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DocuSign Envelope ID: D07924E6-3D94-4398-AO66-B683COBAB33E <br /> 2.2 Pimephales promelas 7-day toxicity test to estimate chronic <br /> toxicity <br /> Method: EPA-821-R-02-013 -- Short-Term Method for Estimating Chronic Toxicity. . . <br /> Test Duration: 7 days <br /> Sample Collection Procedure: grab <br /> Sample Collection Date: 11/4/20, 11/17/20, 1/18/21 <br /> Dilution water: N/A; moderately hard reconstituted water control <br /> Age of organisms at start: <24 hours old <br /> Feeding: 0.15 mL Artemia nauplii 2 times per day <br /> End Point: mortality, growth <br /> Start Date and Time: 0.5 g ethanol/g SO4 11/5/20 13:00 <br /> 1.0 g ethanol/g SO4 11/5/20 13:15 <br /> 0.25 g ethanol/g SO4 11/17/20 15:20 <br /> 0.5 g ethanol/g SO4 <br /> with/without <br /> lime pre-softening 1/19/21 12:00 <br /> End Date and Time: 0.5 g ethanol/g SO4 11/12/20 12:00 <br /> 1.0 g ethanol/g SO4 11/12/20 12:25 <br /> 0.25 g ethanol/g SO4 11/24/20 15:30 <br /> 0.5 g ethanol/g SO4 <br /> with/without <br /> lime pre-softening 1/26/21 11:40 <br /> Type of exposure chamber: 9 oz. disposable plastic cup <br /> Volume of exposure chamber: 230 mL <br /> Number of animals exposed/chamber: 10 <br /> Number of replicates/treatment: 4 <br /> Test temperature: 0.5 g ethanol/g SO4 23.1°C ± 2.0°C <br /> 1.0 g ethanol/g SO4 23.0°C ± 2.0°C <br /> 0.25 g ethanol/g SO4 23.2°C ± 1.0°C <br /> 0.5 g ethanol/g SO4 <br /> with/without <br /> lime pre-softening 23.5°C ± 1.0°C <br /> EPA recommended range: 25.0°C± 1.0°C <br /> Standard toxicant used: NaCl <br /> GEI Consultants,Inc. 5 February 2021 <br /> Ecological Division Whole Effluent Toxicity Test Report <br /> Stantec-Elk Ridge Mining and Reclamation,LLC <br />