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Aqua-Tox, Inc. <br />4880 Robb Street, Suite 8 <br />Wheat Ridge, CO 80033 <br />(303) 621-4880 aqua-tox.com <br />ACUTE TOXICITY REPORT <br />Ceriodaphnia dubia <br />LIENT NAME: Twentymile Coal Company SAMPLE I.D.: 040209-1 <br />% MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />SURVIVAL IN 100% DILUTION: 100% <br />AMPLE DATE: 4-1-09 TIME: 13:35PM CLIENT ID: Site 10 RT <br />RGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />=ST START: 4-2-09 10:40AM TEST FINISH: 4-4-09 10:40AM <br />>'PE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />AMMONIA: ND <br />HARDNESS: 98.15 mg/L <br />ALKALINITY: 55.56 mg/L <br />pH: 8.01 <br />I'ION WATER CONSTITUENI <br />D.O.: 7.40 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.2 deg C <br />URVIVAL DATA DILUTIONS (%) <br />DATE (HOURS)CONTROL 6.25 12.5 25 50 75 100 <br />4/2/2009 INITIAL <br />4/3/2009 24 <br />4/4/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 20 <br />0 0 0 0 0 0 0 <br />100% 100% 100% 100% 100% 100% 100% <br />- (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C) <br />319/329 560/571 796/803 1184/1191 1995/2280 2880/3030 3700/3830 <br />7.2/7.4 7.2/7.4 7.1/7.5 7.1/7.6 7.1/8.0 7.1/8.1 7.1/8.9 <br />8.0/8.1 8.1/8.3 8.1/8.3 8.2/8.4 8.2/8.6 8.2/8.4 8.1/8.3 <br />19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 0.42 mg/L D.O.: 8.88 mg/L <br />HARDNESS: 1093.64 mg/L CHLORINE: ND <br />ALKALINITY: 477.69 mg/L TEMPERATURE: 20.2 deg C <br />pH: 8.12 D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />100% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 K <br />ADDITIONAL COMMENTS: