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M Z & - & - 331% 500 /***% r ARUM - DuPont-11953 TRADE SECRET Study Title H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss f Test Guidelines: U.S. EPA OPPTS, 850.1400 (1996) U.S. EPA TSCA, EG-11 (1982) OECD TG 210 (1992) Author: Robert A. Hoke, Ph.D. Study Completed on: July 28,2004 Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Laboratory Project ED: DuPont-11953 Sponsor: E i. du Pont de Nemours and Company Wilmington, Delaware 19898 . U.S.A. Page 1 of 137 .Company Sanitized. Does not con tai | H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles of Good Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98)17 and MAFF Japan Good Laboratory Practice Standards (59 NohSan Number 3850). Test substance characterization and stability analyses were performed at DuPont Regional Analytical Services (RAS), Deepwater, New Jersey. None o f the aforementioned analyses were performed under Good Laboratory Practice Standards; however, the analyses were conducted in compliance with ISO 9002 regulations. All o f the analyses are considered sufficient for the purposes o f this study. Applicant / Sponsor: E.I. du Pont de Nemours and Company W ihnington, Delaware 19898 U.S.A. Study Director: A & u /" 4L ' Robert A. Hoke, PhJD. Senior Research Ecotoxicologist and Manager Date Applicant / Sponsor:__________________________________________ __________ DuPont Representative Date -2 - J&JIipany anitlzed, TSC& CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss QUALITY ASSURANCE DOCUMENTATION DuPont-l 1953 The conduct of this study has been subjected to periodic Quality Assurance inspections. The dates of inspection are indicated below. Phase Audited Conduct: Report/Records: Audit Dates July 3,23,24,31,2003; August 14,2003; September 3,4,2003; October 3,2003; December 16,22,24,2003; February 4, 2004 May 26, 2004; June 1-3,2004 Dates Reported to Study Director and Management July 23, 2003; October 6, 2003; December 16,22,24,2003; February 5, 2004 June 3, 2004 Reported by: _ 'fU S & J r t. Joseph C. Hamill Quality Assurance Auditor T .C . u& h Date -3 - not contain ^m p X ?anitlzed*53068 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study. Analytical Evaluation by: | BogdanSzostek,Ph.D. SeniorResearchChemist StatisticalAnalysesby: ^ ' JohnW.Green,Ph.D.,Ph.D. PrincipalResearchStatistician iS 'U H L - O o o 'i Date U -U o tf Date Issued bv Stadv Director: Robert A. Hoke, Ph.D. SeniorResearchEcotoxicologist andManager tom not contain TSCA CBi ^empanX Sanitized- Does -4 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 f TABLE OF CONTENTS Page GOOD LABORATORY PRACTICE COMPLIANCE STATEM ENT.............................. 2 QUALITY ASSURANCE DOCUMENTATION.............................................................................. 3 C E R T IF IC A T IO N ....................................................................................................................................4 LIST OF TABLES................................................................................. 6 LIST OF FIGURES............................................................................... 8 LIST OF A PPENDICES.................................................. 8 STUDY INFORM ATION...................................................................................................................... 9 STUDY PERSONNEL......................................... 10 SUM M ARY..............................................................................................................................................11 I N T R O D U C T IO N ...................................................................................................................................13 MATERIALS AND M ETH O D S........................................................................................................ 13 A. Test G uidelines............................................................................................................................ 13 B. Test Substance............. ...................................................................... 13 C. Dilution W ate r............................................................ 14 D. Test Organism Culture........................................ 14 E. Test M ethods............................................................................................................. 15 F. Sample Preparation and Chemical A nalysis............ ............................................. 16 G. Statistical A nalysis................. 18 RESU LTS.................................................................................................................................................19 A. Analytical R eport................................................ 19 B. In-Life Report............................................................................................................................... 20 C. Statistical Analysis R eport......................................................................................................... 21 D IS C U S S I O N ........................................................................................... 21 C O N C L U S IO N ........................................................................................ 21 RECORDS AND SAMPLE STORAGE........................................................................................... 22 REFERENCES........................................................................................................................................ 23 T A B L E S .................................................................................................................................................... 24 F I G U R E S ................................................................................................................................................ 123 APPENDIX............................................................................................................................................ 135 'Company Sanitized. Does not contain TSCA CBt -5 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 LIST OF TABLES Page 1. CHEMICAL CHARACTERISTICS OF HASKELL LABORATORY WELL......................................................25 2. MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS.......................................................... 26 3. WATER CHEMISTRY OF THE DILUTION WATER CONTROL, IPA CONTROL, AND HIGHEST TEST CONCENTRATION WITH SURVIVING ORGANISMS........................................................................... 35 4. TEMPERATURE (C) OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST..................................................................... . 37 5. pH OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST................................................................................................................................. 38 6. DISSOLVED OXYGEN CONCENTRATION (mg/L) OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.....................................39 7. SUMMARY OF HATCHING, SURVIVAL, ABNORMALITIES, AND SWIM-UP FROM HATCHING TO THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.................................................................................................................. 40 8. SUMMARY OF SURVIVAL, ABNORMALITIES, AND GROWTH FROM THINNING TO TEST END FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.......................................................................................... 43 9. DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.................... 45 10. IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST..........................................................51 11. 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.......................................................... 57 12. 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST..........................................................63 13. 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.............. 69 14. 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST......................................................... 75 15. 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST..........................................................81 16. DILUTION WATER CONTROL: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST......................................................................... 87 17. IPA CONTROL: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................... ......................................................................................... 88 18. 0.30 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.................................................................................................................. 89 19. 0.63 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.................................................................................................................. 90 -6 - company not contaIn tSCA CB! DoeB /"--N H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 20. 1.3 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................................................................................................91 21. 2.5 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................................................................................................92 22. 5.0 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................................................................................................93 23. DILUTION WATER CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.................................... 94 24. IPA CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST..........................................................97 25. 0.30 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST........................................................................ 100 26. 0.63 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.........................................................................103 27. 1.3 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, ONCORHYNCHUS MYKISS, EARLY LIFE-STAGE TOXICITY TEST.......................................................... 106 28. 2.5 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST............................................... ,........................109 29. 5.0 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST................. .......... ............................................112 30. DILUTION WATER CONTROL: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST.............................................................................................................................................................................115 31. IPA CONTROL: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................116 32. 0.30 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................117 33. 0.63 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................118 34. 1.3 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................119 35. 2.5 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................120 36. 5.0 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST...................................121 37. SUMMARY TABLE OF STATISTICAL ENDPOINTS.......................................................................................122 Company Sanitized. Does not contain TSCA CBI -7 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 LIST OF FIGURES Page 1. PHOTOGRAPH OF AN EMBRYO AT APPROXIMATELY 22-HOURS POST-FERTILIZATION............ 124 2. 3. R ^ S ^ S S / E ^ O c m O M A r o S S ^ ^ S l S S ^ f e i D A R D S O L U T I O N ..........127 4. REPRESENTATIVE HPLC CHROMATOGRAM OF A DILUTION WATER CONTROL SOLUTION.....129 5. REPRESENTATIVE HPLC CHROMATOGRAM OF AIPA WATER CONTROL SOLUTION...................131 6. REPRESENTATIVE HPLC CHROMATOGRAM OF A H-25436 TEST SOLUTION....................................133 LIST OF APPENDICES A. CERTIFICATE OF ANALYSES Page ...136 .Company Sanitized. Does not contain TSCA CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss STUDY INFORMATION DuPont-11953 Haskell Number: 25436 ,.^99SnsN. Purity: I Known Impurities: Physical Characteristics: Tan to light brown paste Stability: The test substance appeared to be stable under the conditions o f the study; no evidence o f instability was observed. Study Initiated/Completed: June 3,2003 / (see report cover page) In-Life Initiated/Compfeted: November 6, 2003'/ February 4, 2004 .Company Sanitized. Does not contain tsc a CBi -9 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss STUDY PERSONNEL Study Director: Robert A. Hoke, Ph.D. Primary Technician: Jeffrey T. Turner, Sr. Animal Culturist: Barbra D. Ferrell, B.S. Management: Jeanette M. Erhardt, Ph.D. Analytical Associate: Keith B. Prickett, B.S. Analytical Chemist: Bogdan Szostek, Ph.D. Management: S. Mark Kennedy, Ph.D. Statistical Analysis: John W. Green, Ph.D., Ph.D. Management: A. Mike Kaplan, Ph.D. Ecotoxicology Report Preparation: Wanda F. Dinbokowitz Management: Kim D. Birkmeyer, M.S. DuPont-11953 j&oinpany Sanitized. Does not contain TSCA OBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 SUMMARY GUIDELINES: The effect of H -2 5 4 3 a jH H H H J H )n hatching, growth and survival o f rainbow trout^u n co rf^m h u s mykiss, embryos, alevins, and fingerlings was assessed in a flow-through, 90-day early life-stage toxicity test in accordance with the appropriate Good Laboratory Practice standards and test guidelines (U.S. EPA TSCA EG -11; U.S. EPA OPPTS 850.1400, OECD 210). NOMINAL TEST CONCENTRATIONS: 0.30, 0.63,1.3,2.5, and 5.0 mg/L H-25436, dilution water control, isopropyl alcohol (IPA) control MEASURED TEST CONCENTRATIONS:. 0 g/L H-25436 wit one detected in dilution water and IPA controls (LOD, 0.0002 mg7L) ?0 .3 4 ,0.70,1.5,2.7, and 5 j j mg/L H-25436 wit ^ detected in dilution water and IPA coS ols(roA 0^0028^/L ) and 5.4jng/L H-25436 withl one detected in dilution water and IPA ) AGE OF TEST ORGANISMS: Approximately 23 hours post-fertilization at test start SOURCE OF TEST ORGANISMS: Unfertilized eggs and spenn (collected from 3 female and 1 male brood fish) were obtained from Troutlodge Inc., (Sumner, Washington, USA) - TEST DESIGN: A total o f 80 embryos were exposed per concentration (20 embryos per embryo cup, 2 cups per replicate, 2 replicates per concentration) at test start. On day 46, after swim-up had begun in the dilution water control, the fingerlings were thinned to a total o f 30 fish per concentration (15 fish per replicate, 2 replicates per concentration). Analytical verification o f H-25436 concentrations was made on test solutions sampled on day -1 , day 0, once weekly, and on day 90 at test end. Additional samples were submitted for analysis as warranted. Company Sanitized Does not contain CBt H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 Test solutions were supplied to each replicate test chamber by a continuous flow metering system at a rate o f approximately 7.5 L per hour (approximately 24, 7.5-L test solution volume additions per day). Embryos and alevins were held in relative darkness until day 40 and then held under a photoperiod o f 16 hours light and 8 hours darkness (which included 30 minutes o f transitional light between light and dark intervals) for the remainder o f the study. After thinning, the fingerlings were fed newly-hatched brine shrimp (days 46-54) or newly-hatched brine shrimp and trout chow (days 55-89). Test solutions were maintained between 11.7 and 12.0C (mean 11.8C). Daily observations were made for assessment o f number o f dead eggs at hatching, first and last day o f hatching, first day o f swim-up, survival and abnormalities from hatching to thinning, and survival and abnormalities from thinning to test end. Standard length and blotted wet weight of surviving fingerlings were determined at test end. CONCLUSION: The NOEC (No Observable Effect Concentration) for H-25436 in a 90-day early life-stage toxicity study using rainbow trout, Oncorhynchus mykiss, was 1.3 mg/L based on nominal H-25436 concentrations, last day of hatching and the first-day o f swim-up. The LOEC (lowest observed effect concentration) and MATC (maximum acceptable toxicant concentration), for the same endpoints were 2.5 and 1.8 mg/L, respectively. The NOEC value for all other test endpoints was >5.0 mg/L. STUDY COMPLETION: (see report cover page) - 12- , Bpm pm a SMIfced. Does c m w ,, r s n A C|}| H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 INTRODUCTION The purpose o f this study was to assess the early life-stage toxicity o f H-25436 to embryos, alevins, and fingerlings of rainbow trout, Oncorhynchus mykiss, in a flow-through, 90-day test. MATERIALS AND METHODS A. Test Guidelines Except as noted below, the study design complies with the following test guidelines: U.S. Environmental Protection Agency (EPA) (1982). TSCA EG-11, Fish Early Life Stage Toxicity Test/ 1-1 U.S. Environmental Protection Agency (EPA) (1996). OPPTS 850.1400, Fish Early Life Stage Toxicity T est/2) Organisation for Economic Co-Operation and Development (OECD) (1992). TG 210. Fish Early Life Stage Toxicity Test.(3) - A constant test target temperature o f 10 2C was used for the duration o f the test, not 10 2C for embryonic development and 12 2C for alevin and fingerling development (OECD 210). Based on the use o f existing temperature control equipment, which is typical o f that used in most facilities for fish early life-stage studies, the high probability o f exceeding the guideline limit for temperature variation obviated temperature adjustment in the middle o f the study. This exception did not affect the objectives or the validity o f the study. B. Test Substance The testjjubstance, H-25436, was supplied by the sponsor as a tan to light bjgwn paste. H-25436 contain! H-25436 contain; The solubility and stability o f H-25436 in Haskell Laboratory well water (HLWW) were verified under test conditions based on the analytical recoveries o f the target analytes. A master stock solution (Stock A) was prepared at a concentration o f approximately 50 mg/mL H-25436 in a 1:4 solution o f isopropyl alcohokHLWW. Secondary stock solutions were prepared by adding pre-determined vo*lumes o f Stock A to 500 mL volumetric flasks and * -13- Jompany Sanitized. Does not contain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mylass DuPont-11953 bringing the final volume to 500 mL with 100 mL o f isopropyl alcohol and the appropriate remaining volume o f HLWW. All stock solutions were prepared on a weekly basis throughout the study. Throughout the study, 25 and 50 mg/mL stock solutions were cloudy and gold in color with no visible precipitate. The 13 mg/mL stock solution was clear and gold in color with no visible precipitate. The 3 mg/mL and 6.3 mg/mL stock solutions were clear and pale gold in color with no visible precipitate. Test solutions were clear and colorless with no visible precipitate throughout the study. The total amount o f H-25436 used in the definitive study was approximately 700 grams. C. Dilution Water Dilution water originated from the Haskell Laboratory well which is 378-feet deep and is cased and sealed to bedrock. The hardness o f the HLWW is adjusted to approximately 100-140 mg/L as CaCC>3by the flow-proportioned addition o f CaCL- The HLWW is then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 0.45-pm cartridge filters to remove particulates. The water is heated or chilled as appropriate and distributed through aged polyvinyl chloride piping. The dilution water is analyzed twice yearly for major anions and cations, metals, total organochlorine and organophosphate pesticides, and polychlorinated biphenyls (Table 1). The dilution water meets OECD(3) and ASTM(4) criteria and specifications. D. Test Organism Culture Rainbow trout, Oncorhynchus mykiss, unfertilized eggs and sperm were purchased from Troutlodge, Inc., Sumner, Washington. Eggs and sperm (collected from 3 female and 1 male brood fish) were received in separate plastic bags which were kept on ice (approximately 5.6C) during shipping. Eggs and sperm were allowed to equilibrate to approximately 11.2C in a water bath. Approximately 200 mL of the buffer solution containing glycine, TRIS, and sodium chloride was added to the bag containing the eggs. The purpose o f the buffer was to remove any ovarian fluid that could impede fertilization. The bag was placed back into the water bath for approximately 20 minutes. The bag o f eggs was taken out o f the water bath and the buffer solution was removed. The bag o f sperm was removed from the water bath and its contents were added to the bag o f eggs. The bag was gently rocked back and forth to evenly distribute the sperm. Buffer solution (50 mL) was also added to the bag to help activate the sperm. The bag was again gently rocked back and forth and returned to the water bath for about 15 minutes. The embryos were removed from the waterbath and the buffer solution and sperm poured off o f the embryos. The embryos were then rinsed with cold HLWW to remove residual sperm and other debris before being transferred to two stainless steel tanks, each containing 10 L o f aerated HLWW, in a water bath (12.1C) under darkness. Approximately 23 hours later, embryos were randomly placed into embryo cups to begin the test. The embryonic stage at the beginning of exposure was verified by selecting a representative sample o f fertilized eggs, photographing, and comparing with literature sources.(5) Embryos and larvae were identified by labels on aquaria and test chambers. - 14- I, Does not contain TSCA CBf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 E. Test M ethods(1,2,3) Five test substance concentrations, a dilution water control, and an isopropyl alcohol (IPA) control were used in this study. Nominal test substance concentrations were 0.30, 0.63, 1.3, 2.5, and 5.0 mg/L. The concentration o f IPA in all test solutions, with the exception o f the dilution water control which contained no solvent, was <0.1 mL/L. Test chambers (2 replicate chambers per concentration) were 13-L stainless steel aquaria (30 [length] x 14.5 [width] x 30 [height] cm), each holding approximately 7.5 liters o f solution (16.5-cm liquid depth) and fitted with a screen mesh-covered overflow pipe. A recirculating waterbath was used to maintain temperature in the test chambers at 10 2C. Test chambers were positioned in the waterbath using random numbers. Test solutions were supplied to the test chambers under flow-through conditions by a continuous flow metering system. Metering system surfaces exposed to test solutions consisted o f glass, Tygon, and Teflon. The test substance metering system consisted o f a Rainin Rabbit-PlusTM peristaltic pump that delivered the stock solutions and IPA control continuously to the mixing chambers o f the test system at approximately 1.5 mL per hour. Test solutions in the mixing chamber were stirred by a mini mixer. Test solutions were delivered continuously from the mixing chamber to each replicate test chamber at approximately 7.5 L per hour (approximately 24 test solution volume additions per day). Checks o f the flow rate into each replicate chamber were made prior to test start and after test end. Two 212-mL glass embryo cups (5.5-cm diameter) with screen mesh bottoms attached with silicone adhesive were suspended in each replicate. Twenty embryos were placed into each embryo cup (total o f 40 embryos per replicate and 80 embryos per test concentration). Embryo cups were assigned to each replicate and embryos were assigned to embryo cups using random numbers. Embryo cups oscillated at approximately 4 rpm (2 to 7-cm water depth in embryo cup). Embryo cups were inspected daily and unfertilized eggs and damaged or fungus-infected embryos were removed. The criterion for embryo death was opaque appearance. The surviving alevins and fingerlings were thinned to 15 per replicate using random numbers and released into the appropriate test chamber replicate on day 46 after swim-up had begun in the dilution water control. Fish that swim-up are those that occupy and are able to maintain their position in the top half o f the water column and in which the yolk sac has mostly been resorbed. Test organisms thinned from the test chambers were humanely sacrificed with MS-222 and stored frozen for potential residue analysis at a later date. On day 46, feeding was initiated. The fingerlings were fed newly-hatched brine shrimp (Artemia sp., San Francisco Bay Brand, Newark, California) 3 times daily on weekdays and twice daily on weekends and holidays, except on days 46 and 47 when they were fed only once and twice, respectively. On day 55 through day 89, one o f the daily feedings with brine shrimp was replaced by a feeding with Aqua Max Starter Fingerling 300 5D03 (PMI Nutrition International, LLC). The fingerlings were last fed on day 89 approximately 24 hours prior to test conclusion on the following day. Brine shrimp and Aqua Max are analyzed periodically to assure that contaminants are not present at levels that would be expected to affect the scientific integrity o f a study. Uneaten food and/or debris were removed as needed. The number o f live fingerlings was determined daily by actual count. -15- CggPany Sanitized. Does not contain TSCA CSf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 The criteria for alevin and fingerling death were the absence of opercular movement and lack of reaction to gentle prodding. Organisms removed by thinning on day 46 and surviving organisms at test end were humanely sacrificed using MS-222. Measurements were made o f length and blotted wet weight o f surviving organisms at test end. In addition, all test organisms sacrificed at thinning on day 46 and three fish from each test chamber at test end were frozen for potential residue analysis at a later date. Diluter operation was checked twice daily. Test solution concentrations were verified by chemical analysis prior to beginning the test, at test start, weekly, and at test end. Before thinning, water samples were taken from a point between the 2 embryo cups in each replicate test chamber (2 samples per concentration plus back-up samples). After thinning, water samples were taken at mid-depth from each test chamber replicate (2 samples per concentration plus back-up samples). Test solutions were maintained between 11.7 and 12.0C (mean = 11.8C). The diluter was wrapped in black plastic to keep the embryos and alevins in relative darkness until the completion o f hatching. The black plastic was removed from day 40 onwards. A photoperiod o f 16 hours light (310 - 634 Lux) and 8 hours darkness, which included 30 minutes o f transitional light (approximately 2 6 - 198 Lux) between the light and dark intervals, was used throughout the remainder o f the study. Dissolved oxygen, pH, and temperature were measured in both replicates o f the dilution water control, IPA control, and all test concentrations at the beginning o f the test before embryos were added, weekly, and at test end. A continuously-recording thermometer was used to check temperature variations in the test system waterbath. Total alkalinity, EDTA hardness, and conductivity were measured in the "A" replicate of the dilution water control, IPA control, and the highest test substance concentration with surviving test organisms at the beginning o f the test, weekly, and at test end. Test solutions were not aerated during the study. F. Sample Preparation and Chemical Analysis 1. Sample Collection and Treatment Analytical determination of H-25436 concentrations in test solution samples was performed with samples collected from each replicate test chamber prior to test initiation, at test initiation, and weekly after test initiation until test termination. A primary sample plus a back-up sample was collected from each replicate test chamber. All samples were diluted 1:1 with acetonitrile (5 mL of sample added to 5 mL acetonitrile) at the time o f sampling. Aliquots o f all test solutions were transferred to autosampler vials for analysis by LC/MS. Additionally, the following samples were diluted with 50/50% acetonitrile/HLWW before analysis: 2.5 mg/L (diluted 1:2), 5.0 mg/L (diluted 1:4). Backup samples were stored at room temperature. - 16- lllized. Does not contain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss 2. Instrument and Conditions DuPont-11953 HPLC Instrument: MS Instrument: Model 2795, Waters ZQ, Micromass LC Parameters: Column: Mobile Phase: Luna 3u C18 (2), 20 mm x 2mm A - 25 mM ammonium acetate in water B - acetonitrile C - 25 mM ammonium acetate in 20/80% water/acetonitrile Gradient: Time (min) % A %B %C Flow (mL/nain) 0 40 0 60 0.3 5 0 0 100 0.3 6.5 0 0 100 0.3 6.51 0 100 0 0.3 8.5 0 100 0 0.3 8.51 40 0 60 0.3 12 40 0 60 0.3 Column Temperature: Injection Volume: Column Switch: 30C 35 pL 1.25 and 7 min M S Parameters: Ionization mode: Electrospray, positive ions Capillary voltage: 3.5 kV Cone voltage: 30 V Source Temperature: 120C Desolvation Temperature: ,500C Data Acquisition Function: M K S]R o f 690.5, 734.5, 778.6, 822.6, 866.6 m/z; M M P y S IR 0-M n m ^ --- IT C T m /zT o-T m rn* o f 790.5, 834.6, 878.6, 922.7, 966.7 m/z; o f 890.6, 934.6, 978.7, 1022.7, 3. H-25436 i! Quantitation selected for analysis for Ifere ionized in positive ion mode electrospra^reaonnnately 17- Company Sanitized. Does not contain t s c a C W H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 as the ammonium ion adducts when the mobile phase contains a significant concentration of ^ ammonium acetata _Quatfitation was based on the total signal obtained for the fjye selecte Separate calibration curves were made for each groi centration of the test~substance (H-25436) was assigned to each group i at were monitored. A stock solution o f the reference standard, H - 2 5 4 3 w a s made by dissolving approximately 10 m g o f the standard in 10 mL o f acetonitrile. Appropriate aliquots o f the stock solution were diluted with 50/50% acetonitrile/HLWW to prepare calibration standard solutions with concentrations ranging from 0.103 to 1.04 mg/L. Duplicate injections o f test and calibration standard solutions were made and peak areas were determined electronically. The calibration standard curves were generated by regression analysis o f the peak area data from chromatography o f the calibration standard solutions. Data for test solutions were compared to the calibration standard curve to determine concentrations o f H-25436. The limit o f detection (LOD) and limit o f quantitation (LOQ) were determined by calculating the average noise level in chromatograms o f the water and IPA control solution and comparing them to the signal o f a calibration standard o f known concentration. Two chromatograms were examined for noiserelated peaks near the retention time o f the analyte. The LOD was calculated as 3 times the concentration equivalent o f the mean noise level. The LOQ was calculated as 10 times the concentration equivalent o f the mean noise level. G. Statistical Analysis The NOEC for each response was determined through the use o f trend tests (Cochran-Armitage trend test or the Jonckheere trend test) wherever appropriate. These trend tests assume monotonicity in the concentration-response. Parametric, or if appropriate, non-parametric tests, were used to determine if there was significant departure from monotonicity. Other multiple comparison methods were used for non-monotone data. Hatching, mortality and larval abnormality data were evaluated by the Cochran-Armitage test used in a step-down manner with equally-spaced concentration scores. If there was significant departure from monotonicity, then Fisher's exact test was used with a Bonferonni correction. Continuous or reproduction data (length, weight, first and last day o f hatching, first day o f swimup) were evaluated for normality using the Shapiro-Wilk test while homogeneity of variance was assessed by Levene's test. Data found to be both normal and homogenous were evaluated in the context o f ANOVA. Monotonic concentration-response data were evaluated using the Jonckheere-Terpstra trend test/ Non-monotonic concentration-response data were assessed using the Dunnett method if the data were normally distributed and homogeneous; using the Tamhane-Dunnett (or T3) method if the data were normally distributed but heterogeneous. If factors in addition to concentration were considered, such as the number of specimens in the test chamber at test start, then the Jonckheere test was not appropriate and multi-factor ANOVA was used for analysis. Non-normal data were analyzed by alternative -18- K&flipany Smithed!. Does not contain t q c a OBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 methods.(9) If possible, a transformation was found to obtain normality. Otherwise, a KruskalWallis test*105replaced the ANOVA, Dunn's test*75replaced Dunnett's, and the JonckheereTerpstra test was still applicable, except as noted. Outliers were determined by the Tukey outlier rule and, if present, their effect on the conclusions was determined. If massive ties were present in a continuous response, exact*'115permutation methods were used for data analysis An exact version o f the Kruskal-Wallis test was used, followed by exact versions o f Jonckheere-Terpstra or separate Mann-Whitney*105 (or, equivalently, Wilcoxon) comparisons o f each treatment group to the dilution water control, using a Bonferonni correction. Wherever possible, determination o f the EC50 was done by probit analysis*65for all frequency responses, adjusting for dilution water control mortality. Where probit analysis failed, a movingaverage angle or binary estimation procedure*125was used for analysis. Inverse regression*135was used to estimate the LC50 for length and other continuous responses, if appropriate. The No Observable Effect Concentration (NOEC) was defined as the highest measured concentration at or below which no statistically significant effect was observed. The Lowest Observed Effect Concentration (LOEC) was the lowest measured concentration at which a statistically significant effect occurred relative to the dilution water control. The maximum acceptable toxicant concentration (MATC) was defined as the geometric mean o f the NOEC and LOEC. RESULTS Analytical Report Chromatographic Results bluted as a well-resolved peak with a retention time o f approximately 1.97 m i n u t e f l |^ ^ r t H H H ^ ^ l u t e d a s a w e l L r e s o l v e d peak with a retention time o f approximately i .o i n n W t e s .l ^ ^ H H H H H f l H e l u t e d as a well-resolved peak with a retention time o f approxim atelyoA J^nmutes. Figure 2 shows representative calibration curves obtained fox Chromatograms o f a typical calibration standard are shown in Figure 3. Figures 4 through 6 show chromatograms o f a water blank control solution, IPA*blank control solution, and test concentration solution, respectively. The LOD and LOQ were determined to 0.00067 mg/L o f H-25436, respectively. The LOD and LOQ f o i ^ p ^ g g p m p p ^ p v e r e det^mined_to_be 0.0002Sgmg/L and 0.00093 mg/L o f H-25436, respectively. The LOT) and LOQ f o r ^ p p B K H H H H r ere determined to be 0.0004 mg/L and 0.0013 mg/L of H-25436, - 19- JsgWpanjrSagffeed. Does not contain TBCA ca l H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 2. Test Solution Results The measured concntotionsofH -254M for each sampling day are shown in Table 2. Mean measured values o i H | | H H H | ^ H p n g e d fr o n il0^ to |l 17% o f the targeted nominal H I^ cnncenlraiip3rns not adjusted for test substance p u rit} H P ||B [M e a n measured values oi [ranged fron}J 00 to 115% of the ta rg ete^ o m m ai concentrations not adjusted for test `substance p u ritflM H H M e a n measured values o f H ^ | ^ | | H 0 jJBranged fronM 08 to 117% o f the talgetecnimninal concentrations not adjusted for test substance p u rit} J jH ^ H |A ll measured values o f H-25436 were within 1.5 X o f the lowest value for all replicatS am plfs within a concentration with the exception o f a single value on day 34. These data indicate that the H-25436 concentrations were maintained at acceptable levels throughout the definitive test. Control solutions showed no detectable concentrations o f H-25436 (Table 2). B. In-Life Report Selection o f test concentrations for the 90-day trout early-life stage study were based on a previous study.(14) Two initial definitive studies were terminated early due to poor performance o f eggs or control test organisms. Nominal H-25436 concentrations selected for the final 90-day trout early life-stage study were 0.3 0 ,0.61 J .3 , 2.5. and 5.0 m^LL. The corresponding mean, measured concentration o f H-25436 fro m M flflH H H H H M p n a ly se s were 0.35,0.71,1.5, 2.8, -and 5.1 mgflb. The corresponding mean, measmedconcraraations o f H-25436 fr o n J B H B H M H H H ^ H n a l y s e s were 0.34, (L70,1.5.2.7, and 5 .0 m g/L. The corresponding m eanjneasured `concentrations o f H-25436 f r o m g H ^ t i ^ H H M H p a l y s e s were 0.35, 0.71,1.5,2.8, and 5.4 mg/L A dilution water and a n E P A c o n tro ^ ^ ^ H ^ a in this study. The control solutions showed no detectable levels o f H-25436. Dilution water quality was acceptable based on OECD and ASTM dilution water criteria. Based on the most recent semi-annual dilution water analysis (Table 1), contaminant concentrations were below concentrations that could be expected to affect the integrity o f a study. All chemical and physical parameters in the early life-stage study were within expected ranges (Tables 3 to 6). Alkalinity, hardness, and conductivity o f the dilution water control, IPA control, and nominal 5.0 m g/L test concentration ranged from 47-52 mg/L as CaCC>3, 118-132 mg/L as CaCC>3; and 210-282 pmhos/cm, respectively. Temperature and pH o f the test substance, dilution water, and IPA control solutions ranged from 11.7 to 12.0C (mean 11.8C) and 7.1 to 7.7, respectively. Dissolved oxygen concentrations ranged between 8.6 and 10.7 mg/L (80 and 100% o f air saturation based on an air saturation value o f 10.8 mg/L at 12C) over the duration o f the study. Analyses o f brine shrimp and Aqua Max indicated that total organochloride pesticides and PCBs were below concentrations that would be expected to affect the scientific integrity o f the study. A summary o f hatching, swim-up, survival, and abnormalities from hatching to thinning is presented in Table 7. A summary o f survival and abnormalities from thinning to test end, and mean length and wet weight (blotted dry) values at test end is presented in Table 8. Biological observations are presented in Tables 9 to 36. A photograph o f a representative fertilized egg 16 - 20- jfempaoy .Sanitized, Does not contain TSCA CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 used to start the study is presented in Figure 1. Loadings in the A and B replicates o f the dilution water control at test end were 0.1066 and 0.0999 g fish per liter passing through each test chamber in 24 hours based on a daily turnover rate o f 180 L or approximately 24, 7.5-L test solution volumes. C. Statistical Analysis Report The NOECs for number o f eggs hatched, number o f abnormal larvae at time of thinning, number o f abnormal larvae at test end, survival of larvae to thinning, survival o f larvae to test end, all exceeded 5.0 mg/L by the Cochran-Armitage trend test. No EC50 values could be computed since the maximum percent effect for any of these measures was < 50%. First and last day o f hatching and first day of swim-up showed massive ties, so that an exact analysis using StatXact was done. All three measures displayed a monotone trend, so the NOEC was determined by an exact Jonckheere-Terpstra test in each case. The NOEC for first day of hatching exceeded 5 mg/L while the NOECs for the last day o f hatching and the first day of swim-up were 1.3 mg/L. The Jonckheere-Terpstra test determined the NOECs for both length and weight of surviving trout at test end were greater than 5 mg/L. DISCUSSION The mean measured concentrations o f H-25436 were maintained at acceptable concentrations throughout the definitive study. The dilution water control and IPA control contained no detectable concentrations o f H-25436. The m ost sensitive endpoints during the study were the last day o f hatching and the first day o f swim-up (NOEC = 1.3 mg/L) based on the results o f the Jonckheere-Terpstra trend test. CONCLUSION The NOEC (no observable effect concentration) for H-25436 in a 90-day early life-stage toxicity study using rainbow trout, Oncorhynchus myldss, was 1.3 mg/L based on nominal H-25436 concentrations and the last day o f hatching and the first day o f swim-up. The LOEC (lowest observed effect concentration) and MATC (maximum acceptable toxicant concentration), for the same endpoint were 2.5 and 1.8 mg/L, respectively. -21 - -Sanitized. Does not contain TSCA CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 RECORDS AND SAMPLE STORAGE Laboratory-specific or site-specific raw data, such as personnel files and equipment records will be retained by the facility where the work was done. Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. 0as not contain TSCA CBI pompan^ Sanitized. - 22- Sanitized. Does not contain TM>jj| H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 REFERENCES 1. U.S. Environmental Protection Agency (EPA) (1982). TSCA EG -11, Fish Early Life-Stage Toxicity Test. 2. U.S. Environmental Protection Agency (EPA) (1996). OPPTS 850.1400, Fish Early LifeStage Toxicity Test. 3. Organisation for Economic Co-Operation and Development (OECD) (1992). Guideline for the Testing o f Chemicals, Number 210: Early-Life Stage Toxicity Test. 4. American Society for Testing and Materials (ASTM) (1992). Standard Guide for Conducting Early Life-Stage Toxicity Tests with Fishes. E1241-92. Annual Book o fASTM Standards, Vol. 11.04. 5. Leitritz, E. and Lewis, R.C. (1976). Trout and salmon culture (hatchery methods). California Department o f Fish and Game, Fish Bulletin 164. 197 pp. 6. Agresti, A. 1990. Categorical Data Analysis, Wiley, New York. 7. Hochberg, Y. and Tamhane, A.C. (1987). Multiple Comparison Procedures, Wiley, New York. 8. Shapiro, S.S. and Wilk., M.B. (1965). An analysis o f variance test for normality (complete samples). Biometrika 52, 591-611. 9. Box, G.E.P. (1953). Non-normality and tests on variances. Biometrika 40, 318-335. 10. Lehmann, E.I. (1975). Nonparametrics: Statistical Methods Based on Ranks. Holden-Day, San Francisco. 11. Mehta, C. and Patel, N. (1995). StatXact 3fo r Windows. Cytel Software Corporation, Cambridge. 12. Stephan, C. E. (1976). Methods for estimating an LC50, Aquatic Toxicology and Hazard Evaluation, Proceedings of the First Annual Symposium on Aquatic Toxicology, ASTM Special Technical Publication 634, American Society for Testing and Materials, Philadelphia. 13. Draper, N.R. and Smith, H. (1981). Applied Regression Analysis, 2nd edition, pp 266-273. Wiley, New York. 14. DuPont Haskell Laboratory (1991). Static, Acute 96-Hour LC5i Fathead Minnow, (Pimephales promelas). Unpublished report] -23 - f&!pany Sanitized Does not contain TSCA CB1 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLES Doesn oMtccoonntwain TSCfrCBl - 24- ) H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss ) ) DuPont-11953 TABLE 1 CHEMICAL CHARACTERISTICS OF HASKELL LABORATORY WELL3 Parameter MDL b Analytical Value Parameter MDL b Analytical Value BOD, mg/L 2.6 NDc Lead, mg/L 0.0093 ND COD, mg/L 1.7 ND Magnesium, mg/L 0.0183 3.83 DOC, mg/L 0.6 0.82 J d Manganese, mg/L 0.00051 0.103 TOC, mg/L 0.5 ND MBAS/LAS, mg/L 0.035 ND Total Kjeldahl N, mg/L 0.3 ND Mercury, mg/L 0.00016 ND Ammonia N, mg/L 0.03 ND Nickel, mg/L 0.0038 ND Turbidity, NTU 0.09 0.14 J Nitrite, mg/L 0.015 ND Phenolics, mg/L 0.012 ND Nitrate, mg/L 0.04 ND Color, apparent Co/Pte Solids 5.0 ND Ortho-phosphate, mg/L * Potassium, mg/L 0.01 0.0429 0.035 3.55 total suspended, mg/L 1.5 ND Selenium, mg/L 0.0011 ND Aluminum, mg/L 0.0413 ND Silver, mg/L 0.0001 ND Antimony, mg/L 0.0085 ND Sodium, mg/L 0.463 8.08 Arsenic, mg/L 0.0014 ND Sulfate, mg/L 0.3 6.9 Beryllium, mg/L 0.00034 ND Sulfide, mg/L 0.022 ND Boron, mg/L 0.0047 0.0174 J Zinc, mg/L 0.0041 ND Bromide, mg/L 0.4 0.98 Ca/Mg NAf 12.17 Cadmium, mg/L 0.00006 ND Na/K NA 2.28 Calcium, mg/L 0.0494 46.6 Volatile priority Chloride, mg/L 6.0 70.6 pollutants, pg/L 0.5-40 ND Chlorine, residual, mg/L 0.04 ND Acid extractable Chromium, mg/L 0.00025 0.00083 J priority pollutants, pg/L 1.0-19 ND Cobalt, mg/L 0.0016 ND Base/neutral Copper, mg/L 0.00055 0.0061 priority pollutants, pg/L 1.0-19 ND Cyanide, mg/L 0.005 ND Pesticides/PCBs, pg/L 0.0019-0.38 ND-0.0030 J Iron, mg/L 0.0453 ND Organophosphate Fluoride, mg/L 0.08 0.28 pesticides, pg/L 0.38 ND a Sample analyses performed at Lancaster Laboratories, Lancaster, Pennsylvania, date o f sample collection 17 November 2003 unless indicated otherwise, bMDL = method detection limit,cND indicates not detected at the MDL, dA "J" follows analytical values which were greater than the MDL but less than the limit of quantitation,e Units based on cobalt/platinum reference,f NA = not applicable. Sanitized. Does not contain TSCA CBf -25- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration (m z/L f Water Control A Water Control B EPA Control A IPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B Day-T NDC ND ND ND 0.38 0.38 0.71 0.75 1.6 1.5 2.7 2.9 5.3 5.2 Measured^H Concentration fmg/L) Day 0 Day 6 Day 13 Day 20 Day 27 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 0.31 0.28 0.33 0.34 0.33 0.31 0.35 0.34 0.35 0.33 0.61 0.70 0.70 0.72 0.70 0.64 0.72 0.73 0.72 0.70 1.3 1.4 1.5 1.5 1.4 1.3 1.4 1.5 1.4 1.4 2.3 2.7 2.6 2.6 2.5 2.4 2.7 2.7 2.7 2.5 4.4 4.8 5.1 4.8 4.8 4.3 4.7 5.1 4.8 4.8 Day 34 ND ND ND ND 0.33 0.33 0.69 0.47 1.5 1.4 2.5 2.5 4.6 4.7 not contain TSCACB1 gompany Sanitized. Do - 26- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus myMss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration (my/Lf Day 41 Measure Day 48 Day 55 1 Concentration Day 62 Day 69 Day 76 Day 83 Water Control A Water Control B ND ND ND ND ND ND ND ND ND ND ND ND ND ND IPA Control A ND ND ND ND ND ND ND IPA Control B ND ND ND ND ND ND ND 0.30 A 0.30 B 0.34 0.37 0.38 0.37 0.36 0.37 0.38 0.34 0.35 0.33 0.34 0.32 0.36 0.35 0.63 A 0.63 B 0.71 0.73 0.72 0.70 0.71 0.75 0.76 0.69 0.74 0.74 0.73 0.74 0.77 0.77 1.3 A 1.3 B 1.5 1.5 1.5 1.6 1.6 1.6 1.7 1.4 1.4 1.5 1.6 1.6 1.6 1.6 2.5 A 2.5 B 2.8 2.7 2.9 3.0 3.0 3.2 3.2 2.6 2.6 2.7 2.9 2.9 3.1 3.0 5.0 A 5.0 B 4.8 4.9 5.6 5.1 5.8 6.1 5.9 4.8 4.8 5.1 5.2 5.4 5.5 5.7 Sanitized, Does not contain TftCA CBf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration .... (mg/L)a Water Control A Water Control B IPA Control A IPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B rife Concentration (mg/L) u Day 90 Mean. Measured ^ Concentration ** (mg/L)d ND ND ... ND ND -- 0.34 0.35 0.33 0.34 0.73 0.71 0.73 0.71 1.5 1.5 1.6 1.5 2.8 2.8 2.7 2.7 5.4 5.2 5.1 5.0 Mean Percent Recovery6 -- _ ... 117 113 113 113 115 115 112 108 104 100 a Nominal H-25436 concentration based o f l n u r i t y by analysis. b Measured values prior to study start were useeTM verify system function but were not used to calculate mean measured test concentrations o f H - 2 5 4 3 ^ ^ ^ ^ ^ ^ | c ND denotes not detected. The limit of detection for^ ^ ^ ^ ^ ^ ^ M jw as 0.0002 mg/L o f H-25436. d Mean, measured H-25436 concentration was calculated as the average of values obtained from day 0 to day 90. e Based on nominal H-25436 concentration. Company Sanitized. Does not contain TSCA e s i H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration (mg/L)a Water Control A Water Control B IPA Control A IPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B D ay-lb NDC ND ND ND 0.37 0.37 0.70 0.73 1.5 1.4 2.6 2.6 5.2 5.3 m Day 0 ND ND ND ND 0.30 0.31 0.58 0.61 1.3 1.2 2.2 2.1 4.3 4.4 Day 6 ND ND ND ND 0.28 0.34 0.67 0.70 1.4 1.3 2.6 2.4 4.8 4.8 IConcentration Day 13 Day 20 Day 27 ND ND ND ND ND ND ND ND ND ND ND ND 0.31 0.33 0.32 0.33 0.33 0.32 0.67 0.70 0.67 0.69 0.69 0.66 1.4 1.4 1.3 1.4 1.3 1.3 2.6 2.6 2.5 2.4 2.3 2.2 5.0 4.7 4.7 5.2 4.9 4.8 Day 34 ND ND ND ND 0.32 0.32 0.68 0.46 1.4 1.3 2.5 2.2 4.6 4.8 pmpany Sanitized. Does not contain TSCA'CBl -29- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 (mg/L)a Water Control A Water Control B IPA Control A EPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B Day 41 ND ND ND ND 0.33 0.33 0.70 0.65 1.4 1.3 2.6 2.5 4.8 5.0 Day 48 ND ND ND ND 0.34 0.33 0.72 0.67 1.4 1.4 2.5 2.6 4.8 4.9 - Day 55 ND ND ND ND 0.37 0.40 0.72 0.83 1.4 1.5 2.7 2.5 5.2 5.1 Day 62 ND ND ND ND 0.40 0.38 0.72 0.70 1.6 1.6 2.9 2.9 4.9 5.2 loncentration Day 69 Day 76 ND ND ND ND ND ND ND ND 0.39 0.36 0.33 0.34 0.71 0.76 0.71 0.77 1.5 1.7 1.5 1.7 2.9 3.2 2.8 3.1 5.2 5.9 5.0 5.5 Day 8 ND ND ND ND 0.38 0.34 0.76 0.80 1.7 1.6 3.2 3.1 5.7 5.6 Gpmpany Sanitized. Does not contain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration (mg/L)a <| Measured _ Concentration (mg/L) Day 90 ^ Mean, Measured -- ( m concmentratiomn | A* (mg/L)d Mean Percent Recovery6 Water Control A Water Control B ND ND -- -- IPA Control A IPA Control B ND ND -- --- 0.30 A 0.32 0.34 0.30 B 0.31 0.34 113 113 0.63 A 0.71 0.70 0.63 B 0.72 0.69 111 110 1.3 A 1.4 1.5 1.3 B 1.5 1.4 115 108 2.5 A 2.5 2.7 2.5 B 2.6 2.6 108 104 5.0 A 5.2 5.0 5.0 B 5.0 5.0 100 100 a Nominal H-25436 concentration based o^^H Kurity by analysis. b Measured values prior to study start weremsearo verily system function but were not used to calculate mean measured test concentrations of H-2543 c ND denotes not detected. The limit of detection fo: ras 0.00028 mg/L of H-25436. d Mean, measured H-25436 concentration was calculated as the average o f values obtained from day 0 to day 90. e Based on nominal H-25436 concentration. * -31 - fRpmpany Sanitized. Does not contain TSCA CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 Concentration fmg/L)a Water Control A Water Control B IPA Control A EPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B D ay-lb NDC ND ND ND 0.39 0.38 0.75 0.76 1.6 1.5 2.8 2.9 5.6 5.7 Measure! Day 0 Day 6 [Concentration (mg/L) * Day 13 Day 20 Day 27 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 0.30 0.30 0.32 0.33 0.32 0.31 0.35 0.33 0.34 0.32 0.60 0.70 0.69 0.72 0.69 0.61 0.71 0.72 0.71 0.69 1.3 1.5 1.5 1.5 1.4 1.3 1.4 1.5 1.5 1.4 2.4 2.8 2.8 2.8 2.7 2.4 2.7 2.7 2.7 2.6 4.7 5.2 5.5 5.2 5.2 4.7 5.2 5.5 5.3 5.3 Day 34 ND ND ND ND 0.32 0.32 0.70 0.55 1.5 1.4 2.7 2.6 5.1 5.1 .Cwnpany Sanitized. Does not contain TSCA CBl H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus myUss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS Nominal H-25436 ^uuuciiu auDU Cmg/LY Water Control A Water Control B IPA Control A IPA Control B 0.30 A 0.30 B 0.63 A 0.63 B 1.3 A 1.3 B 2.5 A 2.5 B 5.0 A 5.0 B Day 41 ND ND ND ND 0.33 0.33 0.72 0.70 1.5 1.5 2.8 2.8 5.3 5.4 Measure^ Day 48 Day 55 ND ND ND ND ND ND ND ND 0.34 0.36 0.33 0.42 0.72 0.72 0.71 0.79 1.4 1.4 1.4 1.5 2.7 2.8 2.7 2.7 5.2 5.5 5.2 5.4 Day 62 ND ND ND ND 0.45 0.43 0.78 0.70 1.6 1.6 3.0 2.9 5.3 5.5 oncentration Day 69 Day 76 ND ND ND ND ND ND ND ND 0.49 0.36 0.36 0.35 0.76 0.84 0.68 0.76 1.4 1.8 1.5 1.7 2.8 3.2 2.8 3.1 5.2 6.1 5.1 5.8 Day 83 ND ND ND ND 0.36 0.35 0.76 0.76 1.6 1.6 3.2 3.1 5.9 5.9 .Company Sanitized. Does not contain TSCA CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 2 (Continued) MEASURED CONCENTRATIONS OF H-25436 IN TEST SOLUTIONS 1 Measured Nominal H-25436 Concentration (mg/L)a 1 Concentration (mg/L) Day 90 Mean, Measured Concentration (mg/L)" ,, r *1 Mean Percent Recovery6 Water Control A Water Control B P A Control A P A Control B ND ND ND ND -- -- -- -- -- 0.30 A 0.30 B 0.35 0.32 0.35 0.35 117 117 0.63 A 0.63 B 0.70 0.69 0.72 0.70 114 111 1.3 A 1.3 B 1.5 1.5 1.5 1.5 115 115 2.5 A 2.5 B 2.8 2.8 2.8 2.8 112 112 5.0 A 5.0 B 5.5 5.5 5.4 5.4 108 108 a Nominal H-25436 concentration based oi^^HKurity by analysis. b Measured values prior to study start wereusecno verify system function but were not used to calculate mean measured test concentrations ofH-25436. ^ c ND denotes not detected. The limit of detection f o r j B B U t f H j K v a s 0.0004 mg/L ofH-25436. d Mean, measured H-25436 concentration was calculated as the average o f values obtained from day 0 to day 90. *e Based on nominal H-25436 concentration. 4 jQpznpany Sanitized. Does not contain TSCA CB1 - 34- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 3 _ WATER CHEMISTRY OF THE DILUTION WATER CONTROL, IPA CONTROL, AND HIGHEST TEST CONCENTRATION WITH SURVIVING ORGANISMS" Total Alkalinity Day Replicate (mg/L as CaCCh) Dilution Water Control 0A 6A 13 A 20 A 27 A 34 A 41 A 48 A 55 A 62 A 69 A 76 A 83 A 90 A IPA Control 0A 6A 13 A 20 A 27 A 34 A 41 A 48 A 55 A 62 A 69 A 76 A 83 A 90 A 50 50 49 49 47 50 49 49 49 47 47 49 50 47 51 50 50 50 50 51 50 50 49 48 48 49 51 51 EDTA Hardness (mg/L as C aC 0 3) 128 131 130 128 123 124 126 126 127 131 123 126 126 118 128 132 129 131 124 124 128 126 126 130 124 127 126 121 Conductivity (pmhos/cm) 270 270 250 260 240 225 220 240 235 230 220 210 220 220 280 270 250 265 245 225 225 240 235 230 220 210 225 230 .Company Sanitized. Does not contain trca C ii H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 3 (Continued) WATER CHEMISTRY OF THE DILUTION WATER CONTROL, IPA CONTROL, AND HIGHEST TEST CONCENTRATION WITH SURVIVING ORGANISMS" Total Alkalinity EDTA Hardness Day Replicate (mg/L as C aC 0 3) (mg/L as CaCOs) Conductivity (gmhos/cm) Highest Test Concentration with Surviving Organisms - 5.0 mg/L 0A 51 129 6A 47 132 13 A 50 132 20 A 50 130 27 A 48 123 34 A 50 123 41 A 49 126 48 A 51 127 55 A 52 130 62 A 48 131 69 A 47 124 76 A 49 127 83 A 51 127 90 A 49 120 282 270 252 270 250 230 225 250 235 230 220 215 222 230 a Based on nominal H-25436 test concentration. Company Sanitized. Does not contain TSCA CB1 - 36- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 4 TEMPERATURE (C) OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dilution Water EPA Day Replicate Control Control Temperature (C) Nominal Concentrations (mg/L) 0.30 0.63 1.3 2.5 5.0 Water Bath 0A 0B 6A 6B 13 A 13 B 20 A 20 B 27 A 27 B 34 A 34 B 41 A 41 B 48 A 48 B 55 A 55 B 62 A 62 B 69 A 69 B 76 A 76 B 83 A 83 B 90 A 90 B 11.9 11.9 11.9 11.9 11.9 11.9 11.9 12.2 11.9 11.9 11.9 11.9 11.9 11.9 11.9 11.7 11.8 11.9 11.7 11.7 11.7 11.9 12.2 11.7 11.7 11.8 11.7 11.7 11.8 11.7 11.8 11.8 11.9 11.8 11.7 11.7 11.9 12.2 11.8 11.8 11.8 11.8 11.7 11.9 11.8 11.7 11.8 11.9 11.8 11.7 11.7 11.9 12.3 11.7 11.7 11.8 11.8 11.7 12.0 11.8 11.9 11.8 11.9 11.8 11.8 11.7 11.9 12.2 11.8 11.8 11.9 11.8 11.8 11.9 11.8 11.9 11.9 12.0 11.8 11.9 11.8 12.0 12.2 11.9 11.9 11.9 12.0 11.9 11.9 11.9 11.9 11.9 11.9 11.9 11.9 11.8 11.8 12.2 11.8 11.8 11.9 11.8 11.8 11.8 11.8 11.9 11.8 11.9 11.8 11.8 11.8 11.9 12.2 11.8 11.8 11.9 11.9 11.7 11.7 11.9 11.9 11.8 11.9 11.8 11.8 11.8 11.8 12.3 11.8 11.8 11.8 11.8 11.8 11.8 11.8 11.9 11.9 11.9 11.9 11.8 11.9 11.9 12.2 11.9 11.9 11.9 11.9 11.8 11.7 11.8 12.0 11.9 11.9 11.9 11.7 11.9 11.9 12.2 11.9 11.9 11.9 11.9 11.8 11.8 11.8 11.9 11.8 11.9 11.8 11.7 11.9 11.9 12.1 11.9 11.8 11.9 11.9 11.8 11.8 11.9 11.9 11.8 11.9 11.8 11.8 11.9 11.8 12.2 11.9 11.9 11.9 11.8 11.7 11.8 11.8 12.0 11.9 12.0 11.8 11.9 11.9 11.9 12.2 12.0 11.9 12.0 11.9 11.8 11.8 11.9 Temperature (C) Mean for all concentrations, including controls 11.8 Std. Dev. o f for all concentrations, including controls 0.1 Minimum for all concentrations, including controls 11.7 Maximum for all concentrations, including controls 12.0 - 37- Oompany Sanitized. Does not cor>itn tsca OSS H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 5 pH OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dilution Water IPA Day Replicate Control Control pH Nominal Concentrations (mg/L) 0.30 0.63 1.3 2.5 5.0 0A 0B 6A 6B 13 A 13 B 20 A 20 B 27 A 27 B 34 A 34 B 41 A 41 B 48 A 48 B 55 A 55 B 62 A 62 B 69 A 69 B 76 A 76 B 83 A 83 B 90 A 90 B 7.4 7.4 7.4 7.4 7.3 7.3 7.3 7.4 7.4 7.3 7.3 7.3 7.3 7.3 7.5 7.5 7.5 7.5 7.4 7.4 7.4 7.5 7.5 7.4 7.4 7.4 7.4 7.4 7.7 7.7 7.7 7.7 7.7 7.6 7.6 7.7 7.7 7.6 7.7 7.6 7.7 7.6 7.5 7.4 7.5 7.4 7.5 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.3 7.3 7.3 7.3 7.4 7.3 7.3 7.3 7.3 7.3 7.3 7.4 7.4 7.3 7.3 7.3 7.3 7.3 7.4 7.3 7.3 7.4 7.3 7.3 7.3 7.4 7.4 7.4 7.4 7.4 7.4 7.3 7.4 7.4 7.4 7.4 7.4 7.3 7.3 7.4 7.4 7.3 7.3 7.3 7.3 7.3 7.4 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.2 7.2 7.3 7.3 7.3 7.3 7.3 7.2 7.2 7.3 7.3 7.3 7.3 7.3 7.3 7.2 7.3 7.3 7.3 7.3 7.3 7.2 7.2 7.3 7.2 7.3 7.2 7.2 7.2 7.2 7.3 7.2 7.2 7.2 7.2 7.2 7.2 7.3 7.3 7.2 7.2 7.2 7.2 7.1 7.3 7.2 7.2 7.2 7.2 7.2 7.1 7.3 7.3 7.3 7.2 7.2 7.2 7.2 7.3 7.3 7.3 7.2 7.2 7.2 7.2 7.3 7.3 7.3 7.3 7.3 7.3 7.2 7.3 7.3 7.3 7.3 7.3 7.2 7.2 Minimum for all concentrations, including controls 7.1 Maximum for all concentrations, including controls 7.7 - 38- Company Sanitized. Does not contain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 6 DISSOLVED OXYGEN CONCENTRATION (mg/L) OF TEST SOLUTIONS FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dilution Water IPA Day Replicate Control Control Dissolved Oxygen Concentration (mg/L)a Nominal Concentrations (mg/L) 0.30 0.63 1.3 2.5 5.0 0A 10.3 10.4 10.3 10.3 10.1 10.0 10.0 0B 10.3 10.4 10.1 10.3 10.1 10.2 9.9 6A 10.4 10.6 10.4 10.4 10.1 10.0 10.2 6B 10.5 10.5 10.2 10.4 10.3 10.3 10.0 13 A 10.7 10.6 10.6 10.5 10.4 10.3 10.5 13 B 10.6 10.6 10.4 10.6 10.4 10.5 10.3 20 A 10.6 10.5 10.5 10.3 10.2 10.1 10.1 20 B 10.5 10.5 10.2 10.4 10.2 10.3 10.0 27 A 10.3 10.3 10.2 10.2 10.1 9.9 10.0 27 B 10.3 10.3 10.0 10.2 10.1 10.2 9.8 34 A 10.1 10.2 10.0 9.9 9.9 10.0 9.8 34 B 10.2 10.2 10.0 10.0 10.0 10.0 9.8 41 A 10.4 10.2 10.1 10.1 10.0 10.1 9.9 41 B 10.4 10.2 10.1 10.2 10.1 10.1 9.9 48 A 10.4 10.3 10.2 10.1 10.1 10.1 10.0 48 B 10.4 10.2 10.1 10.1 10.1 10.0 9.9 55 A 10.2 10.1 10.0 10.0 10.0 10.0 9.9 55 B 10.2 10.1 10.0 9.9 10.1 10.0 9.8 62 A 10.1 10.0 9.9 10.0 9.9 9.9 9.7 62 B 10.1 10.1 9.9 9.9 9.9 9.9 9.7 69 A 10.1 10.1 9.9 10.0 9.9 9.8 9.6 69 B 10.2 10.1 9.9 10.0 10.0 9.9 9.7 76 A 9.0 9.0 8.8 8.9 8.8 8.8 8.6 76 B 9.1 9.0 8.8 8.8 8.9 8.8 8.6 83 A 9.5 9.4 9.1 9.3 9.3 9.0 9.0 83 B 9.6 9.5 9.3 9.2 9.4 9.3 9.1 90 A 9.8 9.3 8.8 9.3 9.4 8.8 8.8 90 B 9.7 9.6 9.2 8.9 9.4 9.1 9.0 Dissolved Oxygen C oncentration (mg/L) Mean for all concentrations, including controls 9.9 Std. Dev. for all concentrations, including controls 0.5 Minimum for all concentrations, including controls 8.6 Maximum for all concentrations, including controls 10.7 a The theoretical dissolved oxygen concentration at saturation at 12C = 10.8 mg/L. - 39- Coinpany Sanitized. Does not contain TSCA CBI ) H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss ) ) DuPont-11953 TABLE 7 SUMMARY OF HATCHING, SURVIVAL, ABNORMALITIES, AND SWIM-UP FROM HATCHING TO THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Nominal H-25436 Concentration (mg/L) Replicate Hatching Day First Last3 Percent Hatch Number Hatching Total Percent Number Alive Hatching to Thinning Survival Abnormalitiesb Number Number Number Hatching Percent Affected Alive Percent First Day of Swim-Up Dilution Water Control 1A 2A IB 2B Mean0 27 27 26 28 27 30 31 30 30 30 17 20 85% 15 17 88% 2 16 20 80% 15 16 94% 1 18 20 ` 90% 17 18 94% 1 15 20 75% 15 15 100% 0 17 20 83% 16 17 94% 1 15 13% 40 15 7% 40 17 6% 40 15 0% 40 16 6% 40 IPA Control 1A 2A IB 2B Mean0 27 27 28 26 27 30 29 31 31 30 18 20 90% 17 18 94% 1 17 20 85% 16 17 94% 0 17 20 85% 17 17 100% 1 17 20 85% 15 17 88% 0 17 20 86% 16 17 94% 1 17 6% 41 16 0% 40 17 6% 40 15 0% 40 16 6% 40 0.30 1A 27 30 17 20 85% 15 17 88% 0 15 0% 40 2A 27 30 17 20 85% 16 17 94% 1 16 6% 40 IB 27 32 16 20 80% 16 16 100% 1 16 6% 40 2B 27 31 18 20 90% 18 18 100% 4 18 22% 40 Mean0 27 31 17 20 85% 16 17 94% 2 16 13% 40 pompany Sanitized. Does not contain t^c-a ri - 40- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 7 (Continued) SUMMARY OF HATCHING, SURVIVAL, ABNORMALITIES, AND SWIM-UP FROM HATCHING TO THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Nominal H-25436 Concentration (mg/L) Replicate Hatching Day First Last3 Percent Hatch Number Hatching Total Percent Number Alive Hatching to Thinning Survival Abnormalitiesb Number Number Number Hatching Percent Affected Alive Percent First Day of Swim-Up 0.63 1A 26 31 16 20 80% 15 16 94% 2 15 13% 40 2A 27 30 16 20 80% 16 16 100% 1 16 6% 40 IB 27 30 16 20 80% 16 16 100% 0 16 0% 40 2B 26 31 14 20 70% 13 14 93% 1 13 8% 40 Mean0 27 31 16 20 80% 15 16 94% 1 15 7% 40 1.3 1A 27 31 16 20 80% 15 16 94% 1 15 7% 40 2A 25 32 17 20 85% 17 17 100% 1 17 6% 40 IB 26 30 17 20 85% 17 17 100% 0 17 0% 40 2B 27 31 18 20 90% 16 18 89% 0 16 0% 41 Mean0 26 31 17 20 85% 16 17 94% 1 16 7% 40 2.5 1A 27 33 13 20 65% 13 13 100% 0 13 0% 41 2A 28 32 12 20 60% 12 12 100% 1 12 8% 42 IB 27 30 16 20 80% 15 16 94% 0 15 0% 40 2B 27 32 17 20 85% 16 17 94% 0 16 0% 41 Mean0 27 32 15 20 75% 14 15 93% 0 14 0% 41 [Company Sanitized. Does not contain v s c . -41 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 7 (Continued) SUMMARY OF HATCHING, SURVIVAL, ABNORMALITIES, AND SWIM-UP FROM HATCHING TO THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Nominal H-25436 Concentration (mg/L) Replicate Hatching Day First Last3 Percent Hatch Number Hatching Total Percent Hatching t a Thinning Survival Abnormalitiesb Number Number Number Number Alive Hatching Percent Affected Alive Percent First Day of Swim-Up 5.0 1A 26 30 17 20 85% 17 17 100% 0 17 0% 40 2A 25 31 17 20 85% 16 17 94% 2 16 13% 41 IB 28 32 18 20 90% 18 18 100% 2 18 11% 42 2B 28 32 16 20 80% 16 16 100% 0 16 0% 40 Mean0 27 31 17 20 85% 17 17 100% 1 17 6% 41 a Last day o f hatching is defined as the last day that remaining live eggs (or live embryos) hatch to yield live alevins. b Based on abnormalities still present at the time o f thinning, c Calculated as mean o f replicate values, percent is based on mean values. Company Sanitized. Does not c o n ta in tsca O H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 8 SUMMARY OF SURVIVAL, ABNORMALITIES, AND GROWTH FROM THINNING TO TEST END FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Nominal H-25436 Concentration (mg/L) Dilution Water Control IPA Control 0.30 0.63 1.3 Replicate Number Alive Thinning to Test End Survival Abnormalitiesa Number Number Total Percent Affected Alive Percent A B Meanb 15 15 15 A B Meanb 15 15 15 A B Meanb 15 14 15 A B Meanb 13 15 14 A B Meanb 15 15 15 15 100% 0 15 100% 0 15 100% 0 15 100% 0 15 100% 0 15 100% 0 15 100% 0 15 93% 0 15 97% 0 15 87% 0 15 100% 0 15 93% 0 15 100% 1 15 100% 0 15 100% 1 15 0% 15 0% 15 0% 15 0% 15 0% 15 0% 15 0% 14 0% 15 0% 13 0% 15 0% 14 0% 15 7% 15 0% 15 7% Growth of Surviving Fingerlings Mean Mean Standard Length Wet Weight Std. Dev. (cm) Std. Dev. (g) 4.6 0.32 4.5 0.28 4.6 0.30 4.4 0.34 4.3 0.29 4.4 0.32 4.5 0.48 4.4 0.44 4.5 0.46 4.4 0.37 4.4 0.50 4.4 0.44 4.5 rb 0.54 4.3 0.47 4.4 0.51 1.2788 0.2537 1.1987 0.2801 1.2388 0.2672 1.2007 0.3033 1.0568 0.2360 1.1287 0.2717 1.2701 0.4096 1.2061 0.4075 1.2392 0.4086 1.2668 0.3575 1.1690 0.3704 1.2144 0.3645 1.1730 0.4043 1.1151 0.3462 1.1441 0.3764 Company Sanitized, Does not contain TSCA CBf ) H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss ) ) DuPont-11953 TABLE 8 (Continued) SUMMARY OF SURVIVAL, ABNORMALITIES, AND GROWTH FROM THINNING TO TEST END FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Nominal H-25436 Concentration ______ (mg/L) 2.5 5.0 Replicate Number Alive Thinning to Test End Survival Abnormalitiesa Number Number Total Percent Affected Alive Percent A B Mean" 13 15 14 15 87% 0 15 100% 0 15 93% 0 13 0% 15 0% 14 0% A B Meanb 14 15 15 15 93% 0 15 100% 0 15 100% 0 14 0% 15 0% 15 0% Growth o f Surviving Fingerlings Mean Mean Standard Length Wet Weight Std. Dev. (cm) Std. Dev. (g) 4.4 0.43 4.2 0.58 4.3 0.52 1.1420 0.3819 1.0031 0.4461 1.0726 0.4177 4.5 0.22 4.2 0.47 4.3 0.37 1.2207 0.1918 1.0324 0.3628 1.1233 0.2932 a Based on abnormalities still present at test end. b Calculated as mean o f replicate values, percent is based on mean values. I 0o 3 S' *4 r> # - 4 4 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 - - - - - - -45 - Company Sanitized. Does not contain TSCA Cf* H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 (Continued) DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 - - - 46- ' 0mpanjr Similized, Does not contain TSC A CB>. H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 (Continued) DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B 0 0 0 0 0 0 0 0 2 4 2 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 3 17 4 16 2 18 4 16 3 17 4 16 2 18 4 16 3 17 4 16 2 18 4 16 3 17 4 16 2 18 4 16 3 17 4 16 2 18 4 16 3 17 4 16 2 18 4 16 - - - - - - - - - 47- Som paty Sanitized. Does nof co n tain T S C A C B I H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 (Continued) DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 24 2A 24 IB 24 2B 25 1A 25 2A 25 IB 25 2B 26 1A 26 2A 26 IB 26 2B 27 1A 27 2A 27 IB 27 2B 28 1A 28 2A 28 IB 28 2B 29 1A 29 2A 29 IB 29 2B 30 1A 30 2A 30 IB 30 2B 31 1A 31 2A 31 IB 31 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 17 4 16 2 18 4 16 3 17 0 4 16 0 2 18 0 4 16 0 3 17 0 4 16 0 2 16 0 4 16 0 3 16 0 4 15 0 2 15 0 4 16 0 3 12 0 4 12 0 270 4 11 0 33 0 460 220 44 0 3 10 450 2 10 500 3 10 400 2 10 500 - - - 0 0 0 0 0 0 2 0 1 1 3 0 5 4 11 5 14 10 16 12 16 11 17 15 16 16 17 15 - 48- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 (Continued) DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 32 1A 32 2A 32 IB 32 2B 33 1A 33 2A 33 IB 33 2B 34 1A 34 2A 34 IB 34 2B 35 1A 35 2A 35 IB 35 2B 36 1A 36 2A 36 IB 36 2B 37 1A 37 2A 37 IB 37 2B 38 1A 38 2A 38 IB 38 2B 39 1A 39 2A 39 IB 39 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 1 16la 4 0 1 15la 2 0 1 17la 50 0 15 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 1 16la,lb 4 0 1 15la 2 0 1 17la 5 0 0 15la - 49- jCompany Sanitized. Does not contain T<*rA c m H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 9 (Continued) DILUTION WATER CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate1 Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 40 2A 40 IB 40 2B 41 1A 41 2A 41 IB 41 2B 42 1A 42 2A 42 IB 42 2B 43 1A 43 2A 43 IB 43 2B 44 1A 44 2A 44 IB 44 2B 45 1A 45 2A 45 IB 45 2B 46 1A 46 2A 46 IB 46 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 2 152a 4 0 1 15la 2 0 1 17Ia 5 0 0 15la 3 0 2 152a 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 2 152a 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 2 152a 4 0 1 15la 2 0 1 17la 5 0 0 15la 3 0 2 152a 4 0 1 15la 2 0 1 17la 5 00 15 3 0 2 152a 4 0 1 15la 2 0 1 17la 500 15 3 0 2 152a 4 0 1 15la 2 0 1 17la 500 15 f Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color iDjonipany Sanitized. Does not contain TSCA CB1 - 50- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicatet Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 - 0 20 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 0 20 - 1 19 - - -51 - J^npany Sanitized. Does not conW" q H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 (Continued) IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate* Removed Removed Eggs Left Larvae Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 - 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 1 19 1 19 0 20 0 20 1 19 2 18 - - - - -52- felfipany Sanitized. Does not soRfu? **** <*^1 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 (Continued) IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B Dead Eggs Removed 0 0 0 0 0 0 0 0 2 2 2 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Dead Larvae Removed - - - Cumulative Dead Eggs 0 0 1 2 0 0 1 2 2 2 3 3 2 2 3 3 2 2 3 3 2 2 3 3 2 2 3 3 2 2 3 3 Live Eggs Left 20 20 19 18 20 20 19 18 18 18 17 17 18 18 17 17 18 18 17 17 18 18 17 17 18 18 17 17 18 18 17 17 Cumulative Dead Larvae . - - - Live Larvae . - - - .Kggpapf Sanitized. Does not contain TSCA CBi - 53- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 (Continued) IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 24 2A 24 IB 24 2B 25 1A 25 2A 25 IB 25 2B 26 1A 26 2A 26 IB 26 2B 27 1A 27 2A 27 IB 27 2B 28 1A 28 2A 28 IB 28 2B 29 1A 29 2A 29 IB 29 2B 30 1A 30 2A 30 IB 30 2B 31 1A 31 2A 31 IB 31 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 - - - 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 2 18 2 18 3 17 3 17 2 18 0 2 18 0 3 17 0 3 17 0 2 18 0 2 18 0 3 17 0 3 16 0 2 16 0 2 11 0 3 17 0 3 12 1 2 10 0 260 390 382 2 50 2 10 350 342 200 2 10 320 322 200 300 300 302 - - - 0 0 0 0 0 0 0 1 2 7 0 4 8 12 8 7 13 17 12 11 18 17 15 13 18 17 17 15 jGpmpanjf Sanitized. Does not contain TSCA C81 - 54- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 (Continued) IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate* Removed Removed Eggs Left Larvae Larvae 32 1A 0 0 2 0 0 18la,ld 32 2A 32 IB 0 0 0 0 300 17 3 0 0 j-yla,2d 32 2B 33 1A 0 0 0 0 3 0 2 15Id 2 0 0 2g1a,1d 33 2A 33 IB 0 0 0 0 3 0 0 17lb 3 0 0 2^la,2d 33 2B 34 1A 0 0 0 0 3 0 2 15ld 2 0 0 jgla,ld 34 2A 34 IB 0 0 0 0 3 0 0 17lb 3 0 0 2'yla>2d 34 2B 35 1A 0 0 0 0 3 0 2 15ld 2 0 0 2gla,ld 35 2A 35 IB 0 0 1 0 30 1 16 3 0 0 2 la,2d 35 2B 0 0* 3 0 2 15id 36 1A 0 1 2 0 1 17la 36 2A 36 IB 0 0 0 0 30 1 16 3 0 0 j yla,2d 36 2B 37 1A 0 0 0 0 3 0 2 152d 2 0 1 l^la 37 2A 37 IB 0 0 0 0 3 0 1 16 3 0 0 2y la*2d 37 2B 0 0 3 0 2 152d 38 1A 0 0 2 0 1 17la 38 2A 38 IB 0 0 0 0 3 0 1 16 3 0 0 2 la,2d 38 2B 0 0 3 0 2 152d 39 1A 0 0 2 0 1 17la 39 2A 39 IB 0 0 0 0 3 0 1 16 3 0 0 2yla,2d 39 2B 0 0 3 0 2 152bd JS^ppaiif .Sanitized. Does not contain TSCA CBi - 55- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 10 (Continued) IPA CONTROL: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate1 Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 40 2A 40 IB 40 2B 41 1A 41 2A 41 IB 41 2B 42 1A 42 2A 42 IB 42 2B 43 1A 43 2A 43 IB 43 2B 44 1A 44 2A 44 IB 44 2B 45 1A 45 2A 45 IB 45 2B 46 1A 46 2A 46 IB 46 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 1 17la 3 0 1 16 3 0 0 Jrji&y'ld 3 0 2 152bd 2 0 1 17la 3 0 1 16 3 0 0 2rj1a,2d 3 0 2 l 52bd 2 0 1 17la 30 1 16 3 0 0 ^yla,2d 3 0 2 152bd 2 0 1 17la 301 16 3 0 0 ^yla,2d 3 0 2 152bd 2 0 1 17la 30 1 16 3 0 0 2*jla,2d 3 0 2 152bd 2 0 1 17la 3 0 1 16 3 0 0 2^i&jid 302 15 2 0 1 17la 30 1 16 3 0 0 17Ia 302 15 f Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color d Small in size (when compared to appropriate control(s)) Ian2iized. Poes not contain TSCA CBS - 56- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 2 18 0 20 0 20 - _ - Sanitized, Dees not contain TSCA CBI - 57- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 (Continued) 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 - 0 20 . 2 18 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 0 20 1 19 0 20 2 18 0 20 1 19 0 20 2 18 0 20 1 19 - . - ll^PpaSf Sanitized. Does not eon** '***** -58- Sanitized. Does not contain TSCA CBf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 (Continued) 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B Dead Eggs Removed 0 0 0 0 0 0 0 0 3 1 3 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Dead Larvae Removed - - - - - - - - - - - - - - Cumulative Dead Eggs 0 2 0 1 0 2 0 1 3 3 3 2 3 3 3 2 3 3 3 2 3 3 3 2 3 3 3 2 3 3 3 2 Live Eggs Left 20 18 20 19 20 18 20 19 17 17 17 18 17 17 17 18 17 17 17 18 17 17 17 18 17 17 17 18 17 17 17 18 Cumulative Dead Larvae _ - - - - - - - - - - - - - - - Live Larvae - - - - - - - - - - - - - - - - - - 59- fempanjf Sanitized. Does not contain tsca CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 (Continued) 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* 24 1A 24 2A 24 IB 24 2B 25 1A 25 2A 25 IB 25 2B 26 1A 26 2A 26 IB 26 2B 27 1A 27 2A 27 IB 27 2B 28 1A 28 2A 28 IB 28 2B 29 1A 29 2A 29 IB 29 2B 30 1A 30 2A 30 IB 30 2B 31 1A 31 2A 31 IB 31 2B Dead Eggs Removed 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 Dead Larvae Removed - - - 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cumulative Live Dead Eggs Eggs - Left 3 17 3 17 3 17 2 18 3 17 3 17 3 17 2 18 3 17 3 17 3 17 2 18 3 11 3 12 3 16 2 17 34 33 3 13 2 14 31 31 45 23 30 30 43 21 30 30 41 20 Cumulative Dead Larvae - - - 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Live Larvae _ - - - 0 0 0 0 0 0 0 0 6 5 1 1 13 14 4 4 16 16 11 15 17 17 13 17 17 17 15 18 tmpany Sanitized Does not contain rev* P s tio t c o n ta in TSC A C il H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 (Continued) 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 32 1A 32 2A 32 IB 32 2B 33 1A 33 2A 33 IB 33 2B 34 1A 34 2A 34 IB 34 2B 35 1A 35 2A 35 IB 35 2B 36 1A 36 2A 36 IB 36 2B 37 1A 37 2A 37 IB 37 2B 38 1A 38 2A 38 IB 38 2B 39 1A 39 2A 39 IB 39 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 0 0 17Ia 300 17 4 0 0 16la 2 0 0 184a 3 0 0 17Ia 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 0 17la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 0 17la 3 0 0 17Ib 4 0 0 16la 2 0 0 184a 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a &tnp'mx Sanitized. Dees hot contain TSCA c -61 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 11 (Continued) 0.30 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate1 Removed Removed Eggs Left Larvae Larvae 40 1A 0 0 40 2A 0 0 40 IB 0 0 40 2B 0 0 41 1A 0 0 41 2A 0 0 41 IB 0 0 41 2B 0 0 42 1A 0 1 42 2A 0 0 42 IB 0 0 42 2B 0 0 43 1A 0 0 43 2A 0 0 43 IB 0 0 43 2B 0 0 44 1A 0 0 44 2A 0 0 44 IB 0 0 44 2B 0 0 45 1A 0 0 45 2A 0 1 45 IB 0 0 45 2B 0 0 46 1A 0 0 46 2A 0 0 46 IB 0 0 46 2B 0 0 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 3 0 1 16la 3 0 0 17lb 4 0 0 16la 2 0 0 184a 302 15 3 0 0 17lb 4 0 0 16la 2 0 0 184a 302 15 300 4 0 0 16la 2 0 0 184a 302 15 300 4 0 0 16la 200 3 02 15 3 0 1 16la 4 0 0 16la 2 0 0 184a 302 15 3 0 1 16la 4 0 0 16la 2 0 0 184a c*Od?Q f Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color .Sanitized. Boas not contain TSCA H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 12 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 - - Sanlflw a, Does ! contain TSCA cm -63 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus myldss DuPont-11953 TABLE 12 (Continued) 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate1^ Removed Removed Eggs Left Larvae Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 - 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 0 20 1 19 1 19 1 19 0 20 1 19 1 19 1 19 0 20 2 18 1 19 1 19 0 20 2 18 1 19 1 19 0 20 - - JiSWjsaiJ! Sanitized. Does not contain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 12 (Continued) 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate+ Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B 0 0 0 0 0 0 0 0 2 3 2 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 2 18 1 19 1 19 0 20 2 18 1 19 1 19 0 20 4 16 4 16 3 17 4 16 4 16 4 16 3 17 4 16 4 16 4 16 3 17 4 16 4 16 4 16 3 17 4 16 4 16 4 16 3 17 4 16 4 16 4 16 3 17 4 16 - - few pauf Sanitized. Does not contain TSCA CBI - 65- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 12 (Continued) 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 0 24 2A 0 - 24 IB 0 - 24 2B 0 - 25 1A 0 0 25 2A 0 0 25 IB 0 0 25 2B 0 0 26 1A 0 0 26 2A 0 0 26 IB 0 0 26 2B 0 0 27 1A 0 0 27 2A 0 0 27 IB 0 0 27 2B 0 0 28 1A 0 0 28 2A 0 0 28 IB 0 0 28 2B 0 0 29 1A 0 0 29 2A 0 0 29 IB 0 0 29 2B 2 0 30 1A 0 0 30 2A 0 0 30 IB 1 0 30 2B 0 0 31 1A 0 0 31 2A 0 0 31 IB 0 0 31 2B 0 0 4 16 4 16 3 17 4 16 4 16 0 4 16 0 3 17 0 4 16 0 4 15 0 4 16 0 3 17 0 4 15 0 4 14 0 4 15 0 3 15 0 4 13 0 490 460 3 10 0 490 440 420 320 630 420 400 400 620 400 400 400 600 - - - 0 0 0 0 1 0 0 1 2 1 2 3 7 10 7 7 12 14 15 11 14 16 16 12 16 16 16 14 Does not contain TSCA CBl - 66- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 12 (Continued) 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate^ Removed Removed Eggs .. Left Larvae Larvae 32 1A 32 2A 32 IB 32 2B 33 1A 33 2A 33 IB 33 2B 34 1A 34 2A 34 IB 34 2B 35 1A 35 2A 35 IB 35 2B 36 1A 36 2A 36 IB 36 2B 37 1A 37 2A 37 IB 37 2B 38 1A 38 2A 38 IB 38 2B 39 1A 39 2A 39 IB 39 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 4 0 1 15la 4 00 16 4 0 0 16la 6 0 0 14lb 4 0 1 15la 400 16 4 0 0 16la 6 0 0 14lb 4 0 1 152a 400 16 4 0 0 16la 6 0 0 14lb 4 0 1 152a 4 00 16 4 0 0 16la 6 0 0 14lb 4 0 1 152a 400 16 4 0 0 16la 601 13 4 0 1 152a 400 16 4 0 0 16Ia 601 13 4 0 1 152a 400 16 4 0 0 16la 60 1 13 4 0 1 152a 400 16 4 0 0 16la 601 13 Bggggiy Jgaalfteed. Does not contain TSCA CBf -67- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 12 (Continued) 0.63 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate1^ Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 40 2A 40 IB 40 2B 41 1A 41 2A 41 IB 41 2B 42 1A 42 2A 42 IB 42 2B 43 1A 43 2A 43 IB 43 2B 44 1A 44 2A 44 IB 44 2B 45 1A 45 2A 45 IB 45 2B 46 1A 46 2A 46 IB 46 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 4 0 1 152a 400 16 4 0 0 16la 6 0 1 13 4 0 1 152a 400 16 4 0 0 16la 6 0 1 13 4 0 1 152a 400 16 400 16 6 0 1 13Ia 4 0 1 152a 400 16 400 16 6 0 1 13la 4 0 1 152a 400 16 400 16 6 0 1 13la 4 0 1 152a 400 16 400 16 6 0 1 13la 4 0 1 152a 4 0 0 16`dd 400 16 6 0 1 13la f Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color dd Loss of equilibrium ompany Sanitized Does no! contain TSCA CBI -68- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 f TABLE 13 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 - - Company Sanitised. Does not conisti - 69- f*! H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 13 (Continued) 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 1 0 0 0 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 0 20 0 20 0 20 2 18 1 19 0 20 0 20 2 18 1 19 0 20 - - .Company Sanitized. Does not contain TSCA CBf - 70- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 13 (Continued) 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B 0 0 0 0 0 0 0 0 3 1 2 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - - 0 20 2 18 - 1 19 - 0 20 - 0 20 - 2 18 - 1 19 - 0 20 - 3 17 - 3 17 - 3 17 - 2 18 - 3 17 - 3 17 - 3 17 - 2 18 - 3 17 - 3 17 - 3 17 - 2 18 - 3 17 - 3 17 - 3 17 - 2 18 - 3 17 - 3 17 - 3 17 - 2 18 - 3 17 - 3 17 - 3 17 - 2 18 - - -71 - C o m p a n y Sanitized, Does not contain TSCA C m H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 13 (Continued) 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs .... Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 0 24 2A 0 - 24 IB 0 - 24 2B 0 - 25 1A 0 0 25 2A 0 0 25 IB 0 0 25 2B 0 0 26 1A 0 0 26 2A 0 0 26 IB 0 0 26 2B 0 0 27 1A 0 0 27 2A 0 0 27 IB 0 0 27 2B 0 0 28 1A 1 0 28 2A 0 0 28 IB 0 0 28 2B 0 0 29 1A 0 0 29 2A 0 0 29 IB 0 0 29 2B 0 0 30 1A 0 0 30 2A 0 0 30 IB 0 0 30 2B 0 0 31 1A 0 0 31 2A 0 0 31 IB 0 0 31 2B 0 0 3 17 3 17 3 17 2 18 3 17 0 3 16 0 3 17 0 2 18 0 3 17 0 3 16 0 3 16 0 2 18 0 3 15 0 3 15 0 3 16 0 2 16 0 4 10 0 3 11 0 3 12 0 2 12 0 420 330 3 10 260 4 10 320 300 230 400 3 10 300 200 - - - 0 1 0 0 0 1 1 0 2 2 1 2 6 6 5 6 14 14 16 12 15 15 17 15 16 16 17 18 Company Sanitized, Does not confab T sra r* - 72- Do m tsca cw H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss D u P o n t-11953 TABLE 13 (Continued) 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 32 1A 0 0 32 2A 0 0 32 IB 0 0 32 2B 0 0 33 1A 0 0 33 2A 0 0 33 IB 0 0 33 2B 0 0 34 1A 0 0 34 2A 0 0 34 IB 0 0 34 2B 0 0 35 1A 0 1 35 2A 0 0 35 IB 0 0 35 2B 0 2 36 1A 0 0 36 2A 0 0 36 IB 0 0 36 2B 0 0 37 1A 0 0 37 2A 0 0 37 IB 0 0 37 2B 0 0 38 1A 0 0 38 2A 0 0 38 IB 0 0 38 2B 0 0 39 1A 0 0 39 2A 0 0 39 IB 0 0 39 2B 0 0 4 0 0 16la 3 0 0 17la 3 0 0 17la 2 0 0 182a 4 0 0 16lab 3 0 0 17la 3 0 0 17la 2 0 0 l g2ab 4 0 0 16lab 3 0 0 17la 3 0 0 17la 2 0 0 182ab 4 0 1 15 300 17 3 0 0 17la 202 16 4 0 1 15 300 17 3 0 0 17la 202 16 4 0 1 15 300 17 3 0 0 17la 2 02 16 4 0 1 15 3 00 17 3 0 0 17la 2 02 16 4 0 1 15 300 17 3 0 0 17la 2 02 16 Company Sanitized. Does not contain t.sca es H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 13 (Continued) 1.3 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 40 2A 40 IB 40 2B 41 1A 41 2A 41 IB 41 2B 42 1A 42 2A 42 IB 42 2B 43 1A 43 2A 43 IB 43 2B 44 1A 44 2A 44 IB 44 2B 45 1A 45 2A 45 IB 45 2B 46 1A 46 2A 46 IB 46 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 401 15 300 17 3 0 0 17la 202 16 40 1 15 3 0 0 ' 17 3 0 0 17Ia 202 16 4 0 1 15 3 0 0 17la 3 0 0 17la 202 16 4 0 1 15 3 0 0 17la 3 0 0 17la 20 2 16 4 0 1 15 3 0 0 17la 3 0 0 17la 202 16 4 0 1 15 3 0 0 17la 3 0 0 17la 202 16 4 0 1 15id 3 0 0 17la 300 17 202 16 t Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color d Small in size (when compared to appropriate control(s)) CSiBpafjf Sanitized. Does not contain TSCA CBI -74. H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 14 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicatet Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 0 0 - 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0. 0 0 0 0 0 0 0 0 0 0 2 1 0 0 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 20 18 19 20 20 - - - - - Sanitized. Doe no! conia?n t $ga etsi - 75- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 14 (Continued) 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Live Dead Eggs Eggs _ Left Cumulative Dead Larvae Live Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 1 0 0 0 0 0 - - - - - 2 18 1 19 0 20 0 20 2 18 1 19 0 20 0 20 2 18 1 19 0 20 0 20 2 18 1 19 0 20 0 20 2 18 2 18 0 20 0 20 2 18 2 18 0 . 20 1 19 2 18 2 18 1 19 1 19 2 18 2 18 1 19 1 19 ----------. ---------------------- p p m p m y Sanitized Does not contain rftctA H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 14 (Continued) 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B 0 0 0 0 0 0 0 0 2 2 3 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 2 18 2 18 1 19 1 19 2 18 2 18 1 19 1 19 4 16 4 16 4 16 2 18 4 16 5 15 4 16 2 18 4 16 5 15 4 16 2 18 4 16 5 15 4 16 2 18 4 16 5 15 4 16 2 18 4 16 5 15 4 16 2 18 - - Company Sanitized. Doe nnI -77- ""fSOAC81 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 14 (Continued) 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 0 24 2A 0 - 24 IB 0 - 24 2B 0 - 25 1A 0 0 25 2A 0 0 25 IB 0 0 25 2B 0 0 26 1A 0 0 26 2A 0 0 26 IB 0 0 26 2B 0 0 27 1A 1 0 27 2A 0 0 27 IB 0 0 27 2B 0 0 28 1A 0 0 28 2A 0 0 28 IB 0 0 28 2B 0 0 29 1A 2 0 29 2A 0 0 29 IB 0 0 29 2B 0 0 30 1A 0 0 30 2A 0 0 30 IB 0 0 30 2B 0 0 31 1A 0 0 31 2A 0 0 31 IB 0 0 31 2B 0 0 4 16 5 15 4 16 2 18 4 16 0 5 15 0 4 16 0 2 18 0 4 16 0 5 15 0 4 16 0 2 18 0 5 13 0 5 15 0 490 2 16 0 5 10 0 5 13 0 450 2 13 0 740 5 10 0 4 10 250 720 540 400 230 720 530 400 220 - - - 0 0 0 0 0 0 0 0 2 0 7 2 5 2 11 5 9 5 15 13 11 11 16 15 11 12 16 16 Does no? contain TSGACg - 78- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mylass DuPont-11953 TABLE 14 (Continued) 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate1^ Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 32 1A 0 0 32 2A 2 0 32 IB 0 0 32 2B 1 0 33 1A 0 0 33 2A 0 0 33 IB 0 0 33 2B 0 0 34 1A 0 0 34 2A 0 0 34 IB 0 0 34 2B 0 0 35 1A 0 0 35 2A 0 0 35 IB 0 0 35 2B 0 0 36 1A 0 0 36 2A 0 0 36 IB 0 0 36 2B 0 0 37 1A 0 0 37 2A 0 0 37 IB 0 0 37 2B 0 0 38 1A 0 0 38 2A 0 0 38 IB 0 0 38 2B 0 0 39 1A 0 0 39 2A 0 0 39 IB 0 0 39 2B 0 0 710 12 70 0 13 400 16 300 17 70 0 13 700 13 400 16 3 0 0 | fjlbd 70 0 13 7 0 0 13la 400 16 3 0 0 17lbd 70 0 13 7 0 0 13la 400 16 3 0 0 2ylbd 70 0 13 7 0 0 13la 400 16 3 0 0 ^ylbd 70 0 13 7 0 0 13la 400 16 3 0 0 2ylbd 70 0 13 7 0 0 13la 4 0 0 16lb 3 0 0 2ylbd 70 0 13 7 0 0 13la 4 0 0 16lb 3 0 0 2ylbd Company San*etf- o e s not contain TSCA CBi - 79- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 14 (Continued) 2.5 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 0 0 40 2A 0 0 40 IB 0 1 40 2B 0 0 41 1A 0 0 41 2A 0 0 41 IB 0 0 41 2B 0 0 42 1A 0 0 42 2A 0 0 42 IB 0 0 42 2B 0 0 43 1A 0 0 43 2A 0 0 43 IB 0 0 43 2B 0 0 44 1A 0 0 44 2A 0 0 44 IB 0 0 44 2B 0 1 45 1A 0 0 45 2A 0 0 45 IB 0 0 45 2B 0 1 46 1A 0 0 46 2A 0 0 46 IB 0 0 46 2B 0 0 700 13 7 0 0 13la 4 0 1 15 3 0 0 ^ylbd. 700 13 7 0 0 12la* 4 0 1 15 3 0 0 17lbd 700 13 7 0 0 12la 401 15 3 0 0 j ylbd 700 13 7 0 0 12la 4 0 1 15 3 0 0 | y lbd 700 13 7 0 0 12la 4 0 1 15 3 0 1 16 700 13 7 0 0 12la 4 0 1 15 3 0 1 16 700 13 7 0 0 12la 4 0 1 15 3 0 1 16 t Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) b Pale in color d Small in size (when compared to appropriate control(s)) * One larvae missing, possibly escaped from embryo cup and went down standpipe drain. e <pwK*,iirM i. Dossnof contain TSf;. c - 80- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate* Removed Removed Eggs Left Larvae Larvae 0 1A 0 2A 0 IB 0 2B 1 1A 1 2A 1 IB 1 2B 2 1A 2 2A 2 IB 2 2B 3 1A 3 2A 3 IB 3 2B 4 1A 4 2A 4 IB 4 2B 5 1A 5 2A 5 IB 5 2B 6 1A 6 2A 6 IB 6 2B 7 1A 7 2A 7 IB 7 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 - - -81 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 (Continued) 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate* Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 8 1A 8 2A 8 IB 8 2B 9 1A 9 2A 9 IB 9 2B 10 1A 10 2A 10 IB 10 2B 11 1A 11 2A 11 IB 11 2B 12 1A 12 2A 12 IB 12 2B 13 1A 13 2A 13 IB 13 2B 14 1A 14 2A 14 IB 14 2B 15 1A 15 2A 15 IB 15 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 - 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 0 20 1 19 0 20 - - titillili 82 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 (Continued) 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate" Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 16 1A 16 2A 16 IB 16 2B 17 1A 17 2A 17 IB 17 2B 18 1A 18 2A 18 IB 18 2B 19 1A 19 2A 19 IB 19 2B 20 1A 20 2A 20 IB 20 2B 21 1A 21 2A 21 IB 21 2B 22 1A 22 2A 22 IB 22 2B 23 1A 23 2A 23 IB 23 2B 0 0 0 0 0 0 0 0 3 2 1 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 - 0 20 0 20 1 19 0 20 0 20 0 20 1 19 0 20 3 17 2 18 2 18 2 18 3 17 2 18 2 18 2 18 3 17 2 18 2 18 2 18 3 17 2 18 2 18 2 18 3 17 2 18 2 18 2 18 3 17 2 18 2 18 2 18 - - fePiMMf Sanitize.d, Do..e..s..n..o..t..c.o..n..t.a..i.n.--T.S..CH - 83- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 (Continued) 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus myMss, EARLY LIFE-STAGE TOXICITY TEST Dead Eggs Day Replicate1 Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 24 1A 0 24 2A 0 - 24 IB 0 - 24 2B 0 - 25 1A 0 0 25 2A 0 0 25 IB 0 0 25 2B 1 0 26 1A 0 0 26 2A 0 0 26 IB 0 0 26 2B 0 0 27 1A 0 0 27 2A 0 0. 27 IB 0 0 27 2B 0 0 28 1A 0 0 28 2A 0 0 28 IB 0 0 28 2B 0 0 29 1A 0 0 29 2A 0 0 29 IB 0 0 29 2B 0 0 30 1A 0 0 30 2A 0 0 30 IB 0 0 30 2B 1 0 31 1A 0 0 31 2A 0 0 31 IB 0 0 31 2B 0 0 3 17 2 18 2 18 2 18 3 17 0 2 17 0 2 18 0 3 17 0 3 15 0 2 17 0 2 18 0 3 17 0 3 13 0 2 15 0 2 18 0 3 17 0 350 2 12 0 2 15 0 3 13 0 3 10 260 2 10 0 360 300 230 230 450 300 2 10 2 10 4 10 - - - 0 1 0 0 2 1 0 0 4 3 0 0 12 6 3 4 16 12 8 11 17 15 15 11 17 17 17 15 V&mpmy Sanitized. Does not contain TSftft - 84- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 (Continued) 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Dead Dead Cumulative Live Cumulative Eggs Larvae Dead Eggs Dead Live Day Replicate1^ Removed Removed Eggs Left Larvae Larvae 32 1A 0 0 32 2A 0 0 32 IB 0 0 32 2B 0 0 33 1A 0 0 33 2A 1 0 33 IB 0 0 33 2B 0 0 34 1A 0 0 34 2A 0 0 34 IB 0 0 34 2B 0 0 35 1A 0 0 35 2A 0 0 35 IB 0 0 35 2B 0 0 36 1A 0 0 36 2A 0 0 36 IB 0 0 36 2B 0 0 37 1A 0 0 37 2A 0 0 37 IB 0 0 37 2B 0 0 38 1A 0 0 38 2A 0 0 38 IB 0 0 38 2B 0 0 39 1A 0 0 39 2A 0 1 39 IB 0 0 39 2B 0 0 300 17 2 1 0 172a 200 18 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 3 0 0 172a 2 0 0 18la 400 16 300 17 30 1 16la 2 0 0 18la 400 16 Company 'Sanitized. Does not contain TSCA H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 15 (Continued) 5.0 mg/L: EMBRYO SURVIVAL AND HATCHING DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Day Replicate^ Dead Eggs Removed Dead Larvae Removed Cumulative Dead Eggs Live Eggs Left Cumulative Dead Larvae Live Larvae 40 1A 40 2A 40 IB 40 2B 41 1A 41 2A 41 IB 41 2B 42 1A 42 2A 42 IB 42 2B 43 1A 43 2A 43 IB 43 2B 44 1A 44 2A 44 IB 44 2B 45 1A 45 2A 45 IB 45 2B 46 1A 46 2A 46 IB 46 2B 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 300 17 3 0 1 16la 2 0 0 18la 4 00 16 300 17 3 0 1 16la 2 0 0 18la 4 0 0 16 300 17 3 0 1 162a 2 0 0 182a 4 00 16 300 17 3 0 1 162a <3 IN 00 200 400 16 300 17 3 0 1 162a 2 0 0 182a 400 16 300 17 3 0 1 162a 2 0 0 182a 400 16 300 17 3 0 1 162a 2 0 0 182a 4 0 0 16 t Four replicates per concentration (i.e., 1A - 2B). a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) fetttpanjr Sanitized. Does not contain tftn * ^ - 86- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 16 DILUTION WATER CONTROL: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 15 3 40 2A 15 2 40 IB 17 3 40 2B 15 4 41 1A 15 3 41 2A 15 4 41 IB 17 4 41 2B 15 4 42 1A 15 3 42 2A 15 4 42 IB 17 4 42 2B 15 4 43 1A 15 4 43 2A 15 5 43 IB 17 6 43 2B 15 6 44 1A 15 5 44 2A 15 7 44 IB 17 6 44 2B 15 7 45 1A 15 6 45 2A 15 8 45 IB 17 9 45 2B 15 11 46 1A 15 7 46 2A 15 8 46 IB 17 9 46 2B 15 11 gumpiBif Sanitized Does no! contain TSCA CBt - 87- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 17 EPA CONTROL: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 17 40 2A 16 40 IB 17 40 2B 15 41 1A 17 41 2A 16 41 IB 17 41 2B 15 42 1A 17 42 2A 16 42 IB 17 42 2B 15 43 1A 17 43 2A 16 43 IB 17 43 2B 15 44 1A 17 44 2A 16 44 IB 17 44 2B 15 45 1A 17 45 2A 16 45 IB 17 45 2B 15 46 1A 17 46 2A 16 46 IB 17 46 2B 15 0 3 1 4 3 3 3 4 4 6 3 5 4 6 4 5 5 9 5 5 8 9 8 10 8 9 10 10 a B ees 1 contain TSCA CB! H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 18 0.30 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 16 40 2A 17 40 IB 16 40 2B 18 41 1A 16 41 2A 17 41 IB 16 41 2B 18 42 1A 15 42 2A 17 42 IB 16 42 2B 18 43 1A 15 43 2A 17 43 IB 16 43 2B . 18 44 1A 15 44 2A 17 44 IB 16 44 2B 18 45 1A 15 45 2A 16 45 IB 16 45 2B 18 46 1A 15 46 2A 16 46 IB 16 46 2B 18 6 5 3 1 4 4 3 4 5 3 3 4 5 4 3 5 4 3 6 5 8 10 11 11 9 10 11 11 ,Boss m i contain TSCACM - 89- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 19 0.63 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 15 4 40 2A 16 1 40 IB 16 2 40 2B 13 4 41 1A 15 5' 41 2A 16 2 41 IB 16 5 41 2B 13 4 42 1A 15 4 42 2A 16 3 42 IB 16 6 42 2B 13 4 43 1A 15 5 43 2A 16 4 43 IB 16 6 43 2B 13 5 44 1A 15 7 44 2A 16 6 44 IB 16 6 44 2B 13 7 45 1A 15 11 45 2A 16 8 45 IB 16 8 45 2B 13 8 46 1A 15 11 46 2A 16 8 46 IB 16 11 46 2B 13 10 Pompty Sanitized- Do8 contain TSCAC - 90- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 20 1.3 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 15 1 40 2A 17 1 40 IB 17 1 40 2B 16 0 41 1A 15 3 41 2A 17 3 41 IB 17 4 41 2B 16 2 42 1A 15 4 42 2A 17 6 42 IB 17 7 42 2B 16 5 43 1A 15 4 43 2A 17 6 43 IB 17 7 43 2B 16 6 44 1A 15 5 44 2A 17 6 44 IB 17 8 44 2B 16 11 45 1A 15 8 45 2A 17 6 45 IB 17 11 45 2B 16 10 46 1A 15 8 46 2A 17 9 46 IB 17 11 46 2B 16 11 Pem pany B w * - "O' c<"* aI" TSCA CBI -91 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 21 2.5 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 13 40 2A 13 40 IB 15 40 2B 17 41 1A 13 41 2A 12 41 IB 15 41 2B 17 42 1A 13 42 2A 12 42 IB 15 42 2B 17 43 1A 13 43 2A 12 43 IB 15 43 2B 17 44 1A 13 44 2A 12 44 IB 15 44 2B 16 45 1A 13 45 2A 12 45 IB 15 45 2B 16 46 1A 13 46 2A 12 46 IB 15 46 2B 16 0 0 3 0 2 0 3 1 2 1 5 4 4 2 4 6 6 4 5 5 4 8 6 8 6 8 9 9 - 92- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 22 5.0 mg/L: SWIM-UP DATA FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus myJdss, EARLY LIFE-STAGE TOXICITY TEST Total Total Larvae Day Replicate Live Larvae Swimming Up 40 1A 17 40 2A 16 40 IB 18 40 2B 16 41 1A 17 41 2A 16 41 IB 18 41 2B 16 42 1A 17 42 2A 16 42 IB 18 42 2B 16 43 1A 17 43 2A 16 43 IB 18 43 2B 16 44 1A 17 44 2A 16 44 IB 18 44 2B 16 45 1A 17 45 2A 16 45 IB 18 45 2B 16 46 1A 17 46 2A 16 46 IB 18 46 2B 16 1 0 0 1 2 1 0 2 4 1 2 2 6 4 5 5 7 4 7 8 7 4 7 10 8 8 8 10 tfoes Sol contain TSC A CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 23- DILUTION WATER CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 46 B 15 0 47 A 15 0 47 B 15 0 48 A 15 0 48 B 15 0 49 A 15 0 49 B 15 0 50 A 15 0 50 B 15 0 51 A 15 0 51 B 15 0 52 A 15 0 52 B 15 0 53 A 15 0 53 B 15 0 54 A 15 0 54 B 15 0 55 A 15 0 55 B 15 0 56 A 15 0 56 B 15 0 57 A 15 0 57 B 15 0 58 A 15 0 58 B 15 0 59 A 15 0 59 B 15 0 60 A 15 0 60 B 15 0 61 A 15 0 61 B 15 0 62 A 15 0 62 B 15 0 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 0 15 -94- 'g f f lp ig f Rwltteea. Does not contain TSCA Cfl! H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 23 (Continued) DILUTION WATER CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 0 15 63 B 15 0 0 15 64 A 15 0 0 15 64 B 15 0 0 15 65 A 15 0 0 15 65 B 15 0 0 15 66 A 15 0 0 15 66 B 15 0 0 15 67 A 15 0 0 15 67 B 15 0 0 15 68 A 15 0 0 15 68 B 15 0 0 15 69 A 15 0 0 15 69 B 15 0 0 15 70 A 15 0 0 15 70 B 15 0 0 15 71 A 15 0 0 15 71 B 15 0 0 15 72 A 15 0 0 15 72 B 15 0 0 15 73 A 15 0 0 15 73 B 15 0 0 15 74 A 15 0 0 15 74 B 15 0 0 15 75 A 15 0 0 15 75 B 15 0 0 15 76 A 15 0 0 15 76 B 15 0 0 15 77 A 15 0 0 15 77 B 15 0 0 15 78 A 15 0 0 15 78 B 15 0 0 15 Sanitized. Does not contain TSCA CIS - 95- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 23 (Continued) DILUTION WATER CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 79 B 15 80 A 15 80 B 15 81 A 15 81 B 15 82 A 15 82 B 15 83 A 15 83 B 15 84 A 15 84 B 15 85 A 15 85 B 15 86 A 15 86 B 15 87 A 15 87 B 15 88 A 15 88 B 15 89 A 15 89 B 15 90 A 15 90 B 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 8. Boss 1 contain TSCA Cl! -96- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 24 IPA CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 0 15la 46 B 15 0 0 15lff 47 A 15 0 0 15Ia 47 B 15 0 0 15lff 48 A 15 0 0 15la 48 B 15 0 0 15 49 A 15 0 0 15la 49 B 15 0 0 15 50 A 15 0 0 15la 50 B 15 0 0 15 51 A 15 0 0 15la 51 B 15 0 0 15 52 A 15 0 0 15la 52 B 15 0 0 15 53 A 15 0 0 15la 53 B 15 0 0 15 54 A 15 0 0 15la 54 B 15 0 0 15 55 A 15 0 0 15la 55 B 15 0 0 15 56 A 15 0 0 15la 56 B 15 0 0 15 57 A 15 0 0 15la 57 B 15 0 0 15 58 A 15 0 0 15la 58 B 15 0 0 15 59 A 15 0 0 15la 59 B 15 0 0 15 60 A 15 0 0 15la 60 B 15 0 0 15 61 A 15 0 0 15la 61 B 15 0 0 15 62 A 15 0 0 15la 62 B 15 0 0 15 i g p f f f Sanitfe, Does not contain TSCM cm -97- H-25436: Eaxly Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 24 (Continued) IPA CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 0 15la 63 B 15 0 0 15 64 A 15 0 0 15 64 B 15 0 0 15 65 A 15 0 0 15 65 B 15 0 0 15 66 A 15 0 0 15 66 B 15 0 0 15 67 A 15 0 0 15 67 B 15 0 0 15 68 A 15 0 0 15 68 B 15 0 0 15 69 A 15 0 0 15 69 B 15 0 0 15 70 A 15 0 0 15 70 B 15 0 0 15 71 A 15 0 0 15 71 B 15 0 0 15 72 A 15 0 0 15 72 B 15 0 0 15 73 A 15 0 0 15 73 B 15 0 0 15 74 A 15 0 0 15 74 B 15 0 0 15 75 A 15 0 0 15 75 B 15 0 0 15 76 A 15 0 0 15 76 B 15 0 0 15 77 A 15 0 0 15 77 B 15 0 0 15 78 A 15 0 0 15 78 B 15 0 0 15 iilze'ft Does Hofcontain TSCA CBf -98- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 24 (Continued) IPA CONTROL: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 0 79 B 15 0 80 A 15 0 80 B 15 0 81 A 15 0 81 B 15 0 82 A 15 0 82 B 15 0 83 A 15 0 83 B 15 0 84 A 15 0 84 B 15 0 85 A 15 0 85 B 15 0 86 A 15. 0 86 B 15 0 87 A 15 0 87 B 15 0 88 A 15 0 88 B 15 0 89 A 15 0 89 B 15 0 90 A 15 0 90 B 15 0 a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) fif Lying on bottom o f test chamber 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15 %R oes I1 contain TSCA CBI -99- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 25 0.30 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 0 15 46 B 15 0 0 15 47 A 15 0 0 15 47 B 15 0 0 15 48 A 15 0 0 15 48 B 15 0 0 15 49 A 15 0 0 15 49 B 15 0 0 15 50 A 15 0 0 15 50 B 15 0 0 15 51 A 15 0 0 15 51 B 15 0 0 15 52 A 15 0 0 15 52 B 15 0 0 15 53 A 15 0 0 15 53 B 15 0 0 15 54 A 15 0 0 15 54 B 15 0 0 15 55 A 15 0 0 15 55 B 15 0 0 15 56 A 15 0 0 15 56 B 15 0 0 15 57 A 15 0 0 15 57 B 15 0 0 15 58 A 15 0 0 15 58 B 15 0 0 15 59 A 15 0 0 15 59 B 15 0 0 15 60 A 15 0 0 15 60 B 15 0 0 15 61 A 15 0 0 15 61 B 15 0 0 15 62 A 15 0 0 15 62 B 15 0 0 15 te ttfp a f S ittize , D eis m 'Main TS'CAm l - 100- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 25 (Continued) 0.30 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 0 15 63 B 15 0 0 15 64 A 15 0 0 15 64 B 15 0 0 15 65 A 15 0 0 15 65 B 15 0 0 15 66 A 15 0 0 15 66 B 15 0 0 15 67 A 15 0 0 15 67 B 15 0 0 15 68 A 15 0 0 15 68 B 15 0 0 15 69 A 15 0 0 15 69 B 15 0 0 15 70 A 15 0 0 15 70 B 15 0 0 15 71 A 15 0 0 15 71 B 15 0 0 15 72 A 15 0 0 15 72 B 15 0 0 15 73 A 15 0 0 15 73 B 15 0 0 15 74 A 15 0 0 15 74 B 15 0 0 15 75 A 15 0 0 15 75 B 15 0 0 15 76 A 15 0 0 15 76 B 15 0 0 15 77 A 15 0 0 15 77 B 15 0 0 15 78 A 15 0 0 15 78 B 15 1* 1* 14* -101 - D oss nef contain TSCA CM H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 25 (Continued) 0.30 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 79 B 15 80 A 15 80 B 15 81 A 15 81 B 15 82 A 15 82 B 15 83 A 15 83 B 15 84 A 15 84 B 15 85 A 15 85 B 15 86 A 15 86 B 15 87 A 15 87 B 15 88 A 15 88 B 15 89 A 15 89 B 15 90 A 15 90 B 15 One fish missing (escaped) but counted as dead 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 P Js not contain TSCA CBI - 102- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 26 0.63 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 0 15la 46 B 15 0 0 15 47 A 15 0 0 15la 47 B 15 0 0 15 48 A 15 0 0 15la 48 B 15 0 0 15 49 A 15 0 0 15la 49 B 15 0 0 15 50 A 15 0 0 15la 50 B 15 0 0 15 51 A 15 0 0 15la 51 B 15 0 0 15 52 A 15 0 0 15la 52 B 15 0 0 15 53 A 15 0 0 15la 53 B 15 0 0 15 54 A 15 0 0 15la 54 B 15 0 0 15 55 A 15 0 0 15la 55 B 15 0 0 15 56 A 15 0 0 15la 56 B 15 0 0 15 57 A 15 0 0 15la 57 B 15 0 0 15 58 A 15 0 0 15la 58 B 15 0 0 15 59 A 15 0 0 15la 59 B 15 0 0 15 60 A 15 0 0 15la 60 B 15 0 0 15 61 A 15 0 0 15la 61 B 15 0 0 15 62 A 15 0 0 15la 62 B 15 0 0 15 - 103 - .Sanltfxei, Does not contain TSGA CB H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 26 (Continued) 0.63 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 63 B 15 0 64 A 15 0 64 B 15 0 65 A 15 0 65 B 15 0 66 A 15 0 66 B 15 0 67 A 15 0 67 B 15 0 68 A 15 0 68 B 15 0 69 A 15 0 69 B 15 0 70 A 15. 0 70 B 15 0 71 A 15 0 71 B 15 0 72 A 15 0 72 B 15 0 73 A 15 0 73 B 15 0 74 A 15 0 74 B 15 0 75 A 15 1 75 B 15 0 76 A 15 0 76 B 15 0 77 A 15 0 77 B 15 0 78 A 15 0 78 B 15 0 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la,ff 0 15 0 15la,ff 0 15 0 15la,ff 0 15 0 15Ia,ff 0 15 0 l 5 la,ff 0 15 0 15la,ff 0 15 0 151Mr 0 15 0 15la,ff 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 - 104- Dos not contain TSCA C81 H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 26 (Continued) 0.63 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 79 B 15 80 A 15 80 B 15 81 A 15 81 B 15 82 A 15 82 B 15 83 A 15 83 B 15 84 A 15 84 B 15 85 A 15 85 B 15 86 A 15 86 B 15 87 A 15 87 B 15 88 A 15 88 B 15 89 A 15 89 B 15 90 A 15 90 B 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) ff Lying on bottom o f test chamber 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 1 0 2 0 2 0 14 15 14 15 14 15 14 15 14 15 14 15 14 15 14 15 14 15 14 15 13 15 13 15 Ig g p p ip .Sanitized. Does not contain **'* * ***, - 105- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 27 1.3 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 0 15 46 B 15 0 0 15 47 A 15 0 0 15 47 B 15 0 0 15 48 A 15 0 0 15 48 B 15 0 0 15 49 A 15 0 0 15 49 B 15 0 0 15 50 A 15 0 0 15 50 B 15 0 0 15 51 A 15 0 0 15ld 51 B 15 0 0 15 52 A 15 0 0 15ld 52 B 15 0 0 15 53 A 15 0 0 15ld 53 B 15 0 0 15 54 A 15 0 0 15ld 54 B 15 0 0 15 55 A 15 0 0 15ld 55 B 15 0 0 15 56 A 15 0 0 15ld 56 B 15 0 0 15 57 A 15 0 0 15ld 57 B 15 0 0 15 58 A 15 0 0 15ld 58 B 15 0 0 15 59 A 15 0 0 15ld 59 B 15 0 0 15 60 A 15 0 0 15ld 60 B 15 0 0 15 61 A 15 0 0 15ld 61 B 15 0 0 15 62 A 15 0 0 15ld 62 B 15 0 0 15 - 106 - Statfffantr. Doem&t contain TSCA cm H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 27 (Continued) 1.3 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 0 15ld 63 B 15 0 0 15 64 A 15 0 0 j^ld,lz,kk 64 B 15 0 0 15 65 A 15 0 0 j^ld,lz,kk 65 B 15 0 0 15 66 A 15 0 0 66 B 15 0 0 15 67 A 15 0 0 j^ld,lz,kk 67 B 15 0 0 15 68 A 15 0 0 68 B 15 0 0 15 69 A 15 0 0 j^ld,lz,kk 69 B 15 0 0 15 70 A 15 0 0 15ld>lz 70 B 15 0 0 15 71 A 15 0 0 151<Uz 71 B 15 0 0 15 72 A 15 0 0 15lcUz 72 B 15 0 0 15 73 A 15 0 0 15ld,lz 73 B 15 0 0 15 74 A 15 0 0 15ld,lz 74 B 15 0 0 15 75 A 15 0 0 l 5 id,iz 75 B 15 0 0 15 76 A 15 0 0 15lz 76 B 15 0 0 15 77 A 15 0 0 15lz 77 B 15 0 0 15 78 A 15 0 0 15lz 78 B 15 0 0 15 - 107- tew tp lH f SaRRJzetf. Does no! contain H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus myidss DuPont-11953 TABLE 27 (Continued) 1.3 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 79 B 15 80 A 15 80 B 15 81 A 15 81 B 15 82 A 15 82 B 15 83 A 15 83 B 15 84 A 15 84 B 15 85 A 15 85 B 15 86 A 15 86 B 15 87 A 15 87 B 15 88 A 15 88 B 15 89 A 15 89 B 15 90 A 15 90 B 15 d Small in size (when compared to appropriate control(s)) z Deformed gill (right side) kk Swimming at water surface 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 0 15lz 0 15 - 108- m m ol'cehtain TSCA CBI H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 28- 2.5 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 46 B 15 0 47 A 15 0 47 B 15 0 48 A 15 0 48 B 15 0 49 A 15 0 49 B 15 0 50 A 15 0 50 B 15 0 51 A 15 0 51 B 15 0 52 A 15 0 52 B 15 0 53 A .15. 0 53 B 15 . 0 54 A 15 0 54 B 15 0 55 A 15 0 55 B 15 0 56 A 15 0 56 B 15 0 57 A 15 0 57 B 15 0 58 A 15 0 58 B 15 0 59 A 15 0 59 B 15 0 60 A 15 1 60 B 15 0 61 A 15 0 61 B 15 0 62 A 15 0 62 B 15 0 0 15la 0 15 0 15la'lff 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 15la 0 15 0 j^la,laa,ff 0 15 0 J2la,laa,ff 0 15 0 0 15 0 0 15 0 j^la,laa,ff 0 15 0 0 15 0 j ^la,laa,ff 0 15 0 j^la,laa,ff 0 15 1 ^la,aa,ff 0 15 1 j ^l3.,33jfF 0 15 1 0 15 -!9- t h - w t v w u n Ts o t CBi H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 28 (Continued) 2.5 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 1 ^la,aa,ff 63 B 15 64 A 15 0 0 0 15 1 ^la,aa,ff 64 B 15 65 A 15 0 0 0 15 1 ^la,aa,ff 65 B 15 0 0 15 66 A 15 0 1 66 B 15 67 A 15 0 0 0 15 1 ^la,aa,ff 67 B 15 68 A 15 0 0 0 15 1 |^la,aa,ff 68 B 15 0 0 15 69 A 15 0 1 69 B 15 0 0 15 70 A 15 0 1 70 B 15 0 0 15 71 A 15 0 1 ^la,aa,ff 71 B 15 0 0 15 72 A 15 0 1 72 B 15 0 0 15 73 A 15 0 1 j^la,aa,i? 73 B 15 0 0 15 74 A 15 0 1 2^1a,aa,ff 74 B 15 0 0 15 75 A 15 0 1 75 B 15 0 0 15 76 A 15 1 2 13 76 B 15 0 0 15 77 A 15 0 2 13 77 B 15 0 0 15 78 A 15 0 2 13 78 B 15 0 0 15 jSBBBjfttt jjttlHketf, Does not contain TSCA c m - no- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 28 (Continued) 2.5 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 15 79 B 15 80 A 15 80 B 15 81 A 15 81 B 15 82 A 15 82 B 15 83 A 15 83 B 15 84 A 15 84 B 15 85 A 15 85 B 15 86 A 15 86 B 15 87 A 15 87 B 15 88 A 15 88 B 15 89 A 15 89 B 15 90 A 15 90 B 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 a Scoliosis (includes: curved spine, corkscrew tail, and bent spine/tail) aa Rapid respiration ff Lying on bottom o f test chamber 2 0 2 0 2 0 2 0 2 0 2 0 2 0 2 0 2 0 2 0 2 0 2 0 13 15 13 15 13 15 13 15 13 15 13 15 13 15 13 15 13 15 13 15 13 15 13 15 - ill - b e i . D oes Wo!contain TSCA CBf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 29 5.0 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 46 A 15 0 0 15 46 B 15 0 0 15 47 , A 15 0 0 15lff 47 B 15 0 0 15 48 A 15 0 0 15lff 48 B 15 0 0 15 49 A 15 0 0 15lff 49 B 15 0 0 15 50 A 15 0 0 15lff 50 B 15 0 0 15 51 A 15 0 0 15lh,dd 51 B 15 0 0 15 52 A 15 0 0 2 ^1h,aa,ff 52 B 15 0 0 15 53 A 15 0 0 2 ^1h,aa,ff 53 B 15 0 0 15 54 A 15 0 0 j^lh,aa,ff 54 B 15 0 0 15 55 A 15 0 0 55 B 15 0 0 15 56 A 15 0 0 j^lh,aa,ff 56 B 15 0 0 15 57 A 15 0 0 57 B 15 0 0 15 58 A 15 0 0 58 B 15 0 0 15 59 A 15 1 1 14 59 B 15 0 0 15 60 A 15 0 1 14 60 B 15 0 0 15 61 A 15 0 1 14 61 B 15 0 0 15 62 A 15 0 1 14 62 B 15 0 0 15 - 112 - fle e s ! contain TSCA Cis H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 29 (Continued) 5.0 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 63 A 15 0 1 14 63 B 15 0 0 15 64 A 15 0 1 14 64 B 15 0 0 15 65 A 15 0 1 14 65 B 15 0 0 15 66 A 15 0 1 14 66 B 15 0 0 15 67 A 15 0 1 14 67 B 15 0 0 15 68 A 15 0 1 14 68 B 15 0 0 15 69 A 15 0 1 14 69 B 15 0 0 15 70 A 15 0 1 14 70 B 15 0 0 15 71 A 15 0 1 14 71 B 15 0 0 15 72 A 15 0 1 14 72 B 15 0 0 15 73 A 15 0 1 14 73 B 15 0 0 15 74 A 15 0 1 14 74 B 15 0 0 15 75 A 15 0 1 14 75 B 15 0 0 15 76 A 15 0 1 14 76 B 15 0 0 15 77 A 15 0 1 14 77 B 15 0 0 15 78 A 15 0 1 14 78 B 15 0 0 15 - 113 - Siflltfeea lOtfes Wof eftnfafn TSCA C B l H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 29 (Continued) 5.0 mg/L: LARVAL MORTALITY AFTER THINNING FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Larvae Dead Cumulative Live at Larvae Dead Larvae Day Replicate Transfer Removed Larvae Remaining 79 A 79 B 80 A 80 B 81 A 81 B 82 A 82 B 83 A 83 B 84 A 84 B 85 A 85 B 86 A 86 B 87 A 87 B 88 A 88 B 89 A 89 B 90 A 90 B h Yolk sac present (not absorbed) aa Rapid respiration dd Loss o f equilibrium ff Lying on bottom o f test chamber 15 15 15 15 15 15 15 15 15 15 15 15 15 15 15. 15 15 15 15 15 15 15 15 15 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 1 14 0 15 - 114- Does not contain t s c a H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 30 DILUTION WATER CONTROL: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet Weight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B 5.0 4.7 4.2 4.6 4.3 4.8 4.8 4.8 4.8 4.9 4.5 4.9 3.8 4.5 4.7 4.2 4.4 4.1 4.3 4.1 4.5 4.1 4.9 4.5 4.7 4.9 4.7 4.5 4.3 4.8 1.6281 1.2671 0.9640 1.2515 1.0650 1.3542 1.3434 1.4075 1.3933 1.6317 1.2217 1.3862 0.6517 1.1362 1.4809 0.9278 1.1217 0.9130 1.0570 0.8420 1.2287 0.9251 1.5613 1.0699 1.4861 1.7046 1.3841 1.1155 1.0424 1.6013 ilw p ittf S ahlbeC Does not contain TSCA c b ? - 115- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 31 IPA CONTROL: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet Weight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B 4.0 4.1 4.7 4.4 3.9 4.5 4.2 4.3 4.1 4.8 4.8 4.2 4.6 5.0 4.8 4.4 4.7 4.2 4.1 4.0 3.7 3.9 4.3 4.3 4.5 4.6 4.5 4.5 4.7 4.4 0.7917 1.0407 1.3784 1.2848 0.7664 1.1646 1.0193 1.0205 0.8548 1.5978 1.4999 0.9730 1.4529 1.6970 1.4693 1.2643 1.4423 1.0702 0.8778 0.8106 0.5888 0.7406 1.0758 0.9563 1.1270 1.1423 1.1291 1.2498 1.3835 0.9923 fSPilp f Saftftlzecf. Does nof contain TSC.A cflt - 116- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 32 0.30 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B a Fish died prior to test end Standard Length (cm) 4.5 4.6 3.9 4.1 5.0 4.7 4.9 3.8 3.8 5.0 4.2 4.7 4.9 5.2 4.0 4.9 4.1 4.7 4.2 4.1 4.8 4.1 3.9 3.8 4.3 5.0 5.0 4.8 4.0 a Wet W eight (grams) 1.2727 1.2737 0.7769 0.9947 1.7682 1.3858 1.5973 0.6702 0.7024 1.6720 1.1219 1.5625 1.7257 1.7653 0.7628 1.8952 1.0090 1.3474 1.0847 0.9477 1.6443 0.8719 0.7682 0.6331 1.0042 1.6000 1.7710 1.4707 0.8385 a S y p y p y Sanitized, Does no! contain TSCA C81 -117- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss D u P o n t-11953 TABLE 33 0.63 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet Weight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B a Fish died prior to test end 4.1 4.0 4.8 4.8 4.5 4.6 4.7 5.0 4.1 4.0 3.9 4.6 4.2 a a 4.5 4.9 3.7 3.8 5.0 3.4 4.8 4.3 4.7 4.4 4.9 4.7 4.5 4.0 3.9 1.0369 0.9292 1.5758 1.7722 1.1733 1.5429 1.5225 1.7625 0.8883 0.8060 0.8629 1.5206 1.0751 a a 1.2613 1.5593 0.6215 0.7014 1.7190 0.6475 1.4464 1.1015 1.3918 1.0695 1.6121 1.3789 1.3758 0.7730 0.8766 jReinpany Sanitizes Does not contain TSCA CBl - 118- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 34- 1.3 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet W eight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B 5.0 4.9 3.9 4.5 3.5 4.5 4.8 4.8 3.6 4.5 3.6 5.1 4.6 4.5 5.0 3.9 4.6 3.8 3.9 4.3 3.5 4.6 4.7 4.5 3.7 3.6 4.5 4.5 4.9 4.8 1.7466 1.1861 0.6941 1.2027 0.5217 1.2290 1.5022 1.3572 0.5488 1.2535 0.6809 1.7007 1.3366 1.0229 1.6127 0.9927 1.3223 0.7958 1.2228 0.9932 0.4778 1.2850 1.2440 1.3943 0.6192 0.6477 1.3833 1.2095 1.6133 1.5263 -119- oesle t contain TSCA c m H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 35 2.5 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet Weight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B a Fish died prior to test end 4.9 4.1 4.7 3.5 4.5 3.9 4.6 4.3 5.0 4.7 4.4 4.3 3.9 a a 4.9 4.2 3.9 3.6 3.9 3.9 4.1 4.6 5.1 3.5 3.7 4.6 5.0 3.5 5.0 1.6575 0.8647 1.5467 0.4912 1.3753 0.7499 1.2555 0.9677 1.7813 1.2550 1.1513 0.9255 0.8248 a a 1.5774 1.0898 0.7435 0.5565 0.8335 0.6569 0.7471 1.2627 1.9099 0.5225 0.6235 1.3092 0.9864 0.5731 1.6540 f ip p iH f jSatiltizecf. D oes not contain TSCA C8i -120- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 TABLE 36 5.0 mg/L: FISH LENGTHS AND WEIGHTS AT TEST CONCLUSION FOR THE 90-DAY RAINBOW TROUT, Oncorhynchus mykiss, EARLY LIFE-STAGE TOXICITY TEST Number Replicate Standard Length (cm) Wet Weight (grams) 1A 2A 3A 4A 5A 6A 7A 8A 9A 10 A 11 A 12 A 13 A 14 A 15 A 1B 2B 3B 4B 5B 6B 7B 8B 9B 10 B 11 B 12 B 13 B 14 B 15 B Fish died prior to test end 4.7 4.1 4.7 4.6 4.3 4.5 4.5 4.6 4.6 4.7 4.4 4.5 4.0 4.6 a 4.2 4.5 4.1 3.5 4.1 3.7 4.0 3.5 4.7 4.3 4.9 4.2 5.0 3.7 4.5 1.3881 0.8751 1.3667 1.1537 1.0605 1.1634 1.2005 1.2882 1.3625 1.3692 1.3012 1.4060 0.8065 1.3488 a 0.9436 1.2183 1.1129 0.5063 0.9804 0.7074 0.8737 0.5591 1.3672 1.1058 1.4207 1.0398 1.8293 0.5741 1.2472 JSlpRaffif Sanitized. D oes not contain TSCA c w - 121 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 ENDPOINT Egg hatching Larval survival @ thinning Larval abnormality @ thinning Larval survival @ test end Larval abnormality @ test end First day o f hatching Last day o f hatching First day of swimup Length Weight TABLE 37 SUMMARY TABLE OF STATISTICAL ENDPOINTS NOEC (mg/L) >5.0 > 5 .0 >5.0 >5.0 >5.0 >5.0 1.3 1.3 >5.0 >5.0 MATC (mg/L) >5.0 >5.0 >5.0 >5.0 >5.0 >5.0 1.8 1.8 >5.0 >5.0 LOEC (mg/L) >5.0 > 5 .0 >5.0 > 5 .0 >5.0 >5.0 2.5 2.5 >5.0 >5.0 E C 50 (mg/L * * * * * * * * * * STATISTICAL TESTS COMMENT Cochran-Armitage Cochran-Armitage Cochran-Armitage Cochran-Armitage Cochran-Armitage Jonckheere-Terpstra Jonckheere-Terpstra Jonckheere-Terpstra Jonckheere-Terpstra Jonckheere-Terpstra Only one abnormality Exact Analysis Exact Analysis Exact Analysis * Not calculated/could not be adequately determined H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURES - 123 - B ses 1 coirrtaln TSCA CPf H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 1. PHOTOGRAPH OF AN EMBRYO AT APPROXIMATELY 22-HOURS POST-FERTILIZATION - 124- Sanitized. Does no! contain TSCAC H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 2 REPRESENTATIVE ANALYTIC. A: B C ARD CURVE FOR H-25436: Compound namel Coefficient of Detei 399961 Calibration curve: - 1 . 3 5 3 24 e+ 0 0 6 xA2 + 7 . 8 0 66 5 e + 006 x + 3 47 57 . 8 R e s p o n s e type: External Std, Area Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x, Axis trans: None 6665310 Response- T 0.40 Compound name! ____________ Coefficient of Determ nation: K `'2 = u.999'944 Calibration curve: - 8 8 2 4 4 8 * xA2 + 6 . 4 2 4 7 8 e + 0 0 6 * x + 18073.2 R e s p o n s e type: External Std, Area Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x, Axis trans: None 5717656-1 ppm B Response 0.00 0.20 0.40 0.60 0.80 ppm 1.00 CSftpaHf Sanitized. Does not contain TSCA e a t - 125 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynckus mykiss DuPont-11953 FIGURE 2 (Continued) REPRESENTATIVE ANALYTICAL CALIBRATION ST^ Al B: C:i ARD CURVE FOR H-25436: Com pound n a m e ! Coefficient of Detefffl IfiSflfifii = u.yfT9938 Calibration curve: - 2 8 3 1 4 0 * xA2 + 3 . 3 3 6 4 8 e + 0 0 6 * x + -6385.61 Re s po ns e type: External Std, Area Curve type: 2nd Order, Origin: Exclude, Weighting: 1/x, Axis t rans: None 3140718 Response -6386 0.00 1--i--ppm 1.00 ^ ^ ^ ^ Y - S a n U b e d . Does not contain TSCA cbi - 126- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 3 REPRESENTATIVE HPLC CHROMATOGRAM OF A CALIBRATION STANDARD SOLUTION 110703b1 1 0 0 - 1 8251 64.63 3949307 F1:SIR o f5 channels ,ES + TIC 3.961 e+006 % 1 .0 0 2.00 3.0 0 4.00 -j-r - 5.0 0 6.00 lute at a retention time o f approximately 1.97 minutes. Calibration standard solution contains 36 at a concentration of 0.103 mg/L. 110703b19 100- 3.58 672133.1 3 2922611 F2:SIR of5 channels ,ES + TIC 2.948e +006 % m in plute at a retention time o f approximately 3.58 minutes. Calibration standard solution contains H-25436 at a concentration of 0.103 mg/L. SfflpUif Sanitized. Does not contain TSCA l -127- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus myldss DuPont-11953 FIGURE 3 (Continued) REPRESENTATIVE HPLC CHROMATOGRAM OF A CALIBRATION STANDARD SOLUTION lute at a retention time of approximately 5.08 minutes. Calibration standard solution contains H-25436 at a concentration o f 0.103 mg/L. f* i JPpmpany Sanitized. Does not contain TSCA CBI - 128- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 4 REPRESENTATIVE HPLC CHROMATOGRAM OF A DILUTION WATER CONTROL SOLUTION ould elute at a retention time of approximately 3.58 minutes. Company Sanitized. Does not contain TSCA C M - 129- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 4 (Continued) REPRESENTATIVE HPLC CHROMATOGRAM OF A DILUTION WATER CONTROL SOLUTION rould elute at a retention time of approximately 5.08 minutes. <1 ^anjtked. Does not contain TSCAcm H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 5 REPRESENTATIVE HPLC CHROMATOGRAM OF A IPA WATER CONTROL SOLUTION would elute at a retention time o f approximately 1.97 minutes. I ould elute at a retention time of approximately 3.58 minutes. S&lHpsiff Sanitized. Does no! contain tsca cb - 131 - H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 5 (Continued) REPRESENTATIVE HPLC CHROMATOGRAM OF A IPA WATER CONTROL SOLUTION ould elute at a retention time of approximately 5.08 minutes. Cfitopauf Sanitized Does no! contain TSO# " ^ -132- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss______________________________________________________ DuPont-11953 FIGURE 6 REPRESENTATIVE HPLC CHROMATOGRAM OF A H-25436 TEST SOLUTION 1 10703b 100! 1.9.7 12041 90.50 6098132 F1:SIR of5 channels ,ES + TIC 6.1 1 3e + 006 %- 0T d 1.00 2.00 3.00 4.00 5.00 6.00 in L* lute at a retention time of approximately 1.97 minutes. Test solution sample contains H-25436 at a nomini concentration o f 0.3 mg/L. U nominal conce! Jute at a retention time of approximately 3.58 minutes. ration o f 0.3 mg/L. Test solution sample contains H-25436 at a S M nmp Sanilized' 0008 not con,fll,rt T c r * ^ -133- H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 FIGURE 6 (Continued) REPRESENTATIVE HPLC CHROMATOGRAM OF A H-25436 TEST SOLUTION elute at a retention time o f approximately 5.08 minutes. Test solution sample contains H-25436 at a omrnal conce! ration o f 0.3 mg/L. oinpany Sanitized. Does not contai Ten* r H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 APPENDIX felip 'aiif Sanllxetf. Does not r H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont-11953 APPENDIX A Certificate of Analyses 1 :ies not contain r * 11'* 1 -136 - PpMpaitf Sanitized. Does not contain "p* * a H-25436: Early Life-Stage Toxicity to Rainbow Trout, Oncorhynchus mykiss DuPont Specialty Chemicals 1 jg/of Dale Company DuPont-11953 MNMtMiMirttMtftik jmmmUmmiv Wmnn)iMXB( (1M Attention: c ertifica ts o f analysts Product CiMtomar DuPont idm B X PM __^L Total Weight, Lb------------- A aahr Weight. Ib. Drneky 65*C Surbc Tension Appaanmee (2-14-97) 1.312 to 1.WS 23.0 Max. Paste fe a . Lot 6 (MWi.Ml eTM>PSfiy S a n lB a a B . MMaIn T SM CB| -137 - PiM psty Sanftizetf. Dons no! eon m&nm m