Document gDmbyZBYNpQBB9KMg8wbzK63J

CHRONIC TOXICITY TO FISH TEST SUBSTANCE Identity: Perfluorooctanoic acid, ammonium salt; may also be referred to as 78.03, PFOA ammonium salt, Ammonium perfluorooctanoate, PFO, FC-116, FC-126, FC-169, or FC-143. (Octanoic acid, pentadecafluoro-, ammonium salt, CAS # 3825-26-1 ) Remarks: The 3M production lot number was 83. The test sample is FC-143. The testing laboratory refers to it as "78.03". It's purity was not sufficiently characterized, though current information indicates it is a mixture of 96.5 -100% test substance and 0 - 3.5% C, C7, and C9 perfluoro analogue compounds. METHOD: Method Followed: The methodology for the egg and fry exposure closely followed that presented in "Proposed recommended bioassay procedure for egg and fry stages of freshwater fish" (U.S. EPA, 1972). Test Type: Flow-through GLP: No Year study performed: 1978 Species: Fathead minnow (Pimephales promelas) Supplier: U.S. Environmental Protection Agency's Environmental Research Laboratory in Duluth, Minnesota. Test fish age: Eggs within 48-hours after fertilization Analytical monitoring: Temperature, dissolved oxygen concentration, and pH were monitored daily. Weekly samples were taken from each aquarium for determination of ammonium perfluorooctanoate concentration. All samples taken during the test were stored in polyethylene bottles and shipped on May 31, 1978 to the 3M Company. Exposure period: 30 days post-hatch Statistical Methods: Means of measured biological parameters from duplicate aquaria were subjected to analysis of variance (Steel and Torrie, 1960, completely randomized block design, P=0.05). Data for percentage survival and percentage hatch were transformed to arc sin square root of percentage prior to analysis. Test conditions: 003747 Dilution water: Well water pumped to a concrete reservoir where it was aerated before flowing to the exposure system through aged PVC pipe. Dilution water chemistry (0-30 days): Total hardness: 31 - 38 mg/L (as CaC03) Alkalinity: 26 - 32 mg/L (as CaCC>3) pH: 7 .0 -7 .4 Specific conductance: 149-170 pmhos/cm Stock and test solution preparation: A modified, proportional diluter with a 0.50 dilution factor was used. The diluter delivered five nominal concentrations of ammonium perfluorooctanoate ranging from 100 to 6.2 mg/L and control water to duplicate test aquaria. A 4 liter glass mariotte bottle toxicant delivery system was used to deliver 6.6 mL of a nominal ammonium perfluorooctanoate stock concentration of 29.4 mg/mL in distilled water to the mixing chamber of the diluter. Pretreatment: Eggs were placed in a 60 mg/L malachite green solution for 15 seconds to eliminate possible fungus growth. Egg cups: acrylic tubes (3 cm O.D., 7 cm long) with 40 mesh NitexRscreen on one end. An egg cup rocker arm apparatus, as described by Mount (1968), was used to gently oscillate the egg cups in the test water. Fry exposure vessels: Glass test aquarium measuring 30.5 X 30.5 X 30.5 cm with a 17.5 cm high standpipe drain, water volume of 16 liters. Diluter: Delivered 0.50 liters of test water to each aquarium 195 times per day, yielding a 90% test water replacement time of approximately 10 hours. Feeding: fry were fed live brine shrimp nauplii three times daily on weekdays and twice daily on weekends throughout the exposure period. The aquaria were brushed and siphoned to remove excess food and fecal material twice each week. Temperature control: Water bath containing circulating water heated by immersion coil heater and regulated by a mercury column thermoregulator Number of replicates: two Number of eggs for hatchability test: 60 Number of fry for fry exposure test: 40 from each egg cup Number of concentrations: five plus a blank control Water chemistry during the study: Dissolved oxygen: > 95% saturation pH: 7 .0 -7 .3 Temperature: 25 + 1C At the termination of the test, the fry from the control and the high concentration (100 mg/L) were preserved in 10% buffered formalin while the fry from the other test aquaria were frozen. Ten formalin-preserved fry (5 003748 from each replicate) from the control and the high concentration underwent histopathological examination of a transverse section of the nares and cephalic extension of the lateral line (See Table 2). The remaining preserved fry and frozen fry were analyzed at a later date (by 3M Company) for ammonium perfluorooctanoate concentrations. RESULTS___________________________________________________ Nominal concentrations: Bk control, 6.2,12.5, 25, 50, 100 mg/L Element value: Not stated C03749 TABLE 1 PERCENTAGE HATCH OF EGGS, PERCENTAGE SURVIVAL, MEAN AND STANDARD DEVIATION TOTAL LENGTH, AND MEAN WET WEIGHT OF FATHEAD MINNOW (Pimephales promelas) FRY EXPOSED TO AMMONIUM PERFLUOROOCTANOATE FOR 30 DAYS POST HATCH. Nominal concentration (mg/l) Control 6.2 12.5 25 50 100 Replicate A B A B A B A B A B A B Hatch % 98 95 95 94 93 88 98 100 90 95 95 97 30 Days Post-hatch Survival % Mean Mean length in weight in mm mg (+SD) 92 20(2) 62 95 21(3) 75 98 20(2) 59 88 22(2) 79 95 21(3) 70 100 21(2) 72 90 21(2) 74 95 21(2) 70 90 20(2) 60 98 20(3) 65 88 19(2) 59 82 20(2) 60 003750 TABLE 2 RESULTS OF HISTOPATHOLOGICAL EXAMINATION OF PIMEPHALES PROMELAS FRY EXPOSED 30 DAYS TO 100 mg/l OF AMMONIUM PERFLU0R00CTAN0ATE3 Test Material Number of Histopathological Observations Findings Control 10 3/10 Normal 6/10 Liver Fatty Change 3/10 Gill Hyperplasia (Epithelium) I Ammonium 10 5/10 Normal Perfluorooctanoate 5/10 Liver Fatty Change 2/10 Gill Hyperplasia (Epithelium) I a. W ork performed under contract to EG i G Bionomics Laboratory NOTE: "Only those tissues which were missing or contained demonstrable change are listed. The only tissue changes observed were hyperplasia of gill lamellar epithelium and fatty change of the liver. These changes were judged to be minimal and consistent with changes seen routinely in healthy fish. Autolipis of gill tissue was observed in several fish. This change was probably due to the poor penetration of the buffered formalin to the posterior dorsal portion of the gill space." CONCLUSIONS Biological data generated in this study indicate that the nominal concentration of 100 mg/L had no adverse effect upon the hatchability or eggs or upon the survival and growth of fathead minnow fry through 30 days post-hatch. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY Reliability: Klimisch ranking = 2. This study meets all the criteria for quality testing at the time it was conducted, but has several deficiencies. It lacks information on purity of the test substance, and the production lot number from which the test sample was taken. There is no information available on the analysis of the test solution concentrations or on the preserved fry and frozen fry samples. REFERENCES This study was conducted by E G & G, Bionomics, Wareham, Massachusetts, 1978 on the request of 3M Company. 003751 Research Report "THE EFFECTS OF CONTINUOUS AQUEOUS EXPOSURE TO 78.03 (AMMONIUM PERFLUOROOCTANOATE) ON HATCHABILITY OF EGGS AND GROWTH AND SURVIVAL OF FRY OF FATHEAD MINNOW (Pimephales promelas)." Report # BW-78-6-175, E G & G, Bionomics, Aquatic Toxicology Laboratory, 790 Main Street, Wareham, Massachusetts, June 1978. Research Report "SUMMARY OF HISTOPATHOLOGICAL EXAMINATION OF FATHEAD MINNOW (Pimephales promelas) EXPOSED TO 78.03 (AMMONIUM PERFLUOROOCTANOATE) FOR 30 DAYS." Report # BW78-9-301, E G & G, Bionomics, Aquatic Toxicology Laboratory, 790 Main Street, Wareham, Massachusetts, September 1978. OTHER Last changed: 5/25/00 003752