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Coping styles consist of a coherent set of individual physiological and behavioral differences in stress responses that are consistent across time and context. Such consistent inter-individual differences in behavior have already been shown in European sea bass (Dicentrarchus labrax), but the associated mechanisms are still poorly understood. Here, we combine physiological measurements with individual behavioral responses in order to characterize coping styles in fish. Fish were tagged and placed in a tank for group risk-taking tests (GRT) at 8 months of age to evaluate boldness using the proxy latency of leaving a sheltered area towards an open area. A subsample of these fish were individually challenged 16 months later using an open field test (OFT), in which the boldness was assessed after being placed in a shelter within an open arena. Latency to exit the shelter, time spent in the shelter, and distance travelled were recorded for this purpose. The blood and brain were then collected to evaluate plasma cortisol concentration and neurotransmitter levels (dopamine, norepinephrine, serotonin, and related metabolites), as well as brain transcription of key genes involved in stress axis regulation (gr1, gr2 , mr, crf), neurogenesis (neurod1, neurod2, pcna), and neuronal development (egr1). Fish acting bolder in the GRT were not necessarily those acting bolder in the OFT, highlighting the relatively low consistency across different types of tests performed with a 16-months interval. There was, however, a significant correlation between stress markers and boldness. Indeed, mRNA levels of mr, crf, gr2, egr1 , and neurod2 , as well as norepinephrine levels were higher in shy than bold fish, whereas brain serotonergic activity was lower in shy fish. Overall, our study highlights the fact that boldness was not consistent over time when testing context differed (group vs. alone). This is in agreement with previous literature suggesting that social context play a key role in boldness measurement and that the particular life history of each individual may account in shaping the personality fate of a fish. • Boldness is not consistent when characterized using different types of test over an interval period of 16 months (group vs. individual tests). • After open field test, plasma cortisol concentration is similar between bold and shy fish. • Shy fish, however, display higher level of transcription of mr , gr2 , crf , egr1 and neurod2 (trend) in the whole brain. • Shy fish display higher level of norepinephrine and lower turnover ratio between serotonin and s hydroxyindoleacetic acid in the whole brain. [ABSTRACT FROM AUTHOR]

Highlights • Zebrafish (F0) were chronically exposed to a realistic environmental PCB/PBDE mixture via diet. • Molecular and behavioral analyses were performed from F0 to F4 generation. • Environmental PCB/PBDE mixture does not alter behavior in F0 exposed fish. • Gene transcription and behavior are altered in unexposed offspring in F1 to F4 generations. • These results suggest a combination of multi- and transgenerational effects of an environmental PCB/PBDE mixture. Abstract Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants extensively used during the 20th century and still present in aquatic environments despite their ban. Effects of exposure to these compounds over generations are poorly documented. Therefore, our aims were to characterize behavioral responses and underlying molecular mechanisms in zebrafish exposed to an environmentally relevant mixture of PCBs and PBDEs as well as in four unexposed offspring generations. Zebrafish (F0) were chronically exposed from the first meal onward to a diet spiked with a mixture containing 22 PCB and 7 PBDE congeners in proportions and concentrations reflecting environmental situations (ΣPCBs = 1991 and ΣPBDEs = 411 ng/g). Four offspring generations (F1 to F4) were obtained from this F0 and were not further exposed. Behavior was assessed at both larval and adult stages. Mechanisms related to behavioral defects (habenula maturation and c-fos transcription) and methylation (dnmts transcription) were monitored in larvae. Exposed adult F0 as well as F1 and F3 adults displayed no behavioral change while F2 expressed anxiety-like behavior. Larval behavior was also disrupted, i.e. hyperactive after light to dark transition in F1 or hypoactive in F2, F3 and F4. Behavioral disruptions may be related to defect in habenula maturation (observed in F1) and change in c-fos transcription (observed in F1 and F2). Transcription of the gene encoding DNA methyltransferase (dnmt3ba) was also modified in all generations. Our results lead us to hypothesize that chronic dietary exposure to an environmentally relevant mixture of PCB and PBDE triggers multigenerational and transgenerational molecular and behavioral disruptions in a vertebrate model. [ABSTRACT FROM AUTHOR]

Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants that have been shown to affect fish life-history traits such as reproductive success, growth and survival. At the individual level, their toxicity and underlying mechanisms of action have been studied through experimental exposure. However, the number of experimental studies approaching marine environmental situations is scarce, i.e., in most cases, individuals are exposed to either single congeners, or single types of molecules, or high concentrations, so that results can hardly be transposed to natural populations. In the present study, we evaluated the effect of chronic dietary exposure to an environmentally realistic marine mixture of PCB and PBDE congeners on zebrafish life-history traits from larval to adult stage. Exposure was conducted through diet from the first meal and throughout the life cycle of the fish. The mixture was composed so as to approach environmentally relevant marine conditions in terms of both congener composition and concentrations. Life-history traits of exposed fish were compared to those of control individuals using several replicate populations in each treatment. We found evidence of slower body growth, but to a larger asymptotic length, and delayed spawning probability in exposed fish. In addition, offspring issued from early spawning events of exposed fish exhibited a lower larval survival under starvation condition. Given their strong dependency on life-history traits, marine fish population dynamics and associated fisheries productivity for commercial species could be affected by such individual-level effects of PCBs and PBDEs on somatic growth, spawning probability and larval survival. [ABSTRACT FROM AUTHOR]

Polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) are persistent organic pollutants still present in aquatic environments despite their total or partial ban. Previously, we observed that an environmentally realistic mixture of these compounds affects energy balance, growth, and reproduction in exposed zebrafish (F0), and behavior in their unexposed offspring (F1-F4). In the present work, we performed lipidomic and transcriptomic analyses on brains of zebrafish (F0-F2) from exposed and control lineages to identify molecular changes that could explain the observed phenotypes. The use of both technologies highlighted that F0 zebrafish displayed impaired mitochondrial function and lipid metabolism regulation (depletion in triacylglycerols and phospholipids) which can explain disruption of energy homeostasis. A subset of the regulated biological pathways related to energetic metabolism and neurotransmission were inherited in F2. In addition, there were increasing effects on epigenetic pathways from the F0 to the F2 generation. Altogether, we show that the effects of an environmental exposure to PCBs and PBDEs on energetic metabolism as well as neurotransmission extend over 2 generations of zebrafish, possibly due to transgenerational epigenetic inheritance. Unlabelled Image • A mixture of PCBs and PBDEs regulated energetic pathways in zebrafish male brain. • Transcriptomic and lipidomic changes supported the observed phenotypes. • These were partly inherited in the unexposed F2 generation. • Dysregulation of epigenetic mechanisms may be implicated in effect inheritance. [ABSTRACT FROM AUTHOR]

• Groups of fish were exposed to acute stressors (Hypoxia, Hyperoxia, High ammonia). • Behavior was recorded both during stress condition and following stress. • All stress conditions induce a decrease in activity, thigmotaxis and group cohesion. • Novel tank diving test was validated for welfare assessment in European sea bass. • Activity, thigmotaxis and group cohesion are sensitive welfare indicators. In aquaculture, fish are exposed to unavoidable stressors that can be detrimental for their health and welfare. However, welfare in farmed fish can be difficult to assess, and, so far, no standardized test has been universally accepted as a welfare indicator. This work contributes to the establishment of behavioural welfare indicators in a marine teleost in response to different water quality acute stressors. Groups of ten fish were exposed to high Total Ammonia Nitrogen concentration (High TAN, 18 mg.L-1), Hyperoxia (200 % O 2 saturation), Hypoxia (20 % O 2 saturation), or control water quality (100% O 2 saturation and TAN < 2.5 mg.L-1) over 1 hour. Fish were then transferred in a novel environment for a group behaviour test under the same water quality conditions over 2 hours. Videos were recorded to assess thigmotaxis, activity and group cohesion. After this challenge, plasma cortisol concentration was measured in a subsample, while individual behavioural response was measured in the other fish using novel tank diving test. Prior to this study, the novel tank diving test was validated as a behavioural challenge indicative of anxiety state, by using nicotine as anxiolytic drug. Overall, all stress conditions induced a decrease in activity and thigmotaxis and changes in group cohesion while only fish exposed to Hypoxia and High TAN conditions displayed elevated plasma cortisol concentrations. In post -stress condition, activity was still affected but normal behaviour was recovered within the 25 minutes of the test duration. Our work suggests that the activity, thigmotaxis and group cohesion are good behavioural indicators of exposure to degraded water quality, and could be used as standardized measures to assess fish welfare. [ABSTRACT FROM AUTHOR]

• Trout of different personality/competitive ability were exposed to aquaculture stressors (hypoxia and high ammonia). • Hypoxia resulted overall in reduced swimming activity, but induced no physiological stress. • High levels of ammonia were stressful, and induced an increase of avoidance behavior. • Personality-related behavioral differences were evident upon stress (high ammonia). • The data support the use of wireless accelerometer sensors as a promising tool for welfare monitoring in aquaculture. As other vertebrates, fish differ on an individual basis in their responses to disturbance (i.e. stress) and in their capacity for adaptation to environmental change. This individual stress-coping style (SCS) might have an impact on the individual welfare in aquaculture facilities. However, most of the studies about the behavioral and physiological stress responses of fish with different SCS were performed under conditions very unlike usual rearing conditions. Therefore, we aimed in the current study at investigating the behavioral and physiological responses of rainbow trout (Oncorhynchus mykiss) of different SCS to common aquaculture stressors (exposure to hypoxia or high ammonia) under realistic conditions. We first screened the fish according to their competitive ability, as a proxy of SCS, separating the best and worst competitors (winners, W, and losers, L, respectively). Then, we evaluated the behavioral (using both telemetric and video-monitoring approaches) and physiological response of both groups upon exposure to increasing levels of hypoxia and ammonia. Overall, increasing hypoxia induced a slight but progressive decrease in fish activity, independently of the fish SCS. High concentrations of total ammonia induced an increase of certain behavioral displays, such as swimming bursts or approaches to the surface, the latter being overall more frequent in W than in L fish. At the specific stress levels tested (hypoxia: 50 % oxygen saturation; total ammonia: 2.91 mM), the physiological stress markers showed that the behavioral response to ammonia was probably driven by stress, while the behavioral response of the fish to hypoxia was just a behavioral adjustment to accommodate the decrease in oxygen availability. In conclusion, our data show that fish of different competitive ability showed similar activity patterns in routine conditions or when adapting to non-stressful conditions (50 % oxygen saturation) in an aquaculture-like setup, but differed in their behavioral response when exposed to stress (high water ammonia). In addition, our data support the use of acoustic accelerometer transmitters as a promising tool for real-time monitoring of fish welfare in commercial fish farms. [ABSTRACT FROM AUTHOR]