Mild maternal stress disrupts associative learning and increases aggression in offspring.

Maternal stress has been shown to affect behaviour of offspring in a wide range of animals, but this evidence has come from studies that exposed gestating mothers to acute or severe stressors, such as restraint or exposure to synthetic stress hormones. Here we show that exposure of mothers to even a mild stressor reduces associative learning and increases aggression in offspring. Female guppies were exposed to routine husbandry procedures that produced only a minimal, non-significant, elevation of the stress hormone cortisol. In contrast to controls, offspring from mothers that experienced this mild stress failed to learn to associate a colour cue and food reward, and showed a greater amount of inter-individual variation in behaviour compared with control offspring. This mild stress also resulted in offspring that were more aggressive towards their own mirror image than controls. While it is possible that these results could represent the transmission of beneficial maternal characteristics to offspring born into unpredictable environments, the potential for mild maternal stress to affect offspring performance also has important implications for research into the trans-generational effects of stress.

Using physiology and behaviour to understand the responses of fish early life stages to toxicants.

The use of early life stages of fishes (embryos and larvae) in toxicity testing has been in existence for a long time, generally utilizing endpoints such as morphological defects and mortality. Behavioural endpoints, however, may represent a more insightful evaluation of the ecological effects of toxicants. Indeed, recent years have seen a considerable increase in the use of behavioural measurements in early life stages reflecting a substantial rise in zebrafish Danio rerio early life-stage toxicity testing and the development of automated behavioural monitoring systems. Current behavioural endpoints identified for early life stages in response to toxicant exposure include spontaneous activity, predator avoidance, capture of live food, shoaling ability and interaction with other individuals. Less frequently used endpoints include measurement of anxiogenic behaviours and cognitive ability, both of which are suggested here as future indicators of toxicant disruption. For many simple behavioural endpoints, there is still a need to link behavioural effects with ecological relevance; currently, only a limited number of studies have addressed this issue. Understanding the physiological mechanisms that underlie toxicant effects on behaviour so early in life has received far less attention, perhaps because physiological measurements can be difficult to carry out on individuals of this size. The most commonly established physiological links with behavioural disruption in early life stages are similar to those seen in juveniles and adults including sensory deprivation (olfaction, lateral line and vision), altered neurogenesis and neurotransmitter concentrations. This review highlights the importance of understanding the integrated behavioural and physiological response of early life stages to toxicants and identifies knowledge gaps which present exciting areas for future research.

Alarm substance from adult zebrafish alters early embryonic development in offspring.

Alarm substances elicit behavioural responses in a wide range of animals but effects on early embryonic development are virtually unknown. Here we investigated whether skin injury-induced alarm substances caused physiological responses in embryos produced by two Danio species (Danio rerio and Danio albolineatus). Both species showed more rapid physiological development in the presence of alarm substance, although there were subtle differences between them: D. rerio had advanced muscle contraction and heart function, whereas D. albolineatus had advanced heart function only. Hence, alarm cues from injured or dying fish may be of benefit to their offspring, inducing physiological responses and potentially increasing their inclusive fitness.

Chemical excretions of angled bonefish Albula vulpes and their potential use as predation cues by juvenile lemon sharks Negaprion brevirostris.

Bonefish Albula vulpes (n = 7) exercised to exhaustion and air exposed for 1 min as part of a catch-and-release angling event were found to excrete both ammonia and urea, but cortisol and lactate were below detectable levels. Urea made up a greater proportion of total nitrogen excretion from these fish at all time points following an angling event. When captive juvenile lemon sharks Negaprion brevirostris (n = 12) were exposed to a 30 s pulse of these chemicals [ammonia (500 mM), cortisol (20 µg l(-1) ), lactate (6 mM) or urea (3 mM)], they showed a significant reduction in the frequency of resting behaviours when exposed to ammonia and urea than when exposed to control water. It appears that products excreted by A. vulpes, particularly ammonia and urea, may provide an olfactory cue for the post-release predation of A. vulpes by N. brevirostris during catch-and-release angling events.

Physiology and behaviour of Atlantic salmon (Salmo salar) smolts during commercial land and sea transport.

This study examined the physiology (plasma cortisol, glucose, lactate, potassium, sodium and chloride concentrations) and behaviour (underwater video footage) of commercially produced Atlantic salmon (Salmo salar) smolts during transport from freshwater farms to saltwater net pens. Smolts were transported by truck in closed tanks from two freshwater farms to the dock (30-60 min), and then in the flow-through cargo holds of a live-haul vessel, the Sterling Carrier, to the saltwater net pens (~2 h). Some fish were dockside in the vessel for up to 8 h while successive deliveries were loaded into the holds. Fish and water were sampled both before and after truck transport, and then at several time points aboard the vessel. Analysis of plasma constituents showed modest primary and secondary stress responses due to loading and truck transport, and the recovery that occurred dockside in the live-haul vessel was maintained when the vessel was underway. Underwater video footage revealed behavioural differences between fish from the two freshwater facilities that were not evident from the physiological measurements, but the behaviours observed during transport on a live-haul vessel were consistent with a non-stressful environment. Although smolts were subjected to moderately stressful conditions during loading and trucking, they began to recover rapidly aboard the Sterling Carrier. We therefore conclude that smolt transport, as currently conducted by our industry partner, appears to reflect good fish welfare.

The effects of copper on the morphological and functional development of zebrafish embryos.

Waterborne copper exposure can exert a variety of physiological effects in fish, including the disruption of sensory system function, which has wide-reaching implications for fish behaviour. In developing fish larvae, copper is known to affect key parameters, such as survival and growth and more recently has been shown to interfere with the octavolateral system. The present study aimed to take a combined view of morphological (e.g. length, yolk sac area) and functional (e.g. heart beat, behaviour) processes to understand the complex effect of copper on fish development. In the first of two experiments, zebrafish embryos were exposed to a range of copper concentrations (11-1000 microg l(-1)) from fertilisation for a 72 h period. The greatest mortality was seen between 5 and 24h post-fertilisation (hpf) and was more pronounced at the higher copper concentrations. Copper also had an inhibitory effect on hatching. Length and yolk sac area of individuals were recorded across treatments at 72 hpf and elevated copper was found to slow development. Individuals from the higher copper treatments had the fastest heart rates at 28 hpf suggesting that stress responses were induced in the embryos during copper exposure. In the second experiment, embryos were exposed in a similar manner to two copper concentrations, based on those from Experiment 1 that resulted in <50% mortality. At 120 hpf, embryos exposed to both copper concentrations possessed significantly fewer functional neuromasts, an effect which was associated with a reduced ability to orientate in a current. Therefore, although mortality at these copper concentrations was low initially, and then almost non-existent after 24 hpf, the inability of copper-exposed larvae to orientate in a water current as a result of lateral line dysfunction is likely to seriously compromise survival.

The integration of behaviour into comparative physiology.

Comparative physiology has traditionally focused on the physiological responses of animals to their physicochemical environment. In recent years, awareness has increased among physiologists of the potential for behavioural factors, such as the social environment of the animal, to affect physiological condition and responses. This recognition has led to an emerging trend within the field toward using multidisciplinary approaches that incorporate both behavioural and physiological techniques. Research areas in which the integrated study of behaviour and physiology has been particularly fruitful include the physiology of the social environment, sensory physiology and behaviour, and physiological constraints on behavioural ecology. The manner in which incorporating behavioural considerations has informed the physiological data collected is discussed for each of these areas using specific examples.

The effects of trace metal exposure on agonistic encounters in juvenile rainbow trout, Oncorhynchus mykiss.

The effects of five trace metals, copper, cadmium, nickel, zinc and lead (presented as soluble salts) on the ability of juvenile rainbow trout to form social relationships were investigated. Comparable concentrations of the five metals in relation to their acute 96 h LC50s (concentration at which population mortality=50% at 96 h) were used (i.e. 15% of the 96 h LC50) and water quality parameters (hardness=120 mg l(-1) as CaCO(3), pH 8; DOC=3 mg l(-1)) were kept constant throughout. In the first experiment, trout exposed to sublethal concentrations of cadmium for 24 h displayed significantly lower numbers of aggressive attacks during pair-wise agonistic encounters than fish paired in the copper, nickel, zinc, lead and control water. In a second experiment, fish were exposed to the same concentration of metal for 24 h, and then returned to normal water for 24 h. When these metal pre-exposed fish were paired with non-exposed fish only cadmium pre-exposure had a significant effect on social interaction. All of the cadmium pre-exposed fish became subordinate when paired with non-exposed fish, whereas the probability of a fish pre-exposed to copper, nickel, zinc or lead becoming subordinate did not significantly differ from random. Therefore, at around 15% of the 96 h LC50, different metals exert different effects on the social behaviour of fish, suggesting potential implications for social structure and population stability.

Plasma cortisol concentrations before and after social stress in rainbow trout and brown trout.

Two related experiments examined the relationship between plasma cortisol concentrations and the development of social hierarchies in fish. In the first, rainbow trout, Oncorhynchus mykiss, and brown trout, Salmo trutta, were observed for dominance interactions when confined within single-species pairs for 4, 48, or 168 h. Subordinate members of a pair exhibited significantly higher cortisol concentrations than dominant and single fish, but the pattern of cortisol elevation differed between the two species, being quicker to rise and increasing to a higher level in rainbow trout. Cortisol concentrations were correlated with behavioural measurements; the more subordinate the behaviour exhibited by a fish, the higher its cortisol concentration. Social stress was a chronic stressor, and no acclimation to social status occurred during the week. In the second experiment, measurements of plasma cortisol were made before pairing of rainbow trout and then after 48 h of confinement in pairs. Subordinate fish demonstrated significantly higher concentrations of plasma cortisol both before and after social stress. It therefore appears that in addition to cortisol being elevated during periods of social stress, an association may exist between initial cortisol levels and the likelihood of a fish becoming subordinate.

Evidence for a mineralocorticoid-like receptor linked to branchial chloride cell proliferation in freshwater rainbow trout.

Fish acclimated to ion-deficient water exhibit proliferation of branchial chloride cells. The objective of the present study was to investigate the role of cortisol in this response using the corticosteroid receptor antagonists RU486 and spironolactone. RU486 is a potent antagonist of the glucocorticoid actions of cortisol, whereas spironolactone exhibits high-affinity binding to mineralocorticoid receptors, with a resulting blockade of mineralocorticoid properties in mammals. Untreated rainbow trout, as well as rainbow trout given a single intraperitoneal implant of coconut oil alone, coconut oil containing RU486 (0.5 mg g(-1)) or coconut oil containing spironolactone (0.1 mg g(-1)), were exposed to either dechlorinated city-of-Ottawa tapwater or artificial softwater for 7 days. Neither corticosteroid antagonist nor acclimation condition affected circulating plasma cortisol levels, plasma ion concentrations or gill Na(+)-K(+)-ATPase activity. Kidney Na(+)-K(+)-ATPase activity was significantly higher in softwater-acclimated fish than in fish held in dechlorinated tapwater. In addition, whereas RU486 treatment was found to be without effect on gill morphometrics, treatment with spironolactone inhibited the proliferation of chloride cells normally associated with acclimation to ion-deficient water. The results of the present study provide further evidence for the mineralocorticoid actions of cortisol in freshwater fish, specifically in eliciting chloride cell proliferation. Furthermore, these results support the hypothesis that distinct glucocorticoid and mineralocorticoid receptor populations are present in teleost fish, despite the apparent absence of the classic mineralocorticoid hormone, aldosterone.

Socially-induced changes in sodium regulation affect the uptake of water-borne copper and silver in the rainbow trout, Oncorhynchus mykiss.

Subordinate fish take up more copper during water-borne exposure than dominant fish and consequently display higher tissue burdens. The present study demonstrated a similar effect of social status on water-borne silver uptake. We evaluated whether differences in copper and silver accumulation between individuals could be due to differences in metabolic rate, internal concentrations of cortisol or sodium uptake rates. In the absence of social interaction, experimentally increased metabolic rates (via moderate exercise) and elevated whole body cortisol concentrations (via feeding of a cortisol-spiked diet) did not result in increased metal uptake. However, elimination of the difference in sodium uptake rates between dominant and subordinate fish by exposing them to a saturating level of water-borne sodium (50 mM) resulted in an elimination of copper uptake differences. No significant differences in sodium and silver uptake rates were seen between dominant and subordinate fish exposed to elevated silver concentrations. Therefore, it appears that socially-mediated differences in copper and silver accumulation are a result of differences in sodium uptake rates as both silver and copper are known to cross the gill epithelia via sodium transport pathways.