Øyvind Øverli

Short-Term Effects of Fights for Social Dominance and the Establishment of Dominant-Subordinate Relationships on Brain Monoamines and Cortisol in Rainbow Trout

We report changes in brain serotonergic, noradrenergic and dopaminergic activity, along with plasma cortisol concentrations, occurring during the initial 24-h period following the establishment of dominant-subordinate relationships in pairs of rainbow trout. Immediately (within 5 min) after the termination of staged fights for social dominance, a large increase in blood plasma cortisol was observed in both fight losers (future subordinate fish) and winners (future dominant fish). In dominant fish, cortisol decreased rapidly (within 3 h) to the level of unstressed controls, while continuing to increase in subordinate fish. At 3 h following fights, the brain serotonergic system was activated in both dominant fish and subordinate fish, at least in some brain regions (telencephalon). This effect was reversed in dominant individuals within 24 h of social interaction, whereas in subordinate fish a substantial activation of the serotonergic system was manifest in all brain regions by 24 h. Similarly, a strong increase in brain catecholaminergic activation was indicated after 24 h of social interaction in subordinate fish, but not in dominant fish. Relationships between plasma cortisol and brain serotonergic and noradrenergic activity in the various experimental groups suggest that these systems influence cortisol secretion under normal conditions and during moderate or short-term stress.

Evolutionary background for stress-coping styles: relationships between physiological, behavioral, and cognitive traits in non-mammalian vertebrates.

Reactions to stress vary between individuals, and physiological and behavioral responses tend to be associated in distinct suites of correlated traits, often termed stress-coping styles. In mammals, individuals exhibiting divergent stress-coping styles also appear to exhibit intrinsic differences in cognitive processing. A connection between physiology, behavior, and cognition was also recently demonstrated in strains of rainbow trout (Oncorhynchus mykiss) selected for consistently high or low cortisol responses to stress. The low-responsive (LR) strain display longer retention of a conditioned response, and tend to show proactive behaviors such as enhanced aggression, social dominance, and rapid resumption of feed intake after stress. Differences in brain monoamine neurochemistry have also been reported in these lines. In comparative studies, experiments with the lizard Anolis carolinensis reveal connections between monoaminergic activity in limbic structures, proactive behavior in novel environments, and the establishment of social status via agonistic behavior. Together these observations suggest that within-species diversity of physiological, behavioral and cognitive correlates of stress responsiveness is maintained by natural selection throughout the vertebrate sub-phylum.

Behavioral and Neuroendocrine Correlates of Selection for Stress Responsiveness in Rainbow Trout—a Review

Abstract In rainbow trout the magnitude of the cortisol response to stress shows both consistency over time and a moderate to high degree of heritability, and high responding (HR) and low responding (LR) lines of rainbow trout have been generated by individual selection for consistently high or low post-stress cortisol values. Using 2nd and 3rd generation fish, we tested the hypothesis that differential stress responsiveness is associated with behavioral alterations in the HR-LR trout model. LR fish showed a tendency to become socially dominant, a rapid recovery of food intake after transfer to a novel environment, and a reduced locomotor response in a territorial intrusion test. Furthermore, stress induced elevation of brain stem and optic tectum concentrations of the monoamine neurotransmitters serotonin, dopamine, and norepinephrine and their metabolites suggests that both synthesis and metabolism of these transmitters were elevated after stress to a larger degree in HR than in LR trout. A divergent pattern was seen in the hypothalamus, where LR fish displayed elevated levels of 5-hydroxyindoleacetic acid (a serotonin metabolite) and 3-methoxy-4-hydroxyphenylglycol (a norepinephrine metabolite). Thus, selection for a single trait, cortisol responsiveness, in rainbow trout is associated with concurrent changes in both behavior and central signaling systems. The apparent parallel to genetically determined stress coping styles in mammals, and the existence of similar trait associations in unselected populations of rainbow trout, suggests an evolutionarily conserved correlation between multiple traits. Continuing studies on the HR and LR trout lines are aimed at providing the physiological and genetic basis for new marker-assisted selection strategies in the rapidly developing finfish aquaculture industry, as well as increased knowledge of the function and evolution of central neuroendocrine signaling systems.

Stress coping style predicts aggression and social dominance in rainbow trout

Social stress is frequently used as a model for studying the neuroendocrine mechanisms underlying stress-induced behavioral inhibition, depression, and fear conditioning. It has previously been shown that social subordination may result in increased glucocorticoid release and changes in brain signaling systems. However, it is still an open question which neuroendocrine and behavioral differences are causes, and which are consequences of social status. Using juvenile rainbow trout of similar size and with no apparent differences in social history, we demonstrate that the ability to win fights for social dominance can be predicted from the duration of a behavioral response to stress, in this case appetite inhibition after transfer to a new environment. Moreover, stress responsiveness in terms of confinement-induced changes in plasma cortisol was negatively correlated to aggressive behavior. Fish that exhibited lower cortisol responses to a standardized confinement test were markedly more aggressive when being placed in a dominant social position later in the study. These findings support the view that distinct behavioral-physiological stress coping styles are present in teleost fish, and these coping characteristics influence both social rank and levels of aggression.

Effects of cortisol on aggression and locomotor activity in rainbow trout.

Noninvasive administration of cortisol through the diet resulted in relatively rapid (<1.5 h) and highly reproducible increases in plasma cortisol in rainbow trout, comparable to changes seen in fish subjected to substantial stress. Juvenile rainbow trout were reared in isolation for 1 week, before their daily food ration was replaced by a meal of cortisol-treated food corresponding to 6 mg cortisol kg(-1). All fish were observed for 30 min, beginning at 1 or 48 h following the introduction of cortisol-treated food. Additional cortisol (75% of the original dose on Day 2, and 50% on Day 3) was administered to the long-term cortisol-treated group. The resulting blood plasma concentrations of cortisol were similar in short- and long-term treated fish, and corresponded to those previously seen in stressed rainbow trout. Controls were fed similar food without cortisol. Half of the fish from each treatment group (controls and short- and long-term cortisol) were subjected to an intruder test (a smaller conspecific introduced into the aquarium), while half of the fish were observed in isolation. In fish challenged by a conspecific intruder, short-term cortisol treatment stimulated locomotor activity, while long-term treatment inhibited locomotion. Aggressive behavior was also inhibited by long-term cortisol treatment, but not by short-term exposure to cortisol. Cortisol treatment had no effect on locomotor activity in undisturbed fish, indicating that the behavioral effects of cortisol were mediated through interaction with other signal systems activated during the simulated territorial intrusion test. This study demonstrates for the first time that cortisol has time- and context-dependent effects on behavior in teleost fish.

Cortisol and finfish welfare

Previous reviews of stress, and the stress hormone cortisol, in fish have focussed on physiology, due to interest in impacts on aquaculture production. Here, we discuss cortisol in relation to fish welfare. Cortisol is a readily measured component of the primary (neuroendocrine) stress response and is relevant to fish welfare as it affects physiological and brain functions and modifies behaviour. However, we argue that cortisol has little value if welfare is viewed purely from a functional (or behavioural) perspective—the cortisol response itself is a natural, adaptive response and is not predictive of coping as downstream impacts on function and behaviour are dose-, time- and context-dependent and not predictable. Nevertheless, we argue that welfare should be considered in terms of mental health and feelings, and that stress in relation to welfare should be viewed as psychological, rather than physiological. We contend that cortisol can be used (with caution) as a tractable indicator of how fish perceive (and feel about) their environment, psychological stress and feelings in fish. Cortisol responses are directly triggered by the brain and fish studies do indicate cortisol responses to psychological stressors, i.e., those with no direct physicochemical action. We discuss the practicalities of using cortisol to ask the fish themselves how they feel about husbandry practices and the culture environment. Single time point measurements of cortisol are of little value in assessing the stress level of fish as studies need to account for diurnal and seasonal variations, and environmental and genetic factors. Areas in need of greater clarity for the use of cortisol as an indicator of fish feelings are the separation of (physiological) stress from (psychological) distress, the separation of chronic stress from acclimation, and the interactions between feelings, cortisol, mood and behaviour.

Behavioral indicators of stress-coping style in rainbow trout: Do males and females react differently to novelty?

It is becoming increasingly clear that individual differences in the behavioral response to stressful situations are associated with distinct physiological profiles, and stress coping characteristics are of fundamental importance to fitness and life history. Teleost fishes display considerable variation in reproductive strategy, but sex differences in stress-coping style have not been described previously in fish. Prior to sexual maturation, the glucocorticoid response to stress is not affected by sex in salmonid fish. Nevertheless, behavior in novel and stressful situations differed between immature male and female rainbow trout (Oncorhynchus mykiss). When tested 1 week following transport to a new rearing facility, females resumed feeding after transfer to social isolation quicker than males. The locomotor response to acute confinement stress also varied between sexes, with females settling down and ceasing to move in a panic-like manner quicker than males. There was a strong correlation between behavior in the two test situations: individuals that readily resumed feeding behavior in a new environment also moved less in the acute stress test. Thus, the time to resume feeding after a stressful experience is a precise indicator of stress-coping style in salmonid fish, which is likely to reflect the dynamics of neuroendocrine stress responses. Furthermore, these observations could reflect a sex difference in the response to novel and stressful situations, which occur even in the absence of differences in glucocorticoid responsiveness.

Behavioral and neuroendocrine correlates of displaced aggression in trout

In humans and other primates, violent actions performed by victims of aggression are often directed toward an individual or object that is not the source of provocation. This psychological phenomenon is often called displaced aggression. We demonstrate that displaced aggression is either rooted in evolutionarily conserved behavioral and neuroendocrine mechanisms, or represent a convergent pattern that has arisen independently in fish and mammals. Rainbow trout that briefly encountered large, aggressive fish reacted with increased aggression toward smaller individuals. There was a strong negative correlation between received aggression and behavioral change: Individuals subjected to intense aggression were subdued, while moderate assaults induced strong agitation. Patterns of forebrain serotonin turnover and plasma cortisol suggest that the presence of socially subordinate fish had an inhibitory effect on neuroendocrine stress responses. Thus, subordinate individuals may serve as stress-reducing means of aggressive outlet, and displaced aggression toward such individuals appears to be a behavioral stress coping strategy in fishes.

Response to environmental change in rainbow trout selected for divergent stress coping styles.

An extensive literature has documented differences in the way individual animals cope with environmental challenges and stressors. Two broad patterns of individual variability in behavioural and physiological stress responses are described as the proactive and reactive stress coping styles. In addition to variability in the stress response, contrasting coping styles may encompass a general difference in behavioural flexibility as opposed to routine formation in response to more subtle environmental changes and non-threatening novelties. In the present study two different manipulations, relocating food from a previously learned location, and introducing a novel object yielded contrasting responses in rainbow trout selected for high (HR) and low (LR) post stress plasma cortisol levels. No difference was seen in the rate of learning the original food location; however, proactive LR fish were markedly slower than reactive HR fish in altering their food seeking behaviour in response to relocated food. In contrast, LR fish largely ignored a novel object which disrupted feeding in HR fish. Hence, it appears that the two lines appraise environmental cues differently. This observation suggests that differences in responsiveness to environmental change are an integral component of heritable stress coping styles, which in this particular case, had opposite effects on foraging efficiency in different situations. Context dependent fitness effects may thus explain the persistence of stable divergence of this evolutionary widespread trait complex.

Does Serotonin Influence Aggression? Comparing Regional Activity before and during Social Interaction*

Serotonin is widely believed to exert inhibitory control over aggressive behavior and intent. In addition, a number of studies of fish, reptiles, and mammals, including the lizard Anolis carolinensis, have demonstrated that serotonergic activity is stimulated by aggressive social interaction in both dominant and subordinate males. As serotonergic activity does not appear to inhibit agonistic behavior during combative social interaction, we investigated the possibility that the negative correlation between serotonergic activity and aggression exists before aggressive behavior begins. To do this, putatively dominant and more aggressive males were determined by their speed overcoming stress (latency to feeding after capture) and their celerity to court females. Serotonergic activities before aggression are differentiated by social rank in a region‐specific manner. Among aggressive males baseline serotonergic activity is lower in the septum, nucleus accumbens, striatum, medial amygdala, anterior hypothalamus, raphe, and locus ceruleus but not in the hippocampus, lateral amygdala, preoptic area, substantia nigra, or ventral tegmental area. However, in regions such as the nucleus accumbens, where low serotonergic activity may help promote aggression, agonistic behavior also stimulates the greatest rise in serotonergic activity among the most aggressive males, most likely as a result of the stress associated with social interaction.