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_~ s-x 1 i Y `b METHODS A two species acute bioassay was conducted with effluent from the Lone Pine Pipeline, outfall 013. An effluent grab sample was collected on May 1, 2000, stored on ice, and delivered to Commercial Testing and Engineering, Co. (CT&E) on May 2, 2000. The effluent sample was received at a temperature of 14.0 ° C and assigned CT&E sample tracking number 72-4272 ] 5. The bioassay was initiated approximately 27.5 hours after sample collection. Testing was conducted in accordance with the most recent protocols specified by the Water Quality Control Division of the Colorado Department of Health (1998), outlined in Table 1. Basic characteristics of the effluent and dilutions were measured as follows: Characteristic Method/Equipment EPA Method Alkalinity Ammonia Chlorine, Total Residual Conductivity Dissolved Oxygen Hardness pH Temperature Titrimetric (APHAt, 1989) Lachat Analysis Method LaMotte STC-CI Colorimeter Orion Model 160 Cond. Meter Orion Model 850 DO Meter Titrimetric (APHA, 1989) Fisher Mode1925 pH Meter NIST~ calibrated thermometer EPA 310.1 & 130.2 EPA 350.1 EPA accepted EPA 120.1 EPA 360.1 EPA 310.1 & 130.2 EPA 150.1 EPA accepted The tests exposed daphnids, Daphnia magna, to five concentrations of effluent, ] 00%, 75%, 50%, 25%, and 1.9%, prepared using reconstituted laboratory water at 210 mg/L CaC03. Additionally, the tests exposed fathead minnows, Pimephales promelas, to five concentrations of effluent, 100%, 75%, 50°/a, 25%, and 1.9%, prepared using reconstituted laboratory water at 120 mgt CaC03. Control replicates were exposed to 100% laboratory water. The Binomial method determined the LCso's. Significant differences in mortality were analyzed using Steel's Many-One Rank tests. ` 'American Public Health Association 2 National Institute of Standards and Technology .~ u i~ COMMER CIAL TESTING 8 ENGINEERING CO. ~F~ LTA -Original Watertnarketl For Your Protection