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.

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.

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.

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.

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."

Development of G: a test in an amphibious fish.

Heritable variation in, and genetic correlations among, traits determine the response of multivariate phenotypes to natural selection. However, as traits develop over ontogeny, patterns of genetic (co)variation and integration captured by the G matrix may also change. Despite this, few studies have investigated how genetic parameters underpinning multivariate phenotypes change as animals pass through major life history stages. Here, using a self-fertilizing hermaphroditic fish species, mangrove rivulus (Kryptolebias marmoratus), we test the hypothesis that G changes from hatching through reproductive maturation. We also test Cheverud's conjecture by asking whether phenotypic patterns provide an acceptable surrogate for patterns of genetic (co)variation within and across ontogenetic stages. For a set of morphological traits linked to locomotor (jumping) performance, we find that the overall level of genetic integration (as measured by the mean-squared correlation across all traits) does not change significantly over ontogeny. However, we also find evidence that some trait-specific genetic variances and pairwise genetic correlations do change. Ontogenetic changes in G indicate the presence of genetic variance for developmental processes themselves, while also suggesting that any genetic constraints on morphological evolution may be age-dependent. Phenotypic correlations closely resembled genetic correlations at each stage in ontogeny. Thus, our results are consistent with the premise that-at least under common environment conditions-phenotypic correlations can be a good substitute for genetic correlations in studies of multivariate developmental evolution.

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.

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.

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.

Real or fake? Natural and artificial social stimuli elicit divergent behavioural and neural responses in mangrove rivulus, Kryptolebias marmoratus.

Understanding how the brain processes social information and generates adaptive behavioural responses is a major goal in neuroscience. We examined behaviour and neural activity patterns in socially relevant brain nuclei of hermaphroditic mangrove rivulus fish (Kryptolebias marmoratus) provided with different types of social stimuli: stationary model opponent, regular mirror, non-reversing mirror and live opponent. We found that: (i) individuals faced with a regular mirror were less willing to interact with, delivered fewer attacks towards and switched their orientation relative to the opponent more frequently than fish exposed to a non-reversing mirror image or live opponent; (ii) fighting with a regular mirror image caused higher expression of immediate-early genes (IEGs: egr-1 and c-Fos) in the teleost homologues of the basolateral amygdala and hippocampus, but lower IEG expression in the preoptic area, than fighting with a non-reversing mirror image or live opponent; (iii) stationary models elicited the least behavioural and IEG responses among the four stimuli; and (iv) the non-reversing mirror image and live opponent drove similar behavioural and neurobiological responses. These results suggest that the various stimuli provide different types of information related to conspecific recognition in the context of aggressive contests, which ultimately drive different neurobiological responses.

Whole-genome sequencing reveals the extent of heterozygosity in a preferentially self-fertilizing hermaphroditic vertebrate.

The mangrove rivulus, Kryptolebias marmoratus, is one of only two self-fertilizing hermaphroditic fish species and inhabits mangrove forests. While selfing can be advantageous, it reduces heterozygosity and decreases genetic diversity. Studies using microsatellites found that there are variable levels of selfing among populations of K. marmoratus, but overall, there is a low rate of outcrossing and, therefore, low heterozygosity. In this study, we used whole-genome data to assess the levels of heterozygosity in different lineages of the mangrove rivulus and infer the phylogenetic relationships among those lineages. We sequenced whole genomes from 15 lineages that were completely homozygous at microsatellite loci and used single nucleotide polymorphisms (SNPs) to determine heterozygosity levels. More variation was uncovered than in studies using microsatellite data because of the resolution of full genome sequencing data. Moreover, missense polymorphisms were found most often in genes associated with immune function and reproduction. Inferred phylogenetic relationships suggest that lineages largely group by their geographic distribution. The use of whole-genome data provided further insight into genetic diversity in this unique species. Although this study was limited by the number of lineages that were available, these data suggest that there is previously undescribed variation within lineages of K. marmoratus that could have functional consequences and (or) inform us about the limits to selfing (e.g., genetic load, accumulation of deleterious mutations) and selection that might favor the maintenance of heterozygosity. These results highlight the need to sequence additional individuals within and among lineages.

Natural hybridization between divergent lineages in a selfing hermaphroditic fish.

By definition, mating between individuals is infrequent in highly selfing organisms, and so too, therefore, hybridization should be rare between genetically divergent lineages in predominantly self-fertilizing species. Notwithstanding these expectations, here we report a remarkable case of natural hybridization between highly diverged phylogeographic lineages of the mangrove rivulus, a small killifish that reproduces predominantly by self-fertilization and typically is found as highly homozygous lines in most parts of its extensive geographical range. Two distinctive genetic lineages (Kryptolebias marmoratus and a 'Central clade' closely related to K. hermaphroditus) previously were not known in sympatry, but were found by us to co-occur on San Salvador, Bahamas. Genetic analyses of a mitochondrial and multiple nuclear markers determined the direction of a cross producing a hybrid fish. Furthermore, we show that this hybrid individual was viable, as it successfully reproduced by self-fertilization for two generations. Additional sampling of this population will be necessary to determine if backcrossing of hybrids to the parental lineages occurs in nature and to analyse whether such backcross progeny are viable. Application of the biological species concept (BSC) is traditionally difficult in clonally reproducing organisms. Our results show that although mangrove rivulus fish are mostly highly selfing in nature (resulting in isogenic, effectively clonal and homozygous progeny), classification within this taxonomic complex need not be incompatible with the BSC.

Individual variation in ACTH-induced cortisol levels in females of a livebearing fish at different gestational stages.

Individuals vary in their baseline levels of stress hormones (predictive homeostasis) and in their stress responses (reactive homeostasis). Variation in normal reactive scope, both predictive and reactive homeostasis, may be important for understanding how endocrine traits respond to selection. Reactive homeostasis is the increase in glucocorticoid (GCs) hormones above baseline. Individuals at different life history stages, such as gestation in females, may show variation in normal reactive scope. We performed an adrenocorticotropic hormone (ACTH) challenge and measured changes in circulating GCs to estimate the reactive scope of female sailfin mollies (Poecilia latipinna) at different gestational states. We measured cortisol, primary GC in teleost fishes, to obtain baseline release rates prior to injection with either ACTH or saline control. Using water-borne hormones, we measured cortisol release rates at four time intervals post-injection. Females were then sacrificed to determine the developmental stage of embryos, if present, and the number of developing embryos or mature ova. We found that ACTH-injected females had significant increases in cortisol releases rates, whereas cortisol release rates of control females did not change during the 4 h post-injection period. We found high repeatability in predictive homeostasis of cortisol and moderate repeatability in reactive homeostasis and a phenotypic correlation between predictive and reactive homeostasis. Gestational state did not affect female predictive or reactive homeostasis. We applied the reactive scope model to P. latipinna and gained a further understanding of how among- and within-individual variation in both predictive and reactive homeostasis are partitioned and how these traits vary under certain life-history conditions.

Evolution of steroid hormones in reproductive females of the threespine stickleback fish.

Hormones play a prominent role in animal development, mediating the expression of traits and coordinating phenotypic responses to the environment. Their role as physiological integrators has implications for how populations respond to natural selection and can impact the speed and direction of evolutionary change. However, many emerging and established fish models with the potential to be ecologically or evolutionarily informative are small-bodied, making hormone sampling through traditional methods (whole-body or plasma) lethal or highly disruptive. Sampling methodology has thus restricted study design, often limiting sample sizes, and has prevented the study of at-risk/endangered populations. We utilize water-borne hormone sampling, a minimally invasive method of measuring the rate of steroid hormone release across the gills and further validate this method in a novel, evolutionary context. First, we compare water-borne hormone measures of cortisol with those quantified from plasma and whole-body samples collected from the same individuals to establish the relationship between concentrations quantified via the three methods. We then compare the release of steroid hormones in three populations of threespine stickleback to establish the sensitivity of this tool in measuring within-individual and between-individual variation in biologically relevant contexts (reproductive stages), and in assessing differences among populations with distinct evolutionary histories. We demonstrate a strong positive relationship between cortisol concentrations measured with water-borne, plasma, and whole-body collection techniques. Tracking estradiol and testosterone throughout clutch production in females produced anticipated patterns associated with growing and maturing eggs, with divergence in estradiol production in one population. Additionally, differences among populations in cortisol levels at ovulation paralleled the relative presence of a social stressor, and thus expected energetic needs within each population. We confirm that water-borne hormone sampling is sufficiently sensitive to capture biologically relevant fluctuations in steroid hormones between environmental contexts and demonstrate that among-population differences are detectable. This technique can be applied broadly to small fish to answer important ecological and evolutionary questions. By linking population variation in hormones and the multivariate phenotype, this technique will help elucidate both proximate mechanisms underlying phenotypic development and variation, and the way hormone networks alter evolutionary responses to selection.

Heart Rate Variability and Training Load Among National Collegiate Athletic Association Division 1 College Football Players Throughout Spring Camp.

Flatt, AA, Esco, MR, Allen, JR, Robinson, JB, Earley, RL, Fedewa, MV, Bragg, A, Keith, CM, and Wingo, JE. Heart rate variability and training load among National Collegiate Athletic Association Division 1 college football players throughout spring camp. J Strength Cond Res 32(11): 3127-3134, 2018-The purpose of this study was to determine whether recovery of cardiac-autonomic activity to baseline occurs between consecutive-day training sessions among positional groups of a collegiate football team during Spring camp. A secondary aim was to evaluate relationships between chronic (i.e., 4-week) heart rate variability (HRV) and training load parameters. Baseline HRV (lnRMSSD_BL) was compared with HRV after ∼20 hours of recovery before next-day training (lnRMSSDpost20) among positional groups composed of SKILL (n = 11), MID-SKILL (n = 9), and LINEMEN (n = 5) with a linear mixed model and effect sizes (ES). Pearson and partial correlations were used to quantify relationships between chronic mean and coefficient of variation (CV) of lnRMSSD (lnRMSSD_chronic and lnRMSSDcv, respectively) with the mean and CV of PlayerLoad (PL_chronic and PL_cv, respectively). A position × time interaction was observed for lnRMSSD (p = 0.01). lnRMSSD_BL was higher than lnRMSSDpost20 for LINEMEN (p < 0.01; ES = large), whereas differences for SKILL and MID-SKILL were not statistically different (p > 0.05). Players with greater body mass experienced larger reductions in lnRMSSD (r = -0.62, p < 0.01). Longitudinally, lnRMSSDcv was significantly related to body mass (r = 0.48) and PL_chronic (r = -0.60). After adjusting for body mass, lnRMSSDcv and PL_chronic remained significantly related (r = -0.43). The ∼20-hour recovery time between training sessions on consecutive days may not be adequate for restoration of cardiac-parasympathetic activity to baseline among LINEMEN. Players with a lower chronic training load throughout camp experienced greater fluctuation in lnRMSSD (i.e., lnRMSSDcv) and vice versa. Thus, a capacity for greater chronic workloads may be protective against perturbations in cardiac-autonomic homeostasis among American college football players.

Phenotypic differences between the sexes in the sexually plastic mangrove rivulus fish (Kryptolebias marmoratus).

To maximize reproductive success, many animal species have evolved functional sex change. Theory predicts that transitions between sexes should occur when the fitness payoff of the current sex is exceeded by the fitness payoff of the opposite sex. We examined phenotypic differences between the sexes in a sex-changing vertebrate, the mangrove rivulus fish (Kryptolebias marmoratus), to elucidate potential factors that might drive the 'decision' to switch sex. Rivulus populations consist of self-fertilizing hermaphrodites and males. Hermaphrodites transition into males under certain environmental conditions, affording us the opportunity to generate 40 hermaphrodite-male pairs where, within a pair, individuals possessed identical genotypes despite being different sexes. We quantified steroid hormone levels, behavior (aggression and risk taking), metabolism and morphology (organ masses). We found that hermaphrodites were more aggressive and risk averse, and had higher maximum metabolic rates and larger gonadosomatic indices. Males had higher steroid hormone levels and showed correlations among hormones that hermaphrodites lacked. Males also had greater total mass and somatic body mass and possessed considerable fat stores. Our findings suggest that there are major differences between the sexes in energy allocation, with hermaphrodites exhibiting elevated maximum metabolic rates, and showing evidence of favoring investments in reproductive tissues over somatic growth. Our study serves as the foundation for future research investigating how environmental challenges affect both physiology and reproductive investment and, ultimately, how these changes dictate the transition between sexes.

Genetic Subdivision and Variation in Selfing Rates Among Central American Populations of the Mangrove Rivulus, Kryptolebias marmoratus.

We used 32 polymorphic microsatellite loci to investigate how a mixed-mating system affects population genetic structure in Central American populations (N = 243 individuals) of the killifish Kryptolebias marmoratus (mangrove rivulus), 1 of 2 of the world's only known self-fertilizing vertebrates. Results were also compared with previous microsatellite surveys of Floridian populations of this species. For several populations in Belize and Honduras, population structure and genetic differentiation were pronounced and higher than in Florida, even though the opposite trend was expected because populations in the latter region were presumably smaller and highly selfing. The deduced frequency of selfing (s) ranged from s = 0.39-0.99 across geographic locales in Central America. This heterogeneity in selfing rates was in stark contrast to Florida, where s > 0.9. The frequency of outcrossing in a population (t = 1 - s) was tenuously correlated with local frequencies of males, suggesting that males are one of many factors influencing outcrossing. Observed distributions of individual heterozygosity showed good agreement with expected distributions under an equilibrium mixed-mating model, indicating that rates of selfing remained relatively constant over many generations. Overall, our results demonstrate the profound consequences of a mixed-mating system for the genetic architecture of a hermaphroditic vertebrate.

Fighting experience alters brain androgen receptor expression dependent on testosterone status.

Contest decisions are influenced by the outcomes of recent fights (winner-loser effects). Steroid hormones and serotonin are closely associated with aggression and therefore probably also play important roles in mediating winner-loser effects. In mangrove rivulus fish, Kryptolebias marmoratus, individuals with higher testosterone (T), 11-ketotestosterone and cortisol levels are more capable of winning, but titres of these hormones do not directly mediate winner-loser effects. In this study, we investigated the effects of winning/losing experiences on brain expression levels of the receptor genes for androgen (AR), oestrogen α/ß (ERα/ß), glucocorticoid (GR) and serotonin (5-HT1AR). The effect of contest experience on AR gene expression depended on T levels: repeated losses decreased, whereas repeated wins increased AR gene expression in individuals with low T but not in individuals with medium or high T levels. These results lend strong support for AR being involved in mediating winner-loser effects, which, in previous studies, were more detectable in individuals with lower T. Furthermore, the expression levels of ERα/ß, 5-HT1AR and GR genes were higher in individuals that initiated contests against larger opponents than in those that did not. Overall, contest experience, underlying endocrine state and hormone and serotonin receptor expression patterns interacted to modulate contest decisions jointly.