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INTRODUCTION <br />Biomonitoring provides an effective method for testing the toxicity of effluents from <br />municipal and industrial discharges. One of the advantages of Biomonitoring is the ability to <br />test complex effluents containing a broad range of contaminants. These methods generate data <br />that cannot be obtained solely from chemical analyses. <br />Biomonitoring tests were conducted for Mountain Coal Company in April/May 2004. In <br />accordance with accepted Environmental Protection Agency (EPA) and State of Colorado <br />procedures, an accelerated acute test was performed using Daphnia magna. This report <br />details the results of the tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample of the 017 discharge was collected into one-gallon plastic containers at 08:00 <br />on April 28, 2004. The effluent was packed in blue ice and shipped overnight to the lab where it <br />arrived at 09:40 on April 29, 2004. At SeaCrest lab the samples were refrigerated at <br />4° C when not in use. The Chain of Custody forms, documenting sample collection and lab <br />arrival times, are included in Appendix 1. <br />Test Organisms <br />Daphnia magna were chosen as the invertebrate species most able to tolerate the high <br />dissolved solids levels found in Mountain Coal samples. The Daphnia magna used in the test <br />were cultured in the SeaCrest laboratory. The daphnids were less than 24 hours old at the start <br />of the test. The daphnids were fed prior to the test start-up but not during the 48-hour test. <br />Daphnids are fed a mixture of yeast, cereal leaves, and trout chow (YCT); and the green <br />microalga, Selenastrum capricomutum. <br />All in-house organisms are tested at least monthly in a reference toxicant test using <br />sodium chloride to confirm their overall health. <br />Test Procedures <br />Upon receipt at the lab, the water samples were analyzed for alkalinity, hardness, <br />conductivity, dissolved oxygen, chlorine, ammonia and pH. Alkalinity and hardness were <br />determined titrimetrically according to methods described in Hach Chemical Co. (2002). <br />Ammonia was measured by a Thermo Orion ion-specific electrode according to procedures in <br />APHA/AUWVA/WEF (1998). <br />The SeaCrest Group 2 <br />