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.~ <br />Mountain Coal Company CO-0038776 SeaCrest Number: 399272 <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 test <br />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, chemical <br />analyses often go hand in hand with the biomonitoring tests. <br />Biomonitoring tests were conducted for Mountain Coal Company during July, 1999. 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 prome/as). This report details the results of the tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample of the Lone Pine Pipeline (#013) discharge was collected in one-gallon plastic <br />containers at 08:30 on July 14, 1999. The effluent was packed in an ice chest and shipped to the <br />lab the next day. The Chain of Custody form, documenting sample collection and lab arrival <br />times, is included in Appendix 1. In the lab, the sample was 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 cultu2d 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, Se/enastrum capricomutum. <br />Nine day old fathead minnows were also used in the test and came from in-house cultures. <br />Adult fathead minnows are held in ten and twenty gallon glass aquaria where they produce eggs <br />that are collected daily and held under aeration until hatching occurs. Larval fish from one to <br />fourteen days old are used in the acute tests. Larval fish are fed newly-hatched brine shrimp <br />(Anemia sp.) every other day during the 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 />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. (1978). <br />Ammonia was measured by an Orion ion specific electrode according to procedures in <br />APHA/AW1NA/WPCF (1985). <br />The SeaCrest Group <br />