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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 May 2004. Previous <br />testing using Daphnia magna had demonstrated that a high amount of total dissolved solids <br />(TDS) was causing toxicity in the effluent from the mine. A new NPDES permit for the mine <br />discharge required that the effects of the TDS be accounted for in subsequent testing. <br />Accordingly, SeaCrest performed a series of tests in conjunction with the original effluent, as <br />part of the permit requirements, to confirm the effects of TDS against other possible <br />contaminants This report details the results of the acute tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample of the 017 discharge was collected into one-gallon plastic containers at 10:00 <br />AM on May 27, 2004. The effluent was packed in blue ice and shipped overnight to SeaCrest's <br />lab where it arrived at 09:33 AM on May 28, 2004. Upon receipt, the samples were refrigerated <br />at 4° C when not in use. The Chain of Custody forms, documenting sample collection and iab <br />arrival times, are included in Appendix 1. <br />Test Organisms <br />Daphnia magna used in the test were cultured in the SeaCrest laboratory. The daphnids <br />were {ess than 24 hours old at the start of the test. The daphnids were fed prior to the test <br />start-up but not during the 48-hour test. The daphnids were fed a mixture of yeast, cereal <br />leaves, trout chow (YCT); and the green microalga, Selenastrum capricomutum. <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 sample vas 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. (2002). <br />Ammonia was measured by a Thermo Orion ion-specific electrode according to procedures in <br />APHAlA1NWA/WEF (1998). <br />The SeaCrest Group 4 <br />