Parental care behaviour in response to perceived paternity is not mediated by 11-ketotestosterone in bluegill sunfish.

Parental care is critical for the survival of many young animals, but parental care can be costly to the individual providing care. To balance this cost, parents can allocate their care to offspring based on their value, which can be dependent on the offspring's relatedness to the parent. Bluegill (Lepomis macrochirus) is a fish characterized by uniparental male care and high levels of cuckoldry. While parental males of this species have been shown to adaptively adjust their care in response to paternity, the mechanisms for this adjustment are not well understood. Androgens are steroid hormones that are associated with parental care behaviours in many species including bluegill. Here, we test the hypothesis that circulating androgen concentrations mediate the adjustment in care provided by bluegill parental males by manipulating perceived paternity and then measuring circulating 11-ketotestosterone concentration and parental care behaviour. We show that males with higher perceived paternity provide higher levels of nurturing and nest defense behaviour, but contrary to expectations, we found that these males had lower concentrations of 11-ketotestosterone. Furthermore, we found positive correlations between individual circulating plasma 11-ketotestosterone concentrations and nurturing behaviour, but not with the aggressive behaviours that differed between paternity treatments. While bluegill make behavioural changes in response to perceived paternity, these changes do not appear to be modulated by 11-ketotestosterone.

Androgen and prolactin manipulation do not induce changes in immunocompetence measures in a fish with male parental care.

Prolactin and 11-ketotestosterone (11-KT) are important reproductive hormones in fishes, which may also influence immunocompetence. The immunocompetence handicap hypothesis states that higher androgen concentrations that support secondary sex traits are traded off against a decrease in immune system function. To test the relationships between these hormones and immunocompetence, we experimentally manipulated 11-ketotestosterone and prolactin in the freshwater fish, bluegill (Lepomis macrochirus) during parental care using implants that contained either 11-KT, prolactin, or an inert control. We vaccinated individuals to stimulate the acquired immune response, then measured immunocompetence as the number of granulocytes, lymphocytes and monocytes, and the expression of interleukin 8 in each sample. We did not observe any significant differences in the immune measures among the hormone treatments. Our results indicate that in bluegill, there is no trade-off between androgens or prolactin and immunocompetence.

Factors influencing the variation of microplastic uptake in demersal fishes from the upper Thames River Ontario.

Microplastics (plastic particles <5 mm) are abundant in aquatic environments, particularly near urban areas. Little is known, however, about how variations in microplastic abundances within watersheds affect fishes. Microplastics were examined in demersal fishes-white sucker (Catostomus commersonii) and common carp (Cyprinus carpio)-across 11 sites in the Thames River, Ontario, Canada. Microplastics were found in 44% of white sucker, ranging from 0 to 14 particles per fish, and 31% of common carp, ranging from 0 to 128 particles per fish. Across both species, the number of microplastics was higher in urban sites than rural sites, and there was a positive relationship between the number of microplastics in the fish and the abundance of microplastics in the sediment. Body mass was also positively related to number of microplastics in fish. Together these results provide insight into environmental and biological factors that may be influencing microplastic ingestion in demersal fishes.

Proteomic analysis of temperature-dependent developmental plasticity within the ventricle of juvenile Atlantic salmon (Salmo salar).

In teleosts, cardiac plasticity plays a central role in mediating thermal acclimation. Previously, we demonstrated that exposure to elevated temperatures throughout development (+4°C) improved acute thermal tolerance of the heart in juvenile Atlantic salmon. Fish raised in a warmer thermal regime also displayed higher proportions of compact myocardium within their ventricles. In the present study, we investigated the molecular mechanisms supporting this temperature-specific phenotype by comparing relative protein abundance in ventricular tissue from the same experimental fish using mass spectrometry. We provide the first description of the ventricular proteome in juvenile Atlantic salmon and identify 79 proteins displaying differential abundance between developmental treatments. The subset of proteins showing higher abundance in fish raised under elevated temperatures was significantly enriched for processes related to ventricular tissue morphogenesis, and changes in protein abundance support a hypertrophic model of compact myocardium growth. Proteins associated with the vasculature and angiogenesis also showed higher abundance in the warm-developmental group, suggesting capillarization of the compact myocardium in the hearts of these fish. Proteins related to oxidative metabolism and protein homeostasis also displayed substantive shifts in abundance between developmental treatments, underscoring the importance of these processes in mediating thermal plasticity of cardiac function. While rapid growth under warm developmental temperatures has been linked to cardiomyopathies in farmed salmon, markers of cardiac pathology were not implicated in the present study. Thus, our findings offer a molecular footprint for adaptive temperature-dependent plasticity within the ventricle of a juvenile salmonid.

Temperature-dependent plasticity mediates heart morphology and thermal performance of cardiac function in juvenile Atlantic salmon (Salmo salar).

In many fishes, upper thermal tolerance is thought to be limited in part by the heart's ability to meet increased oxygen demands during periods of high temperature. Temperature-dependent plasticity within the cardiovascular system may help fish cope with the thermal stress imposed by increasing water temperatures. In this study, we examined plasticity in heart morphology and function in juvenile Atlantic salmon (Salmo salar) reared under control (+0°C) or elevated (+4°C) temperatures. Using non-invasive Doppler echocardiography, we measured the effect of acute warming on maximum heart rate, stroke distance and derived cardiac output. A 4°C increase in average developmental temperature resulted in a >5°C increase in the Arrhenius breakpoint temperature for maximum heart rate and enabled the hearts of these fish to continue beating rhythmically to temperatures approximately 2°C higher than for control fish. However, these differences in thermal performance were not associated with plasticity in maximum cardiovascular capacity, as peak measures of heart rate, stroke distance and derived cardiac output did not differ between temperature treatments. Histological analysis of the heart revealed that while ventricular roundness and relative ventricle size did not differ between treatments, the proportion of compact myocardium in the ventricular wall was significantly greater in fish raised at elevated temperatures. Our findings contribute to the growing understanding of how the thermal environment can affect phenotypes later in life and identify a morphological strategy that may help fishes cope with acute thermal stress.

Adaptation of a mouse Doppler echocardiograph system for assessing cardiac function and thermal performance in a juvenile salmonid.

Measures of cardiac performance are pertinent to the study of thermal physiology and exercise in teleosts, particularly as they pertain to migration success. Increased heart rate, stroke volume and cardiac output have previously been linked to improved swimming performance and increased upper thermal tolerance in anadromous salmonids. To assess thermal performance in fishes, it has become commonplace to measure the response of maximum heart rate to warming using electrocardiograms. However, electrocardiograms do not provide insight into the hemodynamic characteristics of heart function that can impact whole-animal performance. Doppler echocardiography is a popular tool used to examine live animal processes, including real-time cardiac function. This method allows for nonsurgical measurements of blood flow velocity through the heart and has been used to detect abnormalities in cardiovascular function, particularly in mammals. Here, we show how a mouse Doppler echocardiograph system can be adapted for use in a juvenile salmonid over a range of temperatures and timeframes. Using this compact, noninvasive system, we measured maximum heart rate, atrioventricular (AV) blood flow velocity, the early flow-atrial flow ratio and stroke distance in juvenile Atlantic salmon (Salmo salar) during acute warming. Using histologically determined measures of AV valve area, we show how stroke distance measurements obtained with this system can be used to calculate ventricular inflow volume and approximate cardiac output. Further, we show how this Doppler system can be used to determine cardiorespiratory thresholds for thermal performance, which are increasingly being used to predict the consequences that warming water temperatures will have on migratory fishes.

Effect of isotocin on shoaling behaviour of the Guppy (Poecilia reticulata).

Pro-social effects of oxytocin and its homologues are well-documented in birds and mammals. However, in fishes, the effect of isotocin, the homologue of oxytocin, on social behaviour is less clear. Studies in fishes have generally shown no effect of isotocin on social behaviours or even an anti-social effect. In our study, we measured association preference for conspecifics in 92 adult guppies (46 females and 46 males), half of which were injected with isotocin and the other half with an isotocin antagonist. We found that individuals injected with isotocin spent 29% more time associating with conspecifics than individuals injected with an isotocin antagonist. The effect of isotocin on association time did not differ between males and females. Our study provides some of the first evidence of a pro-social effects of isotocin in a fish and suggests that in fishes, isotocin may have a homologous role to oxytocin, at least in promoting shoaling behaviour.

Androgen and prolactin manipulation induces changes in aggressive and nurturing behavior in a fish with male parental care.

Parental care can include two general types of behavior: (1) aggressive behavior, which is used to defend offspring from predators; and (2) nurturing behavior, which is used to provide offspring with environmental conditions or resources necessary for survival. Many studies have implicated androgens in promoting aggressive behavior and prolactin in promoting nurturing behavior. We experimentally manipulated these hormones to investigate their effects on parental care behavior in bluegill (Lepomis macrochirus). Parental males, which provide sole care to the developing eggs and larvae, received an implant with an androgen (11-ketotestosterone [11-KT]), an androgen antagonist (flutamide), prolactin, a prolactin-release inhibitor (bromocriptine), or castor oil (placebo). We found that 11-KT implants led to a significant increase in the frequency of aggressive behavior directed towards a simulated brood predator, and were associated with a nearly significant decrease in the frequency of nurturing behavior directed towards the developing eggs. In contrast, prolactin implants were associated with a significant increase in the frequency of nurturing behavior, but also reduced the frequency of aggressive behavior directed towards the simulated brood predator. These results suggest a hormone-mediated mechanistic trade-off between nurturing and aggressive behavior, whereby parental males are unable to be both highly nurturing and highly aggressive.

Transcriptome response of Atlantic salmon (Salmo salar) to competition with ecologically similar non-native species.

Non-native species may be introduced either intentionally or unintentionally, and their impact can range from benign to highly disruptive. Non-native salmonids were introduced into Lake Ontario, Canada, to provide recreational fishing opportunities; however, the establishment of those species has been proposed as a significant barrier to the reintroduction of native Atlantic salmon (Salmo salar) due to intense interspecific competition. In this study, we compared population differences of Atlantic salmon in transcriptome response to interspecific competition. We reared Atlantic salmon from two populations (LaHave River and Sebago Lake) with fish of each of three non-native salmonids (Chinook salmon Oncorhynchus tshawytscha, rainbow trout O. mykiss, and brown trout S. trutta) in artificial streams. We used RNA-seq to assess transcriptome differences between the Atlantic salmon populations and the responses of these populations to the interspecific competition treatments after 10 months of competition in the stream tanks. We found that population differences in gene expression were generally greater than the effects of interspecific competition. Interestingly, we found that the two Atlantic salmon populations exhibited similar responses to interspecific competition based on functional gene ontologies, but the specific genes within those ontologies were different. Our transcriptome analyses suggest that the most stressful competitor (as measured by the highest number of differentially expressed genes) differs between the two study populations. Our transcriptome characterization highlights the importance of source population selection for conservation applications, as organisms with different evolutionary histories can possess different transcriptional responses to the same biotic stressors. The results also indicate that generalized predictions of the response of native species to interactions with introduced species may not be appropriate without incorporating potential population-specific response to introduced species.

A test of the effects of androgens on immunity: No relationship between 11-ketotestosterone and immune performance in bluegill (Lepomis macrochirus).

The immunosuppressive effects of androgens are a key component of the immunocompetence handicap hypothesis (ICHH). Here, we use bluegill sunfish (Lepomis macrochirus) to test two predictions arising from this hypothesis: (1) natural circulating concentrations of the androgen 11-ketotestosterone (11-KT) will be negatively related with measures of immunity, and (2) immune stimulation will lower circulating 11-KT concentration. We found no evidence for a relationship between natural circulating 11-KT concentration and measures of immunity (lymphocyte and granulocyte counts, respiratory burst, cytokine mRNA levels), and an immune stimulation with Vibrio vaccine did not affect circulating 11-KT concentration. We also performed a meta-analysis of immune stimulation studies to help interpret our results, and report evidence suggesting that immune stimulation has weaker effects on androgen levels in fishes compared to other vertebrates. These results suggest that the ICHH may not apply to all vertebrates, although it remains premature to state what factors account for the weaker evidence in fishes that androgens are immunosuppressive.

Hematocrit Is Associated with Thermal Tolerance and Modulated by Developmental Temperature in Juvenile Chinook Salmon.

To evaluate whether oxygen-carrying capacity influences thermal tolerance in fishes, we reared four Chinook salmon families in present-day (+0°C) and possible future (+4°C) temperatures and assessed the response of hematocrit (Hct) to acute temperature stress. In the +4°C treatment, Hct increased above control levels when juvenile fish were exposed to their critical thermal maximum (CTmax). Conversely, no effect of temperature stress on Hct was found in the +0°C treatment. Hct was positively associated with CTmax ([Formula: see text]; [Formula: see text]), contributing to the CTmax of the +4°C treatment being significantly higher than that of the +0°C treatment (mean ± SD, [Formula: see text] and [Formula: see text], respectively). The association between CTmax and Hct found here supports the hypothesis that thermal tolerance is affected by oxygen supply to tissue. Moreover, the developmental plasticity of CTmax and Hct could represent an adaptive mechanism for salmon faced with climate change.

Evolution of kin recognition mechanisms in a fish.

Both selection and phylogenetic history can influence the evolution of phenotypic traits. Here we used recently characterized variation in kin recognition mechanisms among six guppy populations to explore the phylogenetic history of this trait. Guppies can use two different kin recognition mechanisms: either phenotype matching, in which individuals are identified based on comparison with a recognition template, or familiarity, in which individuals are remembered based on previous interactions. Across the six populations, we identified four transitions in recognition mechanism: phenotype matching evolved once and was subsequently lost in a single population, whereas familiarity evolved twice. Based on a molecular clock, these transitions occurred among populations that had diverged on a timescale of hundreds of thousands of years, which is two orders of magnitude faster than previously documented transitions in recognition mechanisms. A randomization test provided no evidence that recognition mechanisms were constrained by phylogeny, suggesting that recognition mechanisms have the capacity to evolve rapidly, although the specific selection pressures that may be contributing to variation in recognition mechanisms across populations remain unknown.

Brain Transcriptional Profiles of Male Alternative Reproductive Tactics and Females in Bluegill Sunfish.

Bluegill sunfish (Lepomis macrochirus) are one of the classic systems for studying male alternative reproductive tactics (ARTs) in teleost fishes. In this species, there are two distinct life histories: parental and cuckolder, encompassing three reproductive tactics, parental, satellite, and sneaker. The parental life history is fixed, whereas individuals who enter the cuckolder life history transition from sneaker to satellite tactic as they grow. For this study, we used RNAseq to characterize the brain transcriptome of the three male tactics and females during spawning to identify gene ontology (GO) categories and potential candidate genes associated with each tactic. We found that sneaker males had higher levels of gene expression differentiation compared to the other two male tactics. Sneaker males also had higher expression in ionotropic glutamate receptor genes, specifically AMPA receptors, compared to other males, which may be important for increased spatial working memory while attempting to cuckold parental males at their nests. Larger differences in gene expression also occurred among male tactics than between males and females. We found significant expression differences in several candidate genes that were previously identified in other species with ARTs and suggest a previously undescribed role for cAMP-responsive element modulator (crem) in influencing parental male behaviors during spawning.

Reproductive success in wild and hatchery male coho salmon.

Salmon produced by hatcheries have lower fitness in the wild than naturally produced salmon, but the factors underlying this difference remain an active area of research. We used genetic parentage analysis of alevins produced by experimentally mixed groups of wild and hatchery coho salmon (Oncorhynchus kisutch) to quantify male paternity in spawning hierarchies. We identify factors influencing paternity and revise previously published behavioural estimates of reproductive success for wild and hatchery males. We observed a strong effect of hierarchy size and hierarchy position on paternity: in two-male hierarchies, the first male sired 63% (±29%; s.d.) of the alevins and the second male 37% (±29%); in three-male hierarchies, the first male sired 64% (±26%), the second male 24% (±20%) and the third male 12% (±10%). As previously documented, hatchery males hold inferior positions in spawning hierarchies, but we also discovered that hatchery males had only 55-84% the paternity of wild males when occupying the same position within a spawning hierarchy. This paternity difference may result from inferior performance of hatchery males during sperm competition, female mate choice for wild males, or differential offspring survival. Regardless of its cause, the combination of inferior hierarchical position and inferior success at a position resulted in hatchery males having only half (51%) the reproductive success of wild males.

Indirect genetic effects underlie oxygen-limited thermal tolerance within a coastal population of chinook salmon.

With global temperatures projected to surpass the limits of thermal tolerance for many species, evaluating the heritable variation underlying thermal tolerance is critical for understanding the potential for adaptation to climate change. We examined the evolutionary potential of thermal tolerance within a population of chinook salmon (Oncorhynchus tshawytscha) by conducting a full-factorial breeding design and measuring the thermal performance of cardiac function and the critical thermal maximum (CTmax) of offspring from each family. Additive genetic variation in offspring phenotype was mostly negligible, although these direct genetic effects explained 53% of the variation in resting heart rate (fH). Conversely, maternal effects had a significant influence on resting fH, scope for fH, cardiac arrhythmia temperature and CTmax. These maternal effects were associated with egg size, as indicated by strong relationships between the mean egg diameter of mothers and offspring thermal tolerance. Because egg size can be highly heritable in chinook salmon, our finding indicates that the maternal effects of egg size constitute an indirect genetic effect contributing to thermal tolerance. Such indirect genetic effects could accelerate evolutionary responses to the selection imposed by rising temperatures and could contribute to the population-specific thermal tolerance that has recently been uncovered among Pacific salmon populations.

Alternative male reproductive tactics drive asymmetrical hybridization between sunfishes (Lepomis spp.).

The potential role of alternative reproductive tactics in circumventing premating isolating mechanisms and driving hybridization between species has long been recognized, but to date there is little empirical support from natural systems. Hybridization occurs between bluegill (Lepomis macrochirus) and pumpkinseed sunfish (Lepomis gibbosus) and it is known to be asymmetrical (male bluegill × female pumpkinseed). Here, we test whether this pattern is driven by a recognition failure by pumpkinseed females or by an alternative cuckolder reproductive tactic in bluegill males. Using genetic parentage data, we found that bluegill cuckolders fathered 24.9% of the larvae in bluegill nests, but no evidence that pumpkinseed females spawned in bluegill nests. Pumpkinseed cuckolders fathered 8.7% of the larvae in pumpkinseed nests, whereas bluegill cuckolders fathered 13.6% of the larvae in those nests. Bluegill cuckolders thus frequently spawn in pumpkinseed nests and are responsible for the asymmetrical hybridization between the species. We discuss the evolutionary consequences of interactions between bluegill and pumpkinseed and the role of alternative reproductive tactics in adaptation and introgression.

The effects of elevated temperature on the sexual traits, immunology and survivorship of a tropical ectotherm.

In 2007, the Intergovernmental Panel on Climate Change projected an average global air temperature increase of 1.1-6.4°C by the end of the 21st century. Although the tropics are predicted to experience less extreme temperature increases than regions of higher latitude, tropical ectotherms live close to their thermal limits, and are thus particularly vulnerable to increases in temperature. In this study, we examined how predicted patterns of global warming will affect survival and sexual traits in the Trinidadian guppy (Poecilia reticulata). Guppies were exposed from birth to one of four temperature treatments: 23, 25 (control), 28 or 30°C. We measured brood survival and, at sexual maturity, male ornamentation, sperm traits and immune response. Our results show that increases in temperature result in guppies that have shorter, slower sperm but that there is an optimum temperature for ornamental hue at 28°C. Given the importance of sperm quality for reproduction, these results suggest population viability could be affected by warming. However, we found no difference in brood survival or immune response to a novel antigen across the treatments, indicating that survival may not be as vulnerable as previously thought. Overall, our data suggest that male sexual traits, and in particular sperm performance, are more sensitive than survival to a warming environment.

Polyandry and alternative mating tactics.

Many species in the animal kingdom are characterized by alternative mating tactics (AMTs) within a sex. In males, such tactics include mate guarding versus sneaking behaviours, or territorial versus female mimicry. Although AMTs can occur in either sex, they have been most commonly described in males. This sex bias may, in part, reflect the increased opportunity for sexual selection that typically exists in males, which can result in a higher probability that AMTs evolve in that sex. Consequently, females and polyandry can play a pivotal role in governing the reproductive success associated with male AMTs and in the evolutionary dynamics of the tactics. In this review, we discuss polyandry and the evolution of AMTs. First, we define AMTs and review game theoretical and quantitative genetic approaches used to model their evolution. Second, we review several examples of AMTs, highlighting the roles that genes and environment play in phenotype expression and development of the tactics, as well as empirical approaches to differentiating among the mechanisms. Third, ecological and genetic constraints to the evolution of AMTs are discussed. Fourth, we speculate on why female AMTs are less reported on in the literature than male tactics. Fifth, we examine the effects of AMTs on breeding outcomes and female fitness, and as a source, and possibly also a consequence, of sexual conflict. We conclude by suggesting a new model for the evolution of AMTs that incorporates both environmental and genetic effects, and discuss some future avenues of research.