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Fish models used for chemical exposure in toxicological studies are normally kept in barren tanks without any structural environmental enrichment. Here, we tested the combined effects of environmental enrichment and exposure to two mixtures of endocrine disrupting chemicals (EDCs) in zebrafish. Firstly, we assessed whether developmental exposure to an EDC mixture (MIX G1) combined with rearing the fish in an enriched environment influenced behaviour later in life. This was evaluated using locomotion tracking one month after exposure, showing a significant interaction effect between enrichment and the MIX G1 exposure on the measured locomotion parameters. After three months, we assessed behaviour using custom-made behaviour tanks, and found that enrichment influenced swimming activity. Control fish from the enriched environment were more active than control fish from the barren environment. Secondly, we exposed adult zebrafish to a separate EDC mixture (MIX G0) after rearing them in a barren or enriched environment. Behaviour and hepatic mRNA expression for thyroid-related genes were assessed. There was a significant interaction effect between exposure and enrichment on swimming activity and an effect of environment on latency to approach the group of conspecifics, where enriched fish took more time to approach the group, possibly indicating that they were less anxious. Hepatic gene expression of a thyroid-related gene (thrb) was significantly affected by EDC exposure, while enrichment had no discernible impact on the expression of the measured genes. In conclusion, environmental enrichment is important to consider when studying the effects of EDCs in fish.

Locomotion-based behavioural endpoints have been suggested as suitable sublethal endpoints for human and environmental hazard assessment, as well as for biomonitoring applications. Larval stages of the sand goby (Pomatoschistus minutus) possess a number of attractive qualities for experimental testing that make it a promising species in behavioural ecotoxicology. Here, we present a study aimed at developing a toolkit for using the sand goby as novel species for ecotoxicological studies and using locomotion as an alternative endpoint in toxicity testing. Exposure to three contaminants (copper (Cu), di-butyl phthalate (DBP) and perfluorooctanoic acid (PFOA) was tested in the early life stages of the sand goby and the locomotion patterns of the larvae were quantified using an automatic tracking system. In a photo-motor test, sand goby larvae displayed substantially higher activity in light than in dark cycles. Furthermore, all tested compounds exerted behavioural alterations, such as hypo- and hyperactivity. Our experimental results show that sand goby larvae produce robust and quantifiable locomotive responses, which could be used within an ecotoxicological context for assessing the behavioural toxicity of environmental pollutants, with particular relevance in the Nordic region. This study thus suggests that sand goby larvae have potential as an environmentally relevant species for behavioural ecotoxicology, and as such offer an alternative to standard model species.