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• <br />Table of Contents <br />Abstract Page 2 <br />Methods Page 2 <br />Results Page 3 <br />Narrative & Discussion Page 4 <br />Table 1 - Colorado Test Conditions Page 5 <br />Abstract <br />Table 2 - Methods and Equipment Page 5 <br />References Page 5 <br />Discharge Monitoring Report Form(s) Pg. 6-7 <br />Reference Toxicity Data Chart(s) Pg. 8 <br />Supplement Forms' Pg. 9 <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 outfall007. 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, outfall 007 is in <br />compliance with the chronic WET portion of its Colorado State Discharge Permit for thefourth <br />quarter of 2007 <br />0 Methods <br />A two species chronic bioassay was conducted with effluent from Western Fuels New Horizon Mine, <br />Outfall 007. An effluent composite sample was collected on October 22, 2007, stored on ice, and <br />delivered to SGS North America Inc. on October 23, 2007. The effluent samples were received at a <br />temperature of 3.40 C and 4.3° C (separate coolers) and assigned sample tracking number TA7-JO- <br />P032-001. The bioassay was initiated approximately 24 hours after sample collection. <br />Subsequent samples were collected October 24, 2007. Because of the remote location of the facility, <br />two concurrent samplings events were utilized for this examination. Sufficient sample volume was <br />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 both analyzed for significant differences <br />using Dunnett's test. Ceriodaphnia dubia survival and reproduction data were analyzed for significant <br />differences using Fisher's Exact Test and Wilcoxon's Rank Sum Test, respectively. IC25 values were <br />determined using the Bootstrap Linear Interpolation method. <br />1 Supplemental forms include statistical analysis and chain-of-custody forms <br />2 Inhibition Concentration at 25% reduction <br />2