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Mountain Coal Company CO - 0038776 SCG Project No.: 304204 <br />Site: West Elk Mine <br />INTRODUCTION <br />Biomonitoring provides an effective means by which the toxicity of dischazges from <br />municipal, industrial, and mining operations can be tested. Among the advantages of <br />biomonitoring is the ability to test complex effluents containing a broad range of contaminants. <br />Biomonitoring, when used in conjunction with chemical analyses, can generate data capable of <br />identifying a much wider range of contaminants. <br />In accordance with accepted Environmental Protection Agency (EPA) and State of <br />Colorado procedures, acute tests were performed using Daphnia magna in August 2004. This <br />report details the results of these acute tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample was collected from the discharge system. The sample was delivered chilled to <br />the SeaCrest lab where it was held at 0-6°C. Chain of custody forms showing sample collection <br />and lab arrival times aze included in Appendix 1. <br />Source and Acclimation of Test Organisms <br />The biomonitoring test used Daphnia magna, cultured in the SeaCrest laboratory. The <br />organisms aze batch cultured in 5-gallon aquaria. These organisms are used to supplement brood <br />boards from which individual females aze monitored for reproduction for periods up to two <br />weeks. Neonates are counted daily during this time and young from broods of eight or more are <br />collected for use in tests. The animals aze fed daily with a mixture of Yeast, Cereal Leaves and <br />Trout Chow (YCT). This supplemented with an equal volume of cultured green algae <br />(Selenastrum capricornutum). <br />In-house organisms aze tested at least monthly in a reference toxicant test using sodium <br />chloride to monitor overall health and test reproducibility. <br />Test Procedures <br />Upon receipt at the lab, water samples were analyzed for alkalinity, hardness, <br />conductivity, dissolved oxygen, ammonia, chlorine and pH. Alkalinity and hardness were <br />determined titrimetrically according to methods described in Hach Chemical Company'. <br />Ammonia was measured by a Thermo Orion ion-selective electrode according to the procedures <br />in APHA/AW WA/WEF~. <br />All acute tests were started on August 10, 2004. The tests were performed according to <br />the procedures outlined in USEPA' and the Colorado Department of Public Health and <br />Environment°. They were conducted over a 48-hour period. Exposure concentrations included <br />0%(control), 1.9%, 12.5%, 25%, 50% and 100% effluent mixtures. Moderately hazd laboratory <br />reconstituted water was used as the dilution water and control for the tests. Reconstituted water <br />was prepared by adding sodium bicazbonate, calcium sulfate, magnesium sulfate and potassium <br />chloride to deionized water. <br />The SeaCrest Group, LLC 5 <br />