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Client: Mountain Coal Company CO-0038766 SCG Project No.: 305140 <br />Site: East Sly Gulch <br />INTRODUCTION <br />Biomonitoring provides an effective means by which the toxicity of dischazges 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 June 2005. This report details the results of these acute 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 arrive times are 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 aze batch cultured in 5-gallon aquazia. 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 are 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 mixture of Yeast, Cereal Leaves and <br />Trout Chow (YCT). This supplemented with an equal volume of cultured green algae <br />(Selenastrum capricornutum). <br />Five-day-old Fathead minnows were also obtained from in-house cultures. Adult fish aze <br />maintained in 10-gallon aquaria where females deposit their eggs on the under-surface of split <br />PVC pipe sections. The eggs are collected daily and transferred to aerated containers where they <br />hatch after three to four days. The larval fish aze fed newly hatched Brine shrimp (Artemia sp.) <br />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'. A <br />Thermo Orion ion-selective electrode according to the procedures in APHA/AWWA/WF,F2 <br />measured ammonia. <br />The SeaCrest Group 5 <br />