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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.: 091108-5 <br />D % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />)SURVIVAL IN 100% DILUTION: 100% <br />AMPLE DATE:-9-10-08 TIME: 15:30PM CLIENT ID: Pond D <br />1RGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />EST START: 9-11-08 13:OOPM TEST FINISH: 9-13-08 13:OOPM <br />YPE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />AMMONIA: ND <br />HARDNESS: 90 mg/L <br />ALKALINITY: 54.96 mg/L <br />pH: 8.05 <br />DATE (HOURS) CONTROL 6.25 <br />9/11/2008 INITIAL <br />9/12/2008 24 <br />9/13/2008 48 <br />MORTALITY: <br />% SURVIVAL: <br />D.O.: 7.20 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 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 />41LY CHEMISTRY - (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 />336/352 627/682 960/1014 1535/1607 2730/2840 3850/4030 4820/5080 <br />7.2/7.5 7.2/7.5 7.3/7.5 7.3/7.5 7.4/7.6 7.4/7.5 7.4/7.7 <br />8.1/8.2 7.2/7.5 8.2/8.3 8.3/8.5 8.3/8.6 8.4/8.6 8.3/8.7 <br />19.8/20.2 19.8/20.2 19.8/20.2 19.8/20.2 19.8/20.2 19.8/20.2 19.8/20.2 <br />ISTRY - SAMPLE <br />AMMONIA: .44 mg/L <br />HARDNESS: 745.50 mg/L <br />ALKALINITY: 417.73 mg /L <br />pH: 8.34 <br />D.O.: 7.70 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.0 dec C <br />D.O AFTER AERATION: NA <br />- ANOEC CALCULATION METHOD METHOD-REFERENCE . a . <br />100% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS: <br />Analysis <br />Conducted by: Reviewed by: <br />Date: Date: