Effects of inbreeding and elevated rearing temperatures on strategic sperm investment.

Males often strategically adjust the number of available sperm based on the social context (i.e. sperm priming response), but it remains unclear how environmental and genetic factors shape this adjustment. In freshwater ecosystems, high ambient temperatures often lead to isolated pools of hotter water in which inbreeding occurs. Higher water temperatures and inbreeding can impair fish development, potentially disrupting sperm production. We used guppies (Poecilia reticulata) to investigate how developmental temperature (26 °C, 30 °C) and male inbreeding status (inbred, outbred) influence their sperm priming response. We also tested if sperm priming was affected by whether the female was a relative (sister) and whether she was inbred or outbred. There was no effect of rearing temperature; male inbreeding status alone determined the number of available sperm in response to female presence, her inbreeding status, and her relatedness. Inbred males produced significantly more sperm in the presence of an unrelated, outbred female than when no female was present. Conversely, outbred males did not alter the number of sperm available in response to female presence or relatedness. Moreover, inbred males produced marginally more sperm when exposed to an unrelated female that was outbred rather than inbred, but there was no difference when exposed to an inbred female that was unrelated versus related. Together, a sperm priming response was only observed in inbred males when exposed to an outbred female. Outbred females in our study were larger than inbred females, suggesting that inbred males strategically allocated ejaculate resources toward females in better condition.

Effects of past mating behavior versus past ejaculation on male mate choice and male attractiveness.

Past reproductive effort allows males to assess their ability to acquire mates, but it also consumes resources that can reduce their future competitive ability. Few studies have examined how a male's reproductive history affects his subsequent mate choice, and, to date, no study has determined the relative contribution of past mating behavior and past ejaculate production because these two forms of investment are naturally highly correlated. Here, we disentangled the relative effects of past mating behavior and past ejaculate production in mosquitofish (Gambusia holbrooki) by experimentally preventing some males from ejaculating when trying to mate. We assessed the effect of mating behavior on mate choice by comparing males that had previously been with or without access to females and male rivals for 8 and 16 weeks and assessed the effect of ejaculation on mate choice by comparing males that either could or could not ejaculate when they had access to females for 16 weeks. Reproductive treatment did not affect male attractiveness, but it did affect male mate choice. Somewhat surprisingly, in five of the six treatment-by-age at testing combinations, males preferred a female in the vicinity of a male rival over a solitary female. This preference was marginally stronger for males that had previously engaged in mating behavior but were unaffected by past ejaculate production. We discuss the potential benefits to males of associating with another male when seeking mates. This is the first study to quantify the relative influence of pre- and post-copulatory reproductive investment on male mate choice.

Mitochondrial function and sexual selection: can physiology resolve the 'lek paradox'?

Across many taxa, males use elaborate ornaments or complex displays to attract potential mates. Such sexually selected traits are thought to signal important aspects of male 'quality'. Female mating preferences based on sexual traits are thought to have evolved because choosy females gain direct benefits that enhance their lifetime reproductive success (e.g. greater access to food) and/or indirect benefits because high-quality males contribute genes that increase offspring fitness. However, it is difficult to explain the persistence of female preferences when males only provide genetic benefits, because female preferences should erode the heritable genetic variation in fitness that sexually selected traits signal. This 'paradox of the lek' has puzzled evolutionary biologists for decades, and inspired many hypotheses to explain how heritable variation in sexually selected traits is maintained. Here, we discuss how factors that affect mitochondrial function can maintain variation in sexually selected traits despite strong female preferences. We discuss how mitochondrial function can influence the expression of sexually selected traits, and we describe empirical studies that link the expression of sexually selected traits to mitochondrial function. We explain how mothers can affect mitochondrial function in their offspring by (a) influencing their developmental environment through maternal effects and (b) choosing a mate to increase the compatibility of mitochondrial and nuclear genes (i.e. the 'mitonuclear compatibility model of sexual selection'). Finally, we discuss how incorporating mitochondrial function into models of sexual selection might help to resolve the paradox of the lek, and we suggest avenues for future research.

Long-term environmental stability does not erode plasticity in nest building responses to changing ambient conditions.

The primary function of animal nests is to protect developing offspring from hostile and fluctuating environments. Animal builders have been shown to adjust nest construction in response to changes in their environment. However, the extent of this plasticity, and its dependence on an evolutionary history of environmental variability, is not well understood. To test whether an evolutionary history with flowing water impacts male ability to adjust nests in response to flow regime, we collected three-spined sticklebacks (Gasterosteus aculeatus) from three lakes and three rivers, and brought them into reproductive condition in controlled laboratory aquaria. Males were then allowed to nest under both flowing and static conditions. Nest building behaviour, nest structure and nest composition were all recorded. In comparison to males building nests under static conditions, males building in flowing water took longer to construct their nests and invested more in nesting behaviour. Moreover, nests built in flowing water contained less material, were smaller, more compact, neater and more elongated than nests built under static conditions. Whether males came from rivers or lakes had little impact on nesting activities, or male capacity to adjust behaviours in response to flow treatment. Our findings suggest that aquatic animals which have experienced a stable environment over a long period of time retain plasticity in nest-building behaviours that allow them to adjust nests to ambient flow conditions. This ability may prove crucial in coping with the increasingly unpredictable flow regimes found in anthropogenically altered waterways and those resulting from global climate change. This article is part of the theme issue 'The evolutionary ecology of nests: a cross-taxon approach'.

An innovative approach to using an intensive field course to build scientific and professional skills.

This paper reports on the design and evaluation of Field Studies in Functional Ecology (FSFE), a two-week intensive residential field course that enables students to master core content in functional ecology alongside skills that facilitate their transition from "student" to "scientist." We provide an overview of the course structure, showing how the constituent elements have been designed and refined over successive iterations of the course. We detail how FSFE students: (1) Work closely with discipline specialists to develop a small group project that tests an hypothesis to answer a genuine scientific question in the field; (2) Learn critical skills of data management and communication; and (3) Analyze, interpret, and present their results in the format of a scientific symposium. This process is repeated in an iterative "cognitive apprenticeship" model, supported by a series of workshops that name and explicitly instruct the students in "hard" and "soft" skills (e.g., statistics and teamwork, respectively) critically relevant for research and other careers. FSFE students develop a coherent and nuanced understanding of how to approach and execute ecological studies. The sophisticated knowledge and ecological research skills that they develop during the course is demonstrated through high-quality presentations and peer-reviewed publications in an open-access, student-led journal. We outline our course structure and evaluate its efficacy to show how this novel combination of field course elements allows students to gain maximum value from their educational journey, and to develop cognitive, affective, and reflective tools to help apply their skills as scientists.

Effects of developmental and adult environments on ageing.

Developmental and adult environments can interact in complex ways to influence the fitness of individuals. Most studies investigating effects of the environment on fitness focus on environments experienced and traits expressed at a single point in an organism's life. However, environments vary with time, so the effects of the environments that organisms experience at different ages may interact to affect how traits change throughout life. Here, we test whether thermal stress experienced during development leads individuals to cope better with thermal stress as adults. We manipulated temperature during both development and adulthood and measured a range of life-history traits, including senescence, in male and female seed beetles (Callosobruchus maculatus). We found that thermal stress during development reduced adult reproductive performance of females. In contrast, life span and age-dependent mortality were affected more by adult than developmental environments, with high adult temperatures decreasing longevity and increasing age-dependent mortality. Aside from an interaction between developmental and adult environments to affect age-dependent changes in male weight, we did not find any evidence of a beneficial acclimation response to developmental thermal stress. Overall, our results show that effects of developmental and adult environments can be both sex and trait specific, and that a full understanding of how environments interact to affect fitness and ageing requires the integrated study of conditions experienced during different stages of ontogeny.

The high invasion success of fall armyworm is related to life-history strategies across a range of stressful temperatures.

BACKGROUND: Insects living in unfavorably high or low temperatures are predicted to display a fast or slow life-history strategy. Here, we examined life histories of fall armyworm (FAW), a globally important invasive species with a broad ecological niche, at five constant temperatures of 13, 19, 25, 31 and 37°C, to study life-history responses to different temperatures. RESULTS: In our experiment, FAW had lower lifetime fecundity at unfavorable temperatures, a finding that is consistent with the idea that FAW can shift resources from reproduction to other functions under stressful conditions-such as heat or cold tolerance. Given the adverse effects of stressful temperatures, life-history strategies arise from individuals having limited remaining resources to allocate towards vital functions like survival or reproduction. Here we show plasticity in life-history strategies adopted at different temperatures. Rather than simply varying along a fast-slow continuum, FAW at unfavorably high temperatures exhibited lower daily fecundity and longer reproductive lifespans, and at unfavorably low temperatures showed a shorter peak in reproduction later in life, compared with FAW at 25°C. Such patterns, if adaptive, could suggest a strategy mitigating reproductive and mortality risk in unfavorable environments, however, this remains to be tested. CONCLUSION: Our analysis suggests that the high invasion success of FAW may result from their ability to adjust life-history strategies, across a range of stressful temperatures, in a way that reduces not only mortality, but also fecundity loss. The adoption of such strategies may be instrumental for the global invasion success of FAW. © 2022 Society of Chemical Industry.

Disentangling the effects of male age and mating history: Contrasting effects of mating history on precopulatory mating behavior and paternity success.

Many studies ask whether young or older males are better at acquiring mates. Even so, how age affects reproductive success is still poorly understood because male age and mating history are confounded in most studies: older males usually have more mating experience. To what extent does mating history rather than age explain variation in male mating success? And how do mating history and male age determine paternity when there is also postcopulatory sexual selection? Here, we experimentally manipulated the mating history of old and young males in the eastern mosquitofish (Gambusia holbrooki). We then recorded male mating behavior and share of paternity (1259 offspring from 232 potential sires) when they competed for mates and fertilizations. Old males, and males with no mating experience, spent significantly more time approaching females, and attempting to mate, than did young males and those with greater mating experience. Male age and mating history interacted to affect paternity: old males benefited from having previous mating experience, but young males did not. Our results highlight that the age-related changes in male reproductive traits and in paternity that have been described in many taxa may be partly attributable to male mating history and not simply to age itself.

Male age alone predicts paternity success under sperm competition when effects of age and past mating effort are experimentally separated.

Older males often perform poorly under post-copulatory sexual selection. It is unclear, however, whether reproductive senescence is because of male age itself or the accumulated costs of the higher lifetime mating effort that is usually associated with male age. To date, very few studies have accounted for mating history and sperm storage when testing the effect of male age on sperm traits, and none test how age and past mating history influence paternity success under sperm competition. Here, we experimentally manipulate male mating history to tease apart its effects from that of age on ejaculate traits and paternity in the mosquitofish, Gambusia holbrooki. We found that old, naive males had more sperm than old, experienced males, while the reverse was true for young males. By contrast, neither male age nor mating history affected sperm velocity. Finally, using artificial insemination to experimentally control the number of sperm per male, we found that old males sired significantly more offspring than young males independently of their mating history. Our results highlight that the general pattern of male reproductive senescence described in many taxa may often be affected by two naturally confounding factors, male mating history and sperm age, rather than male age itself.

The role of maternal effects on offspring performance in familiar and novel environments.

Maternal effects are an important evolutionary force that may either facilitate adaptation to a new environment or buffer against unfavourable conditions. The degree of variation in traits expressed by siblings from different mothers is often sensitive to environmental conditions. This could generate a Maternal-by-Environment interaction (M × E) that inflates estimates of Genotype-by-Environment effects (G × E). We aimed to test for environment-specific maternal effects (M × E) using a paternal full-sib/half-sib breeding design in the seed beetle Callosobruchus maculatus, where we split and reared offspring from the same mother on two different bean host types-original and novel. Our quantitative genetic analysis indicated that maternal effects were very small on both host types for all the measured life-history traits. There was also little evidence that maternal oviposition preference for a particular host type predicted her offspring's performance on that host. Further, additive genetic variance for most traits was relatively high on both hosts. While there was higher heritability for offspring reared in the novel host, there was no evidence for G × Es, and most cross-host genetic correlations were positive. This suggests that offspring from the same family ranked similarly for performance on both host types. Our results point to a genetic basis of host adaptation in the seed beetle, rather than maternal effects. Even so, we encourage researchers to test for potential M × Es because, due to a lack of testing, it remains unclear how often they arise.

The effects of competition on fitness depend on the sex of both competitors.

In intraspecific competition, the sex of competing individuals is likely to be important in determining the outcome of competitive interactions and the way exposure to conspecifics during development influences adult fitness traits. Previous studies have explored differences between males and females in their response to intraspecific competition. However, few have tested how the sex of the competitors, or any interactions between focal and competitor sex, influences the nature and intensity of competition. We set up larval seed beetles Callosobruchus maculatus to develop either alone or in the presence of a male or female competitor and measured a suite of traits: development time, emergence weight; male ejaculate mass, copulation duration, and lifespan; and female lifetime fecundity, offspring egg-adult survival, and lifespan. We found effects of competition and competitor sex on the development time and emergence weight of both males and females, and also of an interaction between focal and competitor sex: Females emerged lighter when competing with another female, while males did not. There was little effect of larval competition on male and female adult fitness traits, with the exception of the effect of a female competitor on a focal female's offspring survival rate. Our results highlight the importance of directly measuring the effects of competition on fitness traits, rather than distant proxies for fitness, and suggest that competition with the sex with the greater resource requirements (here females) might play a role in driving trait evolution. We also found that male-male competition during development resulted in shorter copulation times than male-female competition, a result that remained when controlling for the weight of competitors. Although it is difficult to definitively tease apart the effects of social environment and access to resources, this result suggests that something about the sex of competitors other than their size is driving this pattern.

Stress in the city: meta-analysis indicates no overall evidence for stress in urban vertebrates.

As cities continue to grow it is increasingly important to understand the long-term responses of wildlife to urban environments. There have been increased efforts to determine whether urbanization imposes chronic stress on wild animals, but empirical evidence is mixed. Here, we conduct a meta-analysis to test whether there is, on average, a detrimental effect of urbanization based on baseline and stress-induced glucocorticoid levels of wild vertebrates. We found no effect of urbanization on glucocorticoid levels, and none of sex, season, life stage, taxon, size of the city nor methodology accounted for variation in the observed effect sizes. At face value, our results suggest that urban areas are no more stressful for wildlife than rural or non-urban areas, but we offer a few reasons why this conclusion could be premature. We propose that refining methods of data collection will improve our understanding of how urbanization affects the health and survival of wildlife.

Combined effects of rearing and testing temperatures on sperm traits.

Temperature experienced during early development can affect a range of adult life-history traits. Animals often show seemingly adaptive developmental plasticity-with animals reared at certain temperatures performing better as adults at those temperatures. The extent to which this type of adaptive response occurs in gonadal tissue that affects sperm traits is, however, poorly studied. We initially reared male mosquito fish (Gambusia holbrooki) at either 18°C or 30°C, and then measured their sperm reserves as adults. We also looked at the velocity of their sperm, at both the matched and mismatched temperatures. Although males reared at 30°C were larger than those initially reared at 18°C, there was no detectable effect of rearing temperature on absolute sperm number. Sperm swam faster at 30°C than 18°C regardless of the male's rearing temperature. Therefore, we found no evidence of adaptive developmental plasticity. Rearing temperature did, however, significantly influence the relationship between male body size and sperm velocity. Larger males had faster sperm when reared at the warmer temperature and slower sperm when reared at the cooler temperature. This suggests that rearing temperature could alter the relationship between pre-copulatory sexual selection and post-copulatory sexual selection as male size affects mating success. Finally, there was a positive correlation between velocities at the two test temperatures, suggesting that temperature experienced during sperm competition is unlikely to affect a male's relative fertilization success.

Shifts in Reproductive Investment in Response to Competitors Lower Male Reproductive Success.

AbstractIn many species, males exhibit phenotypic plasticity in sexually selected traits when exposed to social cues about the intensity of sexual competition. To date, however, few studies have tested how this plasticity affects male reproductive success. We initially tested whether male mosquitofish, Gambusia holbrooki (Poeciliidae), change their investment in traits under pre- and postcopulatory sexual selection depending on the social environment. For a full spermatogenesis cycle, focal males were exposed to visual and chemical cues of rivals that were either present (competitive treatment) or absent (control). Males from the competitive treatment had significantly slower-swimming sperm but did not differ in sperm count from control males. When two males competed for a female, competitive treatment males also made significantly fewer copulation attempts and courtship displays than control males. Further, paternity analysis of 708 offspring from 148 potential sires, testing whether these changes in reproductive traits affected male reproductive success, showed that males previously exposed to cues about the presence of rivals sired significantly fewer offspring when competing with a control male. We discuss several possible explanations for these unusual findings.

Breaking barriers? Ethnicity and socioeconomic background impact on early career progression in the fields of ecology and evolution.

The academic disciplines of Science, Technology, Engineering and Mathematics (STEM) have long suffered from a lack of diversity. While in recent years there has been some progress in addressing the underrepresentation of women in STEM subjects, other characteristics that have the potential to impact on equality of opportunity have received less attention. In this study, we surveyed 188 early career scientists (ECRs), defined as within 10 years of completing their PhD, in the fields of ecology, evolutionary biology, behaviour, and related disciplines. We examined associations between ethnicity, age, sexual orientation, sex, socioeconomic background, and disability, with measures of career progression, namely publication record, number of applications made before obtaining a postdoc, type of contract, and number of grant applications made. We also queried respondents on perceived barriers to progression and potential ways of overcoming them. Our key finding was that socioeconomic background and ethnicity were associated with measures of career progression. While there was no difference in the number of reported first-authored papers on PhD completion, ethnic minority respondents reported fewer other-authored papers. In addition, ECRs from a lower socioeconomic background were more likely to report being in teaching and research positions, rather than research-only positions, the latter being perceived as more prestigious by some institutions. We discuss our findings in the context of possible inequality of opportunity. We hope that this study will stimulate wider discussion and help to inform strategies to address the underrepresentation of minority groups in the fields of ecology and evolution, and STEM subjects more widely.

No link between nymph and adult coloration in shield bugs: weak selection by predators.

Many organisms use different antipredator strategies throughout their life, but little is known about the reasons or implications of such changes. For years, it has been suggested that selection by predators should favour uniformity in local warning signals. If this is the case, we would expect high resemblance in colour across life stages in aposematic animals where young and adults share similar morphology and habitat. In this study, we used shield bugs (Hemiptera: Pentatomoidea) to test whether colour and colour diversity evolve similarly at different life stages. Since many of these bugs are considered to be aposematic, we also combined multi-species analyses with predation experiments on the cotton harlequin bug to test whether there is evidence of selection for uniformity in colour across life stages. Overall, we show that the diversity of colours used by both life stages is comparable, but adults are more cryptic than nymphs. We also demonstrate that nymphs and adults of the same species do not tend to look alike. Experiments on our model system suggest that predators can generalise among life stages that look different, and exhibit strong neophobia. Altogether, our results show no evidence of selection favouring colour similarity between adults and nymphs in this speciose clade.

Sex in the city: sexual selection and urban colonization in passerines.

Urbanization leads to a rapid and drastic transformation of habitats, forcing native fauna to manage novel ecological challenges or to move. Sexual selection is a powerful evolutionary force, which is sometimes predicted to enhance the ability of species to adapt to novel environments because it allows females to choose high-quality males, but other times is predicted to reduce the viability of populations because it pushes males beyond naturally selected optima. However, we do not know whether or how sexual selection contributes to the likelihood that animals will establish in urban areas. We use a comparative analysis of passerine birds to test whether traits associated with pre- and post-mating sexual selection predict successful colonization of urban areas. We found that plumage dichromatism was negatively associated with urban tolerance, but found no relationship with sexual size dimorphism or testes mass relative to body mass. While we cannot determine the exact reason why species with high plumage dichromatism occur less in cities, it is likely that urban areas increase the costs of expressing bright coloration due, for instance, to dietary constraints, limited male parental care or increased predation.

An experimental test for body size-dependent effects of male harassment and an elevated copulation rate on female lifetime fecundity and offspring performance.

Many studies investigate the benefits of polyandry, but repeated interactions with males can lower female reproductive success. Interacting with males might even decrease offspring performance if it reduces a female's ability to transfer maternal resources. Male presence can be detrimental for females in two ways: by forcing females to mate at a higher rate and through costs associated with resisting male mating attempts. Teasing apart the relative costs of elevated mating rates from those of greater male harassment is critical to understand the evolution of mating strategies. Furthermore, it is important to test whether a male's phenotype, notably body size, has differential effects on female reproductive success versus the performance of offspring, and whether this is due to male body size affecting the costs of harassment or the actual mating rate. In the eastern mosquitofish Gambusia holbrooki, males vary greatly in body size and continually attempt to inseminate females. We experimentally manipulated male presence (i.e., harassment), male body size and whether males could copulate. Exposure to males had strong detrimental effects on female reproductive output, growth and immune response, independent of male size or whether males could copulate. In contrast, there was a little evidence of a cross-generational effect of male harassment or mating rate on offspring performance. Our results suggest that females housed with males pay direct costs due to reduced condition and offspring production and that these costs are not a consequence of increased mating rates. Furthermore, exposure to males does not affect offspring reproductive traits.

Evidence that nonsignificant results are sometimes preferred: Reverse P-hacking or selective reporting?

There is increased concern about poor scientific practices arising from an excessive focus on P-values. Two particularly worrisome practices are selective reporting of significant results and 'P-hacking'. The latter is the manipulation of data collection, usage, or analyses to obtain statistically significant outcomes. Here, we introduce the novel, to our knowledge, concepts of selective reporting of nonsignificant results and 'reverse P-hacking' whereby researchers ensure that tests produce a nonsignificant result. We test whether these practices occur in experiments in which researchers randomly assign subjects to treatment and control groups to minimise differences in confounding variables that might affect the focal outcome. By chance alone, 5% of tests for a group difference in confounding variables should yield a significant result (P < 0.05). If researchers less often report significant findings and/or reverse P-hack to avoid significant outcomes that undermine the ethos that experimental and control groups only differ with respect to actively manipulated variables, we expect significant results from tests for group differences to be under-represented in the literature. We surveyed the behavioural ecology literature and found significantly more nonsignificant P-values reported for tests of group differences in potentially confounding variables than the expected 95% (P = 0.005; N = 250 studies). This novel, to our knowledge, publication bias could result from selective reporting of nonsignificant results and/or from reverse P-hacking. We encourage others to test for a bias toward publishing nonsignificant results in the equivalent context in their own research discipline.

Good parenting may not increase reproductive success under environmental extremes.

For species exhibiting parental care, the way in which parents adjust care behaviour to compensate for environmental change potentially influences offspring survival and, ultimately, population viability. Using the three-spined stickleback (Gasterosteus aculeatus) - a species in which males provide parental care by building and tending a nest and fanning the eggs - we examined how low dissolved oxygen (DO) levels affect paternal care, embryo development and survival. Although levels of nest tending were unaffected by DO level, we found that larger males fanned their embryos more under low oxygen conditions. This resulted in faster rates of embryo development within the clutches of these larger males, but reduced embryo survival at 7 days post-fertilization compared to clutches of smaller males. Our results suggest that although parents may attempt to compensate for environmental change via alterations to care behaviour, their ability to do so can be dependent on parental phenotype. This sets up the potential for oxygen levels to act on the strength and direction of selection within populations. We discuss possible explanations for the surprising result that supposedly adaptive changes in care behaviour by large males (i.e. increased fanning) led to reduced embryo survival at 7 days post-fertilization, and whether, as a consequence, acute environmental conditions may have the potential to overwhelm selection on sexual traits.