Divergent selection between the sexes may enable parasites to better infect one host sex over the other, potentially leading to sex-biased parasitism. Hosts also typically harbor a component community of parasite species that can vary across a spatial scale. We explored spatial variation in sex-biased parasitism in mangrove rivulus (Kryptolebias marmoratus), an androdioecious fish in which simultaneous hermaphrodites can facultatively transition to male. Using cup traps deployed in the mangroves, we sampled populations that varied in sex ratio; three with relatively high and low percentages of males in Belize and the Florida Keys, respectively. Fish were sexed, and their external bodies and all organs were examined for parasites using microscopy. Hermaphrodites had significantly higher parasite species diversity overall (Simpson's index). Sex differences existed for only one parasite species (heart-specific Ascocotyle sp. digenean metacercariae), with significantly male-biased parasitism in one Belizean population and significantly hermaphrodite-biased parasitism in another. Host mass also predicted the abundance of some parasite species independent of sex: larger individuals had higher abundances of gill-specific and heart-specific Ascocotyle sp. digenean metacercariae. Parasite species diversity (Simpson's index) and richness were higher in Belize than the Florida Keys. While males are often the more heavily parasitized sex, our study did not support this, and our results indicate that host-parasite interactions are driven by a complex relationship between host sex, body mass, geography, and parasite species.

Macroparasite species infecting the mangrove rivulus fish (Kryptolebias marmoratus) are relatively unknown and have never been previously described in Florida. We collected 20 specimens from a wild population using cup traps deployed in the mangroves along a transect and then examined each fish for external and internal macroparasites. There was 100

When embryos experience different environments than their parents, plasticity can enable the development of alternate phenotypes that confer higher fitness in the new conditions. Temperature-induced plasticity could be especially critical for species that inhabit areas with considerable thermal variation. We studied transcriptional variation in embryos of mangrove rivulus (Kryptolebias marmoratus)-a self-fertilizing hermaphroditic, eurythermal fish that resides in notoriously spatiotemporally variable mangrove forests-exposed to different thermal regimes during development. To study transcriptional plasticity, we first improved the genome assembly to chromosome length scaffolds (N50 of 28.17 Megabases). Whole transcriptome sequencing revealed that both temperature and developmental timing modulated embryonic gene expression. We found few differences in gene expression between embryos incubated in cold and warm conditions and assessed before the temperature-sensitive period of development, indicating high resistance to stochastic changes in gene expression early in development. Replicate embryos exposed to cold temperatures and sampled after the temperature-sensitive period showed less variation in gene expression than those sampled before, suggesting canalization of the plastic response. DNA replication/repair, organelle, and gas transport pathways were upregulated while nervous system development, cell signaling, and cell adhesion were downregulated in cold-exposed compared to warm-exposed embryos sampled after the temperature-sensitive period. These plastic shifts in gene expression could have major implications for reorganizing the phenotype (e.g., apoptosis, mitosis) in response to environmental changes occurring within a generation.

Behaviours can be adaptively correlated with each other and with other aspects of phenotype. We investigated behaviour across foraging, mating, and risk contexts in females of the poeciliid fish, Girardinus metallicus. We quantified relationships between these behaviours and aggression signals, body size, and reproductive output. Behaviours describing aggression and boldness, some of which were repeatable, were correlated in females. Aggression was signalled by the darkening of a black spot on the extended dorsal fin or by fin flaring. Spot darkening occurred during intra- and intersexual interactions and was positively correlated with interfemale aggression and negatively correlated with the time females spent following males, suggesting that it is an honest indicator of motivational state. In contrast, fin flaring was directed only at females and occurred more frequently in the foraging context. Larger females had fewer offspring and received fewer courtship displays, possibly because they were more aggressive to males. In contrast to studies of males, we found a negative relationship between rank order of boldness and aggression, consistent with either a trade-off or with selection favouring the negative relationship. Our results highlight the importance of studying females in model systems to uncover novel patterns and potentially meaningful departures from what is typically seen in males.

Drift and gene flow affect genetic diversity. Given that the strength of genetic drift increases as population size decreases, management activities have focused on increasing population size through preserving habitats to preserve genetic diversity. Few studies have empirically evaluated the impacts of drift and gene flow on genetic diversity.Kryptolebias marmoratus, henceforth ‘rivulus’, is a small killifish restricted to fragmented New World mangrove forests with gene flow primarily associated with ocean currents. Rivulus form distinct populations across patches, making them a well-suited system to test the extent to which habitat area, fragmentation and connectivity are associated with genetic diversity. Using over 1000 individuals genotyped at 32 microsatellite loci, high-resolution landcover data and oceanographic simulations with graph theory, we demonstrate that centrality (connectivity) to the metapopulation is more strongly associated with genetic diversity than habitat area or fragmentation. By comparing models with and without centrality standardized by the source population’s genetic diversity, our results suggest that metapopulation centrality is critical to genetic diversity regardless of the diversity of adjacent populations. While we find evidence that habitat area and fragmentation are related to genetic diversity, centrality is always a significant predictor with a larger effect than any measure of habitat configuration.

INTRODUCTION/PURPOSE:The purpose of this study was to test the hypothesis that cardiovascular (CV) drift and associated decrements in maximal oxygen uptake (V̇O 2max ) are greater in high-fit compared with low-fit women during exercise at the same %V̇O 2max , but comparable at the same rate of metabolic heat production. METHODS:Six high-fit (HI) and six low-fit (LO) women cycled in 35°C for 15 or 45 min at the same relative intensity (60% V̇O 2max ; 15REL and 45REL) or fixed rate of heat production (500 W; 15FX and 45FX), immediately followed by a graded exercise test to measure V̇O 2max . The separate 15- and 45-min trials permitted measurements of V̇O 2max over the same time interval as CV drift. RESULTS:During 45REL, higher heat production in HI (496 ± 51 vs 364 ± 44 W in LO) resulted in greater end-exercise core temperature (38.7°C ± 0.4°C vs 38.2°C ± 0.1°C, P = 0.03), greater increases in HR (15 bpm (10%) vs 10 bpm (6%), P = 0.03) and decreases in stroke volume (11 mL per beat (16%) vs 5 mL per beat (8%), P = 0.001), and larger reductions in V̇O 2max (16% vs 5%, P = 0.04) compared with LO. During 45FX, temperature responses, CV drift, and decreased V̇O 2max were not different between groups (all P > 0.05), despite differences in %V̇O 2max (60% vs 75% for HI and LO, respectively). CONCLUSIONS:We conclude metabolic heat production modulates the CV drift-V̇O 2max relationship, independent of fitness level. These results support previous findings showing the magnitude of CV drift is proportional to reductions in V̇O 2max .

The amphibian skin microbiome is an important component of anti-pathogen defense, but the impact of environmental change on the link between microbiome composition and host stress remains unclear. In this study, we used radiotelemetry and host translocation to track microbiome composition and function, pathogen infection, and host stress over time across natural movement paths for the forest-associated treefrog, Boana faber . We found a negative correlation between cortisol levels and putative microbiome function for frogs translocated to forest fragments, indicating strong integration of host stress response and anti-pathogen potential of the microbiome. Additionally, we observed a capacity for resilience (resistance to structural change and functional loss) in the amphibian skin microbiome, with maintenance of putative pathogen-inhibitory function despite major temporal shifts in microbiome composition. Although microbiome community composition did not return to baseline during the study period, the rate of microbiome change indicated that forest fragmentation had more pronounced effects on microbiome composition than translocation alone. Our findings reveal associations between stress hormones and host microbiome defenses, with implications for resilience of amphibians and their associated microbes facing accelerated tropical deforestation.

When animals live in long-term groups, the potential for conflict is high. Conflict is costly, so an individual’s decision to engage depends on the information it has about the costs and benefits of fighting. One source of information could be past contest experience, where previous winners/losers typically become more likely to win/lose in the future. However, repeated interactions can familiarize individuals with conflict and provide opportunities to learn to become better fighters, regardless of outcome. We explored how individuals integrate information from previous contests to inform future encounters in a group-living fish, Neolamprologus pulcher. We gave contestants single, reinforcing, and contradictory experiences and measured behavior and post-fight water-borne levels of androgenic steroids (testosterone, 11-ketotestosterone). Contradictory outcomes were associated with reduced investment in fighting. More fighting experience did not lead to greater investment in fighting, as consecutive losses resulted in reduced aggression. Also, there was no effect of fighting treatment on water-borne androgen concentrations. Interestingly, there were sex differences in which behaviors were influenced by experience, and in whether body mass was associated with androgen concentrations, which could indicate that males and females vary in how perceived fighting ability changes with contest experience. Our data reveal the complex ways in which repeated experiences can alter an individual’s propensity to invest in conflict. Repeated interactions associated with predictable changes in behavior can contribute to rank stability in groups and our results indicate that whether and how they do depend on the quality and quantity of interactions plus individual factors such as sex.

Passive dispersal via wind or ocean currents can drive asymmetric gene flow, which influences patterns of genetic variation and the capacity of populations to evolve in response to environmental change. The mangrove rivulus fish (Kryptolebias marmoratus), hereafter "rivulus," is an intertidal fish species restricted to the highly fragmented New World mangrove forests of Central America, the Caribbean, the Bahamas, and Florida. Mangrove patches are biological islands with dramatic differences in both abiotic and biotic conditions compared to adjacent habitat. Over 1,000 individual rivulus across 17 populations throughout its range were genotyped at 32 highly polymorphic microsatellites. Range-wide population genetic structure was evaluated with five complementary approaches that found eight distinct population clusters. However, an analysis of molecular variance indicated significant population genetic structure among regions, populations within regions, sampling locations within populations, and individuals within sampling locations, indicating that rivulus has both broad- and fine-scale genetic differentiation. Integrating range-wide genetic data with biophysical modeling based on 10 years of ocean current data showed that ocean currents and the distance between populations over water drive gene flow patterns on broad scales. Directional migration estimates suggested some significant asymmetries in gene flow that also were mediated by ocean currents and distance. Specifically, populations in the center of the range (Florida Keys) were identified as sinks that received migrants (and alleles) from other populations but failed to export individuals. These populations thus harbor genetic variation, perhaps even from extirpated populations across the range, but ocean currents and complex arrangements of landmasses might prevent the distribution of that genetic variation elsewhere. Hence, the inherent asymmetry of ocean currents shown to impact both genetic differentiation and directional migration rates may be responsible for the complex distribution of genetic variation across the range and observed patterns of metapopulation structure.

Understanding the extent to which behavioural variance is underlain by genotypic, environmental and genotype-by-environment effects is important for predicting how behavioural traits might respond to selection and evolve. How behaviour varies both within and among individuals can change across ontogeny, leading to differences in the relative contribution of genetic and environmental effects to phenotypic variation across ages. We investigated among-individual and among-genotype variation in aggression across ontogeny by measuring, twice as juveniles and twice as adults, both approaches and attacks against a three-dimensional-printed model opponent in eight individuals from each of eight genotypes (N = 64). Aggression was only significantly repeatable and heritabile in juveniles. Additionally, how aggression changed between juvenile and adult life-history stages varied significantly among individuals and genotypes. These results suggest that juvenile aggression is likely to evolve more rapidly via natural selection than adult aggression and that the trajectory of behavioural change across the lifespan has the potential to evolve. Determining when genetic variation explains (or does not explain) behavioural variation can further our understanding of key life-history stages during which selection might drive the strongest or swiftest evolutionary response.

Synopsis The adaptive significance of colorful or exaggerated traits (i.e., ornaments) expressed in females is often unclear. Competing hypotheses suggest that expression of female ornaments arises from maladaptive (or neutral) genetic inheritance from males along with incomplete epigenetic regulation, or from positive selection for ornaments in females under social competition. Whether costly or advantageous, the visibility of such traits can sometimes be behaviorally modulated in order to maximize fitness. Female eastern fence lizards express blue badges that are variable in size and color saturation. These are rudimentary compared to those seen in males and carry important costs such as reduced mating opportunities. Body temperature is a well-established enhancer of badge color, and thus thermoregulation may be one way these animals modulate badge visibility. We quantified realized body temperatures of female lizards paired in laboratory trials and observed that females with larger badges attained higher body temperatures when freely allowed to thermoregulate, sometimes beyond physiological optima. In this association between phenotype and behavior, females with larger badges exhibited thermoregulatory patterns that increase their badges' visibility. This signal-enhancing behavior is difficult to reconcile with the widely held view that female ornaments are maladaptive, suggesting they may carry context-dependent social benefits.

The aim of this study was to examine the potential differences in acute hemodynamic responses and muscular performance outcomes following resistance exercise between traditional blood flow restriction (TRABFR) and a novel band tissue flossing method (BTFBFR). METHODS: Fifteen healthy young adults (23.27 ± 2.69 years) visited the lab for three sessions (≥72 h apart). Each session’s exercise consisted of three sets of 20 maximum-effort seated leg extensions and flexions with one of three conditions: control (CON), TRABFR (50% limb occlusion pressure (LOP)), or BTFBFR. During TRABFR and BTFBFR sessions, occlusion was applied immediately prior to exercise and removed immediately after. Heart rate was collected prior to exercise, after onset of occlusion, immediately after exercise, and one-minute after removal of occlusion. Ultrasonography was performed prior to, and at least 30 s after, occlusion. RESULTS: BTFBFR caused greater reductions in arterial distance (14.28%, p = 0.010) and arterial area (28.43%, p = 0.020) than TRABFR. BTFBFR was able to significantly reduce arterial flow below pre-occlusion values, while TRABFR did not. Both conditions caused significant elevations in heart rate following occlusion (TRABFR: +4.67 bpm, p = 0.046 and BTFBFR: +6.07 bpm, p = 0.034), immediately post-exercise (TRABFR: +56.93 bpm, p < 0.001 and BTFBFR: +52.79 bpm, p < 0.001) and one-minute post-exercise (TRABFR: +15.71, p = 0.003 and BTFBFR: +14.57, p < 0.001). Only BTFBFR caused significant reductions in performance as measured by average power per repetition. CONCLUSIONS: BTFBFR causes a more exaggerated decrease in arterial blood flow as well as muscular power when compared to traditional TRABFR at 50% of LOP.

Colorful traits (i.e., ornaments) that signal quality have well-established relationships with individual condition and physiology. Furthermore, ornaments expressed in females may have indirect fitness effects in offspring via the prenatal physiology associated with, and social consequences of, these signaling traits. Here we examine the influence of prenatal maternal physiology and phenotype on condition-dependent signals of their offspring in adulthood. Specifically, we explore how prenatal maternal testosterone, corticosterone, and ornament color and size correlate with female and male offspring survival to adulthood and ornament quality in the lizard Sceloporus undulatus . Offspring of females with more saturated badges and high prenatal corticosterone were less likely to survive to maturity. Badge saturation and area were negatively correlated between mothers and their male offspring, and uncorrelated to those in female offspring. Maternal prenatal corticosterone was correlated negatively with badge saturation of male offspring in adulthood. Our results indicate that maternal ornamentation and prenatal concentrations of a stress-relevant hormone can lead to compounding fitness costs by reducing offspring survival to maturity and impairing expression of a signal of quality in surviving males. This mechanism may occur in concert with social costs of ornamentation in mothers. Intergenerational effects of female ornamentation and prenatal stress may be interdependent drivers of balancing selection and intralocus sexual conflict over signaling traits.

The vertebrate stress response comprises a suite of behavioural and physiological traits that must be functionally integrated to ensure organisms cope adaptively with acute stressors. Natural selection should favour functional integration, leading to a prediction of genetic integration of these traits. Despite the implications of such genetic integration for our understanding of human and animal health, as well as evolutionary responses to natural and anthropogenic stressors, formal quantitative genetic tests of this prediction are lacking. Here, we demonstrate that acute stress response components in Trinidadian guppies are both heritable and integrated on the major axis of genetic covariation. This integration could either facilitate or constrain evolutionary responses to selection, depending upon the alignment of selection with this axis. Such integration also suggests artificial selection on the genetically correlated behavioural responses to stress could offer a viable non-invasive route to the improvement of health and welfare in captive animal populations.

New World mangrove trees are foundation species, and their range is predicted to expand northward with climate change. Foundation species are commonly prioritized for conservation, with the goal of preserving the entire community that depends on them. However, no studies have explicitly investigated whether mangrove-dependent species' ranges will track the northward expansion of New World mangrove forests. We use the mangrove rivulus fish, Kryptolebias marmoratus, to investigate shifts in habitat suitability in response to various climate change scenarios (Representative Concentration Pathways 2.6, 4.5, 6.0, and 8.5). Niche models for coastal species focus on traditional climatic variables (e.g., precipitation, temperature) even though coastal habitats also are directly influenced by marine variables (e.g., sea surface salinity). We employ a novel data integration method that combines marine and climatic variables, and that accounts for model selection uncertainty using model averaging to provide robust estimates of habitat suitability. Contrary to expectation, suitability of rivulus habitat is predicted to increase in the south and decrease or remain unchanged in the north across all climate change scenarios. Thus, rivulus might experience range contraction, not expansion. Habitat became more suitable with increased salinity of the saltiest month and precipitation of the driest quarter. In laboratory settings, rivulus have higher survival, reproductive success, and growth rates in low salinities. This discrepancy suggests that some combination of the responses of rivulus and its competitors to environmental change will restrict rivulus to habitats that laboratory experiments consider suboptimal. Our models suggest that focusing conservation decisions on foundation species could overestimate habitat availability and resilience of affiliated communities while simultaneously underestimating species declines and extinction risks.

Steroid hormones accumulate in recirculation aquaculture systems (RAS) and may influence the reproductive physiology of farmed fish. Ozone reduces hormone concentrations in freshwater RAS used to rear Atlantic salmon, but its effect on reproductive development is unknown. Accordingly, an 8-month trial was carried out to evaluate the growth, health, and maturation of post-smolt Atlantic salmon (296 +/- 4 g initial weight) reared in six replicated freshwater RAS (9.5 m(3) total volume) operated with or without ozone (N = 3/treatment). Residual ozone was controlled with an oxidation reduction potential (ORP) of 300-320 mV, and mean water temperature was maintained at 14.7 degrees C. Atlantic salmon growth was generally faster in ozonated RAS. Salmon from RAS with and without ozone weighed 2156 +/- 101 and 1810 +/- 15 g, respectively, by the end of the study. Caudal, anal, and pelvic fin damage was greater (P < 0.05) for salmon in ozonated RAS early in the trial but improved thereafter. No statistical differences in gill, skin, and skeletal muscle histopathology were observed between treatments at the end of the study. Waterborne estradiol, testosterone, and 11-ketotestosterone levels were periodically lower (P < 0.05) in ozonated RAS, but maturing salmon were more prevalent in these systems. At the end of the trial, percent maturation of salmon populations reared in RAS with and without ozone was 63 +/- 7 and 48 +/- 1%, respectively; however, maturity appeared to be related to fish size. Improved water quality was observed in ozonated RAS including reduced dissolved copper, iron, and zinc levels, total heterotrophic bacteria counts, and true color, and increased ultraviolet transmittance, which may have supported improved Atlantic salmon growth. Overall, ozone did not inhibit the onset or prevalence of Atlantic salmon maturation, but significant improvements in water quality and salmon growth performance resulted from its use.

Fighting experiences strongly influence aggressive behavior and physiology (winner-loser effects). These effects are conserved from invertebrates to vertebrates, but the underlying mechanisms remain unclear. Recent studies indicate that the brain social decision-making network (SDN) plays a key role in guiding experience-induced behavioral change. Also, while most studies have focused on how winning and losing experiences alter aggression, growing evidence points to these experiences driving multiple behavioral effects, including changes in the ability to learn. In mangrove rivulus fish (Kryptolebias marmoratus), we discovered that single winning experiences significantly improved spatial learning but not risk-avoidance learning, whereas single losing experiences drove the exact opposite to occur. These results provide strong evidence that winning and losing modulate diverse behaviors served by key nodes within the SDN, specifically the dorsolateral pallium (Dl; fish homolog to mammalian hippocampus, which serves spatial learning) and dorsomedial pallium (Dm; fish homolog to mammalian basolateral amygdala, which responds to fear). We therefore quantified whole-proteome expression within the forebrain (where Dm and Dl are located) of adult rivulus with divergent social experiences. We discovered 23 proteins were significantly differentially expressed in the forebrains of winners and losers. Differentially expressed proteins in losers related to modulation of cellular processes, apoptosis and learning while those in winners related to neuronal plasticity, neuroendocrine homeostasis, energy utilization, and learning. These results imply that winner-loser effects might be governed by very different patterns of protein expression, which could explain why winners and losers show such pronounced differences in behavioral performance. Significance Statement Social interactions permeate the daily lives of most animals and often result in changes in behavior for all parties. This implies that social experiences reorganize the brain in ways that promote the expression of alternative behaviors, or that help individuals cope with the outcome of such interactions. But how do aggressive interactions sculpt the brain at the molecular level? We used an emerging model organism, Kryptolebias marmoratus, to examine whether experiences modulate learning ability and then probe the potential neural mechanisms underlying these behavioral changes. We discovered that single winning and losing experiences dramatically altered spatial learning and risk-avoidance learning, respectively, indicating that winning and losing experiences have markedly different effects on the brain and cognitive processes. Classification Biological Sciences, Ecology