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Client; Mountain Coal Company CO - 0038766 SCG Project No.: 305222 <br />Site: west Elk Mine 017 <br />INTRODUCTION <br />Biomonitoring provides an effective means by which the toxicity of discharges from <br />municipal, industrial, and mining operations can be tested. Among the advantages of <br />biomonitoring is the ability to test complex effluents containing a broad range of contaminants. <br />Biomonitoring, when used in conjunction with chemical analyses, can generate data capable of <br />identifying a much wider range of contaminants. <br />In accordance with accepted Environmental Protection Agency (EPA) and State of <br />Colorado procedures, acute tests were performed using Daphnia magna and Fathead minnows <br />(Pimephales promelas) in AugusdSeptember 2005. This report details the results of these acute <br />tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample was collected from the dischazge system. The sample was delivered chilled to <br />the SeaCrest lab where it was held at 0-6°C. Chain of custody forms showing sample collection <br />and lab arrival times aze included in Appendix 1. <br />Source and Acclimation of Test Organisms <br />The biomonitoring test used Daphnia magna, cultured in the SeaCrest laboratory. The <br />organisms are batch cultwed in 5-gallon aquaria. These organisms aze used to supplement brood <br />boards from which individual females aze monitored for reproduction for periods up to two <br />weeks. Neonates aze counted daily during this time and young from broods of eight or more aze <br />collected for use in tests. The animals aze fed daily with a mixtwe of Yeast, Cereal Leaves and <br />Trout Chow (YCT). This supplemented with an equal volume of cultwed green algae <br />(Selenastrum capricornurum). <br />Three-day-old Fathead minnows were also obtained from in-house cultures. Adult fish <br />aze maintained in 10-gallon aquaria where females deposit their eggs on the under-swface of <br />split PVC pipe sections. The eggs aze collected daily and transferred to aerated containers where <br />they hatch after three to fow days. The larval fish aze fed newly hatched Brine shrimp (Anemia <br />sp.) at least twice per day. <br />In-house organisms aze tested at least monthly in a reference toxicant test using sodium <br />chloride to monitor overall health and test reproducibility: <br />Test Procedures <br />Upon receipt at the lab, water samples were analyzed for alkalinity, hazdness, <br />conductivity, dissolved oxygen, ammonia, chlorine and pH. Alkalinity and hazdness were <br />determined titrimetrically according to methods described in Hach Chemical Company'. <br />Ammonia was measwed by a Thermo Orion ion-selective electrode according to the procedwes <br />in APHA/AWWA/wEFz. <br />The acute tests were started on August 31, 2005 for Daphnia magna and August 31, 2005 <br />for the Fathead minnows. The tests were performed according to the procedures outlined <br />The SeaCrest Group 5 <br />