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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 2004. In <br />accordance with accepted Environmental Protection Agency (EPA) and State of Colorado <br />procedures, acute tests were performed using Daphnia magna and the fathead minnow <br />(Pimephales promelas). This report 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 09:00 <br />on April 7, 2004. The effluent was packed in an ice chest and shipped overnight to the lab <br />where it arrived at 09:26 on April 8, 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 capricornutum. <br />The fathead minnow larvae are obtained from adults held in ten and twenty gallon <br />aquaria. The females deposit their eggs on the undersurface of split PVC pipe sections from <br />which they are collected daily. The eggs are transferred from the tiles to aerated containers <br />where they hatch after 3-4 days. The larvae are fed newly hatched brine shrimp (Artemis sp.) <br />at least twice daily. Nine-day-old fish were used in the present test. <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 />The SeaCrest Group 2 <br />