Samuel Péan

Systematic Screening of Behavioral Responses in Two Zebrafish Strains

Wild-type (WT) zebrafish are commonly used in behavioral tests, but the term WT is not a precise description, and corresponds to many different strains (e.g., AB, TU, WIK, and others). Previous studies compared the physiological, behavioral, or metabolic characteristics of different zebrafish strains (indigenous WT populations versus laboratory WT strains). AB and TU are widely used, but at least one study has demonstrated behavioral differences between them. To choose the most appropriate strain for our experiments, we systematically screened behavioral responses of AB and TU fish in several assays. We analyzed the locomotion activity and responses to a light/dark challenge in adults and larvae, and exploratory behavior and color conditioning in adults. Differences were observed for all tests, the strains displaying particular behavior depending on the tests. As larvae, TU displayed a wider activity range than AB larvae at the onset of locomotor behavior; as adults, TU were more reactive to sudden light transitions and recovered the swimming activity faster in T-maze or homebase release in novel tank tests, whereas AB fish had more contrasted circadian rhythms and performed better in color learning. Strain-specific behavior should be considered when designing experiments using behavior.

Long-term dietary-exposure to non-coplanar PCBs induces behavioral disruptions in adult zebrafish and their offspring

The use of polychlorinated biphenyls (PCBs) has been banned for several decades. PCBs have a long biological half-life and high liposolubility which leads to their bioaccumulation and biomagnification through food chains over a wide range of trophic levels. Exposure can lead to changes in animal physiology and behavior and has been demonstrated in both experimental and field analyses. There are also potential risks to high trophic level predators, including humans. A maternal transfer has been demonstrated in fish as PCBs bind to lipids in eggs. In this study, behavioral traits (exploration and free swimming, with or without challenges) of contaminated zebrafish (Danio rerio) adults and their offspring (both as five-day-old larvae and as two-month-old fish reared under standard conditions) were measured using video-tracking. Long-term dietary exposure to a mixture of non-coplanar PCBs was used to mimic known environmental contamination levels and congener composition. Eight-week-old fish were exposed for eight months at 26-28 °C. Those exposed to an intermediate dose (equivalent to that found in the Loire Estuary, ∑(CB)=515 ng g⁻¹ dry weight in food) displayed behavioral disruption in exploration capacities. Fish exposed to the highest dose (equivalent to that found in the Seine Estuary, ∑(CB)=2302 ng g⁻¹ dry weight in food) displayed an increased swimming activity at the end of the night. In offspring, larval activity was increased and two-month-old fish occupied the bottom section of the tank less often. These findings call for more long-term experiments using the zebrafish model; the mechanisms underlying behavioral disruptions need to be understood due to their implications for both human health and their ecological relevance in terms of individual fitness and survival.

Electronic individual identification of zebrafish using radio frequency identification (RFID) microtags.

SUMMARY Although individual electronic tagging using passive integrated acoustic (PIT) tags is established, it is mainly for fish >60 mm in length and is unsuitable for fish of <30 mm, like zebrafish. We used radio frequency identification (RFID) microtags (1 mm in diameter and 6 mm in length, with a mass of ~10 mg) to individually identify juvenile zebrafish (length 16–42 mm, mass 138–776 mg) for the first time, and studied the effects of intracoelomic implantation on fish survival and microtag loss, growth, spawning and exploratory behaviour. After 5.5 months, both high survival (82%) and low microtag loss (11%) were achieved. The smallest surviving fish weighed 178 mg, and success in microtag reading was 73% for the size class 350–450 mg (26 mm). Greater success was achieved when fish were larger at the time of tagging but no negative effects on growth were observed for any size class and some tagged fish spawned. No significant differences in behavioural responses could be detected between tagged fish and untagged controls after 2 months. Overall, the results suggest that the tagging method is highly suitable for fish as small as zebrafish juveniles. We think this method will provide significant advances for researchers of the ever-growing fish model community and more generally for all small-fish users. Tagging is essential when one needs to identify fish (e.g. particular genotypes with no external cue), to run longitudinal monitoring of individual biological traits (e.g. growth) or to repeat assays with the same individual at discrete points in time (e.g. behaviour studies). Such a method will find applications in physiology, genetics, behaviour and (eco)toxicology fields.

Assessment of Genetic Variability of Fish Personality Traits using Rainbow Trout Isogenic Lines

The study of inter-individual variability of personality in fish is a growing field of interest but the genetic basis of this complex trait is still poorly investigated due to the difficulty in controlling fish genetic origin and life history. When available, isogenic lines that allow performing independent tests on different individuals having identical genotype constitute a very relevant experimental material to disentangle the genetic and environmental components of behavioural individuality. We took advantage of heterozygous isogenic lines to investigate the personality in rainbow trout through the analysis of their reactions to different experimental situations. To this end, seven to ten rainbow trout isogenic lines were screened for their spatial exploratory behaviour, their flight response toward a stressor and their risk taking behaviour. Results showed that some lines seemed less sensitive to new events or environmental changes and could be defined as low responsive, while others were very sensitive and defined as high responsive. The use of isogenic lines highlighted the importance of genetic factors, in combination with life history, in the expression of personality in domesticated fish.

Self-feeding behavior changes induced by a first and a second generation of domestication or selection for growth in the European sea bass, Dicentrarchus labrax

Among the strategies that can be used to improve fish welfare in a rearing environment, domestication and/or selective breeding was proposed to minimize fish responsiveness to husbandry practices. To verify this hypothesis on a recently domesticated species, the sea bass Dicentrarchus labrax, two experiments were realized, each using two populations differing according to their level of domestication or selection. For the first experiment, we used one population produced from wild parents (Wild; initial body mass: 106 ± 3 g), and one population from parents selected for growth for one generation (Selected 1; initial body mass: 129 ± 4 g). For the second experiment, we used one population produced from parents domesticated for two generations (Domesticated; initial body mass: 72 ± 3g ), and one produced from parents selected for growth for two generations (Selected 2; initial body mass: 89 ± 4 g). The first experiment was carried out over 112 days with 240 fish (60 fish per tank, 120 fish per population), and the second one over 84 days with 200 fish (50 fish per tank, 100 fish per population). Two variables, self-feeding behavior and growth performance, were measured over the time of the experiments. After a control period, the fish were submitted twice, at three-week intervals, to an acute stress treatment consisting of draining the tank and leaving the fish out of water for one minute. Both self-feeding behavior and growth performance were altered by the acute stress treatment. During the first post-stress period, the Domesticated and Selected (1 and 2) groups showed more pronounced post-stress exposure responses than the Wild fish: they modified their feeding rhythm, their feed intake, and their growth rate. During the second post-stress period, feeding rhythm was still affected (being more diurnal with a well defined peak), but the feed intake and growth rate results showed that the Domesticated and Wild groups seemed less affected than the Selected (1 and 2) populations, which continued to express a high post-stress response. According to these results, it can be concluded that: (1) an application of two acute stress treatments, at three-week intervals, modified fish feeding behavior and growth performance; (2) the domestication process seemed to improve fish adaptation abilities to this kind of stress; and (3) the process of selection for growth led to a final, better growth, but did not seem to improve fish acute stress tolerance.