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Mountain Coal Company SeaCrest Number: 399033 <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 January, 1999. <br />In 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 />(Pimepha/es promelas). 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 January 26, 1999. The effluent was packed in an ice chest and shipped <br />to the lab the next day. The Chain of Custody forth showing 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 />With permission from the Colorado Department of Public Health and Environment, <br />Ceriodaphnia dubia were replaced with Daphnia magna as the invertebrate test organism far <br />Mountain Coal discharge sites. The Daphnia magna were cultured in the laboratory. The <br />daphnids were less than 24 hours old at the start of the test. Eight day old fathead minnows <br />were also used in the test and came from in-house cultures. Adult fathead minnows are held <br />in ten and twenty gallon glass aquaria where they produce eggs that are collected daily and <br />held under aeration until hatching occurs. Larval fish from one to fourteen days old are used <br />in the acute tests. <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 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/AWWANVPCF (1985). <br />The SeaCrest Group 2 <br />