Linking microbiome and stress hormone responses in wild tropical treefrogs across continuous and fragmented forests.

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.

Out to sea: ocean currents and patterns of asymmetric gene flow in an intertidal fish species.

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.

Age-dependent genetic variation in aggression.

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.

Hemodynamic Responses to Resistance Exercise with Blood Flow Restriction Using a Practical Method Versus a Traditional Cuff-Inflation System.

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.

Fitness Costs of Maternal Ornaments and Prenatal Corticosterone Manifest as Reduced Offspring Survival and Sexual Ornament Expression.

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.

Genetic integration of behavioural and endocrine components of the stress response.

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.

Honest signals and sexual conflict: Female lizards carry undesirable indicators of quality.

Sex differences in animal coloration often result from sex-dependent regulatory mechanisms. Still, some species exhibit incomplete sexual dimorphism as females carry a rudimentary version of a costly male trait, leading to intralocus sexual conflict. The underlying physiology and condition dependence of these traits can inform why such conflicts remain unresolved. In eastern fence lizards (Sceloporus undulatus), blue iridophore badges are found in males and females, but melanin pigmentation underneath and surrounding badges is male-exclusive. We track color saturation and area of badges across sexual maturity, and their relationship to individual quality (body condition and immunocompetence) and relevant hormones (testosterone and corticosterone). Saturation and testosterone were positively correlated in both sexes, but hormone and trait had little overlap between males and females. Saturation was correlated with body condition and immunocompetence in males but not in females. Co-regulation by androgens may have released females from resource allocation costs of color saturation, even when in high condition. Badge area was independent of testosterone, but associated with low corticosterone in females, indicating that a nonsex hormone underlies incomplete sexual dimorphism. Given the evidence in this species for female reproductive costs associated with ornamentation, this sex-nonspecific regulation of an honest signal may underlie intralocus sexual conflict.

Extensive hybridization and past introgression between divergent lineages in a quasi-clonal hermaphroditic fish: Ramifications for species concepts and taxonomy.

Extreme inbreeding is expected to reduce the incidence of hybridization, serving as a prezygotic barrier. Mangrove rivulus is a small killifish that reproduces predominantly by self-fertilization, producing highly homozygous lines throughout its geographic range. The Bahamas and Caribbean are inhabited by two highly diverged phylogeographic lineages of mangrove rivulus, Kryptolebias marmoratus and a 'Central clade' closely related to K. hermaphroditus from Brazil. The two lineages are largely allopatric, but recently were found in syntopy on San Salvador, Bahamas, where a single hybrid was reported. To better characterize the degree of hybridization and the possibility of secondary introgression, here we conducted a detailed genetic analysis of the contact zone on San Salvador. Two mixed populations were identified, one of which contained sexually mature hybrids. The distribution of heterozygosity at diagnostic microsatellite loci in hybrids showed that one of these hybrids was an immediate offspring from the K. marmoratus x Central clade cross, whereas the remaining five hybrids were products of reproduction by self-fertilization for 1-3 generations following the initial cross. Two hybrids had mitochondrial haplotypes of K. marmoratus and the remaining four hybrids had a haplotype of the Central clade, indicating that crosses go in both directions. In hybrids, alleles of parental lineages were represented in equal proportions suggesting lack of recent backcrossing to either of the parental lineages. However, sympatric populations of two lineages were less diverged than allopatric populations, consistent with introgression. Results are discussed in terms of applicability of the biological species concept for isogenic, effectively clonal, organisms.

Reconsidering the role of sex hormones in psychopathy development: Estrogen and psychopathy among male justice-involved youth.

Prominent theory suggests that factor one psychopathic traits may develop from increased input from hormones in the hypothalamic pituitary gonadal axis (HPG; i.e., testosterone) and decreased input from the hypothalamic pituitary adrenal axis (HPA; i.e., cortisol). Although there are extensive findings connecting low cortisol to psychopathy, less support has emerged for high levels of testosterone. This study examined whether incorporating the HPG hormone, estradiol, into this model would reveal relationships in line with theory: high levels of estradiol and testosterone in combination with low levels of cortisol would inform psychopathic traits. Baseline and reactive hormone levels were measured and compared to Psychopathy Checklist-Youth Version (PCL-YV) interviews among 66 male justice-involved youth (M age = 15.73) in a Southeastern juvenile detention center. The primary findings of this study were relationships between interacting HPA and HPG axis hormones with facet one and facet two psychopathic traits. Specifically, psychopathy total scores, interpersonal traits, and affective traits related to estradiol and testosterone reactivity, in that psychopathy scores were more likely with decreases in hormone reactivity (i.e., change in hormone level) following a stressor. Moreover, affective traits related to reactivity in all three hormones. These findings support inclusion of estradiol in neurobiological models of psychopathy and consideration of the individual components of psychopathy. This study adds to the growing body of research supporting interactions between variations in functioning of the HPA and HPG axes in relation to psychopathy.

Menstrual cycle effects on cardiovascular drift and maximal oxygen uptake during exercise heat stress.

AIM: Compared to other modulators of physiological strain associated with exercise heat stress, hyperthermia results in the greatest magnitude of cardiovascular (CV) drift and associated decrements in maximal oxygen uptake ([Formula: see text]). PURPOSE: To determine if elevated core temperature in the luteal phase (LP) of the menstrual cycle results in greater CV drift and reductions in [Formula: see text] versus the follicular phase (FP). METHODS: Seven women performed 15- and 45-min cycling bouts on separate occasions (60% [Formula: see text], 35 °C) followed by a [Formula: see text] test during the FP and LP. CV drift was measured between 15 and 45 min during the 45-min bout, and the 15-min bout was for measuring [Formula: see text] over the same time interval that CV drift occurred. RESULTS: Core temperature during LP was ~ 0.3 °C higher than FP (P < 0.05), but changes from rest during exercise were similar between phases (all P > 0.05). Heart rate increased significantly over time but was not different between phases (P = 0.78). Stroke volume decreased more over time during LP compared to FP (P = 0.02), but the values were similar at the end of exercise between phases (both time points P > 0.05). [Formula: see text] decrements for FP (13%) and LP (16%) were also comparable (P = 0.97). CONCLUSIONS: The LP-FP difference in core temperature in this study was not sufficient to amplify CV strain and decrements in [Formula: see text]. Greater differences in core temperature may be required to independently modulate CV drift and accompanying decrements in [Formula: see text] during prolonged exercise heat stress.

Ecdysteroids affect female reproductive status and outcome of contest over hosts in the parasitoid wasp Eupelmus vuilleti.

Ecdysteroids are a family of insect hormones that may play a role in modulating aggressive behavior in reproductive contexts. In Hymenoptera, the few studies investigating the link between ecdysteroid titers, reproduction and aggressiveness during contests concern solely eusocial species. Here, we explored whether ecdysteroid titers influenced female reproductive status as well as aggressiveness and resolution of conflict in a solitary ectoparasitoid, Eupelmus vuilleti (Hymenoptera: Eupelmidae). Eupelmus vuilleti females parasitize and feed upon juvenile stages of Callosobruchus maculatus (Coleoptera: Bruchidae). When two E. vuilleti females are simultaneously present on a patch, they tend to protect the host they exploit by displaying aggressive behaviors towards conspecific competitors. To our knowledge, nothing is known about the association between ecdysteroids and aggressiveness or the outcome of contests for host access in solitary insects. First, we quantified ecdysteroid titers that naturally circulate in females without fighting experience and after a contest over host access. Ecdysteroid titers measured after the contest did not correlate with female aggressiveness during the contest, but winner wasps had higher titers than both losers and females that did not fight. Then, we manipulated hormone titers via injection: ecdysone favored egg maturation (i.e., gonadotropic effect) within 24 h and increased almost immediately the females' probability of winning host access without affecting their aggressiveness. Our results represent an important step in understanding how hormones, such as ecdysteroids, mediate insect behavior during intraspecific competition.

Behavior and gene expression in the brain of adult self-fertilizing mangrove rivulus fish (Kryptolebias marmoratus) after early life exposure to the neurotoxin ß-N-methylamino-l-alanine (BMAA).

ß-N-Methylamino-l-alanine (BMAA), a neurotoxin naturally produced by cyanobacteria, diatoms and dinoflagellates, constitutes a serious environmental and health threat especially during acute blooms, which are becoming more frequent. This neurotoxin is implicated in several neurodegenerative diseases (ND) in humans through contaminated water or food consumption. Even low doses of neurotoxic compounds (NCs) can have lasting effects later in life. In this sense, early stages of development constitute a period of high sensitivity to environmental influence, particularly for the central nervous system. To understand the mechanisms underlying the delayed effects of NCs, newly hatched larvae of the mangrove rivulus fish, Kryptolebias marmoratus, were exposed to two sub-lethal doses of BMAA (20 µg/L and 15 mg/L) for 14 days. This fish naturally produces isogenic lineages due to its self-fertilizing reproduction, which is unique case among vertebrates. It thus provides genetic characteristics that allow scientists to study organisms' true reaction norm, minimizing genetic variability and focusing exclusively on the effects of the environment. Effect assessment was performed at different levels of biological organization to detect inconspicuous effects of BMAA, since this molecule displays long retention in organisms. BMAA effects on life history traits as well as behavioral traits such as boldness and aggressiveness were assessed more than 100 days after exposure. In addition, the relative expression of 7 potential BMAA target genes was studied, given their involvement in neurotransmission or their association with individual variation in boldness and aggressiveness. Selected genes code for reticulon 4 (RTN4), glutamate vesicular transporter 1 (Slc17a7), glutamine synthetase a (Glula), dopamine receptor D4 (DRD4), monoamine oxidase A (MAOA), calmodulin (CaM) and epedymine (Epd). Despite observing no effects of BMAA on growth, reproduction and behavioral traits, BMAA induced a significant increase of the expression of CaM and MAOA genes at 20 µg/L BMAA compared to the control group. A significant decrease of expression was observed between this lowest BMAA dose and 15 mg/L for DRD4, MAOA and CaM genes. Our results suggest disruption of glutamate turnover, intracellular dopamine depletion and activation of astrocyte protective mechanisms, indicating that BMAA might be excitotoxic. Our study revealed that BMAA can have long-lasting effects on the brain that are suspected to affect phenotypic traits with aging. Furthermore, it highlights the importance of studying delayed effects in ecotoxicological studies.

Choice consequences: salinity preferences and hatchling survival in the mangrove rivulus (Kryptolebias marmoratus).

In heterogeneous environments, mobile species should occupy habitats in which their fitness is maximized. Mangrove rivulus fish inhabit mangrove ecosystems where salinities range from 0 to 65 ppt, but are most often collected from areas with salinities of ∼25 ppt. We examined the salinity preference of mangrove rivulus in a lateral salinity gradient, in the absence of predators and competitors. Fish could swim freely for 8 h throughout the gradient with chambers containing salinities ranging from 5 to 45 ppt (or 25 ppt throughout in the control). We defined preference as the salinity in which the fish spent most of their time, and also measured preference strength, latency to begin exploring the arena, and number of transitions between chambers. To determine whether these traits were repeatable, each fish experienced three trials. Mangrove rivulus spent a greater proportion of time in salinities lower (5-15 ppt) than they occupy in the wild. Significant among-individual variation in the (multivariate) behavioral phenotype emerged when animals experienced the gradient, indicating strong potential for selection to drive behavioral evolution in areas with diverse salinity microhabitats. We also showed that mangrove rivulus had a significantly greater probability of laying eggs in low salinities compared with control or high salinities. Eggs laid in lower salinities also had higher hatching success compared with those laid in higher salinities. Thus, although mangrove rivulus can tolerate a wide range of salinities, they prefer low salinities. These results raise questions about factors that prevent mangrove rivulus from occupying lower salinities in the wild, whether higher salinities impose energetic costs, and whether fitness changes as a function of salinity.

Cardiac-Autonomic Responses to In-Season Training Among Division-1 College Football Players.

Flatt, AA, Esco, MR, Allen, JR, Robinson, JB, Bragg, A, Keith, CM, Fedewa, MV, and Earley, RL. Cardiac-autonomic responses to in-season training among Division-1 college football players. J Strength Cond Res 34(6): 1649-1656, 2020-Despite having to endure a rigorous in-season training schedule, research evaluating daily physiological recovery status markers among American football players is limited. The purpose of this study was to determine whether recovery of cardiac-autonomic activity to resting values occurs between consecutive-day, in-season training sessions among college football players. Subjects (n = 29) were divided into groups based on position: receivers and defensive backs (SKILL, n = 10); running backs, linebackers, and tight-ends (MID-SKILL, n = 11) and linemen (LINEMEN, n = 8). Resting heart rate (RHR) and the natural logarithm of the root mean square of successive differences multiplied by 20 (LnRMSSD) were acquired at rest in the seated position before Tuesday and Wednesday training sessions and repeated over 3 weeks during the first month of the competitive season. A position × time interaction was observed for LnRMSSD (p = 0.04), but not for the RHR (p = 0.33). No differences in LnRMSSD between days was observed for SKILL (Tuesday = 82.8 ± 9.3, Wednesday = 81.9 ± 8.7, p > 0.05). Small reductions in LnRMSSD were observed for MID-SKILL (Tuesday = 79.2 ± 9.4, Wednesday = 76.2 ± 9.5, p ≤ 0.05) and LINEMEN (Tuesday = 79.4 ± 10.5, Wednesday = 74.5 ± 11.5, p ≤ 0.05). The individually averaged changes in LnRMSSD from Tuesday to Wednesday were related to PlayerLoad (r = 0.46, p = 0.02) and body mass (r = -0.39, p = 0.04). Cardiac-parasympathetic activity did not return to resting values for LINEMEN or MID-SKILL before the next training session. Larger reductions in LnRMSSD tended to occur in players with greater body mass despite having performed lower workloads, although some individual variability was observed. These findings may have implications for how coaches and support staff address training and recovery interventions for players demonstrating inadequate cardiovascular recovery between sessions.

Does Body Shape in Fundulus Adapt to Variation in Habitat Salinity?

Understanding the ecological pressures that generate variation in body shape is important because body shape profoundly affects physiology and overall fitness. Using Fundulus, a genus of fish that exhibits considerable morphological and physiological variation with evidence of repeated transitions between freshwater and saltwater habitats, we tested whether habitat salinity has influenced the macroevolution of body shape at different stages in development. After accounting for phylogenetic inertia, we find that body shape deviates from the optimal streamlined shape in a manner consistent with different osmoregulatory pressures exerted by different salinity niches at every stage of ontogeny that we examined. We attribute variation in body shape to differential selection for osmoregulatory efficiency because: (1) saline intolerant species developed body shapes with relatively low surface areas more conducive to managing osmoregulatory demands and (2) inland species that exhibit high salinity tolerances have body shapes similar to saline tolerant species in marine environments.

Prolonged survival out of water is linked to a slow pace of life in a self-fertilizing amphibious fish.

Metabolic rate and life-history traits vary widely both among and within species, reflecting trade-offs in energy allocation, but the proximate and ultimate causes of variation are not well understood. We tested the hypothesis that these trade-offs are mediated by environmental heterogeneity, using isogenic strains of the amphibious fish Kryptolebias marmoratus that vary in the amount of time each can survive out of water. Consistent with pace of life theory, the strain that survived air exposure the longest generally exhibited a 'slow' phenotype, including the lowest metabolic rate, largest scope for metabolic depression, slowest consumption of energy stores and least investment in reproduction under standard conditions. Growth rates were fastest in the otherwise slow strain, however. We then tested for fitness trade-offs between 'fast' and 'slow' strains using microcosms where fish were held either with constant water availability or under fluctuating conditions where water was absent for half of the experiment. Under both conditions the slow strain grew larger and was in better condition, and under fluctuating conditions the slow strain produced more embryos. However, the fast strain had larger adult population sizes under both conditions, indicating that fecundity is not the sole determinant of population size in this species. We conclude that genetically based differences in the pace of life of amphibious fish determine survival duration out of water. Relatively slow fish tended to perform better under conditions of limited water availability, but there was no detectable cost under control conditions. Thus, pace of life differences may reflect a conditionally neutral instead of antagonistic trade-off.

Comparison of Lactate and Electromyographical Thresholds After an Exercise Bout.

Snarr, RL, Esco, MR, Tolusso, DV, Hallmark, AV, Earley, RL, Higginbotham, JC, Fedewa, MV, and Bishop, P. Comparison of lactate and electromyographical thresholds after an exercise bout. J Strength Cond Res 33(12): 3322-3331, 2019-The electromyographical threshold (EMGT) has been previously validated as a means to predict the work rate at which lactate threshold (LT) occurs. The reliability of these measures has yet to be examined after a bout of exercise. The purpose was to determine the agreement between electromyography (EMG) and LT after a 30-minute bout of steady-state aerobic exercise. Participants completed 2 graded exercise tests (GXT) on a cycle ergometer separated by 30 minutes of steady-state exercise. Blood lactate was measured the last 45 seconds of each stage during both GXTs, whereas EMG of the vastus lateralis was monitored continuously. Individual agreement demonstrated that pre-exercise and post-exercise LT occurred at the same work rate in 2 of 10 participants, whereas EMGT occurred at the same work rate in 6 of 10 participants. Results showed no mean difference between work rates for LT or EMG threshold for the pre-exercise GXT, but LT was significantly lower (p < 0.01) than EMGT during the post-exercise GXT. Post-GXT LT work rates were also determined to be significantly lower than pre-GXT LT (p = 0.034), whereas no differences existed in EMG thresholds. Although both LT and EMGT testing may display similar properties, they are not interchangeable. The physiological responses to increasing exercise intensity between La and EMG signaling seem to be associated, and their interaction may not be cause-effect. Because of poor individual agreement, caution should be used when determining LT through the use of EMG. Further research is needed to determine the ability of these 2 metrics to prescribe training intensities.

Cryptic Male Phenotypes in the Mangrove Rivulus Fish, Kryptolebias marmoratus.

Alternative male phenotypes exist in many species and impact mating system dynamics, population genetics, and mechanisms of natural and sexual selection that operate within a population. We report on the discovery of a cryptic male phenotype in the mangrove rivulus fish (Kryptolebias marmoratus), one of only two self-fertilizing hermaphroditic vertebrates. In this androdiecious species, males are infrequent, often making up less than 5% of a population; and they have historically been described as having an orange color and lacking or having a very faded outline of the well-defined caudal eyespot (ocellus) that is obvious in hermaphrodites. The cryptic male we describe varies subtly from the hermaphrodite phenotype, without visible orange pigmentation on the body and retention or only minor fading of the ocellus. This male morph was identified by a loss of a defined melanistic "fingerprinting" on the caudal fin seen in hermaphrodites, not previously used as diagnostic for hermaphrodites, and replaced by a diffuse deposition of pigment across the fin. Individuals were identified as male with 85.7% accuracy when using these criteria. We report that in nine populations, spanning three geographically distinct regions in Florida, across two and a half years, 0.3% of the 6057 mangrove rivulus collected exhibited this cryptic male phenotype and were confirmed to have testes via dissection. Overall, 2.3% of the animals were male (normal and cryptic phenotypes), and cryptic males represented 12.9% of all males collected. Even a minor increase in individuals identified as male in a species where males make up such a small portion of the population can have important implications for population genetics. Opportunities for outbreeding are likely enhanced, which has significant evolutionary ramifications.

Correction: Development of G: a test in an amphibious fish.

Figure 3 legend has been corrected to state: "Difference matrices for pairwise-trait phenotypic correlations (rP, below diagonal) and pairwise-trait genetic correlations (rG, above diagonal) from 1, 15, and 100 DPH. Differences are color coded by strength and direction. Differences shown in gray are positive and differences shown in black are negative. When ages are similar, the colored square is small; when ages are very different, the colored square fills the cell. EPL Epural length, EPA epural angle, PHPL parahypural length, PHPA parahypural angle, HYPL hypural length, HYPW hypural width, and SL standard length."

Juvenile hormone manipulation affects female reproductive status and aggressiveness in a non-social parasitoid wasp.

In vertebrates, titers of androgens such as testosterone are known to upregulate aggressive behaviors associated with reproduction. In insects, juvenile hormone (JH) is a good candidate for studying the flexibility of insect endocrine responses because it has important effects on both reproductive processes and behavior. JH has a gonadotropic effect across a broad range of insect species, increasing ovarian development in females, and may have a role in the regulation of aggressive behavior during competition. In Hymenoptera, the functions of JH have been studied in facultatively eusocial species such as polistine wasps, bumblebees, ants and bees. Surprisingly, no work has yet focused on the relationship between JH, reproduction and aggressiveness in a non-social Hymenoptera, although it may help to understand how JH actions have evolved across taxa with different degrees of sociality. Here, we explored how JH treatment influenced: i) female reproductive status, and ii) the intensity (aggressiveness) and resolution of conflict, in Eupelmus vuilleti (Hymenoptera: Eupelmidae), a solitary ectoparasitoid wasp in which females fight over hosts. We demonstrated that intra-abdominal injections of JH increased the number of mature eggs in females after 24 h. In addition, the number of aggressive behaviors displayed by females was affected by the interaction between JH treatment and the number of mature eggs in their abdomen, but mature egg load alone predicted the outcome of staged contests. Wasps were more aggressive when they had more ready-to-lay eggs, with this effect being stronger when females were injected with JH. Moreover, females won more frequently when they had higher mature egg load. Our results highlight how JH affects egg maturation and aggressive behaviors in Eupelmus vuilleti females. To our knowledge, this is the first study showing that hormone manipulation can modulate females' reproduction status and behavior during intraspecific competition over hosts in a non-social hymenopteran parasitoid.