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Animals' selection of environments within a preferred range is key to understanding their habitat selection, tolerance to stressors and responses to environmental change. For aquatic animals, preferred environmental ranges can be studied in so-called shuttle-boxes, where an animal can choose its ambient environment by shuttling between separate choice chambers with differences in an environmental variable. Over time, researchers have refined the shuttle-box technology and applied them in many different research contexts, and we here review the use of shuttle-boxes as a research tool with aquatic animals over the past 50 years. Most studies on the methodology have been published in the latest decade, probably due to an increasing research interest in the effects of environmental change, which underlines the current popularity of the system. The shuttle-box has been applied to a wide range of research topics with regards to preferred ranges of temperature, CO 2 , salinity and O 2   in a vast diversity of species, showing broad applicability for the system. We have synthesized the current state-of-the-art of the methodology and provided best practice guidelines with regards to setup, data analyses, experimental design and study reporting. We have also identified a series of knowledge gaps, which can and should be addressed in future studies. We conclude with highlighting directions for research using shuttle-boxes within evolutionary biology and behavioural and physiological ecology., (© The Author(s) 2021. Published by Oxford University Press and the Society for Experimental Biology.)

Although considered a stenohaline freshwater species, European perch ( Perca fluviatilis ) inhabit brackish waters. The present study determined the maximum salinity tolerance and osmoregulatory capability on individuals originating from brackish water and from freshwater populations. The fish were acclimated for 3 weeks to salinities of 0, 10, 12.5, 15, 17.5 and 20 after an initial stepwise increase to the target salinity. The maximum salinity tolerance was determined as the test salinity below which the fish could not acclimate and lost equilibrium. Blood plasma osmolality was measured if the fish had not lost equilibrium after the acclimation period. The maximum salinity tolerance was 17.5 for brackish water European perch and 10 for fresh water European perch. The high salinity tolerance of the brackish water European perch was caused by their ability to both hyper- and hypo-osmoregulate, whereas the freshwater originating fish could only hyper-osmoregulate. The results showed that maximum salinity tolerances and osmoregulatory capabilities depends on the origin habitat salinity. Due to genetic differentiation between European perch populations in brackish and fresh water, the possibility of brackish water European perch being a subspecies of European perch is discussed, yet vital knowledge concerning heritability of salinity tolerance traits is still missing. Regardless of species status, within-species plasticity in the ability to cope with varying salinities have substantial ecological and conservation implications and underlines the need for managing brackish water and freshwater European perch stocks separately.