Lipid-correction models for δ13C values across small pelagic fishes (Clupeiformes) from the Atlantic Ocean.

The interpretation of δ13C values in trophic ecology requires standardization of the lipid content of organisms estimated through their C:N ratio. To avoid time-consuming lipid extractions, the use of mathematical corrections has been developed for many years, and the conclusions generally point in the direction of species-specific adjustment of the models. This study aimed at defining the maximum taxonomic level required to obtain the best corrected δ13C values in small pelagic fish of the order Clupeiformes. δ13C values of six species were analyzed bulk and lipid-free, and were used to fit and validate linear and mass-balance models at different taxonomic levels. Despite a species effect combined with the C:N ratio effect, the corrected δ13C values produced by a global model for the Clupeiformes were as good as or better when compared to lipid-free samples than those produced by species-specific models, paving the way for possible generalization to other species in this order. At the order level, the linear model outperformed the mass-balance model.

Different mechanisms underpin the decline in growth of anchovies and sardines of the Bay of Biscay.

Declines in individuals' growth in exploited fish species are generally attributed to evolutionary consequences of size-selective fishing or to plastic responses due to constraints set by changing environmental conditions dampening individuals' growth. However, other processes such as growth compensation and non-directional selection can occur and their importance on the overall phenotypic response of exploited populations has largely been ignored. Using otolith growth data collected in European anchovy and sardine of the Bay of Biscay (18 cohorts from 2000 to 2018), we parameterized the breeder's equation to determine whether declines in size-at-age in these species were due to an adaptive response (i.e. related to directional or non-directional selection differentials within parental cohorts) or a plastic response (i.e. related to changes in environmental). We found that growth at age-0 in anchovy declined between parents and their offspring when biomass increased and the selective disappearance of large individuals was high in parents. Therefore, an adaptive response probably occurred in years with high fishing effort and the large increase in biomass after the collapse of this stock maintained this adaptive response subsequently. In sardine offspring, higher growth at age-0 was associated with increasing biomass between parents and offspring, suggesting a plastic response to a bottom-up process (i.e. a change in food quantity or quality). Parental cohorts in which selection favoured individuals with high growth compensation produced offspring high catch up growth rates, which may explain the smaller decline in growth in sardine relative to anchovy. Finally, on non-directional selection differentials were not significantly related to the changes in growth at age-0 and growth compensation at age-1 in both species. Although anchovy and sardine have similar ecologies, the mechanisms underlying the declines in their growth are clearly different. The consequences of the exploitation of natural populations could be long lasting if density-dependent processes follow adaptive changes.

Effects of hunting on genetic diversity, inbreeding and dispersal in Finnish black grouse (Lyrurus tetrix).

Intensive hunting activities such as commercial fishing and trophy hunting can have profound influences on natural populations. However, less intensive recreational hunting can also have subtle effects on animal behaviour, habitat use and movement, with implications for population persistence. Lekking species such as the black grouse (Lyrurus tetrix) may be especially prone to hunting as leks are temporally and spatially predictable, making them easy targets. Furthermore, inbreeding in black grouse is mainly avoided through female-biased dispersal, so any disruptions to dispersal caused by hunting could lead to changes in gene flow, increasing the risk of inbreeding. We therefore investigated the impact of hunting on genetic diversity, inbreeding and dispersal on a metapopulation of black grouse in Central Finland. We genotyped 1065 adult males and 813 adult females from twelve lekking sites (six hunted, six unhunted) and 200 unrelated chicks from seven sites (two hunted, five unhunted) at up to thirteen microsatellite loci. Our initial confirmatory analysis of sex-specific fine-scale population structure revealed little genetic structure in the metapopulation. Levels of inbreeding did not differ significantly between hunted and unhunted sites in neither adults nor chicks. However, immigration rates into hunted sites were significantly higher among adults compared to immigration into unhunted sites. We conclude that the influx of migrants into hunted sites may compensate for the loss of harvested individuals, thereby increasing gene flow and mitigating inbreeding. Given the absence of any obvious barriers to gene flow in Central Finland, a spatially heterogeneous matrix of hunted and unhunted regions may be crucial to ensure sustainable harvests into the future.

Ocean acidification alters the acute stress response of a marine fish.

The absorption of anthropogenic carbon dioxide from the atmosphere by oceans generates rapid changes in seawater carbonate system and pH, a process termed ocean acidification. Exposure to acidified water can impact the allostatic load of marine organism as the acclimation to suboptimal environments requires physiological adaptive responses that are energetically costly. As a consequence, fish facing ocean acidification may experience alterations of their stress response and a compromised ability to cope with additional stress, which may impact individuals' life traits and ultimately their fitness. In this context, we carried out an integrative study investigating the impact of ocean acidification on the physiological and behavioral stress responses to an acute stress in juvenile European sea bass. Fish were long term (11 months) exposed to present day pH/CO2 condition or acidified water as predicted by IPCC "business as usual" (RCP8.5) scenario for 2100 and subjected to netting stress (fish transfer and confinement test). Fish acclimated to acidified condition showed slower post stress return to plasma basal concentrations of cortisol and glucose. We found no clear indication of regulation in the central and interrenal tissues of the expression levels of gluco- and mineralocorticoid receptors and corticoid releasing factor. At 120 min post stress, sea bass acclimated to acidified water had divergent neurotransmitters concentrations pattern in the hypothalamus (higher serotonin levels and lower GABA and dopamine levels) and a reduction in motor activity. Our experimental data indicate that ocean acidification alters the physiological response to acute stress in European sea bass via the neuroendocrine regulation of the corticotropic axis, a response associated to an alteration of the motor behavioral profile. Overall, this study suggests that behavioral and physiological adaptive response to climate changes related constraints may impact fish resilience to further stressful events.

Temporal variations in scale cortisol indicate consistent local-and broad-scale constraints in a wild marine teleost fish.

Environmental changes can alter the nursery function of coastal areas through their impact on juveniles' growth and survival rates, an effect mediated by individuals' chronic stress response. Fish chronic stress can be quantified using scale cortisol but no study has yet been quantified the spatio-temporal variations in scale cortisol and its relationship with growth in wild nurseries. We collected wild sea bass juveniles (Dicentrarchus labrax, four years, three nurseries) and found that scale cortisol levels increased consistently with age and across cohorts in 2019 and 2020 probably due to greater stress history in older fish and/or heatwaves that occurred in summers of 2018 and 2019. Growth was impaired in fish with high scale cortisol in 2019 and 2020, confirming the usefulness of scale cortisol as a biomarker of broad and local constraints in wild fish; longer time series will enable us to identify environmental factors underpinning these temporal variations.

Trace metal elements and organic contaminants are differently related to the growth and body condition of wild European sea bass juveniles.

Chemical contaminants are one of the causes of the ongoing degradation of coastal and estuarine nurseries, key functional habitats in which the juveniles of many marine species grow. As chemical contaminants can cause a decrease in the energy available and induce defence mechanisms reducing the amount of energy allocated to life history traits, quantifying their effect on the fitness of juvenile fish is key to understand their population-level consequences. However, these effects are primarily estimated experimentally or in the wild but on a limited number of contaminants or congeners that do not reflect the wide variety of chemical contaminants to which juvenile fish are exposed. To address this issue, we measured concentrations of 14 trace metal elements (TMEs) and bioaccumulative organic contaminants (OCs) in European sea bass juveniles (1-year-old) from three major French nurseries (Seine, Loire and Gironde estuaries). We tested the hypotheses that (i) levels and profiles of contaminants differed among studied nurseries, and ii) fish growth and body condition (based on morphometric measurements and muscle C:N ratio) were lower in individuals with higher contaminant concentrations. Multivariate analyses showed that each nursery had distinct contaminant profiles for both TMEs and OCs, confirming the specific contamination of each estuary, and the large array of contaminants accumulated by sea bass juveniles. Increasing concentrations in some TMEs were associated to decreased growth, and TMEs were consistently related to lower fish body condition. The effect of OCs was more difficult to pinpoint possibly due to operational constraints (i.e., analyses on pooled fish) with contrasting results (i.e., higher growth and decreased body condition). Overall, this study shows that chemical contaminants are related to lower fish growth and body condition at an early life stage in the wild, an effect that can have major consequences if sustained in subsequent ages and associated with a decline in survival and/or reproductive success.

Spatial and ontogenetic variations in sardine feeding conditions in the Bay of Biscay through fatty acid composition.

Food characteristics are amongst the most influential factors determining the fish life history traits as quantitative and qualitative changes in individuals' diet can lead to a decline in the energy allocated to their growth, and hence influence natural populations' characteristics. The size-at-age and weight of European sardines (Sardina pilchardus) in the Bay of Biscay (BoB) have decreased substantially over the last decade, especially for the youngest age classes, and the factors underlying such changes have not yet been identified. We therefore analysed the fatty acid (FA) composition in the neutral (NL) and polar (PL) lipids in samples collected across the BoB to determine whether the diet of sardines changes with their ages. We found that the total FA contents in both lipid fractions varied mainly with the sampling locations and age. Indeed, sardines aged 1 and 2 years living in South BoB had particularly high contents in FA specific to non-diatom phytoplankton, while older sardines living in the Northern part had higher total FA content and more FA specific to copepods. These differences probably resulted from differences in prey availability and to a lesser extend a change in feeding behaviour with age. The strong dependence of younger sardines' diet to phytoplankton in spring suggests that changes in primary production may explain their decline in size-at-age. Finally, NL clearly reflect finest feeding variations in comparison to PL imprinted by diet variations at longer time scale. Future studies should consider separately NL and PL fractions.

Effects of dispersant treated oil upon exploratory behaviour in juvenile European sea bass (Dicentrarchus labrax).

Accidental spills are pervasive pollution in aquatic ecosystems. Resorting to chemical dispersant is one of the most implemented strategies in response to oil spills, but it results in an increase in the bio-availability of oil compounds known to disturb fish neurosensory capacities and hence fish habitat use. While it has become well established that acute oil exposure can cause a range of physiological defects, sub-lethal consequences on animal behaviour have only received recent attention. Here we investigated the effect of an exposure to a 62 h- dispersant treated oil on the exploration tendency (exploratory activity, and avoidance of unfamiliar open areas) of juvenile European sea bass. Three different concentrations of chemically dispersed oil were tested, low and medium conditions bracketing the range of likely situations that fish encounter following an oil spill, the high dose representing a more severe condition. Fish recovery capacities were also evaluated during 2 weeks post-exposure. Our results suggest a dose-response relationship; the low dose (0.048 ± 0.007 g L-1 of total petroleum hydrocarbons ([TPH])) had no effect on sea bass behavioural response to a novel environment while medium (0.243 ± 0.012 g L-1 [TPH]) and high (0.902 ± 0.031 g L-1 [TPH]) doses altered fish exploratory activity and their typical avoidance of unfamiliar open areas. Our experiment also suggest signs of recovery capacities in the first 10 days following oil exposure even if fish might need more time to fully recover from observed alterations. We discuss the possibility that observed alterations may result from a neurosensory or physiological known defects of oil exposure, causing anaesthetic-like sedative behaviours. Altogether, this study shows that juvenile sea bass exposed to oil spill exhibit transient behavioural impairments that may have major population-level consequences given the high mortality experienced by juveniles.

Age- and quality-dependent DNA methylation correlate with melanin-based coloration in a wild bird.

Secondary sexual trait expression can be influenced by fixed individual factors (such as genetic quality) as well as by dynamic factors (such as age and environmentally induced gene expression) that may be associated with variation in condition or quality. In particular, melanin-based traits are known to relate to condition and there is a well-characterized genetic pathway underpinning their expression. However, the mechanisms linking variable trait expression to genetic quality remain unclear. One plausible mechanism is that genetic quality could influence trait expression via differential methylation and differential gene expression. We therefore conducted a pilot study examining DNA methylation at a candidate gene (agouti-related neuropeptide: AgRP) in the black grouse Lyrurus tetrix. We specifically tested whether CpG methylation covaries with age and multilocus heterozygosity (a proxy of genetic quality) and from there whether the expression of a melanin-based ornament (ultraviolet-blue chroma) correlates with DNA methylation. Consistent with expectations, we found clear evidence for age- and heterozygosity-specific patterns of DNA methylation, with two CpG sites showing the greatest DNA methylation in highly heterozygous males at their peak age of reproduction. Furthermore, DNA methylation at three CpG sites was significantly positively correlated with ultraviolet-blue chroma. Ours is the first study to our knowledge to document age- and quality-dependent variation in DNA methylation and to show that dynamic sexual trait expression across the lifespan of an organism is associated with patterns of DNA methylation. Although we cannot demonstrate causality, our work provides empirical support for a mechanism that could potentially link key individual factors to variation in sexual trait expression in a wild vertebrate.

Stabilising selection on immune response in male black grouse Lyrurus tetrix.

Illnesses caused by a variety of micro- and macro- organisms can negatively affect individuals' fitness, leading to the expectation that immunity is under positive selection. However, immune responses are costly and individuals must trade-off their immune response with other fitness components (e.g. survival or reproductive success) meaning that individuals with intermediate response may have the greatest overall fitness. Such a process might be particularly acute in species with strong sexual selection because the condition-dependence of male secondary sexual-traits might lead to striking phenotypic differences amongst males of different immune response levels. We tested whether there is selection on immune response by survival and reproduction in yearling and adult male black grouse (Lyrurus tetrix) following an immune challenge with a novel antigen and tested the hypothesis that sexual signals and body mass are honest signals of the immune response. We show that yearling males with highest immune response to these challenges had higher survival, but the reverse was true for adults. Adults with higher responses had highest mass loss and adult males with intermediate immune response had highest mating success. Tail length was related to baseline response in adults and more weakly in yearlings. Our findings reveal the complex fitness consequences of mounting an immune response across age classes. Such major differences in the direction and magnitude of selection in multiple fitness components is an alternative route underpinning the stabilising selection of immune responses with an intermediate immune response being optimal.

Simultaneous age-dependent and age-independent sexual selection in the lekking black grouse (Lyrurus tetrix).

Individuals' reproductive success is often strongly associated with their age, with typical patterns of early-life reproductive improvement and late-life senescence. These age-related patterns are due to the inherent trade-offs between life-history traits competing for a limited amount of resources available to the organisms. In males, such trade-offs are exacerbated by the resource requirements associated with the expression of costly sexual traits, leading to dynamic changes in trait expression throughout their life span. Due to the age dependency of male phenotypes, the relationship between the expression of male traits and mating success can also vary with male age. Hence, using longitudinal data in a lekking species with strong sexual selection - the black grouse Lyrurus tetrix - we quantified the effects of age, life span and age of first lek attendance (AFL) on male annual mating success (AMS) to separate the effects of within-individual improvement and senescence on AMS from selective (dis)appearance of certain phenotypes. Then, we used male AMS to quantify univariate and multivariate sexual selection gradients on male morphological and behavioural traits with and without accounting for age and age-related effects of other traits. Male AMS increased with age, and there was no significant reproductive senescence. Most males never copulated, and of the ones that did, the majority had only one successful year. Life span was unrelated to AMS, but early AFL tended to lead to higher AMS at ages 1-3. AMS was related to most morphological and behavioural traits when male age was ignored. Accounting for age and age-specific trait effects (i.e. the interaction between a trait and age) reduced the magnitude of the selection gradients and revealed that behavioural traits are under consistent sexual selection, while sexual selection on morphological traits is stronger in old males. Therefore, sexual selection in black grouse operates primarily on male behaviour and morphological traits may act as additional cues to supplement female choice. These results demonstrate the multifaceted influence of age on both fitness and sexual traits and highlight the importance of accounting for such effects when quantifying sexual selection.

Life-history differences in age-dependent expressions of multiple ornaments and behaviors in a lekking bird.

Age is a major factor explaining variation in life-history traits among individuals with typical patterns of increasing trait values early in life, maximum trait expression, and senescence. However, age-dependent variation in the expressions of sexually selected traits has received less attention, although such variation underpins differences in male competitive abilities and female preference, which are central to sexual selection. In contrast to previous studies focusing on single traits, we used repeated measures of seven sexually selected morphological and behavioral traits in male black grouse (Tetrao tetrix) to quantify the effects of age and life span on their expressions and quantified this variation in relation to male reproductive effort. Trait expression increased with age, but long-lived males had a slower increase and delayed maxima in trait values compared with short-lived males. There was evidence of terminal investment (increasing trait values during the last breeding season) in some traits and senescence in all traits. These trait dynamics were largely explained by the timing of male peak lekking effort. This study shows that fully understanding the variation in sexually selected traits and fitness benefits associated with sexual selection requires accounting for the complex interaction among individual age, life span, and the timing of individuals' investment in reproduction.

Limited indirect fitness benefits of male group membership in a lekking species.

In group living species, individuals may gain the indirect fitness benefits characterizing kin selection when groups contain close relatives. However, tests of kin selection have primarily focused on cooperatively breeding and eusocial species, whereas its importance in other forms of group living remains to be fully understood. Lekking is a form of grouping where males display on small aggregated territories, which females then visit to mate. As females prefer larger aggregations, territorial males might gain indirect fitness benefits if their presence increases the fitness of close relatives. Previous studies have tested specific predictions of kin selection models using measures such as group-level relatedness. However, a full understanding of the contribution of kin selection in the evolution of group living requires estimating individuals' indirect fitness benefits across multiple sites and years. Using behavioural and genetic data from the black grouse (Tetrao tetrix), we show that the indirect fitness benefits of group membership were very small because newcomers joined leks containing few close relatives who had limited mating success. Males' indirect fitness benefits were higher in yearlings during increasing population density but marginally changed the variation in male mating success. Kin selection acting through increasing group size is therefore unlikely to contribute substantially to the evolution and maintenance of lekking in this black grouse population.

Decomposing variation in male reproductive success: age-specific variances and covariances through extra-pair and within-pair reproduction.

Age-specific variances and covariances in reproductive success shape the total variance in lifetime reproductive success (LRS), age-specific opportunities for selection, and population demographic variance and effective size. Age-specific (co)variances in reproductive success achieved through different reproductive routes must therefore be quantified to predict population, phenotypic and evolutionary dynamics in age-structured populations. While numerous studies have quantified age-specific variation in mean reproductive success, age-specific variances and covariances in reproductive success, and the contributions of different reproductive routes to these (co)variances, have not been comprehensively quantified in natural populations. We applied 'additive' and 'independent' methods of variance decomposition to complete data describing apparent (social) and realised (genetic) age-specific reproductive success across 11 cohorts of socially monogamous but genetically polygynandrous song sparrows (Melospiza melodia). We thereby quantified age-specific (co)variances in male within-pair and extra-pair reproductive success (WPRS and EPRS) and the contributions of these (co)variances to the total variances in age-specific reproductive success and LRS. 'Additive' decomposition showed that within-age and among-age (co)variances in WPRS across males aged 2-4 years contributed most to the total variance in LRS. Age-specific (co)variances in EPRS contributed relatively little. However, extra-pair reproduction altered age-specific variances in reproductive success relative to the social mating system, and hence altered the relative contributions of age-specific reproductive success to the total variance in LRS. 'Independent' decomposition showed that the (co)variances in age-specific WPRS, EPRS and total reproductive success, and the resulting opportunities for selection, varied substantially across males that survived to each age. Furthermore, extra-pair reproduction increased the variance in age-specific reproductive success relative to the social mating system to a degree that increased across successive age classes. This comprehensive decomposition of the total variances in age-specific reproductive success and LRS into age-specific (co)variances attributable to two reproductive routes showed that within-age and among-age covariances contributed substantially to the total variance and that extra-pair reproduction can alter the (co)variance structure of age-specific reproductive success. Such covariances and impacts should consequently be integrated into theoretical assessments of demographic and evolutionary processes in age-structured populations.

Physiological costs enforce the honesty of lek display in the black grouse (Tetrao tetrix).

Females may use condition-dependent sexual traits as reliable cues of male "quality" if the costs of the expression of such traits vary with male "quality", and if there is positive genetic correlation between male traits and condition. However, there are multiple ways of measuring the changes in body condition which reflect physiological costs meaning that the multifaceted nature of the physiological costs associated with the expression of sexual traits has rarely been thoroughly examined. In the lekking black grouse (Tetrao tetrix), mating success is highly skewed towards males defending central territories and having high survival rates to the following year, but the mechanisms underpinning such superior performance remain unclear. In this study, we quantified the changes in five measures of body condition before and after the mating season and related these changes to male lek performance (fighting rate, territory centrality and mating success) to understand the physiological costs of male reproductive effort. Between the two capture sessions, male body mass decreased significantly, blood parasite counts and plasma carotenoid concentration increased substantially while the total immunoglobulin concentration tended to increase. There was no overall impairment of individual body condition as the changes in the five measures of body condition were unrelated. Male fighting rate was unrelated to changes in the condition measures but males losing more body mass defended central territories and had high mating success. Therefore, females preferring central, dominant males may select males better able to afford the energetic costs of lek performance thereby effectively enforcing the honesty of male display.

Extra-pair paternity and the variance in male fitness in song sparrows (Melospiza melodia).

The variance in fitness across population members can influence major evolutionary processes. In socially monogamous but genetically polygynandrous species, extra-pair paternity (EPP) is widely hypothesized to increase the variance in male fitness compared to that arising given the socially monogamous mating system. This hypothesis has not been definitively tested because comprehensive data describing males' apparent (social) and realized (genetic) fitness have been lacking. We used 16 years of comprehensive social and genetic paternity data for an entire free-living song sparrow (Melospiza melodia) population to quantify and compare variances in male apparent and realized fitness, and to quantify the contribution of the variances in within-pair reproductive success (WPRS) and extra-pair reproductive success (EPRS) and their covariance to the variance in realized fitness. Overall, EPP increased the variance in male fitness by only 0-27% across different fitness and variance measures. This relatively small effect reflected the presence of socially unpaired males with zero apparent and low realized fitness, small covariance between WPRS and EPRS, and large variance in WPRS that was relatively unaffected by EPP. Therefore, although EPP altered individual males' contributions to future generations, its impact on population-level parameters such as the opportunity for selection and effective population size was limited.

Age-dependent inbreeding risk and offspring fitness costs in female black grouse.

Dispersal is an important mechanism used to avoid inbreeding. However, dispersal may only be effective for part of an individual's lifespan since, post-dispersal individuals that breed over multiple reproductive events may risk mating with kin of the philopatric sex as they age. We tested this hypothesis in black grouse Tetrao tetrix, and show that yearling females never mated with close relatives whereas older females did. However, matings were not with direct kin suggesting that short-distance dispersal to sites containing kin and subsequent overlap of reproductive lifespans between males and females were causing this pattern. Chick mass was lower when kinship was high, suggesting important fitness costs associated with inbred matings. This study shows that increased inbreeding risk might be a widespread yet rarely considered cost of ageing.