How integrated are behavioral and endocrine stress response traits? A repeated measures approach to testing the stress-coping style model

It is widely expected that physiological and behavioral stress responses will be integrated within divergent stress‐coping styles (SCS) and that these may represent opposite ends of a continuously varying reactive–proactive axis. If such a model is valid, then stress response traits should be repeatable and physiological and behavioral responses should also change in an integrated manner along a major axis of among‐individual variation. While there is some evidence of association between endocrine and behavioral stress response traits, few studies incorporate repeated observations of both. To test this model, we use a multivariate, repeated measures approach in a captive‐bred population of Xiphophorus birchmanni. We quantify among‐individual variation in behavioral stress response to an open field trial (OFT) with simulated predator attack (SPA) and measure waterborne steroid hormone levels (cortisol, 11‐ketotestosterone) before and after exposure. Under the mild stress stimulus (OFT), (multivariate) behavioral variation among individuals was consistent with a strong axis of personality (shy–bold) or coping style (reactive–proactive) variation. However, behavioral responses to a moderate stressor (SPA) were less repeatable, and robust statistical support for repeatable endocrine state over the full sampling period was limited to 11‐ketotestosterone. Although post hoc analysis suggested cortisol expression was repeatable over short time periods, qualitative relationships between behavior and glucocorticoid levels were counter to our a priori expectations. Thus, while our results clearly show among‐individual differences in behavioral and endocrine traits associated with stress response, the correlation structure between these is not consistent with a simple proactive–reactive axis of integrated stress‐coping style. Additionally, the low repeatability of cortisol suggests caution is warranted if single observations (or indeed repeat measures over short sampling periods) of glucocorticoid traits are used in ecological or evolutionary studies focussed at the individual level.     

Effects of maternal stress coping style on offspring characteristics in rainbow trout (Oncorhynchus mykiss)

Maternal size, age, and allostatic load influence offspring size, development, and survival. Some of these effects have been attributed to the release of glucocorticoids, and individual variation in these stress hormones is related to a number of traits. Correlated traits are often clustered and used to define the proactive and reactive stress coping styles. Although stress coping styles have been identified in a number of animal groups, little is known about the coupling between stress coping style and offspring characteristics. In the present study, plasma cortisol levels in ovulated mothers and cortisol levels in non-fertilized eggs from two rainbow trout (Oncorhynchus mykiss) strains selected for high (HR) and low (LR) post-stress plasma cortisol levels were compared. Offspring characteristics such as egg size, larval growth, and energy reserves also were compared between the two strains. Maternal plasma and egg cortisol levels were correlated, but no difference between the HR and LR strains was detected in either parameter. LR females produced larger eggs, and larvae with larger yolk sacs compared to HR females, however no differences in larval body size (excluding the yolk) was detected between strains. Considering that the HR and LR strains have a number of correlated behavioral and physiological traits that resemble the reactive and proactive stress coping styles, respectively, the results suggest that proactive mothers invest more energy into their offspring, producing larvae with larger energy reserves. It is possible that larger energy reserves in proactive larvae support the energy requirement for establishing and defending territory in salmonid fish. Furthermore, in the present study we found a positive relationship between mother plasma cortisol and egg cortisol; however neither mother plasma cortisol nor egg cortisol differed between strains. These results indicate that cortisol endowment from the mother to the offspring plays a minor role in the transfer of the behavioral and physiological traits which separates these strains.     

Fitness and behavioral correlates of pre-stress and stress-induced plasma cortisol titers in pink salmon (Oncorhynchus gorbuscha) upon arrival at spawning grounds

Semelparous Pacific salmon (Onchorynchus spp.) serve as an excellent model for examining the relationships between life history, behavior and individual variation in glucocorticoid (GC) stress hormone levels because reproductive behaviors are highly variable between individuals and failure to reproduce results in zero fitness. Pink salmon (O. gorbuscha) were intercepted upon arrival at the spawning grounds across three time periods. Pre-stress and stress-induced plasma cortisol concentrations were assessed in relation to behavior, longevity and reproductive success. Results revealed differences between sexes and with arrival time. The study period marked a year of high reproductive success and only nine females (12% of sample) failed to spawn. Female pre-spawn mortalities were characterized by significantly elevated stress-induced cortisol concentrations and decreased longevity as well as pre-stress cortisol above the normal range in pink salmon from the study area. Interestingly, reproductive behaviors were only associated with pre-stress cortisol levels. For females, aggression and mate interaction time were reduced in individuals with elevated pre-stress cortisol concentrations. In males, a similar negative relationship between pre-stress cortisol concentration and mate interaction time was detected. The observed behavioral correlations are likely a factor of social status where dominant individuals, known to have higher reproductive success, are characterized by lower cortisol levels relative to subordinate conspecifics. Findings show both elevated pre-stress and stress-induced cortisol concentrations at arrival to the spawning grounds to be associated with reduced survival.     

Behavioral plasticity in rainbow trout (Oncorhynchus mykiss) with divergent coping styles: when doves become hawks

Consistent and heritable individual differences in reaction to challenges, often referred to as stress coping styles, have been extensively documented in vertebrates. In fish, selection for divergent post-stress plasma cortisol levels in rainbow trout (Oncorhynchus mykiss) has yielded a low (LR) and a high responsive (HR) strain. A suite of behavioural traits is associated with this physiological difference, with LR (proactive) fish feeding more rapidly after transfer to a new environment and being socially dominant over HR (reactive) fish. Following transport from the UK to Norway, a switch in behavioural profile occurred in trout from the 3rd generation; HR fish regained feeding sooner than LR fish in a novel environment and became dominant in size-matched HR–LR pairs. One year after transport, HR fish still fed sooner, but no difference in social dominance was found. Among offspring of transported fish, no differences in feeding were observed, but as in pre-transported 3rd generation fish, HR fish lost fights for social dominance against size-matched LR opponents. Transported fish and their offspring retained their distinctive physiological profile throughout the study; HR fish showed consistently higher post-stress cortisol levels at all sampling points. Altered risk-taking and social dominance immediately after transport may be explained by the fact that HR fish lost more body mass during transport than did LR fish. These data demonstrate that some behavioural components of stress coping styles can be modified by experience, whereas behavioural plasticity is limited by genetic effects determining social position early in life story.     

Behaviour,physiology and carotenoid pigmentation in Arctic charr Salvelinus alpinus

The behaviour during an exploration task and the response to a confinement stress of Arctic charr Salvelinus alpinus were evaluated. Behaviour of individuals during 90 min of exploration was classified into high and low activity. High‐activity individuals had higher plasma cortisol levels following stress compared to low‐activity individuals. This indicates that high‐ and low‐activity individuals correspond to reactive and proactive stress‐coping styles. Further, a pigmentation analysis showed that high‐activity individuals had a higher number of carotenoid spots cm^?2 than low‐activity individuals. Thus, carotenoid pigmentation, as melanin pigmentation in other salmonids, could be linked to stress‐coping style in S. alpinus.     

Stress coping styles and singing behavior in the short-tailed singing mouse (Scotinomys teguina)

Stress coping styles have been characterized as a proactive/reactive dichotomy in laboratory and domesticated animals. In this study, we examined the prevalence of proactive/reactive stress coping styles in wild-caught short-tailed singing mice (Scotinomys teguina). We compared stress responses to spontaneous singing, a social and reproductive behavior that characterizes this species. To establish proactive/reactive profiles for singing mice, we measured exploratory and anxiety behavior using an open-field behavioral test. We examined correlations between open-field behaviors and fecal corticosterone (CORT) metabolites, baseline plasma CORT, and stress-induced CORT. Mice with proactive behavioral responses in the open-field had higher fecal CORT titers than reactive males, but did not differ in baseline or stress-induced plasma CORT. We suggest that individual differences in CORT metabolism may contribute to this surprising pattern. Males that sang in the open-field were behaviorally proactive and had lower stress-induced CORT, indicating a link between stress responses and singing in this species. Overall, the data demonstrate that singing mice offer an interesting model for exploring how stress reactivity can shape social behaviors.     

Hormonal modulation of the heat shock response: insights from fish with divergent cortisol stress responses

Acute temperature stress in animals results in increases in heat shock proteins (HSPs) and stress hormones. There is evidence that stress hormones influence the magnitude of the heat shock response; however, their role is equivocal. To determine whether and how stress hormones may affect the heat shock response, we capitalized on two lines of rainbow trout specifically bred for their high (HR) and low (LR) cortisol response to stress. We predicted that LR fish, with a low cortisol but high catecholamine response to stress, would induce higher levels of HSPs after acute heat stress than HR trout. We found that HR fish have significantly higher increases in both catecholamines and cortisol compared with LR fish, and LR fish had no appreciable stress hormone response to heat shock. This unexpected finding prevented further interpretation of the hormonal modulation of the heat shock response but provided insight into stress-coping styles and environmental stress. HR fish also had a significantly greater and faster heat shock response and less oxidative protein damage than LR fish. Despite these clear differences in the physiological and cellular responses to heat shock, there were no differences in the thermal tolerance of HR and LR fish. Our results support the hypothesis that responsiveness to environmental change underpins the physiological differences in stress-coping styles. Here, we demonstrate that the heat shock response is a distinguishing feature of the HR and LR lines and suggest that it may have been coselected with the hormonal responses to stress.     

Fitness and behavioral correlates of pre-stress and stress-induced plasma cortisol titers in pink salmon (Oncorhynchus gorbuscha) upon arrival at spawning grounds

Semelparous Pacific salmon (Onchorynchus spp.) serve as an excellent model for examining the relationships between life history, behavior and individual variation in glucocorticoid (GC) stress hormone levels because reproductive behaviors are highly variable between individuals and failure to reproduce results in zero fitness. Pink salmon (O. gorbuscha) were intercepted upon arrival at the spawning grounds across three time periods. Pre-stress and stress-induced plasma cortisol concentrations were assessed in relation to behavior, longevity and reproductive success. Results revealed differences between sexes and with arrival time. The study period marked a year of high reproductive success and only nine females (12% of sample) failed to spawn. Female pre-spawn mortalities were characterized by significantly elevated stress-induced cortisol concentrations and decreased longevity as well as pre-stress cortisol above the normal range in pink salmon from the study area. Interestingly, reproductive behaviors were only associated with pre-stress cortisol levels. For females, aggression and mate interaction time were reduced in individuals with elevated pre-stress cortisol concentrations. In males, a similar negative relationship between pre-stress cortisol concentration and mate interaction time was detected. The observed behavioral correlations are likely a factor of social status where dominant individuals, known to have higher reproductive success, are characterized by lower cortisol levels relative to subordinate conspecifics. Findings show both elevated pre-stress and stress-induced cortisol concentrations at arrival to the spawning grounds to be associated with reduced survival.     

Coping strategies in a strongly schooling fish,the common carp Cyprinus carpio

Individual common carp Cyprinus carpio were screened repeatedly for risk taking (rate of exploration of a novel, potentially dangerous environment) and for competitive ability (success in gaining access to a spatially restricted food source). Marked differences in behaviour were evident, and significant consistency in individual responses across trials was found for both risk taking and competitive ability. In addition, there was a significant positive relationship between individual performance in these two contexts, with fish that explored more quickly in the novel environment tending to be among the first to gain access to restricted food. In two follow‐up studies, resting metabolic rate, blood lactate and glucose and the expression of the cortisol receptor gene in the head kidney and brain were compared in fish from the two extremes of the risk‐taking spectrum. Mass‐specific metabolic rate was significantly higher in risk‐taking than in risk‐avoiding fish, while plasma lactate and glucose concentrations and expression of the cortisol receptor gene were lower. It was concluded that a behavioural syndrome based on boldness and aggression exists in C. carpio, as it does in many other animals, and that this is associated with differences in metabolic and stress physiology (down to the genomic level) similar to those described in animals with different coping strategies.     

A comparative analysis of some life-history traits between cooperatively and non-cooperatively breeding Australian passerines

Summary Comparative analyses were carried out for some life-history traits of cooperatively and non-cooperatively breeding Australian Corvida (i.e. old-endemic passerines). Multivariate statistical analyses at the family and genus levels revealed no significant differences between cooperative and non-cooperative breeders. A matched-pairs analysis between congeneric species showed that cooperatively breeding species lay smaller clutches than non-cooperatively breeding congenerics. Preliminary results also suggest that cooperative breeders have higher probabilities of rearing a second brood in the season and lower probabilities of survival than do non-cooperative breeders. However, the result for survival was significant in only one out of three tests. We conclude that cooperatively and non-cooperatively breeding Australian Corvida cannot be separated into distinct groups showingK- andr-selected life-history traits, respectively. Some life-history traits follow the prediction of ther-K selection model, others show evidence of co-adaptation instead, whereas still others show evidence of trade-offs.     

Three different coping styles in police dogs exposed to a short-term challenge

According to some researchers, animals show different coping styles to deal with stressful situations. In the case of social carnivores, social stress is a substantial part of the overall stress load. Previous research has established two extreme (proactive and reactive) coping styles in several animal species, but means of coping with social stress has not yet been investigated in the case of dogs. The aim of this current study was to examine whether (1) experienced working police dogs adopt different coping strategies during a short-term unexpected social challenge presented by a threatening human, (2) whether this affects post-encounter cortisol levels, and (3) whether there is an association between the cortisol response and the behavior (coping strategy) displayed during the threatening approach. Using factor analysis, we have identified three different group of dogs which were characterized by either fearfulness, aggressiveness, or ambivalence and in parallel showed specific differences in their reaction norm when threatened by an approaching stranger. This grouping also allowed to draw possible parallels between aggressiveness and the proactive behavior style and fearfulness and reactive coping style, respectively. In addition, we have revealed a third group of animals which show ambivalent behavior in a social threatening situation.     

Three different coping styles in police dogs exposed to a short-term challenge

According to some researchers, animals show different coping styles to deal with stressful situations. In the case of social carnivores, social stress is a substantial part of the overall stress load. Previous research has established two extreme (proactive and reactive) coping styles in several animal species, but means of coping with social stress has not yet been investigated in the case of dogs. The aim of this current study was to examine whether (1) experienced working police dogs adopt different coping strategies during a short-term unexpected social challenge presented by a threatening human, (2) whether this affects post-encounter cortisol levels, and (3) whether there is an association between the cortisol response and the behavior (coping strategy) displayed during the threatening approach. Using factor analysis, we have identified three different group of dogs which were characterized by either fearfulness, aggressiveness, or ambivalence and in parallel showed specific differences in their reaction norm when threatened by an approaching stranger. This grouping also allowed to draw possible parallels between aggressiveness and the proactive behavior style and fearfulness and reactive coping style, respectively. In addition, we have revealed a third group of animals which show ambivalent behavior in a social threatening situation.     

Ventilation rates indicate stress-coping styles in Nile tilapia

Behavioural responses to stress can form distinct profiles in a wide range of animals: proactive and reactive profiles or coping styles. Stress responsiveness can also differentiate between the behavioural profiles. The tendency to regain feed intake following transfer to a novel social-isolation tank (the speed of acclimation) can discriminate between proactive or reactive profiles. Consequently, differential stress responsiveness can be linked to this feeding behaviour trait. This study shows that ventilation rates of Nile tilapia, Oreochromis niloticus (L.), correlate with the rate of feeding resumption, following transfer to a novel social-isolation aquarium. Therefore, ventilation rate (VR) indicates coping styles; consequently, VR is a proxy for the way fish will deal with environmental challenges.     

Association of ADHD with reactive and proactive violent behavior in a forensic population

ADHD is associated with social problems and aggressive behavior. As hyperactive-impulsive traits are core symptoms of ADHD, it has been hypothesized that reactive-impulsive violence is more likely related to ADHD psychopathology than proactive-instrumental violence. One hundred and twenty-seven adult violent offenders participated in the study. Diagnosis of ADHD and ratings of reactive and proactive features of the committed crimes were performed using standardized instruments. According to DSM-IV, 16.5% subjects fulfilled diagnostic criteria for ADHD, 23.6% were diagnosed as ADHD in partial remission, and 59.8% had no ADHD. Univariate analyses revealed higher reactive violence ratings in both ADHD groups when compared to subjects without ADHD, whereas the opposite was found regarding proactive violence ratings. Using multivariate analyses of variance controlled for age, gender and comorbid substance use disorders, childhood ADHD psychopathology and current ADHD significantly increased the risk of reactive violence and decreased the risk of proactive violence. Significant impact of male gender on proactive violence was found. The findings suggest that ADHD is associated with reactive but not proactive violence in aggressive offenders.     

Behavioural Interactions and Hormones in Naturally and Hatchery‐Spawned Chinook Salmon

Artificial breeding programmes commonly lead to domestication, which is associated with many behavioural differences that can reduce the success of animals released into natural environments. To better understand the factors contributing to domestication, we used a captive population of Chinook salmon (Oncorhynchus tshawytscha) to partition hormonal and behavioural differences to effects of the breeding method and rearing environment. We compared 9‐mo‐old juveniles from three lines that shared a common genetic background: (1) the Channel line produced by natural spawning and reared in a low‐density environment with a natural substrate for approx. 6 mo before being transferred to the hatchery; (2) the Hatchery line produced by artificial spawning; and (3) the Transfer line produced by natural spawning but reared in the hatchery from the eyed‐egg stage. Plasma concentrations of 11‐ketotestosterone (11‐KT) and cortisol were measured in groups of 150 fish and again after 4 d of social interactions in groups of six fish. There was no difference in 11‐KT among lines in large groups, but in small groups, Transfer fish had lower 11‐KT concentrations and were significantly less aggressive than both Channel and Hatchery fish. Regardless of group size, concentrations of the stress hormone cortisol were nearly twofold higher in Channel fish than in Hatchery and Transfer fish. Furthermore, the elevated cortisol concentrations in Channel fish were associated with 35% lower feeding rates than in the other two lines. Our study details complex behavioural and hormonal responses to breeding method and rearing environment in juvenile salmon.     

Using parentage analysis to estimate rates of straying and homing in Chinook salmon (Oncorhynchus tshawytscha)

We used parentage analysis based on microsatellite genotypes to measure rates of homing and straying of Chinook salmon (Oncorhynchus tshawytscha) among five major spawning tributaries within the Wenatchee River, Washington. On the basis of analysis of 2248 natural‐origin and 11594 hatchery‐origin fish, we estimated that the rate of homing to natal tributaries by natural‐origin fish ranged from 0% to 99% depending on the tributary. Hatchery‐origin fish released in one of the five tributaries homed to that tributary at a far lower rate than the natural‐origin fish (71% compared to 96%). For hatchery‐released fish, stray rates based on parentage analysis were consistent with rates estimated using physical tag recoveries. Stray rates among major spawning tributaries were generally higher than stray rates of tagged fish to areas outside of the Wenatchee River watershed. Within the Wenatchee watershed, rates of straying by natural‐origin fish were significantly affected by spawning tributary and by parental origin: progeny of naturally spawning hatchery‐produced fish strayed at significantly higher rates than progeny whose parents were themselves of natural origin. Notably, none of the 170 offspring that were products of mating by two natural‐origin fish strayed from their natal tributary. Indirect estimates of gene flow based on F_ST statistics were correlated with but higher than the estimates from the parentage data. Tributary‐specific estimates of effective population size were also correlated with the number of spawners in each tributary.     

Relationship between oxidative stress and circulating testosterone and cortisol in pre-spawning female brown trout

Reproduction in vertebrates is an energy-demanding process that is mediated by endogenous hormones and potentially results in oxidative stress. The primary aim of this study was to quantify the relationship between oxidative stress parameters (antioxidant capacity and levels of reactive oxygen metabolites) and circulating testosterone and cortisol in a common and widespread teleost fish, the brown trout (Salmo trutta, L.). Results show that trout with higher testosterone levels prior to spawning have higher levels of oxidative damage at the time that they spawn (although by the time of spawning testosterone levels had dropped, leading to a negative relationship between testosterone and oxidative damage at that time). Cortisol levels were not directly related to oxidative damage or antioxidant capacity, but concentrations of this hormone were positively related to levels of fungal infection, which was itself associated both with lower antioxidant capacity and lower levels of oxidative damage. These results highlight the complexity of interactions between different components of the endocrine system and metabolism and suggest that caution be used in interpreting relationships between a single hormone and indicators of oxidative balance or other fitness proxies.     

Epizootic cutaneous papillomatosis, cortisol and male ornamentation during and after breeding in the roach Rutilus rutilus

The prevalence of epidermal papillomatosis in roach is known to peak during the spawning period and to be higher in males than in females. The high occurrence of papillomatosis in polluted waters suggests that stress may contribute to the outbreak of the disease. However, little is known about breeding-induced stress in fish and its relationship with diseases. In this study, plasma cortisol concentration, hematocrit and the relative size of the spleen were determined in healthy and diseased male and female roach Rutilus rutilus during and shortly after spawning in a wild population. In addition, the sexual ornamentation (breeding tubercles on the lateral sides and on the frontal) of male roach during spawning was examined. Plasma cortisol concentration was higher during than after the spawning period, and higher in males than in females during spawning, indicating a spawning-induced stress and higher spawning stress among males. There was no correlation between cortisol concentration and the intensity of papillomatosis (number of scales under papilloma tumors) among the diseased fish. However, the significant interaction sex x disease status revealed by ANCOVA suggested that diseased males could be more prone to increased cortisol levels than diseased females or healthy males. Hematocrit values (ratio of the volume of red blood cells to total volume of blood) but not condition factor were lowered in papilloma-diseased fish after spawning. The relative size of the spleen was greater in males than in females. The number of frontal breeding tubercles correlated negatively with the intensity of papillomatosis. Experimental studies are needed to investigate the association of papillomatosis with stress and cortisol.