found that two of the oxidative mechanisms co-occur during reproduction. There was evidence of an oxidative challenge associated with reproduction that mothers attempted to mitigate by reducing damage levels during breeding. This mitigation is likely to be of crucial importance, as long-term offspring survival was negatively impacted by maternal oxidative stress. This study demonstrates the value of longitudinal studies of wild animals in order to highlight the interconnected oxidative mechanisms that shape the cost of reproduction.

Rate of egg maturation in marine turtles exhibits 'universal temperature dependence'.

1. The metabolic theory of ecology (MTE) predicts that, after correcting for body mass variation among organisms, the rates of most biological processes will vary as a universal function of temperature. However, empirical support for 'universal temperature dependence' (UTD) is currently equivocal and based on studies of a limited number of traits. 2. In many ectothermic animals, the rate at which females produce mature eggs is temperature dependent and may be an important factor in determining the costs of reproduction. 3. We tested whether the rate of egg maturation in marine turtles varies with environmental temperature as predicted by MTE, using the time separating successive clutches of individual females to estimate the rate at which eggs are formed. We also assessed the phenotypic contribution to this rate, by using radio telemetry to make repeated measurements of interclutch intervals for individual green turtles (Chelonia mydas). 4. Rates of egg maturation increased with seasonally increasing water temperatures in radio-tracked green turtles, but were not repeatable for individual females, and did not vary according to maternal body size or reproductive investment (number and size of eggs produced). 5. Using a collated data set from several different populations and species of marine turtles, we then show that a single relationship with water temperature explains most of the variation in egg maturation rates, with a slope that is statistically indistinguishable from the UTD predicted by MTE. However, several alternative statistical models also described the relationship between temperature and egg maturation rates equally parsimoniously. 6. Our results offer novel support for the MTE's predicted UTD of biological rates, although the underlying mechanisms require further study. The strong temperature dependence of egg maturation combined with the apparently weak phenotypic contribution to this rate has interesting behavioural implications in ectothermic animals. We suggest that maternal thermoregulatory behaviour in marine turtles, and many other reptiles, is consistent with a strategy of adaptively increasing body temperatures to accelerate egg maturation.

Carry-over effects as drivers of fitness differences in animals.

1. Carry-over effects occur when processes in one season influence the success of an individual in the following season. This phenomenon has the potential to explain a large amount of variation in individual fitness, but so far has only been described in a limited number of species. This is largely due to difficulties associated with tracking individuals between periods of the annual cycle, but also because of a lack of research specifically designed to examine hypotheses related to carry-over effects. 2. We review the known mechanisms that drive carry-over effects, most notably macronutrient supply, and highlight the types of life histories and ecological situations where we would expect them to most often occur. We also identify a number of other potential mechanisms that require investigation, including micronutrients such as antioxidants. 3. We propose a series of experiments designed to estimate the relative contributions of extrinsic and intrinsic quality effects in the pre-breeding season, which in turn will allow an accurate estimation of the magnitude of carry-over effects. To date this has proven immensely difficult, and we hope that the experimental frameworks described here will stimulate new avenues of research vital to advancing our understanding of how carry-over effects can shape animal life histories. 4. We also explore the potential of state-dependent modelling as a tool for investigating carry-over effects, most notably for its ability to calculate optimal rates of acquisition of a multitude of resources over the course of the annual cycle, and also because it allows us to vary the strength of density-dependent relationships which can alter the magnitude of carry-over effects in either a synergistic or agonistic fashion. 5. In conclusion carry-over effects are likely to be far more widespread than currently indicated, and they are likely to be driven by a multitude of factors including both macro- and micronutrients. For this reason they could feasibly be responsible for a large amount of the observed variation in performance among individuals, and consequently warrant a wealth of new research designed specifically to decompose components of variation in fitness attributes related to processes across and within seasons.

Phthalate diversity in eggs and associations with oxidative stress in the European herring gull (Larus argentatus).

Phthalates are plastic-derived contaminants that are ubiquitous in natural environments and function as pro-oxidants. The extent to which phthalates bioaccumulate in wild animals and associations with oxidative stress are poorly understood. Here, we describe relationships between maternally-derived phthalates, lipid peroxidation (malondialdehyde, MDA) and the dietary antioxidant α-tocopherol in eggs of European herring gulls (Larus argentatus) in Cornwall, UK. Up to six phthalate parent compounds and four phthalate metabolites were detected. Egg concentrations of MDA were positively associated with dicyclohexyl phthalate (DCHP) and negatively associated with α-tocopherol, suggesting that DCHP is associated with oxidative stress in gulls. The consequences of phthalate exposure in ovo for offspring development warrants study.