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Stress responsiveness differs between individuals and is often categorized into different stress coping styles. Using these stress coping styles for selection in fish farming could be beneficial, since stress is one main factor affecting welfare. In Arctic charr (Salvelinus alpinus) carotenoid pigmentation is associated with stress responsiveness and stress coping styles. Thus this could be an important tool to use for selection of stress resilient charr. However, anaesthetics seem to affect carotenoid pigmentation, and it would be better if the method for selection could be implemented during normal maintenance, which usually includes anaesthetics. Therefore, this study investigated how the use of anaesthetics affected carotenoid pigmentation, i.e. number of spots, over time compared to no-anaesthetic treatment. Additionally, the stress indicators monoamines and glucocorticoids were investigated. The results indicate that the anaesthetic MS-222 affects number of spots on the right side. This anaesthetic also increased dopaminergic activity in the telencephalon. Both brain dopaminergic and serotonergic activity was associated with spottiness. Further, behaviour during anaesthetization was associated with spots on the left side, but not the right side. Repetition of the same treatment seemed to affect spot numbers on the right side. In conclusion, this study shows that inducing stress in charr affects the carotenoid spots. Thus, it is possible to use anaesthetics when evaluating spottiness although careful planning is needed.

Predation is thought to be one of the main structuring forces in animal communities. However, selective predation is often measured on isolated traits in response to a single predatory species, but only rarely are selective forces on several traits quantified or even compared between different predators naturally occurring in the same system. In the present study, we therefore measured behavioral and morphological traits in young-of-the-year Eurasian perch Perca fluviatilis and compared their selective values in response to the 2 most common predators, adult perch and pike Esox lucius. Using mixed effects models and model averaging to analyze our data, we quantified and compared the selectivity of the 2 predators on the different morphological and behavioral traits. We found that selection on the behavioral traits was higher than on morphological traits and perch predators preyed overall more selectively than pike predators. Pike tended to positively select shallow bodied and nonvigilant individuals (i.e. individuals not performing predator inspection). In contrast, perch predators selected mainly for bolder juvenile perch (i.e. individuals spending more time in the open, more active), which was most important. Our results are to the best of our knowledge the first that analyzed behavioral and morphological adaptations of juvenile perch facing 2 different predation strategies. We found that relative specific predation intensity for the divergent traits differed between the predators, providing some additional ideas why juvenile perch display such a high degree of phenotypic plasticity.

Pharmaceuticals as environmental contaminants have received a lot of interest over the past decade but, for several pharmaceuticals, relatively little is known about their occurrence in European surface waters. Benzodiazepines, a class of pharmaceuticals with anxiolytic properties, have received interest due to their behavioral modifying effect on exposed biota. In this study, our results show the presence of one or more benzodiazepine(s) in 86% of the analyzed surface water samples (n = 138) from 30 rivers, representing seven larger European catchments. Of the 13 benzodiazepines included in the study, we detected 9, which together showed median and mean concentrations (of the results above limit of quantification) of 5.4 and 9.6 ng L-1, respectively. Four benzodiazepines (oxazepam, temazepam, clobazam, and bromazepam) were the most commonly detected. In particular, oxazepam had the highest frequency of detection (85%) and a maximum concentration of 61 ng L-1. Temazepam and clobazam were found in 26% (maximum concentration of 39 ng L-1) and 14% (maximum concentration of 11 ng L-1) of the samples analyzed, respectively. Finally, bromazepam was found only in Germany and in 16 out of total 138 samples (12%), with a maximum concentration of 320 ng L-1. This study clearly shows that benzodiazepines are common micro-contaminants of the largest European river systems at ng L-1 levels. Although these concentrations are more than a magnitude lower than those reported to have effective effects on exposed biota, environmental effects cannot be excluded considering the possibility of additive and sub-lethal effects.

Psychoactive substances are used worldwide and constitute one of the most common groups of pharmaceutical contaminants in surface waters. Although these pharmaceuticals are designed to be efficiently eliminated from the human body, very little is known about their trophic-transfer potential in aquatic wildlife. Therefore, the goal of the present study was to quantify and compare uptake of an anxiolytic (oxazepam) from water (bioconcentration) and via the consumption of contaminated diet (trophic transfer) in 2 common freshwater predators: Eurasian perch (Perca fluviatilis) and the dragonfly larvae Aeshna grandis. Bioconcentration and trophic transfer of oxazepam were found in both predator species. However, higher bioconcentrations were observed for perch (bioconcentration factor [BCF], 3.7) than for dragonfly larvae (BCF, 0.5). Perch also retained more oxazepam from consumed prey (41%) than dragonfly larvae (10%), whereas the relative contribution via prey consumption was 14% and 42% for perch and dragonflies, respectively. In addition, bioconcentration was negatively correlated with perch weight, indicating that exposure levels in natural contaminated environments differ between individuals of different size or between different developmental stages. Hence, trophic transfer of pharmaceuticals may indeed occur, and estimates of environmental exposures that do not consider intake via food or size-dependent bioconcentration may therefore lead to wrongful estimations of realized exposure levels in natural contaminated ecosystems. (C) 2016 SETAC

A wide range of biologically active pharmaceutical residues is present in aquatic systems worldwide. As uptake potential and the risk of effects in aquatic wildlife are directly coupled, the aim of this study was to investigate the relationships between stress by isolation, uptake and effects of the psychiatric pharmaceutical oxazepam in fish. To do this, we measured cortisol levels, behavioral stress responses, and oxazepam uptake under different stress and social conditions, in juvenile perch (Percafluviatilis) that were either exposed (1.03 mu gl(-1)) or not exposed to oxazepam. We found single exposed individuals to take up more oxazepam than individuals exposed in groups, likely as a result of stress caused by isolation. Furthermore, the bioconcentration factor (BCF) was significantly negatively correlated with fish weight in both social treatments. We found no effect of oxazepam exposure on body cortisol concentration or behavioral stress response. Most laboratory experiments, including standardized bioconcentration assays, are designed to minimize stress for the test organisms, however wild animals experience stress naturally. Hence, differences in stress levels between laboratory and natural environments can be one of the reasons why predictions from artificial laboratory experiments largely underestimate uptake of oxazepam, and other pharmaceuticals, in the wild.

Many fish spend a large part of their life in groups. The size of the group influences potential costs and benefits of group living, and depending on context a fish may prefer different group sizes. Group-size preference may also depend on personality, with social individuals expected to prefer larger groups than asocial fish. This study investigates context-dependent group size preference in two populations of a highly social fish, young of the year Eurasian perch. The perch were given a choice between a group of two and a group of eight conspecifics under three different situations: the small group was feeding, the small group had access to shelter, and a control treatment with no extra stimuli. In general, the perch associated more with the large group, but significantly less so during the food treatment. Perceived access to shelter did not affect group size preference compared to the control treatment. Consistent individual differences in social attraction were found within each context, but not among all contexts. Also, an individual's sociability did not correlate with its degree of boldness, indicating a lack of a behavioural syndrome between the two personality traits in the studied populations. The results highlight the importance of considering environmental context when studying social behaviour in obligate social fish, and show the complexity of the concept of sociability as a personality trait by demonstrating context dependence in individual consistency in social behaviour.

The individual animal is currently a major focus of behavioural research and an increasing number of studies raise the question of how between-individual behavioural consistency and behavioural plasticity interact. Applying the reaction norm concept on groups, our study addresses both of these aspects in one framework and within an animal's natural social environment. Risk-taking behaviour in 1-year-old perch Perca fluviatilis was assayed in aquarium experiments before and after the fish were subjected to the presence or absence of a piscivorous predator for 3weeks. To analyse the inter-individual behavioural variation across the repeated measurements, we dissected the behavioural change across the predator treatment into individual constant and plastic components using hierarchical mixed-effects models. During the predator treatment, juvenile perch increased in boldness and decreased in vigilance, the magnitude of these behavioural changes was influenced by group composition. However, the behavioural changes were not influenced by the presence of a predator, indicating the difficulties in generating realistic long-term predation pressure in the laboratory. Individuals differed in the relative increase in boldness across the predator treatment and, thus, varied in the shape of their reaction norms. In accordance, the best linear unbiased predictors, extracted from the random effects of separate linear mixed-effects models for the data before and after the predator treatment were only weakly correlated. Hence, between-individual variation seems to change under laboratory conditions and therewith not necessarily represents the initially present natural' variation, giving important implications for the conduction and interpretation of behavioural experiments.

Psychoactive substances are used worldwide and constitute one of the common groups of pharmaceutical contaminants in surface waters. Typically, in field surveys and laboratory studies, muscle or whole - body homogenates are used to quantify pharmaceutical concentrations in biota, although uptake of pharmaceuticals may be tissue - specific. Therefore, the aim of this study was to investigate the tissue - specific (muscle, liver, brain and blood plasma) uptake of the anxiolytic oxazepam in adult Eurasian perch (Perca fluviatilis). In laboratory experiments, perch were exposed to four different concentrations (2, 4, 12 and 20 mu g L-1) of oxazepam for 6 days, and muscle, liver, brain tissue and blood plasma were sampled to determine tissue - specific bioconcentration. We found that the tissue - specific bioconcentration was independent of oxazepam concentration. However, among tissue types, bioconcentration was significantly different, with the concentration in muscle, liver = brain, blood plasma. Hence, it is important to consider the type of tissue used to quantify pharmaceutical uptake in fish, for predictions of species - specific sensitivity and comparisons across studies. Furthermore, our results indicate a somewhat lower transportability (brain/plasma ratio 0.54) of oxazepam from blood to brain in fish compared with in mammals, which should be kept in mind when employing 'read - across' approaches.

Group size, predation risk and habituation are key drivers of behaviour and evolution in gregarious prey animals. However, the extent to which they interact in shaping behaviour is only partially understood. We analyzed their combined effects on boldness and vigilance behaviour in juvenile perch (Perca fluviatilis) by observing individuals in groups of one, two, three and five faced with four different levels of predation risk in a repeated measures design. The perch showed an asymptotic increase in boldness with increasing group size and the highest per capita vigilance in groups of two. With increasing predation risk, individuals reduced boldness and intensified vigilance. The interaction between group size and predation risk influenced vigilance but not boldness. In this context, individuals in groups of two elevated their vigilance compared to individuals in larger groups only when at higher risk of predation. Further, as only group size, they significantly reduced vigilance at the highest level of risk. With increasing habituation, solitary individuals became considerably bolder. Also, predation risk affected boldness only in the more habituated situation. Hence, repeated measures may be essential to correctly interpret certain relationships in behaviour. Our results suggest that perch may adjust boldness behaviour to group size and predation risk independently. This is rather unexpected since in theory, natural selection would strongly favour an interactive adjustment. Finally, vigilance might be particularly effective in groups of two due to the intense monitoring and detailed response to changing levels of risk.

The study of animal behaviour is important for both ecology and ecotoxicology, yet research in these two fields is currently developing independently. Here, we synthesize the available knowledge on drug-induced behavioural alterations in fish, discuss potential ecological consequences and report results from an experiment in which we quantify both uptake and behavioural impact of a psychiatric drug on a predatory fish (Perca fluviatilis) and its invertebrate prey (Coenagrion hastulatum). We show that perch became more active while damselfly behaviour was unaffected, illustrating that behavioural effects of pharmaceuticals can differ between species. Furthermore, we demonstrate that prey consumption can be an important exposure route as on average 46% of the pharmaceutical in ingested prey accumulated in the predator. This suggests that investigations of exposure through bioconcentration, where trophic interactions and subsequent bioaccumulation of exposed individuals are ignored, underestimate exposure. Wildlife may therefore be exposed to higher levels of behaviourally altering pharmaceuticals than predictions based on commonly used exposure assays and pharmaceutical concentrations found in environmental monitoring programmes.