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Mountain Coal Company CO-0038776 SeaCrest Number: 303417 <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 pertormed using Daphnia magna and the <br />fathead minnow (Pimephales promelas). The Daphnia test failed the quarterly test and was <br />required to proceed to accelerated testing to determine persistence of the toxicity. This report <br />details the results of the third accelerated test. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample of the 017 discharge was collected into one-gallon plastic containers at 10:30 <br />on December 1, 2003. The effluent was packed in an ice chest and shipped overnight to the <br />lab where it arrived at 09:58 on December 2, 2003. The Chain of Custody form, documenting <br />sample collection and lab arrival times, is included in Appendix 1. In the lab, the sample was <br />refrigerated at 4° C when not in use. <br />Test Organisms <br />Daphnia magna was the organism that failed the quarterly test and was required to be <br />used in the accelerated testing. The Daphnia magna used in the test were cultured in the <br />SeaCrest laboratory. The daphnids were less than 24 hours old at the start of the test. The <br />daphnids were fed prior to the test start-up but not during the 48 hour test. Daphnids are fed <br />a mixture of yeast, cereal leaves, and trout chow (YCT); and the green microalgae, <br />Selenastrum capricornutum. <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 />• _ _ <br />. to °,'-~^=,u ...~_„':~~._.~:c~m- .:~c*t:.:,v£Siin..,~5~r:..'..tr~v~4~.,rlf`e=: a'Li3._`~~tr*n.rF____._-i <br />APHA/AVWVA/VVEF (1998). <br />The SeaCrest Group 2 <br />