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Table of Contents <br />Abstract Page 2 Table 2 -Methods and Equipment Page 5 <br />Methods Page 2 References Page 5 <br />Results Page 3 Discharge Monitoring Report Form(s) Pg. 6-7 <br />Narrative & Discussion Page 4 Reference Toxicity Da#a Chart(s) Pg. 8 <br />Table 1 -Colorado Test Conditions Page 5 Supplement Forms' Pg. 9 <br />Abstract <br />SGS North America, Inc. conducted a two species chronic bioassay on Western Fuels New Horizon Mine <br />effluent discharge. This discharge has been designated as ou#fall 007. The purpose of this bioassay <br />was to determine if the facility complies with the chronic WET portion of the Colorado State Permit <br />System Authorization to Discharge. The test organisms utilized were the fathead minnow (Pimephales <br />promelas) and cladoceran (Ceriodaphnia dubia). Test results indicate that the effluent was not <br />chronically toxic to fathead minnow or to Ceriodaphnia dubia survival. Therefore, outfa11007 is in <br />compliance with the chronic WET portion of its Colorado State Discharge Permit for the first <br />quarter of 2008. <br />Methods <br />A two species chronic bioassay was conducted with effluent from Westem Fuels New Horizan Mine, <br />Outfall 007. An effluent composite sample was collected on March 3, 2008, stored on ice, and delivered <br />to SGS North America Inc. on March 4, 2008. The effluent samples were received at a temperature of <br />4.3° C and assigned sample tracking number 411-080-B1OA-015. The bioassay was initiated <br />approximately 24 hours after sample collection. <br />Subsequent samples were collected on March 5, 2006, stored on ice, and delivered to SGS North <br />America Inc. on March 6, 2008. The effluent samples were received at a temperature of 3.1 °C. <br />Because of the remote location of the facility, two concurrent samplings events were utilized for this <br />examination. Sufficient sample volume was collected for the entire test. <br />Testing was conducted in accordance with the most recent protocols specified by the Water Quality <br />Control Division of the Colorado Department of Health {1998) outlined in Table 1. Basic characteristics <br />of the effluent and dilutions were measured by the methods and equipment outlined in Table 2. <br />The tests exposed the cladocerans, Ceriodaphnia dubia, and fathead minnows, Pimephales promelas, to <br />five concentrations of effluent, 100%, 75%, 50%, 25%, and 12.5%, prepared using reconstituted <br />laboratory water. Control replicates were exposed to 100% laboratory water. <br />Statistically, fathead minnow survival and growth data were analyzed for significant differences using <br />Steel's Many One Rank Test and Dunnett's test, respectively. Ceriodaphnia dubia survival and <br />reproduction data were analyzed for significant differences using Fisher's Exact Test and Dunnet's Test, <br />respectively. IC252 values were determined using the Bootstrap Linear Interpolation method. <br />`Supplemental forms include statistical anaiysis and chain-of-custody forms <br />2 Inhibition Concentration at 25% reduction <br />