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Client: Western Fuels CO-0000213 SCG Project No.: 310624 <br />Site: New Horizon Mine 007 <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 />The Colorado Water Quality Control Division requires certain NPDES permittees to <br />perform acute and/or chronic biomonitoring tests. The chronic test measures significant <br />differences in lethality and in reproduction (Ceriodaphnia dubia) or growth (Fathead minnow - <br />Pimephales promelas) between control and exposed organisms. <br />The present report discusses the results of chronic biomonitoring tests conducted on <br />effluent from Western Fuels New Horizon Mine Outfall 007 discharge. These tests were <br />conducted in June/July 2010. <br />MATERIALS AND METHODS <br />Sample Collection <br />Two gallons and four gallons of effluent were collected on two separate dates. Samples <br />were delivered chilled to the SeaCrest lab where they were held at 0-60C. Chain of custody <br />forms showing sample collection and lab arrival times are included in Appendix 1. <br />Dilution Water <br />Laboratory reconstituted water was used as both the dilution water source and the control <br />for the tests. Reconstituted water for the Ceriodaphnia dubia test was produced by adding <br />sodium bicarbonate, calcium sulfate, magnesium sulfate, potassium chloride, and sodium <br />selenate to deionized water. Reconstituted water for the Fathead minnow test was produced by <br />adding sodium bicarbonate, calcium sulfate, magnesium sulfate and potassium chloride to <br />deionized water. <br />Test Organisms <br />The tests were conducted with Ceriodaphnia dubia and Fathead minnow. <br />Ceriodaphnia dubia is cultured in the SeaCrest laboratory. Stock cultures are maintained in 5- <br />gallon aquaria. Brood females are cultured in individual plastic beakers on brood boards for a <br />period of up to 14-days. Neonates less than 24-hours old released from third or subsequent <br />broods of eight or more within an 8-hour period are removed from the brood chambers and used <br />in tests. Brood and stock organisms are fed daily with a mixture of Yeast, Cereal Leaves and <br />Trout Chow (YCT). This is supplemented with an equal volume of green algae (Selenastrum <br />capricornutum). <br />Less than one-day-old Fathead minnow, cultured in the laboratory, were also used in the <br />test. Adult fish are maintained in 10-gallon aquaria where females deposit their eggs on the <br />under-surface of split PVC pipe sections. The eggs are collected daily and transferred to aerated <br />containers where they hatch after three to four days. The larval fish are fed newly hatched Brine <br />shrimp (Artemia sp.) at least twice per day. <br />SeaCrest Group