The role of oxidative stress as an underlying mechanism in life-history trade-offs

A key aspect underpinning life-history theory is the existence of trade-offs. Trade-offs occur because resources are limited, meaning that individuals cannot invest in all traits simultaneously, leading to costs for traits such as growth and reproduction. Such costs may be the reason for the sub-maximal growth rates that are often observed in nature, though the fitness consequences of these costs would depend on the effects on lifetime reproductive success. Recently, much attention has been given to the physiological mechanism that might underlie these life-history trade-offs, with oxidative stress (OS) playing a key role. OS is characterised by a build-up of oxidative damage to tissues (e.g. protein, lipids and DNA) from attack by reactive species (RS). RS, the majority of which are by-products of metabolism, are usually neutralised by antioxidants, however OS occurs when there is an imbalance between the two. There are two main theories linking OS with growth and reproduction. The first is that traits like growth and reproduction, being metabolically demanding, lead to an increase in RS production. The second involves the diversion of resources away from self-maintenance processes (e.g. the redox system) when individuals are faced with enhanced growth or reproductive expenditure. Previous research investigating trade-offs involving growth or reproduction and self-maintenance has been equivocal. One reason for this could be that associations among redox biomarkers can vary greatly so that the biomarker selected for analysis can influence the conclusion reached about an individual’s oxidative status. Therefore the first aim of my thesis was to explore the strength and pattern of integration of five biomarkers of OS (three antioxidants, one damage and one general oxidation measure) in wild blue tit (Cyanistes caeruleus) adults and nestlings (Chapter 2). In doing so, I established that all five biomarkers should be included in future analyses, thus using this collection of biomarkers I explored my next aims; whether enhanced growth (Chapters 3 and 4) or reproductive effort (Chapter 5) can lead to increased OS levels, if these traits are traded off against self-maintenance. I accomplished these aims using both a meta-analytic and experimental approach, the latter involving manipulation of brood size in wild blue tits in order to experimentally alter growth rate of nestlings and provisioning rate (a proxy for reproductive expenditure) of adults. I also investigated the potential for redox integration to be used as an index of body condition (Chapter 2), allowing predictions about future fitness consequences of changes to oxidative state to be made. A growth – self-maintenance trade off was supported by my meta-analytic results (Chapter 4) which found OS to be a constraint on growth. However, when faced with experimentally enhanced growth, animals were typically not able to adjust this trade-off so that oxidative damage resulted. This might support the idea that energetically expensive growth causes resources to be diverted away from the redox system; however, antioxidants did not show an overall reduction in response to growth in the meta-analysis suggesting that oxidative costs of growth may result from increased RS production due to the greater metabolism needed for enhanced growth. My experimental data (Chapter 3) showed a similar pattern, with raised protein damage levels (protein carbonyls; PCs) in the fastest growing blue tit chicks in a brood, compared with their slower growing sibs. These within-brood differences in OS levels likely resulted from within-brood hierarchies and might have masked any between-brood differences, which were not observed here. Despite evidence for a growth – self-maintenance trade off, my experimental results on blue tits found no support for the hypothesis that self-maintenance is also traded off against reproduction, another energetically demanding trait. There was no link between experimentally altered reproductive expenditure and OS, nor was there a direct correlation between reproductive effort and OS (Chapter 5). However, there are various factors that likely influence whether oxidative costs are observed, including environmental conditions and whether such costs are transient. This emphasises the need for longitudinal studies following the same individuals over multiple years and across a wide range of habitats that differ in quality. This would allow investigation into how key life events interact; it might be that raised OS levels from rapid early growth have the potential to constrain reproduction or that high parental OS levels constrain offspring growth. Any oxidative costs resulting from these life-history trade-offs have the potential to impact on future fitness. Redox integration of certain biomarkers might prove to be a useful tool in making predictions about fitness, as I found in Chapter 2, as well as establishing how the redox system responds, as a whole, to changes to growth and reproduction. Finally, if the tissues measured can tolerate a given level of OS, then the level of oxidative damage might be irrelevant and not impact on future fitness at all.

Genetic evidence for polygynandry in the black-striped pipefish syngnathus abaster: a microsatellite-based parentage analysis

Sexual selection theory predicts that, in organisms with reversed sex roles, more polyandrous species exhibit higher levels of sexual dimorphism. In the family Syngnathidae (pipefish, seahorses, and seadragons), males provide all parental care by carrying developing embryos on their ventral surfaces, and females develop secondary sex characters. Syngnathids exhibit a variety of genetic mating patterns, making them an ideal group to test predictions of sexual selection theory. Here, we describe the mating system of the black-striped pipefish Syngnathus abaster, using 4 highly variable microsatellites to analyze parentage of 102 embryos. Results revealed that 1) both sexes mate multiple times over the course of a pregnancy (polygynandrous mating system), 2) eggs are spatially segregated by maternity within each brood pouch, and 3) larger females have higher mating success (Kolmogorov–Smirnov test; P < 0.05). Together with similar studies of other syngnathid species, our results support the hypothesis that the mating system is related to the intensity of sexual dimorphism.

Feeding ecology and life-history strategy of nesting males in a fish with long parental care, Lusitanian toadfish (Halobatrachus didactylus, Batrachoididae)

The Lusitanian toadfish, Halobatrachus didactylus, like other batrachoidids, is a benthic fish species with nesting behaviour during the breeding season. During this prolonged period it engages in mating activities and remains in the nest providing parental care. It is not known whether males feed while providing parental care but it is likely that their limited mobility may restrict their diet and influence their fitness. As a consequence, egg cannibalism could occur as a life-history strategy. The aim of the present study is to ascertain the feeding behaviour of nesting males, in comparison to mature non-nesting males, and to identify potential life-history traits related to egg cannibalism. Nest-holders were sampled from artificial nests placed in an intertidal area of the Tagus estuary, only exposed during spring low tides. The diet of nest-holders was compared with that of non-nesting mature males from the same area, captured by otter trawl. The present study demonstrates that despite their constrained mobility nest-holders feed during the breeding season, although in a more opportunistic fashion than non-nesting males. Nest-holders showed a generalist feeding behaviour, with a more heterogeneous diet. Egg cannibalism was not related to male condition, paternity or brood size but showed a higher incidence early in the season when water temperatures were lower. The results suggest a possible seasonal trade-off strategy between care and energy recovery, triggered by environmental factors, where under unfavourable conditions to sustain viable eggs the male may recover energy by eating eggs, thus benefiting future reproductive success, later in the season.

Preservation of Honey Bee (Apis mellifera L.) Semen

L’abeille domestique (Apis mellifera Linnaeus) joue un rôle crucial comme pollinisateur dans l’industrie de l’agriculture. Cependant, durant les dernières décennies, une mortalité des colonies d’abeilles a été observée partout à travers le monde. La conservation du sperme d’abeille est un outil efficace pour sauvegarder la diversité génétique. Sa conservation est possible à température pièce, mais la cryoconservation serait une meilleure méthode pour la conservation à long terme. Notre objectif général est de développer une méthode de cryoconservation de la semence d’abeille. L’hypothèse no.1 était que la cryoconservation de la semence d’abeille est plus efficace à long terme que les températures au-dessus de 0 °C. Nous avons évalué l’efficacité, basé sur la viabilité des spermatozoïdes, de deux températures de conservation: -196 °C et 16 °C. Après un an de conservation, la semence congelée avait une meilleure viabilité comparée à 16°C (76% ± 5% vs 0%; p < 0,05). Par la suite, la spermathèque des reines inséminées avec la semence cryoconservée a été évaluée par la migration des spermatozoïdes ainsi que la viabilité des spermatozoïdes. Il y avait beaucoup de variabilités dans nos résultats. Nous n’avons pas été en mesure de vérifier si l’ajout de la centrifugation après la conservation améliore la fertilité des reines après insémination. Toutefois, nos résultats confirment que la cryoconservation est une technique efficace pour conserver la semence d’abeille à long terme.

Reticence or vigilance at the nest: a cruel bind for the endangered Black-capped Vireo

Breeding birds vocalize to find mates and establish and defend territories, but these same critical communications may also attract predators or brood parasites, placing birds in a cruel bind. Although vigilant birds may better maintain social relationships with mates and neighbors through frequent vocalizations, reticent birds may reduce risk to their nests by being relatively quiet and making infrequent vocalizations. Selection for vocalization patterns that minimize brood parasitism might be particularly strong for birds that are unable to fledge both their own young and the parasite. Temporal plasticity in the frequency of vocalizations near nests, however, may allow birds to balance trade-offs and optimize nest-defense strategies. The Black-capped Vireo (Vireo atricapilla) is an endangered songbird that faces intensive brood parasitism in areas where Brown-headed Cowbirds (Molothrus ater) are present. Vireo nests that produce cowbird fledglings always fail to fledge vireo young. We recorded vocalizations at vireo nests across three nesting stages (building, laying, and early incubation) and three periods of the day (morning, midday, and evening) and compared vocalization frequency with eventual depredation or parasitism fate as well as local cowbird density to test two hypotheses. The predator-attraction hypothesis predicts that predators will be attracted by frequent vocalizations, whereas cowbirds will parasitize nests with relatively quiet parents and less predation risk; thus, vireos will experience trade-offs between reticence and vigilance in mediating specific risks. The parasite-assessment hypothesis predicts that vireos will become more secretive as local cowbird densities increase. Vireo vocalization response to nest predation and parasitism risk interacted with nest stage, and we found little evidence of risk mediation through vocalizations except during the building stage. Vireos, however, did benefit overall by optimizing temporal patterns in vocalizations. Vireo nests were less likely to be depredated or parasitized if males vocalized most during laying and least during the middle of the day. Birds vocalized more during the midday and less during the laying period when local cowbird densities were higher, however, perhaps demonstrating limited plasticity in social communication.