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Mountain Coal Company CO-0038776 Sea Crest Number: 303369 303385 <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 October and <br />November, 2003. In accordance with accepted Environmental Protection Agency (EPA) and <br />State of Colorado procedures, acute tests were performed using Daphnia magna and the <br />fathead minnow (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 October 16, 2003. The effluent was packed in an ice chest and shipped overnight to the lab <br />where it arrived at 09:30 on October 17, 2003. This sample was used to start acute tests with <br />both Daphnia magna and fathead minnows. The fathead minnow test had excessive control <br />failure and the test had to be repeated. Another sample of the discharge was collected at 10:00 <br />on November 3, 2003. The sample was shipped overnight and arrived at 09:30 on November <br />4, 2003. The Chain of Custody forms, documenting sample collection and lab arrival times, are <br />included in Appendix 1. In the lab, the samples were refrigerated at 4° C when not in use. <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 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 (Artemia sp.) <br />at least twice daily. Eleven day old fish were used in th}e, present test. <br />~.. ~_ ..4~e...a..... ~~~~-' _: lC ~._ .__...... ~: ........ ,,,v, i',j „ u ,~=..iGi G~n.G ivn`iialll IGJI lJ J'lllg <br />sodium chloride to confirm their overall health. Purchased organisms are tested in a separate <br />roup 2 <br />