Much research has focused on the population-level effects of climate change on Eastern Brook Trout (). While some studies have considered here sub-lethal stress caused by warming waters, the role of multiple, interacting stressors remains largely unexplored. We used inducible heat shock protein 70 (HSP70) as a molecular biomarker to assess response of Eastern Brook Trout in headwater streams to multiple potential stressors, including temperature. Over 7 sampling events during 2018 and 2019, we sampled 141 fish and found that HSP70 expression and 3-day mean water temperature exhibited a quadratic relationship ( -adj = 0.68). Further analyses showed that HSP70 expression was explained by temperature, relative water level and their interaction ( -adj = 0.75), while fish size and capture location were not factors. We observed a significant increase in HSP70 expression during periods of low relative water level with warm temperatures (~18°C) and also during high relative water level with cold temperatures (~8°C). Our results suggest that temperatures at the edges of the preferred range coupled with relative water level might act together to trigger the cellular stress response in Eastern Brook Trout and that there is greater variation in response at colder temperatures. These findings reinforce the need to consider complex, interactive stressors in influencing the health and persistence of Eastern Brook Trout populations into the future.
