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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 />CLIENT NAME: Twentymile Coal Company SAMPLE I.D.: 070809-3 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 100% <br />SAMPLE DATE: 7-7-09 TIME: 11:25AM CLIENT ID: Site 84 Pond D <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 7-8-09 11:45AM TEST FINISH: 7-10-09 11:45AM <br />TYPE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />RATELY <br />AMMONIA: ND <br />HARDNESS: 93.25 mg/L <br />ALKALINITY: 55.96 mg/L <br />pH: 8.00 <br />D.O.: 8.53 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.0 deg C <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 />DILUTIONS (%) <br />12.5 95 5n 7..S inn <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 ion <br />CONDUCTIVITY (microS, <br />D.O. (mg/L; <br />pH <br />TEMPERATURE (deg C; <br />312/321 605/616 851/876 1348/1391 12620/2720 13620/3790 4680/4840 <br />7.0/7.4 7.1/7.4 7.2/7.4 7.3/7.4 7.3/7.5 7.4/7.9 7.3/8.3 <br />8.0/8.1 8.2/8.2 8.2/8.4 8.2/8.5 8.3/8.7 8.3/8.8 8.2/8.8 <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 />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 0.16 mg/L D.O.: 8.31 mg/L <br />HARDNESS: 730.37 mg /L CHLORINE: ND <br />ALKALINITY: 505.68 mg /L TEMPERATURE: 20.0 deg C <br />pH: 8.23 D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />100% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />MENTS: <br />Analysis <br />Conducted by: <br />Date: <br />Reviewed by: <br />Date: