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Client: Western Fuels CO-0000213 SCG Project No.: 310547 <br />Site: New Horizon Mine NPDES 011 <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 Ceriodaphnia dubia and Fathead minnow <br />(Pimephales promelas) in May 2010. This report details the results of these acute tests. <br />MATERIALS AND METHODS <br />Sample Collection <br />A sample was collected from the discharge system. The sample was delivered chilled to <br />the SeaCrest lab where it was held at 0-60C. 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 Ceriodaphnia dubia, cultured in the SeaCrest laboratory. <br />The organisms are batch cultured in 5-gallon aquaria. These organisms are used to supplement <br />brood boards from which individual females are monitored for reproduction for periods up to <br />two weeks. Neonates are counted daily during this time and young from broods of eight or more <br />are collected for use in tests. The animals are fed daily with a mixture of Yeast, Cereal Leaves <br />and. Trout Chow (YCT). This supplemented with an equal volume of cultured green algae <br />(Selenastrum capricornutum). <br />Fourteen-day-old Fathead minnow were also obtained from in-house cultures. Adult fish <br />are maintained in 10-gallon aquaria where females deposit their eggs on the under-surface of <br />split PVC pipe sections. The eggs are collected daily and transferred to aerated containers where <br />they hatch after three to four days. The larval fish are fed newly hatched Brine shrimp (Artemia <br />sp.) at least twice per day. <br />In-house organisms are 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, hardness, <br />conductivity, dissolved oxygen, ammonia, chlorine and pH. Alkalinity and hardness were <br />determined titrimetrically according to methods described in Hach Chemical Company'. <br />Ammonia was measured by a Thermo Orion ion-selective electrode according to the procedures <br />in APHA/AWWA/WEF'. <br />The acute tests were started on 05-25-2010 for the Ceriodaphnia dubia and on 05-25- <br />2010 for the Fathead minnow. The tests were performed according to the procedures outlined in <br />USEPA' and the Colorado Department of Public Health and Envirorunent'. The <br />SeaCrest Group