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Aggression is a crucial behavior that impacts access to limited resources in different environmental contexts. Androgens synthesized by the gonads promote aggression during the breeding season. However, aggression can be expressed during the non-breeding season, despite low androgen synthesis by the gonads. The brain can also synthesize steroids ("neurosteroids"), including androgens, which might promote aggression during the non-breeding season. Male song sparrows, Melospiza melodia, are territorial year-round and allow the study of seasonal changes in the steroid modulation of aggression. Here, we quantified steroids following a simulated territorial intrusion (STI) for 10 min in wild adult male song sparrows during the breeding and non-breeding seasons. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we examined 11 steroids: pregnenolone, progesterone, corticosterone, dehydroepiandrosterone, androstenedione, testosterone, 5α-dihydrotestosterone, 17β-estradiol, 17α-estradiol, estriol, and estrone. Steroids were measured in blood and 10 microdissected brain regions that regulate social behavior. In both seasons, STI increased corticosterone in the blood and brain. In the breeding season, STI had no rapid effects on androgens or estrogens. Intriguingly, in the non-breeding season, STI increased testosterone and androstenedione in several behaviorally relevant regions, but not in the blood, where androgens remained non-detectable. Also in the non-breeding season, STI increased progesterone in the blood and specific brain regions. Overall, rapid socially modulated changes in brain steroid levels are more prominent during the non-breeding season. Brain steroid levels vary with season and social context in a region-specific manner and suggest a role for neuroandrogens in aggression during the non-breeding season. [ABSTRACT FROM AUTHOR]

Testosterone promotes mating effort, which involves intraspecific aggression for males of many species. Therefore, males with higher testosterone levels are often thought to be more aggressive. For mammals living in multimale groups, aggression is hypothesized to link male social status (i.e. dominance rank) and testosterone levels, given that high status predicts mating success and is acquired partly through aggressive intragroup competition. In male chimpanzees, Pan troglodytes , dominance rank has been repeatedly linked to interindividual variation in testosterone levels, but evidence directly linking interindividual variation in testosterone and aggression is lacking. In the present study, we test both aggression levels and lean muscle mass, as measured by urinary creatinine, as links between dominance rank and testosterone levels in a large sample of wild male chimpanzees. Multivariate analyses indicated that dominance rank was positively associated with total rates of intragroup aggression, average urinary testosterone levels and average urinary creatinine levels. Testosterone was positively associated with creatinine levels but negatively associated with total aggression rates. Furthermore, mediation analyses showed that testosterone levels facilitated an association between dominance rank and creatinine levels. Our results indicate that (1) adult male chimpanzees with higher average testosterone levels are often higher ranking but not more aggressive than males with lower testosterone and (2) lean muscle mass links dominance rank and testosterone levels in Ngogo males. We assert that aggression rates are insufficient to explain links between dominance rank and testosterone levels in male chimpanzees and that other social variables (e.g. male–male relationship quality) may regulate testosterone's links to aggression. • We examined rank, testosterone, aggression and creatinine in male chimpanzees. • Urinary testosterone levels were positively associated with male dominance rank. • Urinary testosterone levels were negatively associated with aggression rates. • Urinary creatinine was positively associated with dominance rank and testosterone. • Urinary testosterone mediated dominance rank's association with urinary creatinine. [ABSTRACT FROM AUTHOR]

Simple Summary: In most primates, the "challenge hypothesis" predicts increases in male–male aggressive behavior and testosterone during reproductive challenges, i.e., mating competition, or during periods of social instability, especially in seasonal breeders. However, few studies have investigated the role of stress hormones, such as glucocorticoids, and social rank in non-seasonal primates. In this study, we tested some predictors of the "challenge hypothesis" during a period of social stability in a non-seasonal primate. We collected data on aggressive behavior (male-to-male and male-to-female), copulation, and fecal samples (n = 700) to quantify testosterone and glucocorticoid metabolites (fTm and fGCm, respectively) in seven adults of stumptail macaques (Macaca arctoides) living in semi-captivity. We found that fTm and fGCm increased during times of copulation, although neither of the two hormones were associated with aggression between males. Besides, fGCm levels were positively associated with male-to-female aggression, indicating a possible social stress challenge. Both fTm and fGCm levels were higher in higher- and middle-ranking males. Our findings partially support the "challenge hypothesis" in stumptail males and highlight the importance of glucocorticoid secretion during challenging situations; our results also indicate that during periods of social stability, testosterone is not associated with aggression, at least in a non-seasonal primate. The "challenge hypothesis" predicts higher male–male aggressive behavior along with increases in testosterone levels during times of reproductive challenges and social instability. In addition, in some primate species, higher glucocorticoid levels can be observed as well, but this is usually modulated by dominance rank. We studied rank-related aggressive behavior, mating activity, and fecal testosterone and glucocorticoid metabolites (fTm and fGCm) in male stumptail macaques (Macaca arctoides) in order to test some predictions of the "challenge hypothesis". Over a 20-month period, we collected data on aggressive behavior and copulation, as well as fecal samples (n = 700) to quantify fTm and fGCm in seven adult stumptail males living in captivity. During periods of mating activity, male-to-male aggression increased in higher- and middle-ranking males. Neither fTm nor fGCm levels predicted male-to-male aggression. fGCm levels (but not fTm) were positively associated with male-to-female aggression; however, this association was pronounced during periods of mating activity. fGCm levels differed according to social rank, with middle-ranking males having the highest levels. Both hormones were higher during periods of mating activity, but only in higher- and middle-ranking males. Taken together, our findings partially support the challenge hypothesis in a non-seasonal primate and shed some light on the unique social and mating system of the stumptail macaque. [ABSTRACT FROM AUTHOR]

Our review focuses on findings from mammals as part of a Special Issue "30th Anniversary of the Challenge Hypothesis". Here we put forth an integration of the mechanisms through which testosterone controls territorial behavior and consider how reproductive experience may alter these mechanisms. The emphasis is placed on the function of socially induced increases in testosterone (T) pulses, which occur in response to social interactions, as elegantly developed by Wingfield and colleagues. We focus on findings from the monogamous California mouse, as data from this species shows that reproductive status is a key factor influencing social interactions, site fidelity, and vigilance for offspring defense. Specifically, we examine differences in T pulses in sexually naïve versus sexually experienced pair bonded males. Testosterone pulses influence processes such as social decision making, the winner-challenge effect, and location preferences through rewarding effects of T. We also consider how social and predatory vigilance contribute to T pulses and how these interactions contribute to a territory centered around maximizing reproduction. Possible underlying mechanisms for these effects include the nucleus accumbens (rewarding effects of testosterone), hippocampus (spatial memories for territories), and the bed nucleus of the stria terminalis (social vigilance). The development of the challenge effect has provided an ideal framework for understanding the complex network of behavioral, environmental, physiological and neural mechanisms that ultimately relates to competition and territoriality across taxa. The opportunity to merge research on the challenge effect using both laboratory and field research to understand social behavior is unparalleled.

For decades, avian endocrinology has been informed bymale perspectives andmalefocused research, leaving significant gaps in our understanding of female birds. Male birds have been favored as research subjects because their reproductive behaviors are considered more conspicuous and their reproductive physiology is presumably less complex than female birds. However, female birds should not be ignored, as female reproductive behavior and physiology are essential for the propagation of all avian species. Endocrine research in female birds has made much progress in the last 20 years, but a substantial disparity in knowledge between male and female endocrinology persists. In this perspective piece, we provide examples of why ornithology has neglected female endocrinology, and we propose considerations for field and laboratory techniques to facilitate future studies. We highlight recent advances that showcase the importance of female avian endocrinology, and we challenge historic applications of an oversimplified, male-biased lens. We further provide examples of species for which avian behavior differs from the stereotypically described behaviors of male and female birds, warning investigators of the pitfalls in approaching endocrinology with a binary bias. We hope this piece will inspire investigators to engage in more comprehensive studies with female birds, to close the knowledge gap between the sexes, and to look beyond the binary when drawing conclusions about what is 'male' versus 'female' biology. [ABSTRACT FROM AUTHOR]

For decades, avian endocrinology has been informed by male perspectives and male-focused research, leaving significant gaps in our understanding of female birds. Male birds have been favored as research subjects because their reproductive behaviors are considered more conspicuous and their reproductive physiology is presumably less complex than female birds. However, female birds should not be ignored, as female reproductive behavior and physiology are essential for the propagation of all avian species. Endocrine research in female birds has made much progress in the last 20 years, but a substantial disparity in knowledge between male and female endocrinology persists. In this perspective piece, we provide examples of why ornithology has neglected female endocrinology, and we propose considerations for field and laboratory techniques to facilitate future studies. We highlight recent advances that showcase the importance of female avian endocrinology, and we challenge historic applications of an oversimplified, male-biased lens. We further provide examples of species for which avian behavior differs from the stereotypically described behaviors of male and female birds, warning investigators of the pitfalls in approaching endocrinology with a binary bias. We hope this piece will inspire investigators to engage in more comprehensive studies with female birds, to close the knowledge gap between the sexes, and to look beyond the binary when drawing conclusions about what is ‘male’ versus ‘female’ biology.

The “challenge hypothesis” predicts higher male–male aggressive behavior along with increases in testosterone levels during times of reproductive challenges and social instability. In addition, in some primate species, higher glucocorticoid levels can be observed as well, but this is usually modulated by dominance rank. We studied rank-related aggressive behavior, mating activity, and fecal testosterone and glucocorticoid metabolites (fTm and fGCm) in male stumptail macaques (Macaca arctoides) in order to test some predictions of the “challenge hypothesis”. Over a 20-month period, we collected data on aggressive behavior and copulation, as well as fecal samples (n = 700) to quantify fTm and fGCm in seven adult stumptail males living in captivity. During periods of mating activity, male-to-male aggression increased in higher- and middle-ranking males. Neither fTm nor fGCm levels predicted male-to-male aggression. fGCm levels (but not fTm) were positively associated with male-to-female aggression; however, this association was pronounced during periods of mating activity. fGCm levels differed according to social rank, with middle-ranking males having the highest levels. Both hormones were higher during periods of mating activity, but only in higher- and middle-ranking males. Taken together, our findings partially support the challenge hypothesis in a non-seasonal primate and shed some light on the unique social and mating system of the stumptail macaque.

The Challenge Hypothesis, designed originally to explain the patterning of competitive behavior and androgen levels in seasonally breeding birds, predicts that males will increase their androgen levels in order to become more competitive in reproductive contexts. Here we test predictions derived from the Challenge Hypothesis in white-faced capuchin monkeys (Cebus capucinus), a species that has somewhat seasonal reproduction. We analyzed demographic and hormonal data collected over a 5.25-year period, from 18 males in nine social groups living in or near Lomas Barbudal Biological Reserve, Costa Rica. Alpha males had higher androgen levels than subordinates. Contrary to our predictions, neither the number of breeding-age males nor the number of potentially fertile females was obviously associated with androgen levels. Furthermore, male androgen levels were not significantly linked to social stability, as measured by stability of male group membership or recency of change in the alpha male position. Androgen levels changed seasonally, but not in a manner that had an obvious relationship to predictions from the Challenge Hypothesis: levels were generally at their lowest near the beginning of the conception season, but instead of peaking when reproductive opportunities were greatest, they were at their highest near the end of the conception season or shortly thereafter. This lack of correspondence to the timing of conceptions suggests that there may be ecological factors not yet identified that influence ifA levels. We expected that the presence of offspring who were young enough to be vulnerable to infanticide during an alpha male takeover might influence androgen levels, at least in the alpha male, but this variable did not significantly impact results.

A contribution to a special issue on Hormones and Human Competition. The relationship between androgenic hormones, like testosterone (T), and aggression is extensively studied in human populations. Yet, while this work has illuminated a variety of principals regarding the behavioral and phenotypic effects of T, it is also hindered by inherent limitations of performing research on people. In these instances, animal research can be used to gain further insight into the complex mechanisms by which T influences aggression. Here, we explore recent studies on T and aggression in numerous vertebrate species, although we focus primarily on males and on a New World rodent called the California mouse (Peromyscus californicus). This species is highly territorial and monogamous, resembling the modern human social disposition. We review (i) how baseline and dynamic T levels predict and/or impact aggressive behavior and disposition; (ii) how factors related to social and physical context influence T and aggression; (iii) the reinforcing or "rewarding" aspects of aggressive behavior; and (iv) the function of T on aggression before and during a combative encounter. Included are areas that may need further research. We argue that animal studies investigating these topics fill in gaps to help paint a more complete picture of how androgenic steroids drive the output of aggressive behavior in all animals, including humans.

The Challenge Hypothesis is an influential framework for understanding how androgens are involved in the promotion of competitive behavior during mating-related challenges and has been tested extensively in studies across scientific disciplines. Mixed support in psychological research led scholars to develop the Dual Hormone Hypothesis as a potential path forward, which argues that glucocorticoids moderate the relationship between androgens and status-striving. In the current study, we examine the Challenge Hypothesis and the Dual Hormone Hypothesis in wild male mountain gorillas, representing the first time the latter hypothesis has been tested in a non-human primate. In a sample of 30 adult males comprising over 600 days of observation, we find some limited support for the Challenge Hypothesis. Greater daily rates of targeted aggression toward other adult males corresponded to higher fecal androgen metabolites 1–2 days following observations, though this pattern did not fully generalize to dominance rank or other competitive behaviors examined. However, we find no support for the Dual Hormone Hypothesis: neither dominance rank nor any category of competitive behavior was predicted by the interaction between androgens and glucocorticoids. We close by discussing how this initial investigation might be leveraged toward the development of an expanded Dual Hormone Hypothesis that draws on the large evidence base in primate behavioral ecology. • We test the Challenge and Dual Hormone Hypotheses (DHH) in wild male mountain gorillas. • Androgens predicted some measures of competitive behavior, but support was inconsistent. • Associations with androgens were not moderated by glucocorticoids, contrary to the DHH. • Considering socioecology is key for adapting the DHH to non-human primates. [ABSTRACT FROM AUTHOR]

It has been hypothesized that androgens respond to the social interactions as a way to adjust the behavior of individuals to the challenges of the social environment in an adaptive manner. Therefore, it is expected that transient changes in circulating androgen levels within physiological scope should impact the state of the brain network that regulates social behavior, which should translate into adaptive behavioral changes. Here, we examined the effect that a transient peak in androgen circulating levels, which mimics socially driven changes in androgen levels, has on the forebrain state, which harbors most nuclei of the social decision-making network. For this purpose, we successfully induced transient changes in circulating androgen levels in an African cichlid fish (Mozambique tilapia, Oreochromis mossambicus) commonly used as a model in behavioral neuroendocrinology by injecting 11-ketotestosterone or testosterone, and compared the forebrain transcriptome of these individuals to control fish injected with vehicle. Forebrain samples were collected 30 min and 60 min after injection and analyzed using RNAseq. Our results showed that a transient peak in 11-ketotestosterone drives more accentuated changes in forebrain transcriptome than testosterone, and that transcriptomic impact was greater at the 30 min than at the 60 min post-androgen administration. Several genes involved in the regulation of translation, steroid metabolism, ion channel membrane receptors, and genes involved in epigenetic mechanisms were differentially expressed after 11-ketotestosterone or testosterone injection. In summary, this study identified specific candidate genes that may regulate socially driven changes in behavioral flexibility mediated by androgens.

Testosterone (T) is a sexual hormone capable of modulating several traits in birds, including aggressiveness and reproductive behavior. Although variation in T-related traits is well-known for temperate zone birds, this variation has not been extensively studied in tropical species. The campo miner (Geositta poeciloptera) is a threatened bird endemic to the grasslands of the South American Cerrado. We investigated the seasonal variation in plasma T levels and associated behavior in the campo miner, addressing the following questions: (1) Does the species exhibit seasonal variation in T profile? (2) Do males have higher plasma T levels than females, irrespective of season? (3) Are males with higher plasma T levels more aggressive than males with lower T levels? (4) Do males' plasma T levels decrease after females lay eggs? We found that T levels are higher during the breeding season than during the nonbreeding season and that males present higher T levels than females throughout the year. Such high T levels are associated with a higher probability to engage in aggressive behavior; however, T levels decline toward the egg-laying date and keep decreasing afterward. Higher T levels before egg laying are apparently related to territorial defense against invaders and extrapair copulations. With the beginning of parental care, T levels decrease, which is in line with previous observations that the species becomes less aggressive after egg laying. This study contributes to the understanding of environmental endocrinology of tropical birds, filling some knowledge gaps about the diverse Neotropical avifauna. [ABSTRACT FROM AUTHOR]

Testosterone plays an important role in mediating male reproductive trade-offs in many vertebrate species, augmenting muscle and influencing behavior necessary for male-male competition and mating-effort. Among humans, testosterone may also play a key role in facilitating male provisioning of offspring as muscular and neuromuscular performance are deeply influenced by acute changes in testosterone. This study examines acute changes in salivary testosterone among 63 Tsimane men ranging in age from 16-80 (mean 38.2) years during one-hour bouts of tree-chopping while clearing horticultural plots. The Tsimane forager-horticulturalists living in the Bolivian Amazon experience high energy expenditure associated with food production, have high levels of parasites and pathogens, and display significantly lower baseline salivary testosterone than age-matched US males. Mixed-effects models controlling for BMI and time of specimen collection reveal increased salivary testosterone (p

It has been hypothesized that androgens respond to the social interactions as a way to adjust the behavior of individuals to the challenges of the social environment in an adaptive manner. Therefore, it is expected that transient changes in circulating androgen levels within physiological scope should impact the state of the brain network that regulates social behavior, which should translate into adaptive behavioral changes. Here, we examined the effect that a transient peak in androgen circulating levels, which mimics socially driven changes in androgen levels, has on the forebrain state, which harbors most nuclei of the social decision-making network. For this purpose, we successfully induced transient changes in circulating androgen levels in an African cichlid fish (Mozambique tilapia, Oreochromis mossambicus) commonly used as a model in behavioral neuroendocrinology by injecting 11-ketotestosterone or testosterone, and compared the forebrain transcriptome of these individuals to control fish injected with vehicle. Forebrain samples were collected 30 min and 60 min after injection and analyzed using RNAseq. Our results showed that a transient peak in 11-ketotestosterone drives more accentuated changes in forebrain transcriptome than testosterone, and that transcriptomic impact was greater at the 30 min than at the 60 min post-androgen administration. Several genes involved in the regulation of translation, steroid metabolism, ion channel membrane receptors, and genes involved in epigenetic mechanisms were differentially expressed after 11-ketotestosterone or testosterone injection. In summary, this study identified specific candidate genes that may regulate socially driven changes in behavioral flexibility mediated by androgens. [ABSTRACT FROM AUTHOR]

The association between androgens and competition in women has been understudied compared with men. The current study examined the link between testosterone (T) and competition in elite female athletes, using a sample of female wrestlers that included athletes competing at both the national and international level. In a repeated-measures design, saliva samples were collected before and after wrestling bouts, with comparable samples of wins and losses, and subsequently analyzed for T. Study results showed a 22% increase in circulating bioavailable T from pre- to postbout, F(1, 12) = 9.71, P = .009. There was no significant difference in T between win or loss outcomes. These findings—showing a link between individual head-to-head competition and T in women—demonstrate that women's androgenic responses to environmental contexts are dynamic and may be an important factor to address in research on competitive performance. [ABSTRACT FROM AUTHOR]

This study examines factors affecting androgen and cortisol levels in wild, male golden lion tamarins (Leontopithecus rosalia). Golden lion tamarins are a cooperatively breeding species in which groups often contain two potentially breeding males. Brothers frequently emigrate together and develop a clear dominance hierarchy, but interactions between them are primarily affiliative. Duos in which the males are not related are less stable. In addition, reproductive skew theory predicts that dominant males will be less likely to share reproduction with related subordinates. As such, we predicted that both androgens and cortisol would be higher in subordinate males unrelated to the dominant male. We also predicted that androgens in breeding males would be higher during the mating season than the birth/infant care season, as per Wingfield's "challenge hypothesis" (1990). Fecal samples were collected from 24 males in 14 social groups and assayed by enzyme immunoassay. Androgen levels were higher in breeding males during the mating season, thus supporting the challenge hypothesis. However, while subordinate males unrelated to the dominant male had significantly lower androgens than any other group, cortisol levels were not correspondingly higher. These results suggest that unrelated subordinate males show measurable reproductive suppression and may use strategies such as infantilization to avert aggression from dominant males.

Social living animals have to adjust their behavior to rapid changes in the social environment. It has been hypothesized that the expression of social behavior is better explained by the activity pattern of a diffuse social decision-making network (SDMN) in the brain than by the activity of a single brain region. In this study, we tested the hypothesis that it is the assessment that individuals make of the outcome of the fights, rather than the expression of aggressive behavior per se, that triggers changes in the pattern of activation of the SDMN which are reflected in socially driven behavioral profiles (e.g., dominant vs. subordinate specific behaviors). For this purpose, we manipulated the perception of the outcome of an agonistic interaction in an African cichlid fish (Oreochromis mossambicus) and assessed if either the perception of outcome or fighting by itself was sufficient to trigger rapid changes in the activity of the SDMN. We have used the expression of immediate early genes (c-fos and egr-1) as a proxy to measure the neuronal activity in the brain. Fish fought their own image on a mirror for 15 min after which they were allocated to one of three conditions for the two last minutes of the trial: (1) they remained fighting the mirror image (no outcome treatment); (2) the mirror was lifted and a dominant male that had just won a fight was presented behind a transparent partition (perception of defeat treatment); and (3) the mirror was lifted and a subordinate male that had just lost a fight was presented behind a transparent partition (perception of victory treatment). Results show that these short-term social interactions elicit distinct patterns in the SDMN and that the perception of the outcome was not a necessary condition to trigger a SDMN response as evidenced in the second treatment (perception of defeat treatment). We suggest that the mutual assessment of relative fighting behavior drives these acute changes in the state of the SDMN.

Objectives This study examined potentially bidirectional connections of older U.S. women's salivary testosterone with their social network connections. Methods Data were from the 2005–2006 and 2010–2011 waves of the National Social Life, Health and Aging Project (NSHAP), a national probability sample of older U.S. adults. Autoregressive cross-lagged panel models tested linkages of women's testosterone with their social networks. Results Consistent with recent biological theory suggesting social modulation of hormones, a higher kin proportion in one's egocentric (person-centered) network, arguably a stable compositional feature, negatively predicted women's testosterone levels. In contrast, findings for tie strength were consistent with hormonal regulation of women's sociality—with both perceived support from friends and family, and closeness to network members, negatively influenced by testosterone. Discussion Rather than being a static and exogenous biological factor, older women's testosterone levels seem partly an outcome of their social context. Implications for sexual health and hormone therapy are discussed. However, this androgen also influences dimensions of their intimate networks critical to successful aging. Findings suggest the need for social scientists to engage with the neuroendocrine literature, which offers suggestions on linkages of hormones with specific network patterns. [ABSTRACT FROM AUTHOR]

Social living animals have to adjust their behavior to rapid changes in the social environment. It has been hypothesized that the expression of social behavior is better explained by the activity pattern of a diffuse social decision-making network (SDMN) in the brain than by the activity of a single brain region. In this study, we tested the hypothesis that it is the assessment that individuals make of the outcome of the fights, rather than the expression of aggressive behavior per se , that triggers changes in the pattern of activation of the SDMN which are reflected in socially driven behavioral profiles (e.g., dominant vs. subordinate specific behaviors). For this purpose, we manipulated the perception of the outcome of an agonistic interaction in an African cichlid fish (Oreochromis mossambicus) and assessed if either the perception of outcome or fighting by itself was sufficient to trigger rapid changes in the activity of the SDMN. We have used the expression of immediate early genes (c-fos and egr-1) as a proxy to measure the neuronal activity in the brain. Fish fought their own image on a mirror for 15 min after which they were allocated to one of three conditions for the two last minutes of the trial: (1) they remained fighting the mirror image (no outcome treatment); (2) the mirror was lifted and a dominant male that had just won a fight was presented behind a transparent partition (perception of defeat treatment); and (3) the mirror was lifted and a subordinate male that had just lost a fight was presented behind a transparent partition (perception of victory treatment). Results show that these short-term social interactions elicit distinct patterns in the SDMN and that the perception of the outcome was not a necessary condition to trigger a SDMN response as evidenced in the second treatment (perception of defeat treatment). We suggest that the mutual assessment of relative fighting behavior drives these acute changes in the state of the SDMN. [ABSTRACT FROM AUTHOR]

Highlights • The Challenge Hypothesis (CH) concerns the dynamics of male life-history strategies. • The CH predicts that testosterone (T) mediates shifts between mating and parenting. • In humans, pair-bonding and fatherhood generally correspond to lower T. • We meta-analyze 114 effects to quantify magnitudes of predicted CH effects in men. • We find both robust effects and evidence of selective reporting. Abstract Males of many species must allocate limited energy budgets between mating and parenting effort. The Challenge Hypothesis provides a framework for understanding these life-history trade-offs via the disparate roles of testosterone (T) in aggression, sexual behavior, and parenting. It predicts that males pursuing mating opportunities have higher T than males pursuing paternal strategies, and in humans, many studies indeed report that men who are fathers and/or pair-bonded have lower T than childless and/or unpaired men. However, the magnitude of these effects, and the influence of methodological variation on effect sizes, have not been quantitatively assessed. We meta-analyzed 114 effects from 66 published and unpublished studies covering four predictions inspired by the Challenge Hypothesis. We confirm that pair-bonded men have lower T than single men, and fathers have lower T than childless men. Furthermore, men more oriented toward pair-bonding or offspring investment had lower T. We discuss the practical meaningfulness of the effect sizes we estimate in relation to known factors (e.g., aging, geographic population) that influence men's T concentrations. [ABSTRACT FROM AUTHOR]

Testosterone affects human social behavior in various ways. While testosterone effects are generally associated with muscular strength and aggressiveness, human studies also point towards enhanced status–seeking motives after testosterone administration. The current study tested the causal influence of exogenous testosterone on male behavior during a competitive provocation paradigm. In this double blind, randomized, placebo (PL)-controlled study, 103 males were assigned to a PL or testosterone group receiving a colorless PL or testosterone gel. To induce provocation, males played a rigged reaction time game against an ostensible opponent. When participants lost, the opponent subtracted money from the participant who in return could subtract money from the ostensible opponent. Participants subjectively indicated anger and self-estimated treatment affiliation (testosterone or PL administration). A trial-by-trial analysis demonstrated that provocation and success during the repeated games had a stronger influence on participants’ choice to reduce money from the opponent if they had received testosterone. Participants who believed to be in the testosterone group were angrier after the experiment and increased monetary reductions during the task course. In line with theories about mechanisms of testosterone in humans, provocation is shown to be necessary for the agency of exogenous testosterone. Thus, testosterone reinforces the conditional adjustment of aggressive behavior but not aggressive behavior per se. In contrast undirected frustration is not increased by testosterone but probably interferes with cognitive appraisals about biological mechanisms of testosterone.

Objective: Cross‐culturally, men's T declines in response to pairbonding and fatherhood, but less is known about what happens to T during and after life history transitions that theoretically lead to renewed mating effort. We tested whether men's T rises (or declines less with age) as their children age, or when pairbonds end, independent of changes in fatherhood‐related variables such as co‐residence with children. Methods: We used demographic, behavioral, and salivary hormone data (waking and pre‐bed T) collected in 2009 and 2014 for the Cebu Longitudinal Health and Nutrition Survey (n = 571 men). Results: Fathers with older children tended to have attenuated decline in pre‐bedtime T between 2009 and 2014 compared to men with younger children, after controlling for pairbonding (ß = 1.58, SE = 0.88, P = 0.074). Separated men had higher pre‐bedtime T than pairbonded men, controlling for fatherhood‐related variables (ß = 11.74, SE = 4.33, P = 0.007). Change in T did not significantly differ for men who separated between the two surveys, relative to men who remained pairbonded throughout. Conclusion: We found modest support for the prediction that men experience less of an age‐related drop in T as their youngest child ages, a trend that might strengthen as children age further. We also replicate the finding that separated men have higher T, although longitudinal changes in the hormone were not significantly different in these men. Our data suggest that, of two life history transitions that may predict renewed mating effort, pair bond loss is more strongly endocrine mediated than potential mating effort shifts related to the aging of children. [ABSTRACT FROM AUTHOR]

Territorial aggression has been widely studied in males and it has been historically suggested that androgens are key mediators of this behavior. However, more recent evidence suggests that it is the aromatization to estrogens, rather than androgens themselves, that is key to regulating this behavior. Females also display aggressive behaviors, but the physiological regulation of female aggression is still understudied when compared to males. In this context, the challenge hypothesis postulates that male-male aggressive interactions stimulate the production of androgens in males in periods of social instability. Here we determine plasma sex steroid levels in Cichlasoma dimerus to assess whether estrogens are related to aggressive behavior and to test the challenge hypothesis in both males and females. We set-up challenge trials as intrasexual dyadic encounters and determined androgen and estrogen levels before and after the trial in both winners and losers. Even though there were no differences in initial estradiol-17β plasma levels between male winners and losers, initial levels were higher ( p  = .046) in female winners than in losers, while there were no differences in testosterone or 11-ketotestosterone levels. After trials, both males and females showed elevated levels of estradiol-17β and both androgens, but only males exhibited a significant 1.45, 5.42 and 3.2-fold increase in estradiol-17β, testosterone and 11-ketotestosterone, respectively ( p  = .023, p  = .016, p  = .018). Moreover, changes in circulating levels of estradiol-17β in females after the trials do not depend on their reproductive status or on the outcome of the contest. We suggest that female aggression is associated with initial levels of estradiol-17β, and that the challenge hypothesis, originally defined for androgens, could also be extended to estrogens. [ABSTRACT FROM AUTHOR]

In many vertebrates, males increase circulating testosterone (T) levels in response to seasonal and social changes in competition. Females are also capable of producing and responding to T, but the full extent to which they can elevate T across life history stages remains unclear. Here we investigated T production during various breeding stages in female tree swallows ( Tachycineta bicolor ), which face intense competition for nesting sites. We performed GnRH and saline injections and compared changes in T levels 30 min before and after injection. We found that GnRH-injected females showed the greatest increases in T during territory establishment and pre-laying stages, whereas saline controls dramatically decreased T production during this time. We also observed elevated rates of conspecific aggression during these early stages of breeding. During incubation and provisioning, however, T levels and T production capabilities declined. Given that high T can disrupt maternal care, an inability to elevate T levels in later breeding stages may be adaptive. Our results highlight the importance of saline controls for contextualizing T production capabilities, and they also suggest that social modulation of T is a potential mechanism by which females may respond to competition, but only during the period of time when competition is most intense. These findings have broad implications for understanding how females can respond to their social environment and how selection may have shaped these hormone-behavior interactions. [ABSTRACT FROM AUTHOR]

Urban songbirds of several species more vigorously defend their territories in response to conspecific song playback than do their rural counterparts, but the hormonal basis of this behavioral difference is unclear. It is well established in vertebrates that both testosterone and corticosterone affect the intensity of territoriality. Previous studies have found no evidence that initial (i.e., immediately following territorial challenge, but prior to restraint) plasma testosterone accounts for the elevated territorial aggression of urban birds. Determining if testosterone still contributes to urban-rural differences in territoriality requires also assessing males' abilities to transiently increase plasma testosterone (in response to an injection of gonadotropin-releasing hormone). We tested whether these hormones are correlated with the territorial response to conspecific song playback in urban and rural male Song Sparrows ( Melospiza melodia ) in Montgomery County, Virginia. We found that the elevated territorial aggression of urban sparrows was not related to variation in either initial plasma testosterone or the ability to transiently increase testosterone. In contrast, despite no overall habitat difference in initial corticosterone, levels of this hormone were positively correlated with territoriality in urban and rural sparrows. Furthermore, for a given level of corticosterone, urban sparrows were more territorially aggressive. Our findings suggest that initial corticosterone may either play a role in the regulation of persistent differences in territorial behavior between free-ranging urban and rural male Song Sparrows or be affected by the intensity of behavioral response to territorial challenge. [ABSTRACT FROM AUTHOR]

Historic bias toward study of sex hormones and sexual ornamentation in males currently constrains our perspective of hormone—behavior—phenotype relationships. Resolving how ornamented female phenotypes evolve is particularly important for understanding the diversity of social signals across taxa. Studies of both males and females in taxa with variable female phenotypes are needed to establish whether sexes share mechanisms underlying expression of signaling phenotypes and behavior. White-shouldered Fairywren (Malurus alboscapulatus) subspecies vary in female ornamentation, baseline circulating androgens, and response to territorial intrusion. The moretoni ornamented female subspecies is characterized by higher female, but lower male baseline androgens, and a stronger pair territorial response relative to pairs from the lorentzi unornamented female subspecies. Here we address whether subspecific differences in female ornamentation, baseline androgens, and pair territoriality are associated with ability to elevate androgens following gonadotropin releasing hormone (GnRH) challenge and in response to simulated territorial intrusion. We find that subspecies do not differ in their capacity to produce androgens in either sex following GnRH or simulated territorial intrusion (STI) challenges. STI-induced androgens were predictive of degree of response to territorial intrusions in females only, but the direction of the effect was mixed. GnRH-induced androgens did not correlate with response to simulated intruders, nor did females sampled during intrusion elevate androgens relative to flushed controls, suggesting that increased androgens are not necessary for the expression of territorial defense behaviors. Collectively, our results suggest that capacity to produce androgens does not underlie subspecific patterns of female ornamentation, territoriality, and baseline plasma androgens. • White-shouldered Fairywren subspecies vary in female ornamentation and androgens. • Degree of androgen elevation following GnRH challenge does not differ by subspecies. • Capacity to produce androgens does not appear to explain subspecific variation in territoriality. • Baseline, but not STI-induced androgens, correlate with STI response in females. [ABSTRACT FROM AUTHOR]

Investigating causes and consequences of variation in hormonal expression is a key focus in behavioral ecology. Many studies have explored patterns of secretion of the androgen testosterone in male vertebrates, using the challenge hypothesis (Wingfield, Hegner, Dufty, & Ball, 1990; The American Naturalist, 136(6), 829-846) as a theoretical framework. Rather than the classic association of testosterone with male sexual behavior, this hypothesis predicts that high levels of testosterone are associated with male-male reproductive competition but also inhibit paternal care. The hypothesis was originally developed for birds, and subsequently tested in other vertebrate taxa, including primates. Such studies have explored the link between testosterone and reproductive aggression as well as other measures of mating competition, or between testosterone and aspects of male behavior related to the presence of infants. Very few studies have simultaneously investigated the links between testosterone and male aggression, other aspects of mating competition and infant-related behavior. We tested predictions derived from the challenge hypothesis in wild male Barbary macaques ( Macaca sylvanus), a species with marked breeding seasonality and high levels of male-infant affiliation, providing a powerful test of this theoretical framework. Over 11 months, 251 hr of behavioral observations and 296 fecal samples were collected from seven adult males in the Middle Atlas Mountains, Morocco. Fecal androgen levels rose before the onset of the mating season, during a period of rank instability, and were positively related to group mating activity across the mating season. Androgen levels were unrelated to rates of male-male aggression in any period, but higher ranked males had higher levels in both the mating season and in the period of rank instability. Lower androgen levels were associated with increased rates of male-infant grooming during the mating and unstable periods. Our results generally support the challenge hypothesis and highlight the importance of considering individual species' behavioral ecology when testing this framework. [ABSTRACT FROM AUTHOR]

A contribution to a special issue on Hormones and Human Competition. The relationship between androgenic hormones, like testosterone (T), and aggression is extensively studied in human populations. Yet, while this work has illuminated a variety of principals regarding the behavioral and phenotypic effects of T, it is also hindered by inherent limitations of performing research on people. In these instances, animal research can be used to gain further insight into the complex mechanisms by which T influences aggression. Here, we explore recent studies on T and aggression in numerous vertebrate species, although we focus primarily on males and on a New World rodent called the California mouse ( Peromyscus californicus ). This species is highly territorial and monogamous, resembling the modern human social disposition. We review (i) how baseline and dynamic T levels predict and/or impact aggressive behavior and disposition; (ii) how factors related to social and physical context influence T and aggression; (iii) the reinforcing or “rewarding” aspects of aggressive behavior; and (iv) the function of T on aggression before and during a combative encounter. Included are areas that may need further research. We argue that animal studies investigating these topics fill in gaps to help paint a more complete picture of how androgenic steroids drive the output of aggressive behavior in all animals, including humans. [ABSTRACT FROM AUTHOR]

A contribution to a special issue on Hormones and Human Competition. Since Archer's (2006) influential meta-analysis, there has been a major increase in the number of studies investigating the effect of competition outcome on testosterone reactivity patterns in humans. Despite this increased research output, there remains debate as to whether competition outcome modulates testosterone concentrations. The present paper examines this question using a meta-analytic approach including papers published over the last 35 years. Moreover, it provides the first meta-analytic estimate of the effect of competition outcome on testosterone concentrations in women. Results from a meta-analysis involving 60 effect sizes and > 2500 participants indicated that winners of a competition demonstrated a larger increase in testosterone concentrations relative to losers ( D = 0.20)—an effect that was highly heterogeneous. This ‘winner-loser’ effect was most robust in studies conducted outside the lab (e.g., in sport venues) ( D = 0.43); for studies conducted in the lab, the effect of competition outcome on testosterone reactivity patterns was relatively weak ( D = 0.08), and only found in studies of men ( D = 0.15; in women: D = − 0.04). Further, the 'winner-loser' effect was stronger among studies in which pre-competition testosterone was sampled earlier than ( D = 0.38, after trim and fill correction) rather than within ( D = 0.09) 10 min of the start of the competition. Therefore, these results also provide important insight regarding study design and methodology, and will be a valuable resource for researchers conducting subsequent studies on the 'winner loser' effect. [ABSTRACT FROM AUTHOR]

Hormones can facilitate the expression of behavior, but relatively few studies have considered individual variation and repeatability in hormone-behavior relationships. Repeated measures of hormones are valuable because repeatability in hormone levels might be a mechanism that drives repeatability in behavior (“personality”). Testosterone is predicted to promote territorial aggression and suppress parental behaviors. In our population of eastern bluebirds ( Sialia sialis ), parental care and nest defense aggression toward a heterospecific are repeatable. We tested the hypothesis that repeatability of testosterone levels within individuals underlies repeatable behaviors observed in our population. We measured nestling provisioning and aggressive nest defense against a heterospecific. After behavioral observations we captured either the male or female bluebird, and determined initial testosterone levels and maximum capacity of the gonads to secrete testosterone by injecting gonadotropin-releasing hormone (GnRH). We found among-individual variation in initial testosterone levels for males and females. Individual males were repeatable in both initial and GnRH-induced testosterone levels across behavioral contexts, while individual females were repeatable in GnRH-induced testosterone levels. However, testosterone levels were not significantly related to parental or nest defense behaviors, suggesting that repeatable testosterone levels may not drive repeatable parental and heterospecific nest defense behaviors in this population. The absence of a relationship between testosterone and parental and heterospecific nest defense behaviors might be due to among-individual variation in testosterone levels. Considering the sources of variation in testosterone levels may reveal why some populations exhibit high individual variation in hormone levels. [ABSTRACT FROM AUTHOR]

Testosterone is an important mediator of behavior, morphology and physiology. A cascade of signals regulates the amount of testosterone (T) circulating in the plasma; in response to stimulus the hypothalamus releases gonadotropin-releasing hormone (GnRH), which triggers secretion of gonadotropins from the pituitary, stimulating the synthesis and release of T from the gonads. Previous work has shown that changes to the social environment can alter circulating T-levels, which may have important fitness consequences, but it is currently unclear whether these changes are due to alterations in the signal from the brain, or changes in the ability of the pituitary and gonads to respond to this signal. Further, the strength and direction of response to a changing environment may differ according to life-history strategy. Species with genetically determined alternative strategies offer a pathway for examining these differences. Here we use a finch with a genetically determined polymorphism, the Gouldian finch ( Erythrura gouldiae ), to determine whether T-levels change in response to social environment. We also use injections of GnRH to determine whether these changes are due to alterations in the ability of the pituitary and gonads to respond to this signal. We found that social environment (presence of females) had a rapid effect on male circulating T-levels, and that this difference was reflected in responsiveness to GnRH. We observed no overall morph differences in T-levels, but we did observe morph differences in the pattern of T secretion across environments, and morph differences in the repeatability of T-levels across time and environment. [ABSTRACT FROM AUTHOR]

How and why individuals differ from each other is a central question in behavioral and evolutionary ecology, because selection particularly acts on this among-individual variation. It is therefore important to accurately partition phenotypic variances into their within- and between-individual components. Partitioning covariances into both components can also inform about underlying mechanistic pathways that potentially interlink trait expression. In the current study, we applied such a (co)variance partitioning approach to test key predictions of two central hypotheses in behavioral ecology, namely the immunocompetence handicap hypothesis and the challenge hypothesis. To this end, we assessed potential pleiotropic effects of testosterone on male sexual signaling, immune function, and parental care. We here repeatedly measured a set of relevant traits in 47 breeding pairs of captive canaries ( Serinus canaria). We found that a within-individual increase in female testosterone level suppressed immune function. Furthermore, testosterone levels were positively related to male song repertoire size as an important component of sexual signaling at the between-male level. These were, however, the only relevant significant correlations. Overall, our data do therefore not convincingly support the hypotheses tested and suggest rather limited hormonal pleiotropic effects of testosterone on immune function, parental care, and male sexual signaling, at least in our study system. Significance statement: Phenotypic variances and covariances can nowadays be partitioned easily into within- and between-individual components. These components inform about trait repeatability and the extent to which multiple traits form phenotypic suites of traits, as well as about their joint underlying mechanistic pathways. Testosterone for example, may be key to mediate the expression of suites of traits in many vertebrate species. We here used captive canaries and repeatedly measured male and female traits, relevant in the context of endocrinology, sexual signaling, immune function, and parental investment. For both sexes, we report particularly weak correlations between testosterone and all other measured traits at both within- and between-individual levels. Our case study thus questions the pleiotropic effects of testosterone, but exemplifies the applicability and relevance of (co)variance partitioning in behavioral ecological research. [ABSTRACT FROM AUTHOR]

Testosterone mediates reproductive behaviours in male vertebrates. For example, breeding season territoriality depends on testosterone in many species of birds and in some, territorial interactions feed back on testosterone concentrations. However, the degree to which territorial behaviour and testosterone are associated differs even between species with seemingly similar life histories, especially between species that also defend territories outside the breeding season. Here, we investigate the link between territorial behaviour and testosterone in European stonechats. Previous studies found that territorial aggression in stonechats depends on testosterone in a breeding, but not in a non-breeding context. We investigated whether stonechats show a rise in testosterone during simulated territorial intrusions (STI) during the breeding season. Post-capture testosterone concentrations of males caught after an STI were not higher than those of males caught in a control situation regardless of breeding stage. However, most of the males would have been able to mount a testosterone response because the same individuals that did not increase testosterone during the STI showed a substantial increase in testosterone after injections of gonadotropin-releasing hormone (GnRH). GnRH-induced and post-capture testosterone concentrations were positively correlated and both decreased with successive breeding stages. Further, territory owners with a short latency to attack the decoy expressed higher post-capture testosterone concentrations than males with a longer latency to attack the decoy. Thus, there is no evidence for behavioural feedback on testosterone concentrations during male-male interactions in stonechats. In combination with previous studies our data suggest that testosterone functions as an on/off switch of high intensity territorial aggression during the breeding season in stonechats. The among-species variation in the androgen control of territorial behaviour may be only partly a result of environmental differences. Instead, potential differences in how territoriality evolved in different species may have influenced whether and how a reproductive hormone such as testosterone was co-opted into the mechanistic control of territorial behaviour. [ABSTRACT FROM AUTHOR]

This study used population-representative longitudinal data from the 2005-2006 and 2010-2011 waves of the National Social Life, Health and Aging Project-a probability sample of US adults aged 57-85 at baseline (N = 650 women and 620 men)-to examine the causal direction in linkages of endogenous testosterone (T) with sexual activity and relationship quality. For both genders, our autoregressive effects indicated a large amount of temporal stability, not just in individual-level attributes (T, masturbation) but also dyadic ones (partnered sex, relationship quality)-indicating that a need for more nuanced theories of relational processes. Cross-lagged results suggested gender-specific effects-generally more consistent with sexual or relational modulation of T than with hormonal causation. Specifically, men's findings indicated their T might be elevated by their sexual (masturbatory) activity but not vice versa, although androgen levels did lower men's subsequent relationship quality. Women's T, in contrast, was negatively influenced not just by their higher relationship quality but also by their more frequent partnered sex-perhaps reflecting a changing function of sexual activity in late life. [ABSTRACT FROM AUTHOR]

Decades of behavioral endocrinology research have shown that hormones and behavior have a bidirectional relationship; hormones both influence and respond to social behavior. In contrast, hormones are often thought to have a unidirectional relationship with ornaments. Hormones influence ornament development, but little empirical work has tested how ornaments influence hormones throughout life. Here, we experimentally alter a visual signal of fighting ability in Polistes dominulus paper wasps and measure the behavioral and hormonal consequences of signal alteration in signalers and receivers. We find wasps that signal inaccurately high fighting ability receive more aggression than controls and receiving aggression reduces juvenile hormone (JH) titers. As a result, immediately after contests, inaccurate signalers have lower JH titers than controls. Ornaments also directly influence rival JH titers. Three hours after contests, wasps who interacted with rivals signaling high fighting ability have higher JH titers than wasps who interacted with rivals signaling low fighting ability. Therefore, ornaments influence hormone titers of both signalers and receivers. We demonstrate that relationships between hormones and ornaments are flexible and bidirectional rather than static and unidirectional. Dynamic relationships among ornaments, behavior, and physiology may be an important, but overlooked factor in the evolution of honest communication. [ABSTRACT FROM AUTHOR]

Androgens respond to social challenges and this response has been interpreted as a way for males to adjust androgen-dependent behavior to social context. However, the androgen responsiveness to social challenges varies across species and a conceptual framework has been developed to explain this variation according to differences in the mating system and parental care type, which determines the regimen of challenges males are exposed to, and concomitantly the scope (defined as the difference between the physiological maximum and the baseline levels) of response to a social challenge. However, this framework has been focused on territorial species and no clear predictions have been made to gregarious species (e.g. shoaling fish), which although tolerating same-sex individuals may also exhibit intra-sexual competition. In this paper we extend the scope of this conceptual framework to shoaling fish by studying the endocrine response of zebrafish ( Danio rerio ) to social challenges. Male zebrafish exposed to real opponent agonistic interactions exhibited an increase in androgen levels (11-ketotestosterone both in Winners and Losers and testosterone in Losers). This response was absent in Mirror-fighters, that expressed similar levels of aggressive behavior to those of winners, suggesting that this response is not a mere reflex of heightened aggressive motivation. Cortisol levels were also measured and indicated an activation of the hypothalamic–pituitary–interrenal axis in Winners of real opponent fighters, but not Losers or in Mirror-fighters. These results confirm that gregarious species also exhibit an endocrine response to an acute social challenge. [ABSTRACT FROM AUTHOR]

Animals exhibit continuous seasonal changes in physiological, morphological and behavioral traits associated to their natural annual cycles. One of the most important changes in the organism occurs at the initiation of breeding season. In males these changes include activation of spermatogenesis, enhance of sexual secondary characters and an increase of aggressiveness. All of theses changes are basically regulated by testosterone, but the physiological basis of this testosterone regulation and the effective hormone proportion that acts in target tissues are unknown. In this work I evaluated the relationship between total testosterone, free testosterone associated to aggressive behavior in wild males of Octodon degus. I compared hormonal levels and aggressive behavior during pre, middle and post breeding periods. Results showed that behavioral aggressiveness was correlated with both total and free testosterone in June, during the beginning of breeding season, but not at other times. Results also indicated a lack of relationship between free and total testosterone in most of breeding period. I discuss the importance of this behavior-hormonal regulation (determined by free testosterone) and the physiological importance of this mechanism to the organismLos animales presentan variaciones estacionales continuas que incluyen cambios fisiológicos, morfológicos y conductuales, asociados al ciclo anual. Estos cambios generalmente están activados por factores endógenos o exógenos, pero son regulados por el sistema endocrino de organismo. Uno de los cambios más importantes ocurre en el inicio del periodo reproductivo. Durante este periodo los machos presentan cambios asociados a la producción de esperatozoides, exacerbación de caracteres sexuales secundarios y un aumento de la agresividad. Todos estos cambios son regulados en parte por los niveles de testosterona, sin embargo, los mecanismos fisiológicos de esta regulación son desconocidos. En este trabajo evalué la relación que existe entre los niveles de testosterona libre y totales, y examiné su posible asociación con los niveles de agresividad en machos de una población natural de Octodon degus. Mis resultados indicaron que la agresividad se correlaciona directamente con los niveles de testosterona libre y totales solo durante el mes de junio, al inicio del periodo reproductivo, pero no durante el resto del ciclo reproductivo de esta especie. Esto indicaría que la agresividad solo se correlaciona con los niveles de testosterona libre ya que en el medio y el final del periodo reproductivo los niveles de testosterona se mantienen altos sin que se manifiesten dichas conductas. Finalmente, discuto la importancia de la regulación fina de la conducta determinada por la testosterona libre y la importancia fisiológica de este mecanismo de regulación dentro del organismo

[Display omitted] • We investigate the androgen response to territorial challenges in a poison frog. • Acoustic playbacks induce an increase in androgen levels in Allobates femoralis. • We provide novel support for the challenge hypothesis in a territorial frog. • We quantified repeatedly androgen concentrations via a non-invasive technique. Territoriality has been widely described across many animal taxa, where the acquisition and defence of a territory are critical for the fitness of an individual. Extensive evidence suggests that androgens are involved in the modulation of territorial behaviours in male vertebrates. Short-term increase of androgen following a territorial encounter appears to favour the outcome of a challenge. The "Challenge Hypothesis" proposed by Wingfield and colleagues outlines the existence of a positive feedback relationship between androgen and social challenges (e.g., territorial intrusions) in male vertebrates. Here we tested the challenge hypothesis in the highly territorial poison frog, Allobates femoralis , in its natural habitat by exposing males to simulated territorial intrusions in the form of acoustic playbacks. We quantified repeatedly androgen concentrations of individual males via a non-invasive water-borne sampling approach. Our results show that A. femoralis males exhibited a positive behavioural and androgenic response after being confronted to simulated territorial intrusions, providing support for the Challenge Hypothesis in a territorial frog. [ABSTRACT FROM AUTHOR]