Social buffering of oxidative stress and cortisol in an endemic cyprinid fish.

Fish exhibit complex social behaviours that can influence their stress levels and well-being. However, little is known about the link between social interactions and stress in wild fish, especially in running water environments. While many studies have explored the stress axis in fish, most have focused on specific social contexts, leaving gaps in understanding stress responses to social changes. Our study investigated collective behaviour and stress in wild Italian riffle dace (Telestes muticellus) in a controlled experimental setup simulating a natural river system. Results reveal that group-living fish have lower cortisol and oxidative stress levels in muscle tissue compared to solitary counterparts, suggesting a calming effect of conspecific presence. Additionally, we observed upregulated expression of antioxidant enzymes in group-living fish, indicating potential benefits to antioxidant defence systems. These insights shed light on the dynamic relationship between group behaviour and stress in wild fish within running water habitats and emphasise the use of multidisciplinary approaches.

Female reproductive fluid attracts more and better sperm: implications for within-ejaculate cryptic female choice.

Mounting evidence shows that the female reproductive fluid (FRF) can differently affect sperm performance of different males by biasing paternity share among competing males. Here, we tested for the first time the potential of 'within-ejaculate cryptic female choice' mediated by the FRF in the zebrafish (Danio rerio). Using a recently developed sperm selection chamber, we separated and collected FRF-selected from non-selected sperm to compare the two subpopulations of sperm in terms of sperm number, viability, DNA integrity and fertilizing ability. We showed that the sperm attracted by FRF are more numerous, more viable and with higher DNA integrity. In addition, FRF-selected sperm fertilized more eggs, but if this is due to fertilization ability per se or numerical advantage remains to be tested. Our results suggest that FRF can select sperm with a better phenotype, highlighting the crucial and impactful role that FRF might play in the process of fertilization and post-mating sexual selection dynamics, along with the potential implications for sperm selection in assisted reproductive techniques.

Resource-dependent investment in male sexual traits in a viviparous fish.

Exaggerated and conspicuous sexually selected traits are often costly to produce and maintain. Costly traits are expected to show resource-dependent expression, since limited resources prevent animals from investing maximally in multiple traits simultaneously. However, there may be critical periods during an individual's life where the expression of traits is altered if resources are limited. Moreover, costly sexual traits may arise from sexual selection acting both before (pre-copulatory) and after mating (post-copulatory). Gaining a robust understanding of resource-dependent trait expression therefore requires an approach that examines both episodes of sexual selection after resource limitation during different times in an individual's life. Yet few studies have taken such an approach. Here, we examine how resource restriction influences a set of pre- and post-copulatory traits in male pygmy halfbeaks (Dermogenys collettei), which invest in sexual ornaments and routinely engage in male-male contests and sperm competition. Critically, we examined responses in males when resources were restricted during development and after reaching sexual maturity. Both pre- and post-copulatory traits are resource-dependent in male halfbeaks. Body size, beak size, courtship behavior, and testes size were reduced by diet restriction, while, unexpectedly, the restricted-diet group had a larger area of red color on the beak and fins after diet treatment. These patterns were generally consistent when resources were restricted during development and after reaching sexual maturity. The study reinforces the role of resource acquisition in maintaining variation among sexual traits.

Immediate predation risk alters the relationship between potential and realised selection on male traits in the Trinidad guppy Poecilia reticulata.

Imminent predation risk affects mating behaviours in prey individuals in a multitude of ways that can theoretically impact the strength of sexual selection, as well as its operation on traits. However, empirical studies of the effects of imminent predation risk on sexual selection dynamics are still scarce. Here we explore how perceived predation affects: (1) the relationship between the opportunity for selection and the actual strength of selection on male traits; and (2) which traits contribute to male fitness and the shape of selection on these traits. We simulate two consecutive reproductive episodes, under control conditions and perceived predation risk using experimental populations of Trinidad guppies. The opportunity for selection is higher under predation risk compared to the control condition, but realised selection on traits remains unaffected. Pre- and postcopulatory traits follow complex patterns of nonlinear selection in both conditions. Differences in selection gradients deviate from predictions based on evolutionary and non-lethal effects of predation, the most notable being strong disruptive selection on courtship rate under predation risk. Our results demonstrate that sexual selection is sensitive to imminent predation risk perception and reinforce the notion that both trait-based and variance-based metrics should be employed for an informative quantification.

Does brain size affect mate choice? An experimental examination in pygmy halfbeaks.

Choosing a mate is one of the most important decisions in an animal's lifetime. Female mate choice is often guided by the presence or intensity of male sexual ornaments, which must be integrated and compared among potential mates. Individuals with greater cognitive abilities may be better at evaluating and comparing sexual ornaments, even when the difference in ornaments is small. While brain size is often used as a proxy for cognitive ability, its effect on mate choice has rarely been investigated. Here, we investigate the effect of brain size on mate preferences in the pygmy halfbeak Dermogenys collettei, a small freshwater fish that forms mixed-sex shoals where mating takes place. Pygmy halfbeaks are ideal models as their semi-transparent heads allow for external brain measurements. After validating the use of external measurements as a proxy for internal brain size, we presented females with large or small brains (relative to body length) with two males that had either a large or small difference in sexual ornamentation (measured by the total area of red coloration). Unexpectedly, neither total relative brain size nor relative telencephalon size affected any measured aspect of mate preference. However, the difference in male sexual ornamentation did affect preference, with females preferring males with a smaller area of red coloration when the difference in ornaments was large. This study highlights the complexities of mate choice and the importance of considering a range of stimuli when examining mate preferences.

Sperm Accumulation Induced by the Female Reproductive Fluid: Putative Evidence of Chemoattraction Using a New Tool.

There is considerable evidence that female reproductive fluid (FRF) interacts intimately with sperm, affecting several sperm traits, including sperm motility and longevity, and ultimately fertilization success. One of the first documented interactions between FRF and sperm is the ability of FRF to attract and guide sperm towards the eggs. However, most of the evidence of FRF's chemoattraction proprieties comes from a limited number of taxa, specifically mammals and invertebrate broadcasting spawners. In other species, small FRF volumes and/or short sperm longevity often impose methodological difficulties resulting in this gap in chemoattraction studies in non-model species. One of the outcomes of sperm chemotaxis is sperm accumulation towards high chemoattractant concentrations, which can be easily quantified by measuring sperm concentration. Here, we tested sperm accumulation towards FRF in the zebrafish, Danio rerio, using an ad hoc developed, 3D printed, device ('sperm selection chamber'). This easy-to-use tool allows to select and collect the sperm that swim towards a chemical gradient, and accumulate in a chemoattractant-filled well thus providing putative evidence for chemoattraction. We found that sperm accumulate in FRF in zebrafish. We also found that none of the sperm quality traits we measured (sperm swimming velocity and trajectory, sperm motility, and longevity) were correlated with this response. Together with the 3D printable project, we provide a detailed protocol for using the selection chamber. The chamber is optimized for the zebrafish, but it can be easily adapted for other species. Our device lays the foundation for a standardized way to measure sperm accumulation and in general chemoattraction, stimulating future research aimed at understanding the role and the mechanisms of sperm chemoattraction by FRF.

Corrigendum to: Male-male behavioral interactions drive social-dominance mediated differences in ejaculate traits.

[This corrects the article DOI: 10.1093/beheco/araa118.].

Male-male behavioral interactions drive social-dominance-mediated differences in ejaculate traits.

Higher social status is expected to result in fitness benefits as it secures access to potential mates. In promiscuous species, male reproductive success is also determined by an individual's ability to compete for fertilization after mating by producing high-quality ejaculates. However, the complex relationship between a male's investment in social status and ejaculates remains unclear. Here, we examine how male social status influences ejaculate quality under a range of social contexts in the pygmy halfbeak Dermogenys collettei, a small, group-living, internally fertilizing freshwater fish. We show that male social status influences ejaculate traits, both in the presence and absence of females. Dominant males produced faster swimming and more viable sperm, two key determinants of ejaculate quality, but only under conditions with frequent male-male behavioral interactions. When male-male interactions were experimentally reduced through the addition of a refuge, differences in ejaculate traits of dominant and subordinate males disappeared. Furthermore, dominant males were in a better condition, growing faster, and possessing larger livers, highlighting a possible condition dependence of competitive traits. Contrary to expectations, female presence or absence did not affect sperm swimming speed or testes mass. Together, these results suggest a positive relationship between social status and ejaculate quality in halfbeaks and highlight that the strength of behavioral interactions between males is a key driver of social-status-dependent differences in ejaculate traits.

Contrasting female mate preferences for red coloration in a fish.

Understanding how animals select their mates requires knowing the factors that shape mate preferences. Recent theoretical and empirical considerations suggest that female mating status can influence the degree to which a female engages in mate choice, with virgin females predicted to be less choosy than mated females. In this study, we investigated mate choice in both virgin and mated females in the pygmy halfbeak Dermogenys collettei. Halfbeaks are small, live-bearing, internally fertilizing freshwater fish that live in mixed-sex groups where females have ample opportunity to engage in mate choice. Using a dichotomous choice assay, we quantified and contrasted in virgin and mated females mate preferences for differences in male body size, beak size, and area of yellow and red coloration. We also examined how mating status influenced the amount of time a female associated with the first male encountered and the relative amount of time a female associated with each male. We demonstrate that mate preferences of female halfbeaks are driven primarily by the size of red coloration present on males. Females showed contrasting preferences based on mating status, with virgin females preferentially associating with drab males whereas mated females preferentially associate with males possessing large areas of red. Contrary to expectations, female mating status did not influence how females associate with the first males encountered or how females biased their association time among males. Although the precise drivers of these effects need further studying, our finding highlights a possible explanation for how variation in male ornamentation can be maintained.

Female sperm storage mediates post-copulatory costs and benefits of ejaculate anticipatory plasticity in the guppy.

Males of many species evolved the capability of adjusting their ejaculate phenotype in response to social cues to match the expected mating conditions. When females store sperm for a prolonged time, the expected fitness return of plastic adjustments of ejaculate phenotype may depend on the interval between mating and fertilization. Although prolonged female sperm storage (FSS) increases the opportunity for sperm competition, as a consequence of the longer temporal overlap of ejaculates from several males, it may also create variable selective forces on ejaculate phenotype, for example by exposing trade-offs between sperm velocity and sperm survival. We evaluated the relationship between the plasticity of ejaculate quality and FSS in the guppy, Poecilia reticulata, a polyandrous live-bearing fish in which females store sperm for several months and where stored sperm contribute significantly to a male's lifelong reproductive success. In this species, males respond to the perception of future mating opportunities by increasing the quantity (number) and quality (swimming velocity) of ready-to-use sperm (an anticipatory response called 'sperm priming'). Here we investigated (a) the effect of sperm priming on in vitro sperm viability at stripping and its temporal decline (as an estimate of sperm survival), and (b) the in vivo competitive fertilization success in relation to female sperm storage using artificial insemination. As expected, sperm-primed males produced more numerous and faster sperm, but with a reduced in vitro sperm viability at stripping and after 4 hr, compared with their counterparts. Artificial insemination revealed that the small (nonsignificant) advantage of primed sperm when fertilization immediately follows insemination is reversed when eggs are fertilized by female-stored sperm, weeks after insemination. By suggesting a plastic trade-off between sperm velocity and viability, these results demonstrate that prolonged female sperm storage generates divergent selection pressures on ejaculate phenotype.

Chemical signals from eggs facilitate cryptic female choice in humans.

Mate choice can continue after mating via chemical communication between the female reproductive system and sperm. While there is a growing appreciation that females can bias sperm use and paternity by exerting cryptic female choice for preferred males, we know surprisingly little about the mechanisms underlying these post-mating choices. In particular, whether chemical signals released from eggs (chemoattractants) allow females to exert cryptic female choice to favour sperm from specific males remains an open question, particularly in species (including humans) where adults exercise pre-mating mate choice. Here, we adapt a classic dichotomous mate choice assay to the microscopic scale to assess gamete-mediated mate choice in humans. We examined how sperm respond to follicular fluid, a source of human sperm chemoattractants, from either their partner or a non-partner female when experiencing a simultaneous or non-simultaneous choice between follicular fluids. We report robust evidence under these two distinct experimental conditions that follicular fluid from different females consistently and differentially attracts sperm from specific males. This chemoattractant-moderated choice of sperm offers eggs an avenue to exercise independent mate preference. Indeed, gamete-mediated mate choice did not reinforce pre-mating human mate choice decisions. Our results demonstrate that chemoattractants facilitate gamete-mediated mate choice in humans, which offers females the opportunity to exert cryptic female choice for sperm from specific males.

Male mate choice for large gravid spots in a livebearing fish.

Male mate choice occurs in a wide range of species, and males can increase their reproductive success by distinguishing between females based on their fecundity (e.g., large body size) or their expected sperm competition risk (e.g., virgins). However, patterns of male mate choice could be mitigated by variation in female physiological receptivity, as males can benefit by directing their mating efforts toward females that are at a point in their reproductive cycle when fertilization probability is highest. Here, we perform three experiments to assess whether male mate choice is influenced by cues of female physiological receptivity, fecundity, or sperm competition risk in the pygmy halfbeak (Dermogenys collettei), a small livebearing fish. Female halfbeaks possess a "gravid spot"-an orange abdominal marking that is caused by pigmentation of the females' skin and variation in embryo development and pigmentation during pregnancy. We show that gravid spot size increases toward parturition and is largest right before giving birth, independent of abdominal width or body size. Males consistently chose females with large gravid spots over females with small gravid spots. In contrast, males did not prefer larger females over smaller females or virgin females over mated females. As female halfbeaks store sperm prior to fertilizations, we suggest that males use the size of the gravid spot as a cue to direct their mating efforts to those females where the chance of fertilization is highest.

Differential gene regulation in selected lines for high and low sperm production in male guppies.

In species where females mate with more than one male during the same reproductive event, males typically increase the number of sperm produced to boost their fertilization share. Sperm is not limitless, however, and theory predicts that their production will come at the cost of other fitness-related traits, such as body growth or immunocompetence, although these evolutionary trade-offs are notoriously difficult to highlight. To this end, we combined artificial selection for sperm production with a transcriptome analysis using Poecilia reticulata, a fish characterized by intense sperm competition in which the number of sperm transferred during mating is the most important predictor of fertilization success, yet sperm production is highly variable among males. We compared the brain and testes transcriptome in male guppies of lines artificially selected for high and low sperm production by identifying pivotal differentially expressed gene sets that may regulate spermatogenesis and immune function in this species. Despite the small differences in single genes' expression, gene set enrichment analysis showed coordinated gene expression differences associated with several pathways differentially regulated in the two selection lines. High sperm production males showed an upregulation of pathways related to immunosuppression and development of spermatozoa indicating a possible immunological cost of sperm production.

Predation shapes sperm performance surfaces in guppies.

Sperm velocity is a key determinant of competitive fertilization success in many species. Selection is therefore expected to favour the evolution of faster sperm when the level of sperm competition is high. However, several aspects can determine the direction and strength of selection acting on this key performance trait, including ecological factors that influence both sperm competition and the strength of selection acting on correlated traits that may constrain evolutionary responses in sperm velocity. Here, we determine how a key ecological variable, the level of predation, shapes sperm swimming speed across 18 Trinidadian populations of guppies ( Poecilia reticulata). We use performance analysis, a statistical tool akin to the familiar methods of multivariate selection analyses, to determine how the level of predation influences sperm velocity (modelled as a performance trait) when accounting for correlated pre- and postcopulatory traits that are also impacted by predation. We show that predation affects the combination of pre- and postcopulatory traits that ultimately predict sperm performance. Overall, we report evidence for disruptive relationships between sperm performance and combinations of ornaments and sperm morphology, but the specific combinations of traits that predict sperm velocity depended on the level of predation. These analyses underscore the complex nonlinear interrelationships among pre- and postcopulatory traits and the importance of considering ecological factors that may ultimately change the way in which multiple traits interact to determine a trait's performance value. As such, our results are likely to be broadly applicable across systems where selection is influenced by ecological conditions.

Sexual selection and ageing: interplay between pre- and post-copulatory traits senescence in the guppy.

Traits associated with mating and fertilization success are expected to senesce with age, but limited information is available on their relative rates of senescence. In polyandrous species, male reproductive fitness depends on both mating and fertilization success. Because successful mating is a prerequisite for post-copulatory sexual selection, ejaculate traits are expected to senesce faster than pre-copulatory traits, as pre-copulatory sexual selection is often deemed to be stronger than post-copulatory sexual selection. This pattern has generally been found in the few empirical studies conducted so far. We tested this prediction in the guppy ( Poecilia reticulata), a livebearing fish characterized by intense sperm competition, by comparing the expression of male sexual traits at two ages (four and nine months). Contrary to prediction, we found that post-copulatory traits senesced at a significantly slower rate than pre-copulatory traits. We also looked at whether early investment in those sexual traits affects longevity, and the interaction between sperm age (duration of sperm storage inside the male) and male age. Our results suggest that the relative senescence rate of pre- and post-copulatory sexual traits may vary among species with different mating systems and ecology.

Population demography and heterozygosity-fitness correlations in natural guppy populations: An examination using sexually selected fitness traits.

Heterozygosity-fitness correlations (HFCs) have been examined in a wide diversity of contexts, and the results are often used to infer the role of inbreeding in natural populations. Although population demography, reflected in population-level genetic parameters such as allelic diversity or identity disequilibrium, is expected to play a role in the emergence and detectability of HFCs, direct comparisons of variation in HFCs across many populations of the same species, with different genetic histories, are rare. Here, we examined the relationship between individual microsatellite heterozygosity and a range of sexually selected traits in 660 male guppies from 22 natural populations in Trinidad. Similar to previous studies, observed HFCs were weak overall. However, variation in HFCs among populations was high for some traits (although these variances were not statistically different from zero). Population-level genetic parameters, specifically genetic diversity levels (number of alleles, observed/expected heterozygosity) and measures of identity disequilibrium (g2 and heterozygosity-heterozygosity correlations), were not associated with variation in population-level HFCs. This latter result indicates that these metrics do not necessarily provide a reliable predictor of HFC effect sizes across populations. Importantly, diversity and identity disequilibrium statistics were not correlated, providing empirical evidence that these metrics capture different essential characteristics of populations. A complex genetic architecture likely underpins multiple fitness traits, including those associated with male fitness, which may have reduced our ability to detect HFCs in guppy populations. Further advances in this field would benefit from additional research to determine the demographic contexts in which HFCs are most likely to occur.

Directional postcopulatory sexual selection is associated with female sperm storage in Trinidadian guppies.

Female sperm storage (FSS) is taxonomically widespread and often associated with intense sperm competition, yet its consequences on postcopulatory sexual selection (PCSS) are poorly known. Theory predicts that FSS will reduce the strength of PCSS, because sperm characteristics favored before and after FSS may be traded-off, and opportunities for nondirectional PCSS should increase. We explored these questions in the guppy (Poecilia reticulata), by allowing females to mate multiply and by comparing the paternity pattern in two successive broods. Contrary to predictions, the variance in male fertilization success increased after FSS, driven by a change in male paternity share across broods. This change was positively associated with sperm velocity (measured before FSS) but not with the duration of FSS, indirectly suggesting that faster sperm were better in entering female storage organs, rather than in persisting within them. Other male traits, such as male size and orange color, heterozygosity, and relatedness to the female, did not influence paternity after FSS. These results indicate that processes associated with FSS tend to reinforce the strength of PCSS in guppies, rather than weaken it. Further work is necessary to test whether this pattern changes in case of more prolonged FSS.

Multivariate selection drives concordant patterns of pre- and postcopulatory sexual selection in a livebearing fish.

In many species, females mate with multiple partners, meaning that sexual selection on male traits operates across a spectrum that encompasses the competition for mates (that is, before mating) and fertilizations (after mating). Despite being inextricably linked, pre- and postcopulatory sexual selection are typically studied independently, and we know almost nothing about how sexual selection operates across this divide. Here we bridge this knowledge gap using the livebearing fish Poecilia reticulata. We show that both selective episodes, as well as their covariance, explain a significant component of variance in male reproductive fitness. Moreover, linear and nonlinear selection simultaneously act on pre- and postcopulatory traits, and interact to generate multiple phenotypes with similar fitness.

Assessment of left ventricular function by pulse wave analysis in critically ill patients.

PURPOSE: Left ventricular (LV) performance is often quantified by echocardiography in critically ill patients. Pulse wave analysis (PWA) systems can also monitor cardiac function but in a continuous fashion. We compared echocardiographic and PWA-derived indices of LV function. METHODS: We enrolled 70 critically ill patients equipped with invasive arterial pressure monitoring who required echocardiography. We simultaneously assessed LV ejection fraction (LVEF), the rate of LV pressure rise during systole (dP/dt MAX) obtained with echocardiography (EC-dP/dt MAX), the ratio of effective arterial elastance to LV end-systolic elastance (E a/E es) determined by echocardiography, the dP/dt MAX estimated from the arterial pressure waveform (AP-dP/dt MAX) and the cardiac cycle efficiency (CCE) using PWA. RESULTS: Mean LVEF was 53 ± 18 % and CCE 0.16 ± 0.26. CCE was correlated linearly with LVEF (r = 0.88, 95 % CI 0.81 to 0.92, P < 0.001), and the dP/dt MAX values from the two techniques were linearly correlated (r = 0.93, 95 % CI 0.87 to 0.96, P < 0.001). There was minimal bias between the techniques for measurement of dP/dt MAX (23.7 mmHg/ms; 95 % CI -23.6 to 71.0). E a/E es and CCE were inversely correlated (r = -0.81, 95 % CI -0.88 to -0.71, P < 0.001). A CCE value of <0.07 predicted LVEF <40 % with a sensitivity of 0.93 and a specificity of 0.96 (AUC 0.98, 95 % CI 0.90 to 1.0, P < 0.001). A CCE value of >0.12 predicted LVEF ≥50 % with a sensitivity of 0.96 and a specificity of 0.82 (AUC 0.94, 95 % CI 0.87 to 1.0, P < 0.001). A CCE value <0.12 predicted E a/E es ≥1.3 with a sensitivity of 0.93 and a specificity of 0.89 (AUC 0.94, 95 % CI 0.83 to 1.0, P < 0.001). CONCLUSIONS: PWA-derived variables provide relevant information on cardiac contractility and performance in critically ill patients. PWA provides an easy method for online hemodynamic evaluation in critically ill patients.

Pattern of inbreeding depression, condition dependence, and additive genetic variance in Trinidadian guppy ejaculate traits.

In polyandrous species, a male's reproductive success depends on his fertilization capability and traits enhancing competitive fertilization success will be under strong, directional selection. This leads to the prediction that these traits should show stronger condition dependence and larger genetic variance than other traits subject to weaker or stabilizing selection. While empirical evidence of condition dependence in postcopulatory traits is increasing, the comparison between sexually selected and 'control' traits is often based on untested assumption concerning the different strength of selection acting on these traits. Furthermore, information on selection in the past is essential, as both condition dependence and genetic variance of a trait are likely to be influenced by the pattern of selection acting historically on it. Using the guppy (Poecilia reticulata), a livebearing fish with high levels of multiple paternity, we performed three independent experiments on three ejaculate quality traits, sperm number, velocity, and size, which have been previously shown to be subject to strong, intermediate, and weak directional postcopulatory selection, respectively. First, we conducted an inbreeding experiment to determine the pattern of selection in the past. Second, we used a diet restriction experiment to estimate their level of condition dependence. Third, we used a half-sib/full-sib mating design to estimate the coefficients of additive genetic variance (CVA) underlying these traits. Additionally, using a simulated predator evasion test, we showed that both inbreeding and diet restriction significantly reduced condition. According to predictions, sperm number showed higher inbreeding depression, stronger condition dependence, and larger CVA than sperm velocity and sperm size. The lack of significant genetic correlation between sperm number and velocity suggests that the former may respond to selection independently one from other ejaculate quality traits. Finally, the association between sperm number and condition suggests that this trait may mediate the genetic benefits of polyandry which have been shown in this species.