Michael J. Lawrence

Stress, nutrition and parental care in a teleost fish: Exploring mechanisms with supplemental feeding and cortisol manipulation

ABSTRACT Parental care is an essential life-history component of reproduction for many animal species, and it entails a suite of behavioural and physiological investments to enhance offspring survival. These investments can incur costs to the parent, reducing their energetic and physiological condition, future reproductive capabilities and survival. In fishes, relatively few studies have focused on how these physiological costs are mediated. Male smallmouth bass provide parental care for developing offspring until the brood reaches independence. During this energetically demanding life stage, males cease active foraging as they vigorously defend their offspring. Experimental manipulation of cortisol levels (via implantation) and food (via supplemental feeding) in parental males was used to investigate the fitness consequences of parental care. Improving the nutritional condition of nest-guarding males increased their reproductive success by reducing premature nest abandonment. However, supplemental feeding and cortisol treatment had no effect on parental care behaviours. Cortisol treatment reduced plasma lymphocyte numbers, but increased neutrophil and monocyte concentrations, indicating a shift in immune function. Supplemental feeding improved the physiological condition of parental fish by reducing the accumulation of oxidative injury. Specifically, supplemental feeding reduced the formation of 8-hydroxy-2′-deoxyguanosine (8-OHdG) on DNA nucleotides. Increasing the nutritional condition of parental fish can reduce the physiological cost associated with intensive parental activity and improve overall reproductive success, illustrating the importance of nutritional condition as a key modulator of parental fitness. Summary: Stress and nutritional condition are key modulators of physiological condition and reproductive success in a parental teleost fish.

An experimental evaluation of the role of the stress axis in mediating predator-prey interactions in wild marine fish

The hypothalamic-pituitary-interrenal (HPI) axis, through corticosteroid secretion, is an integral mechanism regulating internal homeostasis when vertebrates are faced with a stressor. However, continued HPI-axis stimulation can produce homeostatic overload, where corticosteroids are detrimental to organismal function. This overload condition may play an important role in mediating predator-prey interactions, because chronically/previously stressed animals may have higher rates of predator-induced mortality. However, the mechanism(s) underlying this observation are unknown. Using fish as models, we hypothesized that chronic stress would increase predation susceptibility owing to a poor physiological state (e.g. homeostatic overload) with corresponding sub-optimal changes in predator-avoidance behaviour. As cortisol is also required in low quantities to help regulate basic metabolic functions in fish, we expected that a glucocorticoid receptor antagonist (GR; e.g. homeostatic failure) may produce similar effects. Schoolmaster snapper (Lutjanus apodus) were given intraperitoneal implants of cocoa butter impregnated with nothing (sham; 5ml/kg body weight (BW)), cortisol (50mg/kg BW) or the GR antagonist RU486 (100mg/kg BW). At 24-h post-implantation, fish were tethered to the seafloor and observed for behavioural metrics associated with predation. Blood samples were collected from a subset of fish to assess the physiological consequences of the implants. Cortisol- and RU486-implanted fish both had significantly higher plasma cortisol concentrations than sham fish, with blood glucose and plasma urea being elevated only in the former. Further, anti-predator behaviours and predation mortality did not differ significantly among treatments. Despite changes in physiological state, predation susceptibility was unaffected, a finding that may reflect the complex relationships linking the physiology and behaviour of an organism as well as potential tethering artefacts.

Cortisol treatment affects locomotor activity and swimming behaviour of male smallmouth bass engaged in paternal care: A field study using acceleration biologgers

Paternal care, where the male provides sole care for the developing brood, is a common form of reproductive investment among teleost fish and ubiquitous in the Centrarchidae family. Throughout the parental care period, nesting males expend energy in a variety of swimming behaviours, including routine and burst swimming, vigilantly monitoring the nest area and protecting the brood from predators. Parental care is an energetically demanding period, which is presumably made even more difficult if fish are exposed to additional challenges such as those arising from human disturbance, resulting in activation of the hypothalamic-pituitary-interrenal axis (i.e., elevation of cortisol). To study this situation, we examined the effects of experimental manipulation of the stress hormone cortisol on locomotor activity and behaviour of nest guarding male smallmouth bass (Micropterus dolomieu). We exogenously elevated circulating cortisol levels (via intracoelomic implants) and attached tri-axial accelerometers to wild smallmouth bass for three days. During the recovery period (i.e., ≤4h post-release), cortisol-treated fish exhibited significantly reduced locomotor activity and performed significantly less burst and routine swimming relative to control fish, indicating cortisol uptake was rapid, as were the associated behavioural responses. Post-recovery (i.e., >4h post-release), fish with high cortisol exhibited lower locomotor activity and reduced routine swimming relative to controls. Fish were less active and reduced routine and burst swimming at night compared to daylight hours, an effect independent of cortisol treatment. Collectively, our results suggest that cortisol treatment (as a proxy for anthropogenic disturbance and stress) contributed to altered behaviour, and consequently cortisol-treated males decreased parental investment in their brood, which could have potential fitness implications.

Influence of supraphysiological cortisol manipulation on predator avoidance behaviours and physiological responses to a predation threat in a wild marine teleost fish

The stress axis in teleost fish attempts to maintain internal homeostasis in the face of allostatic loading. However, stress axis induction has been associated with a higher predation rate in fish. To date, the physiological and behavioral factors associated with this outcome are poorly understood. The purpose of the present study was to investigate the impact of experimental cortisol elevation on anti-predator behavior and physiological responses to predator presence. We hypothesized that semi-chronic cortisol elevation would increase susceptibility to predation by increasing stress-induced risk-taking behaviors. To test this hypothesis, schoolmaster snapper were given cocoa butter implants without cortisol (sham) or with cortisol (50 mg/kg body weight) and tethered to cover. Fish were exposed to either a lemon shark or control conditions for 15-min. Space use and activity were recorded throughout and fish were terminally sampled for blood. Cortisol implantation, relative to shams, resulted in higher blood glucose and plasma cortisol concentrations with a lower plasma lactate concentration. Shark exposure, relative to controls, elicited higher blood glucose and lactate concentrations but had no effect on plasma cortisol concentration. No interactions were detected between shark exposure and cortisol treatment for any physiological trait. Behavioral metrics, including shelter use and activity, were unaffected by either cortisol implantation or shark exposure. Physiological responses to cortisol implantation likely resulted from enhanced gluconeogenic activity, whereas alterations under predator exposure may have been the product of catecholamine mobilization. Further work should address context-specific influences of stress in mediating behavioral responses to predation.

Does experimental cortisol elevation mediate risk-taking and antipredator behaviour in a wild teleost fish?

The hypothalamic-pituitary-interrenal (HPI) axis is centrally implicated in stressor mitigation in teleost fishes. Sustained HPI axis activation can be detrimental to the physiological functioning of an organism and can result in fitness-related trade-offs. Predator-induced mortality is known to be higher in stressed fish than in unstressed conspecifics, suggesting a role for the HPI axis in mediating fish behaviour. However, the underlying specific mechanism(s) for this phenomenon is(are) unknown. The purpose of the current study was to address how the HPI axis influences risk-taking, and antipredator behaviours in a wild teleost, the pumpkinseed sunfish (Lepomis gibbosus). Here, individual juvenile pumpkinseed were implanted either with cocoa butter as a sham control or with a biologically-relevant concentration of cortisol. Forty-eight hours post-implantation, fish were assessed for behavioural metrics associated with boldness and risk taking in three sequential behavioural tests: (i) a predation-risk test, (ii) an exploration tendency test, and (iii) a shoaling tendency test, with test order randomized among different trials. Cortisol treatment had no influence on antipredator, exploratory, or shoaling behaviours. However, post-attack swimming duration (in predation-risk test) and exploratory activity (in Z-maze exploration test) were significantly affected by body mass. Collectively, our results indicate that cortisol may not have a role in mediating sociability, boldness, and risk-taking behaviours in pumpkinseed sunfish, at least under the current laboratory conditions. However, cortisol may nonetheless play a role in mediating predator-prey interactions in fishes in more natural environmental settings that were not considered here.