Detection of relatedness in chemical alarm cues by a selfing vertebrate depends on population and the life stage producing the alarm cue.

Aquatic prey have impressive abilities to extract information from a variety of chemical cues. For example, they can use the alarm cues released by wounded individuals during a predator attack to learn about predation risk, and they can also distinguish kin from non-kin individuals during interactions. However, it remains unclear whether animals can combine this information on predation risk with kin recognition of the particular individuals under threat. To examine how the relatedness of the individuals in alarm cue affects behaviour we used the self-fertilizing hermaphroditic mangrove rivulus (Kryptolebias marmoratus), in which lineages produce genetically identical offspring through selfing. We explored this in two populations that differ in their level of outcrossing. We measured activity before and after exposure to alarm cue made from individuals (either adults or embryos) from their own lineage or an unrelated lineage from the same population. Fish responded weakly to embryo alarm cues, but tended to reduce their activity more when the alarm cues were from an unrelated lineage compared to alarm cues from their own lineage, particularly in fish from the outcrossing population. In contrast, there was no effect of cue relatedness on the response to adult alarm cues but there was a strong population effect. Specifically, individuals from the outcrossing population tended to react more strongly to alarm cues compared to individuals from the predominantly selfing population. We discuss the potential roles of the major histocompatibility complex in cue detection, differences between adult vs embryo alarm cues in terms of concentration and information, and underlying differences among populations and genetic lineages in their production and detection of chemical cues. Whether this kin recognition offers adaptive benefits or is simply a consequence of being able to detect relatedness in living individuals would be an exciting area for future research.

Predator-induced transgenerational plasticity in animals: a meta-analysis.

There is growing evidence that the environment experienced by one generation can influence phenotypes in the next generation via transgenerational plasticity (TGP). One of the best-studied examples of TGP in animals is predator-induced transgenerational plasticity, whereby exposing parents to predation risk triggers changes in offspring phenotypes. Yet, there is a lack of general consensus synthesizing the predator-prey literature with existing theory pertaining to ecology and evolution of TGP. Here, we apply a meta-analysis to the sizable literature on predator-induced TGP (441 effect sizes from 29 species and 49 studies) to explore five hypotheses about the magnitude, form and direction of predator-induced TGP. Hypothesis #1: the strength of predator-induced TGP should vary with the number of predator cues. Hypothesis #2: the strength of predator-induced TGP should vary with reproductive mode. Hypothesis #3: the strength and direction of predator-induced TGP should vary among offspring phenotypic traits because some traits are more plastic than others. Hypothesis #4: the strength of predator-induced TGP should wane over ontogeny. Hypothesis #5: predator-induced TGP should generate adaptive phenotypes that should be more evident when offspring are themselves exposed to risk. We found strong evidence for predator-induced TGP overall, but no evidence that parental predator exposure causes offspring traits to change in a particular direction. Additionally, we found little evidence in support of any of the specific hypotheses. We infer that the failure to find consistent evidence reflects the heterogeneous nature of the phenomena, and the highly diverse experimental designs used to study it. Together, these findings set an agenda for future work in this area.

Can age-related changes in parental care modulate inbreeding depression? A test using the burying beetle, Nicrophorus orbicollis.

Parental care has been shown to reduce the magnitude of inbreeding depression in some species with facultative care. However, parents often vary in the quality or amount of care they provide to their offspring, and it is less clear whether this variation also impacts the magnitude of inbreeding depression. Here, we tested whether age-related changes in parental care modulate the expression of inbreeding depression in the burying beetle, Nicrophorus orbicollis. Consistent with previous studies, we found that older parents produced larger broods of offspring than younger parents without sacrificing mean larval mass. Inbreeding depression was evident in several fitness-related traits: brood size at dispersal, the proportion of the brood that survived to eclosion, and mean age at death were all reduced in inbred broods compared with outbred broods. Surprisingly, inbred offspring were heavier at dispersal than outbred offspring. This was likely due to reduced sibling competition in inbred broods. Despite evidence for age-related changes in parental investment and the existence of inbreeding depression, there was no evidence that an interaction between the two influenced any of the traits we measured. Our results suggest that age-related changes in parental care may be too slight to influence the expression of inbreeding depression.

Polyphenisms and polymorphisms: Genetic variation in plasticity and color variation within and among bluefin killifish populations.

The presence of stable color polymorphisms within populations begs the question of how genetic variation is maintained. Consistent variation among populations in coloration, especially when correlated with environmental variation, raises questions about whether environmental conditions affect either the fulcrum of those balanced polymorphisms, the plastic expression of coloration, or both. Color patterns in male bluefin killifish provoke both types of questions. Red and yellow morphs are common in all populations. Blue males are more common in tannin-stained swamps relative to clear springs. Here, we combined crosses with a manipulation of light to explore how genetic variation and phenotypic plasticity shape these patterns. We found that the variation in coloration is attributable mainly to two axes of variation: (1) a red-yellow axis with yellow being dominant to red, and (2) a blue axis that can override red-yellow and is controlled by genetics, phenotypic plasticity, and genetic variation for phenotypic plasticity. The variation among populations in plasticity suggests it is adaptive in some populations but not others. The variation among sires in plasticity within the swamp population suggests balancing selection may be acting not only on the red-yellow polymorphism but also on plasticity for blue coloration.

Effects of predation risk on egg steroid profiles across multiple populations of threespine stickleback.

Predation often has consistent effects on prey behavior and morphology, but whether the physiological mechanisms underlying these effects show similarly consistent patterns across different populations remains an open question. In vertebrates, predation risk activates the hypothalamic-pituitary-adrenal (HPA) axis, and there is growing evidence that activation of the maternal HPA axis can have intergenerational consequences via, for example, maternally-derived steroids in eggs. Here, we investigated how predation risk affects a suite of maternally-derived steroids in threespine stickleback eggs across nine Alaskan lakes that vary in whether predatory trout are absent, native, or have been stocked within the last 25 years. Using liquid chromatography coupled with mass spectroscopy (LC-MS/MS), we detected 20 steroids within unfertilized eggs. Factor analysis suggests that steroids covary within and across steroid classes (i.e. glucocorticoids, progestogens, sex steroids), emphasizing the modularity and interconnectedness of the endocrine response. Surprisingly, egg steroid profiles were not significantly associated with predator regime, although they were more variable when predators were absent compared to when predators were present, with either native or stocked trout. Despite being the most abundant steroid, cortisol was not consistently associated with predation regime. Thus, while predators can affect steroids in adults, including mothers, the link between maternal stress and embryonic development is more complex than a simple one-to-one relationship between the population-level predation risk experienced by mothers and the steroids mothers transfer to their eggs.

Boldness is affected by recent experience with predation cues and body size in mosquitofish.

Adjusting behaviour can be crucial to prey surviving a predator encounter. How any one individual modifies their behaviour in response to predation risk might be affected by their previous experience with danger and their own vulnerability. Using western mosquitofish, we examined how boldness in different contexts was affected by an individual's recent experience with predation risk. Individuals were repeatedly chased by a largemouth bass model and encountered alarm cue to mimic conditions of high risk (cues twice on 2 days), low risk (cues twice on 1 day), or no risk (water only). We then measured boldness and avoidance behaviour under three different contexts: in a novel tank, with a shoal of conspecifics, and with alarm cues and a model predator. We found that how recent experiences influenced boldness in a novel tank depended on body size. Smaller fish from the no and low risk treatments were more likely to emerge from a shelter into a novel environment than larger individuals. When individuals had recently experienced high levels of risk however, this pattern was reversed. We also found that individuals who had experienced any recent risk (low and high) were more likely to leave the safety of a shoal and approach a novel object compared to individuals who had not experienced any recent danger. Avoidance behaviour across the three assays was not affected by recent experiences but was affected by body size to varying degrees. For example, larger fish were more likely to stay in the plants, away from the cues of predation compared to smaller fish. Overall, our results suggest that how recent experiences with risk influence subsequent behaviour can depend on a variety of interacting factors including the intensity of recent experiences, the particular behaviour examined, and an individual's body size.

Integrating Ecological and Evolutionary Context in the Study of Maternal Stress.

Maternal stress can prenatally influence offspring phenotypes and there are an increasing number of ecological studies that are bringing to bear biomedical findings to natural systems. This is resulting in a shift from the perspective that maternal stress is unanimously costly, to one in which maternal stress may be beneficial to offspring. However, this adaptive perspective is in its infancy with much progress to still be made in understanding the role of maternal stress in natural systems. Our aim is to emphasize the importance of the ecological and evolutionary context within which adaptive hypotheses of maternal stress can be evaluated. We present five primary research areas where we think future research can make substantial progress: (1) understanding maternal and offspring control mechanisms that modulate exposure between maternal stress and subsequent offspring phenotype response; (2) understanding the dynamic nature of the interaction between mothers and their environment; (3) integrating offspring phenotypic responses and measuring both maternal and offspring fitness outcomes under real-life (either free-living or semi-natural) conditions; (4) empirically testing these fitness outcomes across relevant spatial and temporal environmental contexts (both pre- and post-natal environments); (5) examining the role of maternal stress effects in human-altered environments-i.e., do they limit or enhance fitness. To make progress, it is critical to understand the role of maternal stress in an ecological context and to do that, we must integrate across physiology, behavior, genetics, and evolution.

Effects of mothers' and fathers' experience with predation risk on the behavioral development of their offspring in threespined sticklebacks.

Stressors experienced by parents can influence the behavioral development of their offspring. Here, we review recent studies in threespined sticklebacks (a species in which males are the sole providers of parental care) showing that when parents are exposed to an ecologically relevant stressor (predation risk), there are consequences for offspring. For example, female sticklebacks exposed to predation risk produce eggs with higher concentrations of cortisol, a stress hormone, and offspring with altered behavior and physiology. Male sticklebacks exposed to predation risk produce offspring that are less active, smaller, and in poorer condition. The precise mechanisms by which maternal and paternal experiences with predators affect offspring phenotypes are under investigation, and could include steroid hormones, olfactory cues and/or parental behavior. As in other species, some of the consequences of parental exposure to predation risk for offspring in sticklebacks might be adaptive, but depend on the stressor, the reliability of the parental and offspring environments and the evolutionary history of the population.

A female's past experience with predators affects male courtship and the care her offspring will receive from their father.

Differential allocation occurs when individuals adjust their reproductive investment based on their partner's traits. However, it remains unknown whether animals differentially allocate based on their partner's past experiences with predation risk. If animals can detect a potential mate's experience with predators, this might inform them about the stress level of their potential mate, the likelihood of parental effects in offspring and/or the dangers present in the environment. Using threespined stickleback (Gasterosteus aculeatus), we examined whether a female's previous experience with being chased by a model predator while yolking eggs affects male mating effort and offspring care. Males displayed fewer conspicuous courtship behaviours towards females that had experienced predation risk in the past compared with unexposed females. This differential allocation extended to how males cared for the resulting offspring of these matings: fathers provided less parental care to offspring of females that had experienced predation risk in the past. Our results show for the first time, to our knowledge, that variation among females in their predator encounters can contribute to behavioural variation among males in courtship and parental care, even when males themselves do not encounter a predator. These results, together with previous findings, suggest that maternal predator exposure can influence offspring development both directly and indirectly, through how it affects father care.

No apparent benefits of allonursing for recipient offspring and mothers in the cooperatively breeding meerkat.

Cooperative behaviours by definition are those that provide some benefit to another individual. Allonursing, the nursing of non-descendent young, is often considered a cooperative behaviour and is assumed to provide benefits to recipient offspring in terms of growth and survival, and to their mothers, by enabling them to share the lactation load. However, these proposed benefits are not well understood, in part because maternal and litter traits and other ecological and social variables are not independent of one another, making patterns hard to discern using standard univariate analyses. Here, we investigate the potential benefits of allonursing in the cooperatively breeding Kalahari meerkat, where socially subordinate females allonurse the young of a dominant pair without having young of their own. We use structural equation modelling to allow us to account for the interdependence of maternal traits, litter traits and environmental factors. We find no evidence that allonursing provides benefits to pups or mothers. Pups that received allonursing were not heavier at emergence and did not have a higher survival rate than pups that did not receive allonursing. Mothers whose litters were allonursed were not in better physical condition, did not reconceive faster and did not reduce their own nursing investment compared to mothers who nursed their litters alone. These patterns were not significantly influenced by whether mothers were in relatively good, or poor, condition. We suggest that allonursing may persist in this species because the costs to allonurses may be low. Alternatively, allonursing may confer other, more cryptic, benefits to pups or allonurses, such as immunological or social benefits.

Effect of maternal predator exposure on the ability of stickleback offspring to generalize a learned colour-reward association.

Maternal stress can have long-term negative consequences for offspring learning performance. However, it is unknown whether these maternal effects extend to the ability of offspring to apply previously learned information to new situations. In this study, we first demonstrate that juvenile threespine sticklebacks, Gasterosteus aculeatus, are indeed capable of generalizing an association between a colour and a food reward learned in one foraging context to a new foraging context (i.e. they can apply previously learned knowledge to a new situation). Next, we examined whether this ability to generalize was affected by maternal predator stress. We manipulated whether mothers were repeatedly chased by a model predator while yolking eggs (i.e. before spawning) and then assessed the learning performance of their juvenile offspring in groups and pairs using a colour discrimination task that associated a colour with a food reward. We found that maternal predator exposure affected the tendency of offspring to use social cues: offspring of predator-exposed mothers were faster at copying a leader's behaviour towards the rewarded colour than offspring of unexposed mothers. However, once the colour-reward association had been learned, offspring of predator-exposed and unexposed mothers were equally able to generalize their learned association to a new foraging task. These results suggest that offspring of predator-exposed mothers might be able to overcome learning deficits caused by maternal stress by relying more on social cues.

Paternal care in a fish: epigenetics and fitness enhancing effects on offspring anxiety.

In many animals, including humans, interactions with caring parents can have long-lasting effects on offspring sensitivity to stressors. However, whether these parental effects impact offspring fitness in nature is often unclear. In addition, despite evidence that maternal care can influence offspring behaviour via epigenetic alterations to the genome, it remains unclear whether paternal care has similar effects. Here, we show in three-spined sticklebacks, a fish in which fathers are the sole provider of offspring care, that the direct care provided by fathers affects offspring anxiety and the potential for epigenetic alterations to the offspring genome. We find that families are differentially vulnerable to early stress and fathers can compensate for this differential sensitivity with the quality of their care. This variation in paternal care is also linked to the expression in offspring brains of a DNA methyltransferase (Dnmt3a) responsible for de novo methylation. We show that these paternal effects are potentially adaptive and anxious offspring are unlikely to survive an encounter with a predator. By supplying offspring care, fathers reduce offspring anxiety thereby increasing the survival of their offspring-not in the traditional sense through resource provisioning but through an epigenetic effect on offspring behavioural development.

Individual variation in foraging behavior reveals a trade-off between flexibility and performance of a top predator.

There is increasing evidence that behavioral flexibility is associated with the ability to adaptively respond to environmental change. Flexibility can be advantageous in some contexts such as exploiting novel resources, but it may come at a cost of accuracy or performance in ecologically relevant tasks, such as foraging. Such trade-offs may, in part, explain why individuals within a species are not equally flexible. Here, we conducted a reversal learning task and predation experiment on a top fish predator, the Northern pike (Esox lucius), to examine individual variation in flexibility and test the hypothesis that an individual's behavioral flexibility is negatively related with its foraging performance. Pikes were trained to receive a food reward from either a red or blue cup and then the color of the rewarded cup was reversed. We found that pike improved over time in how quickly they oriented to the rewarded cup, but there was a bias toward the color red. Moreover, there was substantial variation among individuals in their ability to overcome this red bias and switch from an unrewarded red cup to the rewarded blue cup, which we interpret as consistent variation among individuals in behavioral flexibility. Furthermore, individual differences in behavioral flexibility were negatively associated with foraging performance on ecologically relevant stickleback prey. Our data indicate that individuals cannot be both behaviorally flexible and efficient predators, suggesting a trade-off between these two traits.

Heritable variation underlies behavioural types in the mating context in male bluefin killifish.

In many species, consistent behavioural differences among individuals are linked to fitness variation. Determining the environmental and genetic factors that mould these behavioural types is crucial to understanding how behaviours might respond to selection. Male bluefin killifish, Lucania goodei, show extensive consistent behavioural variation in their levels of courtship, male-directed aggression and female-directed aggression, resulting in a range of fitness-related behavioural types coexisting within a population. To determine whether the behavioural components underlying a male's stable behavioural type in the mating context are heritable and genetically correlated, we performed paternal half-sib crosses. Using animal models, we found that all three of these mating behaviours were moderately heritable (h2 = 0.17-0.29) and courtship behaviour was also heritable as a binomial trait (court yes/no: h2 = 0.50). Including effects of dam identity/common rearing environment experienced by full sibs decreased model fit, suggesting that early social interactions might contribute to behavioural types. In addition, we found evidence consistent with the possibility that the positive phenotypic correlations among mating behaviours are underlain by positive genetic correlations. Thus, it is possible that the seemingly maladaptive aggression that males direct towards females during social interactions persist due to genetic constraints and direct selection on both male-directed aggression and courtship behaviour.

Reciprocal behavioral plasticity and behavioral types during predator-prey interactions.

How predators and prey interact has important consequences for population dynamics and community stability. Here we explored how predator-prey interactions are simultaneously affected by reciprocal behavioral plasticity (i.e., plasticity in prey defenses countered by plasticity in predator offenses and vice versa) and consistent individual behavioral variation (i.e., behavioral types) within both predator and prey populations. We assessed the behavior of a predator species (northern pike) and a prey species (three-spined stickleback) during one-on-one encounters. We also measured additional behavioral and morphological traits in each species. Using structural equation modeling, we found that reciprocal behavioral plasticity as well as predator and prey behavioral types influenced how individuals behaved during an interaction. Thus, the progression and ultimate outcome of predator-prey interactions depend on both the dynamic behavioral feedback occurring during the encounter and the underlying behavioral type of each participant. We also examined whether predator behavioral type is underlain by differences in metabolism and organ size. We provide some of the first evidence that behavioral type is related to resting metabolic rate and size of a sensory organ (the eyes). Understanding the extent to which reciprocal behavioral plasticity and intraspecific behavioral variation influence the outcome of species interactions could provide insight into the maintenance of behavioral variation as well as community dynamics.

Maternal predator-exposure has lifelong consequences for offspring learning in threespined sticklebacks.

Learning is an important form of phenotypic plasticity that allows organisms to adjust their behaviour to the environment. An individual's learning performance can be affected by its mother's environment. For example, mothers exposed to stressors, such as restraint and forced swimming, often produce offspring with impaired learning performance. However, it is unclear whether there are maternal effects on offspring learning when mothers are exposed to ecologically relevant stressors, such as predation risk. Here, we examined whether maternal predator-exposure affects adult offsprings' learning of a discrimination task in threespined sticklebacks (Gasterosteus aculeatus). Mothers were either repeatedly chased by a model predator (predator-exposed) or not (unexposed) while producing eggs. Performance of adult offspring from predator-exposed and unexposed mothers was assessed in a discrimination task that paired a particular coloured chamber with a food reward. Following training, all offspring learned the colour-association, but offspring of predator-exposed mothers located the food reward more slowly than offspring of unexposed mothers. This pattern was not driven by initial differences in exploratory behaviour. These results demonstrate that an ecologically relevant stressor (predation risk) can induce maternal effects on offspring learning, and perhaps behavioural plasticity more generally, that last into adulthood.

Maternal exposure to predation risk decreases offspring antipredator behaviour and survival in threespined stickleback.

1. Adaptive maternal programming occurs when mothers alter their offspring's phenotype in response to environmental information such that it improves offspring fitness. When a mother's environment is predictive of the conditions her offspring are likely to encounter, such transgenerational plasticity enables offspring to be better-prepared for this particular environment. However, maternal effects can also have deleterious effects on fitness.2. Here, we test whether female threespined stickleback fish exposed to predation risk adaptively prepare their offspring to cope with predators. We either exposed gravid females to a model predator or not, and compared their offspring's antipredator behaviour and survival when alone with a live predator. Importantly, we measured offspring behaviour and survival in the face of the same type of predator that threatened their mothers (Northern pike).3. We did not find evidence for adaptive maternal programming; offspring of predator-exposed mothers were less likely to orient to the predator than offspring from unexposed mothers. In our predation assay, orienting to the predator was an effective antipredator behaviour and those that oriented, survived for longer.4. In addition, offspring from predator-exposed mothers were caught more quickly by the predator on average than offspring from unexposed mothers. The difference in antipredator behaviour between the maternal predator-exposure treatments offers a potential behavioural mechanism contributing to the difference in survival between maternal treatments.5. However, the strength and direction of the maternal effect on offspring survival depended on offspring size. Specifically, the larger the offspring from predator-exposed mothers, the more vulnerable they were to predation compared to offspring from unexposed mothers.6. Our results suggest that the predation risk perceived by mothers can have long-term behavioural and fitness consequences for offspring in response to the same predator. These stress-mediated maternal effects can have nonadaptive consequences for offspring when they find themselves alone with a predator. In addition, complex interactions between such maternal effects and offspring traits such as size can influence our conclusions about the adaptive nature of maternal effects.

Coevolutionary constraints? The environment alters tripartite interaction traits in a legume.

Third party species, which interact with one or both partners of a pairwise species interaction, can shift the ecological costs and the evolutionary trajectory of the focal interaction. Shared genes that mediate a host's interactions with multiple partners have the potential to generate evolutionary constraints, making multi-player interactions critical to our understanding of the evolution of key interaction traits. Using a field quantitative genetics approach, we studied phenotypic and genetic correlations among legume traits for rhizobium and herbivore interactions in two light environments. Shifts in plant biomass allocation mediated negative phenotypic correlations between symbiotic nodule number and herbivory in the field, whereas positive genetic covariances suggested shared genetic pathways between nodulation and herbivory response. Trait variance-covariance (G) matrices were not equal in sun and shade, but nevertheless responses to independent and correlated selection are expected to be similar in both environments. Interactions between plants and aboveground antagonists might alter the evolutionary potential of traits mediating belowground mutualisms (and vice versa). Thus our understanding of legume-rhizobium genetics and coevolution may be incomplete without a grasp of how these networks overlap with other plant interactions.

Mechanisms of reproductive isolation among sympatric broadcast-spawning corals of the Montastraea annularis species complex.

Many coral species spawn simultaneously and have compatible gametes, leading to controversy over the nature of species boundaries and the frequency with which hybridization occurs. Three western Atlantic corals, Montastraea annularis, M. faveolata, and M. franksi, typify this controversy; they all spawn sympatrically on the same evenings after the fall full moons. Here we show, in both Panama and the Bahamas for multiple years, how a variety of mechanisms may act in concert to reproductively isolate all three species. Field studies indicate that M. franksi spawns two hours earlier than the other two species, and the eggs released during this earlier period disperse an average of 500 m by the time the other species spawn. Field measures of fertilization indicate that peak fertilization occurs when spawning synchrony is high and that corals that spawn at the tails of the spawning distributions have greatly reduced fertilization success. Laboratory studies indicate that there is a gametic incompatibility between M. faveolata and the other two species. There are regional differences in the gametic compatibility of M. franksi and M. annularis. In Panama, the two species are completely compatible, whereas in the Bahamas, M. franksi sperm can fertilize M. annularis eggs but the reciprocal cross often fails. Gamete age influences patterns of fertilization, such that very young eggs seem resistant to fertilization and old sperm lose viability after two hours. In sum, the combination of temporal differences in spawning, sperm aging, gamete dispersal and dilution, and gametic incompatibility act in various combinations among the three species, making it unlikely that hybrid fertilization would occur.