Supplementary material and data for ''Interactive effects of sedimentary turbidity and elevated water temperature on the Pugnose Shiner (<i>Miniellus anogenus</i>), a Threatened freshwater fish''
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This supplementary material and data is associated with: Fortin-Hamel, L and Chapman, LJ ''Interactive effects of turbidity and elevated water temperature on the Pugnose Shiner (<i>Miniellus anogenus</i>), a Threatened freshwater fish'' (<i>under revision</i>).See our methods for detailed information on the collection of each dataset. Please contact Liana Fortin-Hamel (liana.fortin-hamel[at]mail.mcgill.ca) for any questions or comments.<b>Abstract</b>High turbidity and elevated water temperature are environmental stressors that can co-occur in freshwater ecosystems such as when deforestation increases solar radiation and sedimentary runoff. However, we have limited knowledge about their combined impacts on fish behaviour and physiology. We explored independent and interactive effects of sedimentary turbidity and temperature on the swimming activity and both thermal- and hypoxia tolerance of the Pugnose Shiner (<i>Miniellus anogenus</i>, formerly<i> Notropis anogenus</i>), a small leuciscid fish listed as Threatened under Canada’s Species at Risk Act (SARA). Fish underwent a 15-week acclimation to two temperatures (16 °C or 25 °C) crossed with two turbidities (~0 NTU or 8.5 NTU). Swimming activity was measured during the first 8 weeks of acclimation. Fish in warm water were more active compared to those in cold water, but turbidity had no effect on activity. Behavioural response to hypoxia was measured after 12 weeks of acclimation, as the oxygen level at which fish used aquatic surface respiration (ASR). Fish in warm water engaged in ASR behaviour at higher oxygen thresholds, indicating less tolerance to hypoxia. Turbidity had no effect on ASR thresholds. Finally, thermal tolerance was measured as the critical thermal maximum (CT<sub>max</sub>) after 13 to 15 weeks of acclimation. Acclimation to warm water increased fish CT<sub>max</sub> and T<sub>ag</sub> (agitation temperature) but reduced the agitation window (°C difference between T<sub>ag</sub> and CT<sub>max</sub>) and thermal safety margin (°C difference between acclimation temperature and CT<sub>max</sub>). Furthermore, fish in warm, turbid water had a lower CT<sub>max</sub> and smaller thermal safety margin than fish in warm, clear water, indicating an interaction between turbidity and temperature. This reduced thermal tolerance observed in Pugnose Shiner in warm, turbid water highlights the importance of quantifying independent and interactive effects of multiple stressors when evaluating habitat suitability and conservation strategies for imperiled species.<b>Code/Software</b>R scripts and codes used for statistical analysis are provided in our supplementary material (R Core Team, 2022; v.4.2.1). Essential packages for statistical analysis include:<i>lme4</i> v.1.1.30<i>car</i> v.3.1-0<i>emmeans</i> v.1.8.1-1<br>
