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ENSR Consulting and Engineering <br />1.1 Objective <br />To determine if the effluent is chronically toxic to Ceriodaphnia dubia under static renewal test <br />conditions. <br />1.2 Test Effluent <br />Three composite effluent samples will be collected by the sponsor. The samples will be <br />collected in disposable cubitainers, placed on ice, and delivered to ENSR via overnight <br />delivery. <br />2.1 Basis <br />1.0 INTRODUCTION <br />This protocol is designed to comply with USEPA method 1002.0, Ceriodaphnia dubia Survival <br />and Reproduction Test (USEPA 1994), and Colorado Department of Health (1993) guidelines. <br />2.2 Test Organism <br />2.0 MATERIALS AND METHODS <br />ENSR Project No.: 8503 -096 -193 <br />Protocol No.: CD4C0.193.001 <br />Effective: 08/96 <br />Page: 2 of 5 <br />1. Species - Ceriodaphnia dubia <br />2. Age - Ceriodaphnia dubia will be < 24 hours old (all within 8 hours of the same <br />age) at test initiation. <br />3. Source - Ceriodaphnia dubia will be obtained from ENSR's in -house culture or <br />from a commercial supplier. <br />4. Feeding - Ceriodaphnia dubia will be fed 0.1 ml of a yeast -trout chow - cereal <br />leaves suspension (YTC. USEPA 1994) and 0.1 ml of Se /enastrum <br />capricornutum suspension (3.0 X 10 to 3.5 X 10' cells /ml) per 15 -m1 exposure <br />chamber daily. <br />3.1 Dilution Water <br />3.2 Temperature <br />3.0 TEST SYSTEM <br />Dilution water will be from the receiving stream collected from a point upstream of, and <br />unaffected by, the effluent discharge. If the receiving stream water is proven to be toxic, a <br />reconstituted laboratory water, prepared to match (± 15 %) the hardness of the receiving <br />stream, will be,substituted. <br />Test temperature will be 25 ± 1 °C. Testing will be conducted in an environmental chamber <br />or temperature - controlled water bath. <br />