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Mountain Coal Company CO-0038776 SeaCrest Number: 303251 <br />INTRODUCTION <br />Biomonitoring provides an effective method for testing the toxicity of effluents from <br />municipal and industrial discharges. Among the advantages of biomonitoring is the ability to <br />test complex effluents containing a broad range of contaminants. The biomonitoring methods <br />generate data that cannot be obtained solely from chemical analyses. At the same time, <br />chemical analyses often go hand in hand with the biomonitoring tests. <br />Biomonitoring tests were conducted for Mountain Coal Company during July, 2003. 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 in one-gallon plastic containers at 08:15 <br />on July 15, 2003. The efFluent was packed in an ice chest and shipped overnight to the lab <br />where it arrived at 09:40 on July 16, 2003, The Daphnia magna test run with this sample failed <br />for control animal survival and the test had to be rerun. A second sample was requested, which <br />was collected at 08:00 on July 29, 2003. The sample was shipped overnight to the SeaCrest <br />lab and arrived at 10:00 on July 30, 2003. The Chain of Custody forms, documenting sample <br />collection and lab arrival times, are included in Appendix 1. In the lab, the samples were <br />refrigerated at 4°C until used. <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 />microalgae, Selenastrum capricomutum. <br />One day old fathead minnows were also used in the test and came from Aquatic <br />BioSystems, Inc, in Ft. Collins, Colorado. The animals were acclimated to test temperature <br />prior to use and were fed newly-hatched brine shrimp (Artemis sp.) at least twice a day during <br />holding. <br />All in-house organisms are tested at least monthly in a reference toxicant test using <br />sodium chloride to confirm their overall health. Purchased organisms are tested in a separate <br />reference toxicant test, conducted during the effluent test or as soon as possible afterwards. <br />The SeaCrest Group <br />