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Twentymile Coal Company 7 bite 62 Chronic Whole Effluent Toxicity Results <br />2nd Quarter 2013 AT[ Lot # 042313 -1abc: <br />Summary of Laboratory Conditons & Bioassay Results <br />Parameter <br />Ceriodaphnia dubia <br />Pimephales promelas <br />Test Type <br />7 -day static renewal <br />7 -day static renewal <br />Temperature (dea C) <br />25 +/ - 1 deg C <br />25 +/ - 1 deg C <br />Light QualitvV <br />ambient lab levels <br />ambient lab levels <br />Light Intensity (footcandles) <br />50 -100 ft -c <br />50 -100 ft-c <br />Photoperiod <br />16 hr light/ 8 hr dark <br />16 hr light/ 8 hr dark <br />Test Chamber Size <br />40 mL <br />500 mL <br />Test Solution Volume <br />30 mL <br />300 mL <br />Renewal of Test Solutions <br />every 24 hours <br />every 24 hours <br />Aqe of Test Orqanisms <br />less than 24 hours <br />less than 24 hours <br /># of neonates per test chamber <br />1 <br />10 <br /># replicate test chambers per conc. <br />10 <br />4 <br />Feeding regime <br />0 2 mL algae(YTC per day <br />0.15 mL A. nauplh 2xdady <br />Dilution Water <br />MHFSW <br />MHFSW <br />Test Duration <br />7 -8 days <br />7 -days <br />Effluent Sampling Events and MHFSW Inorganic Chemistries <br />Parameter <br />Sampling Event 1 <br />Sampling Event 2 <br />Sampling Event 3 <br />MHFSW <br />pH <br />7.88 <br />7.91 <br />7.99 <br />7.88 <br />Dissolved Oxygen (mg /L) <br />6.29 <br />7.44 <br />7.25 <br />6.90 <br />Conductivity (pmhos /cm) <br />3090 <br />3180 <br />3390 <br />313 <br />Ammonia (mg /L) <br />1 10 <br />0.84 <br />0.65 <br />ND <br />Alkalinity (mg /L) <br />360.54 <br />370.59 <br />356.52 <br />50.06 <br />Hardness (mg /L) <br />1749.43 <br />1859.27 <br />1771.40 <br />8527 <br />Residual Chlorine (mg /L) <br />ND <br />ND <br />ND <br />ND <br />Ceriodaphnia dubia & Pimephales promelas Chronic Statistical Endpoints <br />Species <br />Parameter <br />IC25 <br />NOEC <br />LOEC <br />Ceriodaphnia dubia <br />Survival <br />S >100% <br />100% <br />>100% <br />Ceriodaphnia dubia <br />Re roduction <br />59.28% <br />50% <br />75% <br />Pimephales promelas <br />Survival <br />) >100% <br />100% <br />>100% <br />Pimephales promelas <br />Growth <br />>100% <br />50% <br />75% <br />Ceriodaphnia dubia Survival and Reproduction Data <br />Concentration <br />Number Exposed <br />Percent Survival <br />Mean Averaqe Number of Neonates <br />Control <br />10 <br />100% <br />32.8 <br />12.5% <br />10 <br />100% <br />39.0 <br />25% <br />10 <br />100% <br />36.7 <br />50% <br />10 <br />11000/0 <br />30.8 <br />75% <br />10 <br />100% <br />20.9 <br />100% <br />10 <br />100% <br />10.2 <br />