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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 />.TENT NAME: Twentymile Coal Company SAMPLE I.D.: 070809-2 <br />% MORTALITY TEST: Fail TEST RESULT LC50: 85.7% <br />SURVIVAL IN 100% DILUTION: 15% <br />kMPLE DATE: 7-7-09 TIME: 10:23AM CLIENT ID: Site 62 FCT <br />?GANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />ST START: 7-8-09 11:25AM TEST FINISH: 7-10-09 11:25AM <br />PE OF TEST: LC50. 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />AMMONIA: ND <br />HARDNESS: 93.25 mg/L <br />ALKALINITY: 55.96 mg/L <br />pH: 8.00 <br />DATE HOURS CONTROL 6.25 <br />7/8/2009 INITIAL <br />7/9/2009 24 <br />7/10/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />D.O.: 8.53 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.0 deg C <br />DILUTIONS (%) <br />12.5 25 50 75 100 <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 19 6 <br />19 20 20 20 20 16 3 <br />1 0 0 0 0 4 17 <br />95% 100% 100% 100% 100% 80% 15% <br />LY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6 _25 12.5 PS 5n 7.5 1no <br />CONDUCTIVITY (micros; <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C) <br />312/327 625/639 920/938 1466/1503 2660/2710 3570/3630 4450/4560 <br />7.0/7.4 7.1/7.4 7.2/7.4 7.3/7.5 7.3/7.7 7.3/8.2 7.3/8.5 <br />8.0/8.1 8.1/8.2 8.2/8.3 8.1/8.5 8.1/8.3 8.0/8.2 8.0/8.1 <br />19.6/20.2 19.6/20.2 19.6/20.2 19.6/20.2 19.6/20.2 19.6/20.2 19.6/20.2 <br />CRY - SAMPLE <br />AMMONIA: 0.02 mg/L <br />HARDNESS: 2686.34 mg/L <br />ALKALINITY: 456.91 mg /L <br />pH: 7.99 <br />D.O.: 8.53 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.0 deg C <br />D.O AFTER AERATION: NA <br />II ANOEC I CALCULATION METHOD I METHOD REFERENCE II <br />50% <br />Analysis <br />Conducted by: <br />Date: <br />Reviewed by: <br />Date: <br />SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78