PEER REVIEW DRAFT - DO NOT CITE OR QUOTE United States Office of Chemical Safety and Environmental Protection Agency Pollution Prevention Draft Risk Evaluation for Carbon Tetrachloride Systematic Review Supplemental File: Data Quality Evaluation of Ecological Hazard Studies CASRN: 56-23-5 ci i ci-"crci CI January 2020 ------- 1 1 3 6 9 11 14 17 19 21 23 25 27 Table of Contents Data Type Acute (0-96 hour); Aquatic; Inver- tebrates Chronic (>21 days); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; other Amphibians Chronic (>21 days); Aquatic; Fish Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; other Photobacteriae Acute (0-96 hour); Aquatic; Plants Acute (0-96 hour); Aquatic; Inver- tebrates Other; Aquatic; other Bacteria Reference Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daph- nia magna). Bulletin of Environmental Contamination and Toxicology 24:684- 691 Barrows, M. E.,Petrocelli, S. R.,Macek, K. J.,Carroll, J. J.. 1980. Bioconcen- tration and elimination of selected water pollutants by bluegill sunfish (Lepomis macrochirus). Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of Environmental Con- tamination and Toxicology 26:446-452 Dawson, G. W.,Jennings, A. L.,Drozdowski, D.,Rider, E.. 1977. The acute toxicity of 47 industrial chemicals to fresh and saltwater fishes. Journal of Hazardous Materials 1:303-318 Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo- larval stages of fish and amphibians. 133 Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo- larval stages of fish and amphibians. 133 Lee, S. M.,Lee, S. B.,Park, C. H.,Choi, J.. 2006. Expression of heat shock protein and hemoglobin genes in Chironomus tentans (Diptera, chironomidae) larvae exposed to various environmental pollutants: A potential biomarker of freshwater monitoring. Chemosphere 65:1074-1081 Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 ------- 29 33 35 37 40 42 44 46 48 50 52 54 Acute (0-96 hour); Aquatic; Plants Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Inver- tebrates Other; Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Other; Aquatic; Fish Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Brack, W.,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228 Martins, J.,Soares, M. L.,Saker, M. L.,01ivateles, L.,Vasconcelos, V. M.. 2007. Phototactic behavior in Daphnia magna Straus as an indicator of toxicants in the aquatic environment. Ecotoxicology and Environmental Safety 67:417-422 Martins, J. C.,Saker, M. L.,Teles, L. F.,Vasconcelos, V. M.. 2007. Oxygen consumption by Daphnia magna Straus as a marker of chemical stress in the aquatic environment. Environmental Toxicology and Chemistry 26:1987-1991 Yoshioka, Y.,Ose, Y.,Sato, T.. 1985. Testing for the toxicity of chemicals with Tetrahymena pyriformis. Science of the Total Environment 43:149-157 Bauder, M. B.,Palace, V. P.,Hodson, P. V.. 2005. Is oxidative stress the mech- anism of blue sac disease in retene-exposed trout larvae?. Environmental Toxi- cology and Chemistry 24:694-702 Jia, R.,Cao, L. P.,Du, J. L.,Wang, J. H.,Liu, Y. J.,Jeney, G.,Xu, P.,Yin, G. J.. 2014. Effects of carbon tetrachloride on oxidative stress, inflammatory response and hepatocyte apoptosis in common carp (Cyprinus carpio). Aquatic Toxicology 152 de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Ore- ochromis mossambicus. Science of the Total Environment 214:193-202 de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Ore- ochromis mossambicus. Science of the Total Environment 214:193-202 Khangarot, B. S.,Das, S.. 2009. Acute toxicity of metals and reference toxicants to a freshwater ostracod, Cypris subglobosa Sowerby, 1840 and correlation to EC(50) values of other test models. Journal of Hazardous Materials 172:641-649 Jia, R.,Cao, L.,Du, J.,Xu, P.,Jeney, G.,Yin, G.. 2013. The protective effect of silymarin on the carbon tetrachloride (CC14)-induced liver injury in common carp (Cyprinus carpio). In Vitro Cellular and Developmental Biology 49:155- 161 Y. Liu, L. Cao, J. Du, R. Jia, J. Wang, P. Xu, G. Yin. 2015. Protective ef- fects of Lycium barbarum polysaccharides against carbon tetrachloride-induced hepatotoxicity in precision-cut liver slices in vitro and in vivo in common carp (Cyprinus carpio L.). Comparative Biochemistry and Physiology - Part C: Tox- icology and Pharmacology 169:65-72 Chen, C. Y.,Wooster, G. A.,Bowser, P. R.. 2004. Comparative blood chemistry and histopathology of tilapia infected with Vibrio vulnificus or Streptococcus iniae or exposed to carbon tetrachloride, gentamicin, or copper sulfate. Aqua- culture 239:421-443 ------- 56 58 62 65 68 71 73 75 77 79 81 83 85 87 89 Acute (0-96 hour); Aquatic; other Amphibians Other; Aquatic; Invertebrates Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Plants Other; Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Inver- tebrates Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Fish Acute (0-96 hour); Aquatic; Inver- tebrates Chronic (>21 days); Aquatic; Fish Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction. Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxi- cants Derived by a Closed-System Technique. Environmental Toxicology and Chemistry 26:1931-1939 Schell, J. D. J.. 1987. Interactions of Halogenated Hydrocarbon Mixtures in the Embryo of the Japanese Medaka (Oryzias latipes). Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Com- parisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Com- parisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Geiger, D. L.,Brooke, L. T.,Call, D. J.. 1990. Acute toxicities of organic chem- icals to fathead minnows (Pimephales promelas): Volume V. Weber, L. J.,Gingerich, W. H.,Pfeifer, K. F.. 1979. Alterations in Rainbow Trout Liver Function and Body Fluids Following Treatment with Carbon Tetra- chloride or Monochlorobenzene. 99:401-413 Richie, J. P., Jr.,Mills, B. J.,Lang, C. A.. 1984. The Verification of a Mam- malian Toxicant Classification Using a Mosquito Screening Method. 4:1029- 1035 Koskinen, H.,Pehkonen, P.,Vehniainen, E.,Krasnov, A.,Rexroad, C.,Afanasyev, S.,Molsa, H.,Oikari, A.. 2004. Response of Rainbow Trout Transcriptome to Model Chemical Contaminants. 320:745-753 Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. ------- 3684293 4338225 Chronic (>21 days); Aquatic; In- vertebrates Chronic (>21 days); Aquatic; Fish Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Kotsanis, N.,Metcalfe, C. D.. 1988. Accelerating an in vivo trout carcinogenesis assay with carbon tetrachloride and partial hepatectomy. 15th Annual Aquatic Toxicity Workshop 91 93 < ------- Study Citation: Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daphnia magna). Bulletin of Environmental Contamination and Toxicology 24:684-691 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 7508 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High x 2 2 Metric 2: Test Substance Source Medium X 1 2 Obtained from commercial supplier, but details were omitted. Metric 3: Test Substance Purity Medium X 1 2 Study reports a minimum purity of 80 percent Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 ^ Metric 6: Randomized Allocation High X 1 1 Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion High X 2 2 Metric 8: Consistency of Exposure Administration High X 1 1 Metric 9: Measurement of Test Substance Concentra- tion Medium X 1 2 While CC14 is volatile and the not measured, the researchers did attempt to have a closed system. Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels Medium X 1 2 5-8 test concentrations were reported to be used for each chemical, but the actual values were not avail- able. Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High x 2 2 Metric 14: Acclimitization and Pretreatment Conditions Low x 1 3 Study didn't report whether test organisms were ac- climatized. Continued on next page ------- . continued from previous page Study Citation: Leblanc, G. A.. 1980. Acute toxicity of priority pollutants to water flea (Daphnia magna). Bulletin of Environmental Contamination and Toxicology 24:684-691 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 7508 Domain Metric Rating^ MWF* Score Comments"^ It appears there were 15 daphnia in each test con- centration for CC14 and no replicates to avoid losing CC14 to volatilization. OECD TG 202recommends at least 20 total daphnids and separated into 4 dif- ferent test vessels. Metric 15: Number of Organisms and Replicates per Medium x 1 Group Metric 16: Adequacy of Test Conditions High x 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Metric 23: Explanation of Unexpected Outcomes High Medium High x 1 x 2 x 1 Data for most but not all outcomes by study group were reported but these minor uncertainties or limi- tations are unlikely to have a substantial impact on results. Overall Quality Determination^ High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = J]. (Metric Score; x MWF;) / ^ MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Barrows, M. E.,Petrocelli, S. R.,Macek, K. J.,Carroll, J. J.. 1980. Bioconcentration and elimination of selected water pollutants by bluegill sunfish (Lepomis macrochirus). Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 18050 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High x 2 2 Metric 2: Test Substance Source High x 1 1 Metric 3: Test Substance Purity Low X 1 3 No purity of test chemical was reported, but liquid gas chromatography was performed during the ex- periment and purity of the chemical could be de- termined then, although it wasn't reported in the paper. Domain 2: Test Design Metric 4: Negative Controls High X 2 2 co Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Method for allocation was not reported. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion High X 2 2 Metric 8: Consistency of Exposure Administration High X 1 1 Metric 9: Measurement of Test Substance Concentra- tion High X 1 1 Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels High X 1 1 Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium x 2 4 Minor reservations about the source of fish. Three populations of bluegill sunfish (Lepomis macrochirus) were obtained from a commercial fish farmer in Connecticut, one population obtained from a commercial fish farmer in Nebraska. Age not reported, but length and weight was documented, and age may not be a big factor in determining BCF. Continued on next page ------- . continued from previous page Study Citation: Barrows, M. E.,Petrocelli, S. R. ,Macek, bluegill sunfish (Lepomis macrochirus). Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 18050 K. J.,Carroll, J. J.. 1980. Bioconcentration and elimination of selected water pollutants by Domain Metric Rating^ MWF* Score Comments^ Metric 14: Acclimitization and Pretreatment Conditions High x 1 Metric 15: Number of Organisms and Replicates per Medium x 1 Group Metric 16: Adequacy of Test Conditions Low x 1 Study started with 100 organisms per exposure group, and took out 5 fish on each sampling day. OECD recommends having enough to remove at least 4. Number of replicates not reported. Recommended water temperature for bluegill is 20- 25 degrees C and this study was conducted at 16 degrees C which could have lowered metabolism in fish. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Low Metric 18: Consistency of Outcome Assessment x 2 Medium x 1 BCFs and half-lives were reported, but assessment was not as sensitive as it should be for calculating a BCF. OECD recommends noting if both sexes are used, and ensuring that differences in growth and lipid content between sexes is not significant before the start of the exposure, in particular if it is antic- ipated that pooling of male and female fish will be necessary to ensure detectable substance concentra- tions and/or lipid content. This was not noted. Incomplete reporting of minor details of outcome as- sessment protocol execution Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and Low x 2 Procedures Metric 20: Outcomes Unrelated to Exposure Medium x 1 OECD recommends noting if both sexes are used, and ensuring that differences in growth and lipid content between sexes is not significant before the start of the exposure, in particular if it is anticipated that pooling of male and female fish will be neces- sary to ensure detectable substance concentrations and/or lipid content. This was not noted. Data on attrition and health outcomes unrelated to exposure were not reported for each study group. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data High x 1 Medium x 2 Not all regressions, lipid content, and weights were reported, but BCFs and half-lives were reported for all chemicals. Continued on next page ------- . continued from previous page Study Citation: Barrows, M. E.,Petrocelli, S. R.,Macek, K. J.,Carroll, J. J.. 1980. Bioconcentration and elimination of selected water pollutants by bluegill sunfish (Lepomis macrochirus). Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 18050 Domain Metric Rating^ MWF* Score Comments^ Metric 23: Explanation of Unexpected Outcomes High x 1 Overall Quality Determination"'" High 1.7 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. ( 4 if any metric is Unacceptable Overall rating = I V. (Metric Score,- x MWF,-) I V .. J]. (Metric Score; x MWF;) / J] . MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of Environmental Contamination and Toxicology 26:446-452 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 18064 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Metric 2: Test Substance Identity Test Substance Source Metric 3: Test Substance Purity High x 2 Medium x 1 Medium x 1 Study says all chemicals tested were purchased from commercial chemical suppliers, but does not specify where CC14 came from. Study does state "were pro- cured from those commercial sources able to provide the purest grade available. All chemicals tested were greater than or equal to 80 percent pure..." Study reports a minimum purity of 80 percent for all chemicals tested, but does not specify what the purity is for CC14. Domain 2: Test Design Metric 4: Negative Controls Metric 5: Negative Control Response Metric 6: Randomized Allocation High Low High x 2 x 1 x 1 Many chemicals tested and no details provided about negative control response, although it says control mortality was recorded. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Medium x 2 tion Metric 8: Consistency of Exposure Administration High Metric 9: Measurement of Test Substance Concentra- Low tion Metric 10: Exposure Duration and Frequency High x 1 x 1 x 2 Volatile chemicals were capped, but paper does not specify headspace in the capped jars. The jars capped could have had low DO content, but DO was measured at 0 and at 96 hours.. Nominal concentrations were used and were not measured. CC14 is volatile, but test jars were capped immediately following addition of test chem- ical. Precipitate was observed in test jars indicating test concentrations may have been above water sol- ubility Continued on next page ------- . continued from previous page Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of Environmental Contamination and Toxicology 26:446-452 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 18064 Domain Metric Rating^ MWF* Score Comments^ Metric 11: Number of Exposure Groups/Spacing of Ex- Low posure Levels Metric 12: Testing at or Below Solubility Limit Low x 1 x 1 Study says that the test was conducted according to EPA's "Methods for acute toxicity tests with fish, macroinvertebrates, and amphibians" which says for static tests you must have 10 organisms in each treatment divided into at least two test chambers; not sure how they got the exposure concentrations used of what the exposure concentrations were. Test substance concentration was not reported. Pa- per states " The acute toxicity of most of the chem- icals tested was at concentrations above their water solubility and therefore, the test material or one or more of its constituents precipitated ..." Precipitate was observed for CC14 Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium x 2 Metric 14: Acclimitization and Pretreatment Conditions Medium x 1 Metric 15: Number of Organisms and Replicates per Medium x 1 2 Group Metric 16: Adequacy of Test Conditions Low x 1 3 Test animals utilized were young of the year bluegill (L. macrochirus) obtained from commercial fish sup- pliers within the continental United States. Ag e and weight reported, sex not reported Acclimation period not stated, but does state tests followed "Methods for acute toxicity tests with fish, macroinvertebrates and amphibians" which specifies a 14 day acclimation period for fish. Study does report a 48 hour time prior to test where fish were not fed and observed; fish were not used if had >3 percent mortality, Number of fish per test jar reported, but number of replicates not reported Minor uncertainties around housing conditions (headspace in jar) DO cones for all chemicals ranged from 9.7 mg/L at start of test to 0.3 mg/L at 96 hours. Low DO can impact survival; DO at end of test for CCL4 not reported. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High High x 2 x 1 Domain 6: Confounding / Variable Control Continued on next page ------- . continued from previous page Study Citation: Buccafusco, R. J.,Ells, S. J.,Leblanc, G. A.. 1981. Acute toxicity of priority pollutants to bluegill (Lepomis macrochirus). Bulletin of Environmental Contamination and Toxicology 26:446-452 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 18064 Domain Metric Rating^ MWF* Score Comments^ Metric 19: Confounding Variables in Test Design and Low X 2 6 Study did not provide enough information to allow Procedures a comparison of environmental conditions Metric 20: Outcomes Unrelated to Exposure Low X 1 3 Do not provide information about health outcomes of each study group Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Medium X 1 2 Not clear what method was used to calculate LC50 for CC14: "The LC50s and 95 percent confidence in- tervals were calculated, where possible, by the mov- ing average angle method (HARRIS 1959). The nominal test concentrations were transformed to log- arithms and corresponding percentage mortalities to angles. Each group of these successive angles was then averaged and the LCSO was estimated by lin- ear interpolation, between the successive concentra- tions whole average angles bracketed 45" . When the test data did not meet Harris' method requirements, the LC50s were calculated by the log probit method, a modification of the LITCHFIELD + WILCOXON (1949) method." Metric 22: Reporting of Data Low X 2 6 The data for the static test were not presented in full, and no information was reported for controls. Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination"'" Medium 2.0 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Dawson, G. W.,Jennings, A. L.,Drozdowski, D.,Rider, E.. 1977. The acute toxicity of 47 industrial chemicals to fresh and saltwater fishes. Journal of Hazardous Materials 1:303-318 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 18670 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Metric 2: Test Substance Identity Test Substance Source Metric 3: Test Substance Purity Low X 2 6 Analytical confirmation of CC14 was not reported. Low X 1 3 CC14 was either research or chemically pure grade quality from commercial sources. Low X 1 3 Purity was not reported. Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation High X 1 1 Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion High X 2 2 Metric 8: Consistency of Exposure Administration High X 1 1 Metric 9: Measurement of Test Substance Concentra- tion Low X 1 3 Did not report whether or not CC14 was measured. Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels High X 1 1 Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13 Metric 14 Metric 15 Metric 16: Test Organism Characteristics Acclimitization and Pretreatment Conditions Number of Organisms and Replicates per Group Adequacy of Test Conditions High X 2 2 High X 1 1 Low X 1 3 The number of organisms/replicates was not re- ported. Medium X 1 2 Minor uncertainties and will not have substantial impact on the results. Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Dawson, G. W.,Jennings, A. L.,Drozdowski, D.,Rider, E.. 1977. The acute toxicity of 47 industrial chemicals to fresh and saltwater fishes. Journal of Hazardous Materials 1:303-318 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 18670 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data Low x 2 6 Metric 23: Explanation of Unexpected Outcomes Low x 1 3 Data for exposure-related findings were not shown for each study group. The study did not report any measures of variability and/or insufficient information was provided. Overall Quality Determination"'" High ¥ Medium 1.6 Downgrade from high to medium: The purity of CC14 and number of replicates is absent from the paper. It is also unclear if the researchers analyti- cally quantified CC14. Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Acute (0-96 hour); Aquatic; other Amphibians Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 The test substance was identified as carbon tetra- chloride. Metric 2: Test Substance Source Low X 1 3 The toxicant source was not identified in the publi- cation. Metric 3: Test Substance Purity High X 1 1 All test substances used in the toxicity tests were reagent grade quality. Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Amphibian controls were used in the study. Metric 5: Negative Control Response High X 1 1 The control survival ranged from 84-99 percent. Metric 6: Randomized Allocation Low X 1 3 There was no mention of randomized allocation of test organisms. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High X 2 2 Flow-through testing with a closed vessel was devoid tion of air space to minimize volatilization. Metric 8: Consistency of Exposure Administration High X 1 1 The researchers administrated the test solutions (ex- posure scenario) consistently across the toxicity test. Metric 9: Measurement of Test Substance Concentra- High X 1 1 Gas-liquid chromatography was used to measure test tion concentrations daily. Metric 10: Exposure Duration and Frequency High X 2 2 Amphibian embryo-larvae were exposed up to 4 days post-hatch, sufficient to determine effects in embryos and larvae. Metric 11 Number of Exposure Groups/Spacing of Ex- High X 1 1 There were 6 exposure concentrations with appro- posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 priate spacing used fore each amphibian tested. All exposure concentrations were below the water solubility of carbon tetrachloride. Domain 4: Test Organism Metric 13: Test Organism Characteristics High x 2 2 Amphibians used were appropriate for this study, with the exception of the African Clawed frog, which is not endemic to the U.S. Continued on next page ------- . continued from previous page Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Acute (0-96 hour); Aquatic; other Amphibians Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments^ Metric 14: Acclimitization and Pretreatment Conditions Medium x 1 Metric 15: Number of Organisms and Replicates per High x 1 Group Metric 16: Adequacy of Test Conditions Medium x 1 2 Controls and exposed organisms were appeared to be treated identical with the exception of CC14 in the controls. After re-reading, I did not see any ac- climatization and pretreatment conditions reported, but if there were adverse effects from this, it would have shown up in the controls and it did not. 1 Single replicates of 50 to 125 eggs were used per test concentration. 2 A loading rate of up to 125 eggs per test concen- tration was used, which did not appear to impact test results. Environmental conditions were within acceptable ranges, and control mortality was accept- able. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High X 2 2 Test vessels observed daily to assess development and remove dead test organisms. High X 1 1 LC50, LC10, LCls were assessed adjusted for control mortality, but detailed control mortality data were not provided. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 Procedures Metric 20: Outcomes Unrelated to Exposure Medium x 1 Environmental conditions appeared consistent across test concentrations and control mortality ranged from 1-16 percent. Teratogenesis was reportedly infrequently in the controls ( percent teratogenicity not reported) and control mortality ranged from 1 to 16 percent, which is acceptable. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Medium x 1 Medium x 2 Survival data was reported as percent of total organ- isms at each exposure concentration after corrected for control mortality, but detailed control data were not reported. LC50s, LClOs, and LCls were calcu- lated using log-probit analysis. Most, but not all, data endpoints were reported. You could not re-create the statistics in the paper. Continued on next page ------- . continued from previous page Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Acute (0-96 hour); Aquatic; other Amphibians Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments"^ Metric 23: Explanation of Unexpected Outcomes High X 1 1 Unexpected outcomes were not reported in the study. Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 The test substance was identified as carbon tetra- chloride. Metric 2: Test Substance Source Low X 1 3 The toxicant source was not identified in the publi- cation. Metric 3: Test Substance Purity High X 1 1 All test substances used in the toxicity tests were reagent grade quality. Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Fish control eggs were used in the study. Metric 5: Negative Control Response High X 1 1 The control survival ranged from 84-99 percent. Metric 6: Randomized Allocation Low X 1 3 There was no mention of randomized allocation of test organisms. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion High X 2 2 Flow-through testing with closed vessel devoid of air space was used to minimize volatilization. Metric 8: Consistency of Exposure Administration High X 1 1 The researchers administrated the test solutions (ex- posure scenario) consistently across the toxicity test. Metric 9: Measurement of Test Substance Concentra- tion High X 1 1 Gas-liquid chromatography was used to measure test concentrations daily. Metric 10: Exposure Duration and Frequency High X 2 2 Fish embryo-larvae were exposed up to 4 days post- hatch , sufficient to determine effects in embryos and larvae. Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels High X 1 1 There were 6 exposure concentrations with appro- priate spacing used for each fish tested. Metric 12: Testing at or Below Solubility Limit High X 1 1 All exposure concentrations were below the water solubility of carbon tetrachloride. Domain 4: Test Organism Continued on next page ------- . . . continued from previous page Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments^ Metric 13: Test Organism Characteristics High x 2 Metric 14: Acclimitization and Pretreatment Conditions Medium x 1 Metric 15: Number of Organisms and Replicates per High x 1 Group Metric 16: Adequacy of Test Conditions Medium x 1 Rainbow trout and fathead minnow are well known species. The trout were obtained from a hatchery and freshly fertilized fathead minnow eggs were ob- tained from the EPA Newtown Fish Toxicology Lab- oratory. Controls and exposed organisms were appeared to be treated identical with the exception of CC14 in the controls. I did not see any acclimatization and pretreatment conditions reported, but if there were adverse effects from this, it would have shown up in the controls and it did not. Single replicates of 50 to 125 eggs were used per test concentration. A loading rate of up to 125 eggs per test concen- tration was used, which did not appear to impact test results. Environmental conditions were within acceptable ranges, and control mortality was accept- able. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Test vessels observed daily to assess development and remove dead test organisms. Metric 18: Consistency of Outcome Assessment High x 1 1 LC50, LC10, LCls were assessed adjusted for control mortality, but detailed control mortality data were not provided. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 Procedures Metric 20: Outcomes Unrelated to Exposure Medium x 1 Environmental conditions appeared consistent across test concentrations and control mortality ranged from 1-16 percent. Teratogenesis was reportedly infrequent in controls ( percent teratogenicity not reported) and control mortality ranged from 1 to 16 percent, which is ac- ceptable. Domain 7: Data Presentation and Analysis Continued on next page ------- . continued from previous page Study Citation: Black, J. A.,Birge, W. J.,McDonnell, W. E.,Westerman, A. G.,Ramey, B. A.,Bruser, D. M.. 1982. The aquatic toxicity of organic compounds to embryo-larval stages of fish and amphibians. 133 Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 93660 Domain Metric Rating^ MWF* Score Comments^ Metric 21: Statistical Methods High x 1 Metric 22: Reporting of Data Medium x 2 Metric 23: Explanation of Unexpected Outcomes High x 1 Survival data was reported as percent of total organ- isms at each exposure concentration after corrected for control mortality, but detailed control data were not reported. LC50s, LClOs, and LCls were calcu- lated using log-probit analysis. Most, but not all, data endpoints were reported. You could not re-create the statistics in the paper. Unexpected outcomes were not reported in the study. Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Lee, S. M.,Lee, S. B.,Park, C. H.,Choi, J.. 2006. Expression of heat shock protein and hemoglobin genes in Chironomus tentans (Diptera, chironomidae) larvae exposed to various environmental pollutants: A potential biomarker of freshwater monitoring. Chemosphere 65:1074-1081 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 492760 Domain Metric Rating^ MWF* Score Comments"^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High x 2 2 tion Metric 8: Consistency of Exposure Administration High x 1 1 Metric 9: Measurement of Test Substance Concentra- Low x 1 3 tion Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 NOMINAL 24 HR EXP Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Lee, S. M.,Lee, S. B.,Park, C. H.,Choi, J.. 2006. Expression of heat shock protein and hemoglobin genes in Chironomus tentans (Diptera, chironomidae) larvae exposed to various environmental pollutants: A potential biomarker of freshwater monitoring. Chemosphere 65:1074-1081 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 492760 Domain Metric Rating^ MWF* Score Comments"^ Metric 17: Outcome Assessment Methodology High X 2 2 Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. ^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; X MWF;) / J] . MWF, if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Source/Information not reported Metric 3: Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion Metric 8: Consistency of Exposure Administration Metric 9: Measurement of Test Substance Concentra- tion Metric 10: Exposure Duration and Frequency Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels Metric 12: Testing at or Below Solubility Limit High High Medium High Medium x 2 x 1 x 1 x 2 x 1 Medium x 1 Concentrations were measured using gas chromatog- raphy, but concentrations were not reported in the paper Number of exposure groups and spacing of exposure levels not reported, though followed OECD guideline 203 Solvent concentrations were not discussed; closed containers to minimize volatility used Domain 4: Test Organism Metric 13 Metric 14 Metric 15 Test Organism Characteristics Acclimitization and Pretreatment Conditions Number of Organisms and Replicates per Group Metric 16: Adequacy of Test Conditions Medium High High High x 2 x 1 x 1 x 1 Source of fish not reported Continued on next page ------- . continued from previous page Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High High x 2 x 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Metric 23: Explanation of Unexpected Outcomes Medium x 1 Medium x 2 High x 1 No details on statistical methods were reported. Just reported 48-hr LC50 as mortality ( percent) vs concentration Reported 48 hr LC50, but no additional details in- cluded Overall Quality Determination^ High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; other Photobacteriae Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Source/Information not reported Metric 3: Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4: Metric 5: Metric 6: Negative Controls Negative Control Response Randomized Allocation Low Low Low X 2 6 Used Microtox test, which includes negative con- trols, but controls were not described X 1 3 Negative control response not described X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Medium x 2 tion Metric 8: Consistency of Exposure Administration Metric 9: Measurement of Test Substance Concentra- tion Metric 10: Exposure Duration and Frequency Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels Metric 12: Testing at or Below Solubility Limit Medium x 1 High Medium High Medium x 1 x 1 x 2 x 1 Experimental system and test media were described, but not in great detail. Cite "Microtox test" and German standard DIN 38412 L 34. Concentrations were measured using gas chromatog- raphy, but concentrations were not reported in the paper Number of exposure groups and spacing of exposure levels not reported, though EC50 was reported Solvent concentrations were not discussed Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium X 2 4 Source of organisms not reported Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per Medium X 1 2 Replicates were not discussed Group Metric 16: Adequacy of Test Conditions High X 1 1 Continued on next page ------- . continued from previous page Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; other Photobacteriae Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Medium X 1 2 No details on statistical methods were reported Metric 22: Reporting of Data Medium X 2 4 Reported EC50, but no additional details included Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ Medium 1.8 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Source/Information not reported Metric 3: Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion Metric 8: Consistency of Exposure Administration Metric 9: Measurement of Test Substance Concentra- tion Metric 10: Exposure Duration and Frequency Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels Metric 12: Testing at or Below Solubility Limit High High Medium High Medium x 2 x 1 x 1 x 2 x 1 Medium x 1 Concentrations were measured using gas chromatog- raphy, but concentrations were not reported in the paper Number of exposure groups and spacing of exposure levels not reported, though followed modified OECD guideline 201 Solvent concentrations were not discussed; used modified test containers to minimize volatility with- out causing growth inhabitation or death merely due to closed containers Domain 4: Test Organism Metric 13 Metric 14 Metric 15 Test Organism Characteristics Acclimitization and Pretreatment Conditions Number of Organisms and Replicates per Group Medium x 2 High x 1 High x 1 Metric 16: Adequacy of Test Conditions High x 1 Source of algae not reported Continued on next page ------- . continued from previous page Study Citation: Ereitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Metric 23: Explanation of Unexpected Outcomes Medium x 1 Medium x 2 High x 1 No details on statistical methods were reported. Just reported EC50/72 hours as percentage of growth inhibition versus concentration Reported EC50/72hrs, but no additional details in- cluded Overall Quality Determination"'" High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/Information not reported Metric 3 Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- Medium X 2 4 Specific methodology not reported in paper, cites tion OECD guidelines Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Medium X 1 2 Concentrations were measured using gas chromatog- tion raphy, but concentrations were not reported in the paper Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 Number of exposure groups and exposure levels not posure Levels reported, though EC50 was reported Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium X 2 4 Source of organisms not reported Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Freitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Other; Aquatic; other Bacteria Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 chemical name and structure Metric 2 Test Substance Source Low X 1 3 Source/Information not reported Metric 3 Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response Low X 1 3 Negative control response not described Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 Used an automatic test apparatus (Sapromat) tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Medium X 1 2 Concentrations were measured using gas chromatog- tion raphy, but concentrations were not reported in the paper Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Medium X 1 2 Number of exposure groups and exposure levels not posure Levels reported, though EC50 was reported Metric 12: Testing at or Below Solubility Limit Medium X 1 2 Solvent concentrations were not discussed Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium X 2 4 Source of organisms not reported Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per Medium X 1 2 Replicates were not discussed Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Ereitag, D.,Ballhorn, L.,Behechti, A.,Fischer, K.,Thumm, W.. 1994. Structural configuration and toxicity of chlorinated alkanes. Chemosphere 28:253-259 Data Type: Other; Aquatic; other Bacteria Hero ID: 660810 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High x 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Medium x 1 Medium x 2 Metric 23: Explanation of Unexpected Outcomes High x 1 2 No details on statistical methods were reported 4 Reported EC50/5 days, but no additional details in- cluded Overall Quality Determination"'" High 1.6 Extracted Yes * MWF = Metric Weighting Factor I High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. ^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; X MWF;) / J] . MWF, if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. U Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Brack, W. ,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 661061 Domain Metric Rating^ MWF* Score Comments"^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Test substance was identified by name. Metric 2: Test Substance Source High X 1 1 Authors identified Merck as the source of the test substance. Metric 3: Test Substance Purity Low X 1 3 "p.a." is reported for CC14, which is analytical grade quality. Domain 2: Test Design Metric 4: Negative Controls Metric 5: Negative Control Response Metric 6: Randomized Allocation High Low Low x 2 x 1 x 1 "Each test series contained three controls without toxicant and two controls with 0.8 mg/L Cu2+ (CuS04). This concentration reduces algal growth to50 percent and is used to check normal sensitivity of the organisms." The biological responses of the negative control groups were not reported It was not reported whether there was random place- ment of flasks. Domain 3: Exposure Characterization Continued on next page ------- . continued from previous page Study Citation: Brack, W. ,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 661061 Domain Metric Ratingt MWF* Score Commentstt Metric 7: Experimental System/Test Media Prepara- High tion x 2 Metric 8: Consistency of Exposure Administration High Metric 9: Measurement of Test Substance Concentra- High tion Metric 10: Exposure Duration and Frequency High Metric 11: Number of Exposure Groups/Spacing of Ex- High posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 x 1 x 2 x 1 x 1 The purpose of the test was to determine a way of doing algae tests with volatile chemicals, as the OECD guidelines recommends using a permeable stopper in the flask to allow C02 to pass through so as not to impede algae growth. However with volatile chemicals this is not possible because of loss of test substance through vitalization. Therefore in test, they used a closed system that still provided a source of C02 for the algae. Authors reported, "Deviations between the duplicates, extracted from the same test culture were less than 5 percent . To estimate recovery of this analytical method, 20 mL headspace vials were filled completely with water or alga suspension. The vials were sealed gas" tight with septa. Gravimetrically defined amounts of the volatile chlorinated hydrocarbons were injected via syringe through the Septa into the liquids and dis- solved. From these solutions samples were taken and extracted as explained above. Recovery of the method amounted to 90 " S percent and was inde- pendent from cell density." Exposures were administered consistently across study groups. Analytical measurments by gas chromatography/ electron capture detector (GC/ECD) following liquid-liquid microextraction were taken at test ini- tiation and end. The test was 72 hours in duration, which is recom- mended by OECD Guideline 201. Test concentrations are reported in figure 3 and show a dose response for growth inhibition. The figure shows at least 5 concentrations tested which is rec- ommended by OECD Guideline 201. The test cone for CC14 shown in figure 3 (highest cone is <10 mg/1) are well below CC14's solubility level of 793 mg/1. Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium x 2 This is not a commonly used algal species. Not a TG species. Continued on next page ------- . . . continued from previous page Study Citation: Brack, W. ,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 661061 Domain Metric Ratingt MWF* Score Commentstt Metric 14: Acclimitization and Pretreatment Conditions High x 1 Metric 15: Number of Organisms and Replicates per Medium x 1 2 Group Metric 16: Adequacy of Test Conditions High x 1 1 Pretreatment conditions included, "Precultures and test cultures were grown in the medium for unicel- lular algae according to KUflL (1962) (Table 2). In- cubation of all cultures was done in a Orbital In- cubator (Gallenkamp). The cultures were shaken permanently with a frequency of 120 rpm. They wereilluminated from above with 130 "E/m2s with- out light dark cycle. The photosynthetically effec- tive light was determined with a Quantum Sensor from Licor Inc. The temperature was maintained at 20 " 1 deg C." Two replicates per test concentration (8 concentra- tions). Three replicates are preferred. Glass flasks which are recommended in OECD 201. Temp and pH were within recommended ranges. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High x 2 High x 1 Biomass assessed using fluorometric measurement of total chlorophyll for controls and treatment groups to determined EClOs and EC50s. No inconsistencies were reported, and both positive and negative controls performed as expected. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 There were no reported differences among study groups in environmental conditions or other factors that would influence the outcome assessment. Positive and negative controls performed as ex- pected and no outcomes unrelated to exposures were reported. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data High x 1 Medium x 2 1 Probit analysis was used to assess significant differ- ences in biomass. 4 Figure 3 shows the results of the tests at each cone for each chemical but it's difficult to determine the exact concentrations from the figure, so some minor uncertainties remain. Continued on next page ------- . continued from previous page Study Citation: Brack, W. ,Rottler, H.. 1994. Toxicity testing of highly volatile chemicals with green algae: A new assay. 1:223-228 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 661061 Domain Metric Ratingt MWF* Score Commentstt Metric 23: Explanation of Unexpected Outcomes Medium x 1 2 SDs were provided, but it was unclear whether or not there were any unexpected outcomes. Overall Quality Determination^ High 1.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = J]. (Metric Score; x MWF;) / J] . MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Martins, J.,Soares, M. L.,Saker, M. L.,01ivateles, L.,Vasconcelos, V. M.. 2007. Phototactic behavior in Daphnia magna Straus as an indicator of toxicants in the aquatic environment. Ecotoxicology and Environmental Safety 67:417-422 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 661491 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High x 2 2 Metric 2 Test Substance Source High x 1 1 Metric 3 Test Substance Purity High x 1 1 Domain 2: Test Design Metric 4 Negative Controls High x 2 2 Metric 5 Negative Control Response High x 1 1 Metric 6 Randomized Allocation Low x 1 3 Did not report randomization. Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High x 2 2 tion Metric 8 Consistency of Exposure Administration High x 1 1 Metric 9 Measurement of Test Substance Concentra- Medium x 1 2 It is not clear, but it appears that nominal concen- tion trations were used in the study. Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High x 2 2 Metric 14: Acclimitization and Pretreatment Conditions High x 1 1 Metric 15: Number of Organisms and Replicates per High x 1 1 Group Metric 16: Adequacy of Test Conditions High x 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Continued on next page ------- . continued from previous page Study Citation: Martins, J.,Soares, M. L.,Saker, M. L.,01ivateles, L.,Vasconcelos, V. M.. 2007. Phototactic behavior in Daphnia magna Straus as an indicator of toxicants in the aquatic environment. Ecotoxicology and Environmental Safety 67:417-422 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 661491 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Martins, J. C.,Saker, M. L.,Teles, L. F.,Vasconcelos, V. M.. 2007. Oxygen consumption by Daphnia magna Straus as a marker of chemical stress in the aquatic environment. Environmental Toxicology and Chemistry 26:1987-1991 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 661492 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Randomization was not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High tion Metric 8: Consistency of Exposure Administration High Metric 9: Measurement of Test Substance Concentra- Medium tion Metric 10: Exposure Duration and Frequency High Metric 11: Number of Exposure Groups/Spacing of Ex- N/A posure Levels Metric 12: Testing at or Below Solubility Limit High x 2 x 1 x 1 x 2 x 1 It is unclear if the test concentration was measured. N/A Only one concentration was reported and is accept- able for this type of test. Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions Medium X 1 2 Metric 15: Number of Organisms and Replicates per High X 1 1 It was not clear, but was described in another paper on CC14 from the same laboratory/test group. Group Metric 16: Adequacy of Test Conditions High x 1 Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Martins, J. C.,Saker, M. L.,Teles, L. F.,Vasconcelos, V. M.. 2007. Oxygen consumption by Daphnia magna Straus as a marker of chemical stress in the aquatic environment. Environmental Toxicology and Chemistry 26:1987-1991 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 661492 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data High x 2 2 Metric 23: Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.2 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1985. Testing for the toxicity of chemicals with Tetrahymena pyriformis. Science of the Total Environ- ment 43:149-157 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 676758 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Source of test chemicals not reported Metric 3: Test Substance Purity Medium X 1 2 Purity not reported; study states "all other reagents were of analytical grade" Domain 2: Test Design Metric 4: Negative Controls Low x 2 6 Metric 5: Negative Control Response N/A N/A Metric 6: Randomized Allocation Low x 1 3 The study states "The relative growth rate was cal- culated as the ratio of the number of cells cultured with a chemical against the number cultivated in a blank", which implies the blank is a control but this is not stated. Very little information is presented about what is in the blank. This is an acute study with lots of chemicals re- ported, and they did not report on the control re- sponse for each chemical. No mention of random allocation Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Medium x 2 4 The methods section does not state test chambers ¦j-Jq-q were closed for CC14, but page 155 states "The au- thors adopted 24 h for the test time and the con- ditions of No. 4 for culturing. The EC50 values of 57 chemicals were determined by themethod and are shown in Table 1." Test condition 4 on Figure 2 indicates "cultured in vertical vessel with a silicone rubber stopper"The study also states "the air space of 20 ml in the test tube is sufficient to determine the EC50 value of a chemical for a short cultivation period; volatile chemicals can therefore be tested in the sealed vessel." Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1985. Testing for the toxicity of chemicals with Tetrahymena pyriformis. Science of the Total Environ- ment 43:149-157 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 676758 Domain Metric Rating^ MWF* Score Comments^ Metric 8: Consistency of Exposure Administration Low Metric 12: Testing at or Below Solubility Limit High x 1 Metric 9: Measurement of Test Substance Concentra- Low x 1 tion Metric 10: Exposure Duration and Frequency High x 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Unacceptable x 1 posure Levels x 1 There were differences in how exposure was adminis- tered but because the point of the study was to figure out what housing conditions were best for this type of protozoa. These differences could have effected the EC50 reported. Authors report that some of the temperatures, and amount of food changed the growth rate of the protozoa. Study does not state whether exposure concentra- tions are nominal or measured No information was provided on number of expo- sure groups or spacing of exposures for CC14. Figure 2 shows five exposure concentrations used to deter- mine the EC50 value for aniline. Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium Metric 14: Acclimitization and Pretreatment Conditions High Metric 15: Number of Organisms and Replicates per Low Group Metric 16: Adequacy of Test Conditions High x 2 x 1 x 1 x 1 Tetrahymena pyriformis was preserved in a sterile medium of 2 percent proteasepeptone at 20" C which was renewed at 2-4 week intervals. Unsure but it sounds like they cultured their own animals in the lab from descriptions of previous studies in this pa- per. Acknowledgements say "Pr. Nozawa of Gifu University for providing T. pyriformis in germ-free condition" Number of test organisms and replicates were not reported for the test groups. Each test solution was inoculated with 0.2 ml of pre-cultures T. pyriformis, but pre-exposure numbers in that 0.2 ml were not counted. Number of replicates not stated.It was re- ported that 20 cells per slide were counted using one method of counting, but that was the only number provided. Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1985. Testing for the toxicity of chemicals with Tetrahymena pyriformis. Science of the Total Environ- ment 43:149-157 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 676758 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology Medium x 2 4 They describe two different methods for counting the cells. Some uncertainty regarding the method se- lected to calculate the EC50 values, but the correla- tion coeffieicnt between the two methods was 0.998. Metric 18: Consistency of Outcome Assessment Medium X 1 2 Assessment protocol was reported with minor uncer- tainties. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High Procedures Metric 20: Outcomes Unrelated to Exposure High x 2 x 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data Metric 23: Explanation of Unexpected Outcomes High Low High x 1 x 2 x 1 Data for exposure related findings were not shown for each study group. Overall Quality Determination"'" Unacceptable 4.0 Metric mean score : 2.0. Extracted No ** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4), EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score is presented solely to increase transparency. * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = I V. (Metric Score,- x MWF,-) I V .. J]. (Metric Score; x MWF;) / J] . MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Bauder, M. B.,Palace, V. P.,Hodson, P. V.. 2005. Is oxidative stress the mechanism of blue sac disease in retene-exposed trout larvae?. Environmental Toxicology and Chemistry 24:694-702 Data Type: Other; Aquatic; Fish Hero ID: 1617737 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/information not reported Metric 3 Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Not measured tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 1 concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions Low X 1 3 Acclimation not reported Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Bauder, M. B.,Palace, V. P.,Hodson, P. V.. 2005. Is oxidative stress the mechanism of blue sac disease in retene-exposed trout larvae?. Environmental Toxicology and Chemistry 24:694-702 Data Type: Other; Aquatic; Fish Hero ID: 1617737 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Jia, R.,Cao, L. P.,Du, J. L.,Wang, J. H.,Liu, Y. J.,Jeney, G.,Xu, P.,Yin, G. J. 2014. Effects of carbon tetrachloride on oxidative stress, inflammatory response and hepatocyte apoptosis in common carp (Cyprinus carpio) . Aquatic Toxicology 152 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 2366621 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source High X 1 1 Metric 3 Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Not measured; nominal tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 1 Concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions Low X 1 3 Acclimation not reported Metric 15: Number of Organisms and Replicates per Low X 1 3 Number of organisms and replicates not reported Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Jia, R.,Cao, L. P.,Du, J. L.,Wang, J. H.,Liu, Y. J.,Jeney, G.,Xu, P.,Yin, G. J.. 2014. Effects of carbon tetrachloride on oxidative stress, inflammatory response and hepatocyte apoptosis in common carp (Cyprinus carpio). Aquatic Toxicology 152 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 2366621 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Oreochromis mossambicus. Science of the Total Environment 214:193-202 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 2468140 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source High X 1 1 Metric 3 Test Substance Purity Low X 1 3 Grade/purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Not measured tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 1 concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Oreochromis mossambicus. Science of the Total Environment 214:193-202 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 2468140 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Oreochromis mossambicus. Science of the Total Environment 214:193-202 Data Type: Other; Aquatic; Fish Hero ID: 2468140 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source High X 1 1 Metric 3 Test Substance Purity Low X 1 3 Grade/purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Not measured tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 1 concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: de Vera, M. P.,Pocsidio, G. N.. 1998. Potential protective effect of calcium carbonate as liming agent against copper toxicity in the African tilapia Oreochromis mossambicus. Science of the Total Environment 214:193-202 Data Type: Other; Aquatic; Fish Hero ID: 2468140 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Khangarot, B. S.,Das, S.. 2009. Acute toxicity of metals and reference toxicants to a freshwater ostracod, Cypris subglobosa Sowerby, 1840 and correlation to EC(50) values of other test models. Journal of Hazardous Materials 172:641-649 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 2592033 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Medium X 1 2 Purchased from SRL (India) and E. Merck (India) Metric 3 Test Substance Purity Low X 1 3 Purity not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Researchers did not report how organisms were al- located to study groups. Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- tion High X 2 2 Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Only nominal concentrations were reported in the tion paper. EC50 values were based on nominal concen- trations. Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit Medium X 1 2 Solvent was discussed for some chemicals, but not for CC14. Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Continued on next page ------- . continued from previous page Study Citation: Khangarot, B. S.,Das, S.. 2009. Acute toxicity of metals and reference toxicants to a freshwater ostracod, Cypris subglobosa Sowerby, 1840 and correlation to EC(50) values of other test models. Journal of Hazardous Materials 172:641-649 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 2592033 Domain Metric Rating^ MWF* Score Comments^ Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21 Metric 22 Metric 23 Statistical Methods High x 1 1 Reporting of Data High x 2 2 Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. ^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. 4 if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. U Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Jia, R.,Cao, L.,Du, J.,Xu, P.,Jeney, G.,Yin, G.. 2013. The protective effect of sifymarin on the carbon tetrachforide (CCf4)-induced fiver injury in common carp (Cyprinus carpio). fn Vitro Ceffufar and Devefopmentaf Biofogy 49:155-161 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3481018 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Metric 3: Test Substance Purity High X 1 1 Commercial source not specified Domain 2: Test Design Metric 4: Negative Controis High X 2 2 Metric 5: Negative Controi Response High X 1 1 Metric 6: Randomized Afiocation High X 1 1 Domain 3: Exposure Characterization Metric 7: Experimentaf System/Test Media Prepara- High x 2 2 tion Metric 8: Consistency of Exposure Administration High x 1 1 Metric 9: Measurement of Test Substance Concentra- Low x 1 3 tion Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low x 1 3 posure Leveis Metric 12: Testing at or Beiow Soiubiiity Limit High x 1 1 nominal injection Only one concentration Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acciimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Repiicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodoiogy High x 2 2 Continued on next page ------- . continued from previous page Study Citation: Jia, R.,Cao, L.,Du, J.,Xu, P.,Jeney, G.,Yin, G.. 2013. The protective effect of silymarin on the carbon tetrachloride (CC14)-induced liver injury in common carp (Cyprinus carpio). In Vitro Cellular and Developmental Biology 49:155-161 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3481018 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Y. Liu, L. Cao, J. Du, R. Jia, J. Wang, P. Xu, G. Yin. 2015. Protective effects of Lycium barbarum poiysaccharides against carbon tetrachforide-induced hepatotoxicity in precision-cut fiver sfices in vitro and in vivo in common carp (Cyprinus carpio L.). Comparative Biochemistry and Physiofogy - Part C: Toxicofogy and Pharmacofogy 169:65-72 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3481539 Domain Metric Rating^ MWF* Score Comments"^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity Low X 1 3 Grade/Purity not reported Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High x 2 2 tion Metric 8: Consistency of Exposure Administration High x 1 1 Metric 9: Measurement of Test Substance Concentra- Low x 1 3 tion Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low x 1 3 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 Not measured 1 concentration Domain 4: Test Organism Metric 13 Metric 14 Metric 15 Test Organism Characteristics Acclimitization and Pretreatment Conditions Number of Organisms and Replicates per Group Metric 16: Adequacy of Test Conditions High X 2 2 Low X 1 3 Acclimation not reported Low X 1 3 Number of organisms and replicates not reported High x 1 Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Y. Liu, L. Cao, J. Du, R. Jia, J. Wang, P. Xu, G. Yin. 2015. Protective effects of Lycium barbarum poiysaccharides against carbon tetrachforide-induced hepatotoxicity in precision-cut fiver sfices in vitro and in vivo in common carp (Cyprinus carpio L.). Comparative Biochemistry and Physiofogy - Part C: Toxicofogy and Pharmacofogy 169:65-72 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3481539 Domain Metric Rating^ MWF* Score Comments"^ Metric 17: Outcome Assessment Methodology High X 2 2 Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overaff Quafity Determination"'" High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. ^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; X MWF;) / J] . MWF, if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. It Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Chen, C. Y.,Wooster, G. A.,Bowser, P. R.. 2004. Comparative blood chemistry and histopathology of tilapia infected with Vibrio vulnificus or Streptococcus iniae or exposed to carbon tetrachloride, gentamicin, or copper sulfate. Aquaculture 239:421-443 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3568343 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Medium X 1 2 Manufacturer identified, but not certified by manu- facturer Metric 3: Test Substance Purity Low X 1 3 Purity/grade not identified Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High X 2 2 tion Metric 8: Consistency of Exposure Administration Medium X 1 2 Did not specify if the controls were also injected Metric 9: Measurement of Test Substance Concentra- Low X 1 3 Not measured tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 Only 1 concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per Medium X 1 2 Number of organisms reported, but not replicates Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Chen, C. Y.,Wooster, G. A.,Bowser, P. R.. 2004. Comparative blood chemistry and histopathology of tilapia infected with Vibrio vulnificus or Streptococcus iniae or exposed to carbon tetrachloride, gentamicin, or copper sulfate. Aquaculture 239:421-443 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3568343 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data High x 2 2 Metric 23: Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction. Data Type: Acute (0-96 hour); Aquatic; other Amphibians Hero ID: 3616521 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response Medium X 1 2 Data were not shown beyond stating that the control survival ranged from 82 to 98 percent. Metric 6: Randomized Allocation Low X 1 3 Randomized allocation was not reported, which is a deficiency. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High X 2 2 tion Metric 8: Consistency of Exposure Administration High X 1 1 Metric 9: Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions Low X 1 3 Acclimatization and pretreatment conditions were not reported. Metric 15: Number of Organisms and Replicates per Medium X 1 2 Number of replicates were reported, but not number Group of organisms per replicate. Metric 16: Adequacy of Test Conditions Medium X 1 2 All organisms were purchased from suppliers and control mortality was acceptable. As a result, this is not a major flaw. Continued on next page ------- . continued from previous page Study Citation: Birge, W. J.,Black, J. A.,Kuehne, R. A.. 1980. Effects of Organic Compounds on Amphibian Reproduction. Data Type: Acute (0-96 hour); Aquatic; other Amphibians Hero ID: 3616521 Domain Metric Ratingt MWF* Score Commentstt Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High High x 2 x 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data High x 1 Medium x 2 Metric 23: Explanation of Unexpected Outcomes High x 1 P/chem and statistics such as LC50 were reported, but not all the unmodified data necessary to re- create the statistics. Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. 4 if any metric is Unacceptable Overall rating = J]. (Metric Score; x MWF;) / J] . MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Other; Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Metric 2: Test Substance Identity Test Substance Source Metric 3: Test Substance Purity High Low Low x 2 x 1 x 1 Source of CC14 was not reported, but it was noted that analytical grade CC14 was used. Purity not reported Domain 2: Test Design Metric 4: Negative Controls Low Metric 5: Negative Control Response Metric 6: Randomized Allocation x 2 Low x 1 Low x 1 The study refers to a blank but doesn't say what's in the blank for CC14. Figure 1 notes that the blank concentration for nitrobenzene is 0 mg/L. Notes re- generation rate determined on Day 7 as most D. japonica in the blank test could normally regener- ate. Study reports that "In the blank tests, the average abnormal regeneration rate was 10 percent and no dead D. japonica were observed through the tests", but does not discuss CC14 specifically It's not reported whether animals were randomly al- located. Domain 3: Exposure Characterization Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Other; Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Metric 7: Experimental System/Test Media Prepara- Low x 2 tion Metric 8: Consistency of Exposure Administration Low x 1 3 Metric 9: Measurement of Test Substance Concentra- Low x 1 3 tion Metric 10: Exposure Duration and Frequency Medium x 2 It's unclear whether the experiement was conducted in a closed or open system using static or flow through methods. The study reports, "The breed- ing liquid for Dugesia japonica was prepared by dis- solving 3.74 g of NaCl, 0.49 g of KC1, and 8.5 5 g of CaC12 into distilled water to make 500 ml. This was diluted 100 times and neutralized by NaHC03 before use. Dugesiajaponica were collected from a stream around which there was no source of pollution and left without food for over 7 days in the breeding liquid to excrete alimentary canal contents. Those of about _2 cm long were used. Dugesia japonica was cut into two parts (head and body part) at the nearest section to the eyes of the trisected part be- tween pharynx and eyes. The body part was used for the head regeneration test. Ten body parts were put in 100 ml ofa test solution, and this was left at 20 " 1" C for 7 days. Observation for head re- generation was carried out with a stereomicroscope on Days 3, 4, 5, 6, and 7 after head cutting, and the test solution was replaced at every observatiort. The degree of regeneration was classified as normal, eye spot, tetratophthalmic, anophthalmic, aciphthalmic, and death. The total number of eye spot, tetratoph- thalmic, anophthalmic, aciphthalmic, and death was regarded as the abnormal regeneration number. The ratio of the number to 10 on Day 7 was defined as the abnormal regeneration rate. The concentration of the chemical, at which the abnormal regenera- tion rate reached 50 percent, was defined as EC50" LC50 of D. japonica was determined at the same time. LC50 and EC50 values of the test mentioned above were determined on semilogarithmic paper." Exposure methods were not reported for each study group it was not reported whether nominal or measured cone were used. CC14 is volatile, and study does not report whether test container was closed or open Exposure occurred over 7 days, and observation was carried out on days 3, 4, 5, 6, and 7 after head cut- ting, and the test solution was replaced at every ob- servation. Continued on next page . . . ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Other; Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Metric 11: Number of Exposure Groups/Spacing of Ex- Low x 1 3 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 Not reported for CC14, but for nitrobenzene reports 4 exposure groups used plus control. Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium x 2 Metric 14: Acclimitization and Pretreatment Conditions Low x 1 Metric 15: Number of Organisms and Replicates per Low x 1 Group Metric 16: Adequacy of Test Conditions Medium x 1 Minor uncertainties about the quality of the test or- ganisms given they were collected from the field and no acclimation is mentioned. Study reports, "Du- gesia japonica were collected from a stream around which there was no source of pollution and left with- out food for over 7 days in the breeding liquid to excrete alimentary canal contents. Those of about 2 cm long were used." Did not report whether they were acclimatized and they were collected from the field. Organisms were left without food for 7 days in the breeding liquid to excrete alimentary canal contents before exposure. The study says "Dugesia japonica was cut into two parts (headand body part) at the nearest section to the eyes of the trisected part between pharynx and eyes. The body part was used for the head regenera- tion test.Ten body parts were put in 100 ml of a test solution, and this was left at 20 " 1"C for 7 days." n = 10 body parts per test concentration. Number of replicates not reported. Body parts were put in 100 ml of a test solution and this was left 20 " 1"C for 7 days. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High x 2 Medium x 1 Observation for head regeneration was carried out with a stereomicroscope on Days 3, 4, 5, 6, and 7 af- ter head cutting, and the test solution was replaced at every observation. Outcomes for CC14 not specif- ically reported. Domain 6: Confounding / Variable Control Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Other; Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Metric 19: Confounding Variables in Test Design and Medium x 2 Procedures Metric 20: Outcomes Unrelated to Exposure Low x 1 Confounding variables are discussed for planarian in terms of comparability of results with results from other species, the study says that confounding may occur due to the cutting of the head (stress of cutting of the head). Data on health and attrition were not reported for each study group. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Medium X 1 2 Methods for calculating LC50 not described clearly Metric 22: Reporting of Data Low X 2 6 Data for exposure related findings not reported for each study group for CC14 Metric 23: Explanation of Unexpected Outcomes Medium x 1 2 They did report unexpected outcomes and explained relatively sufficiently, e.g. the planarian LC50 num- bers being very different than the other two species. Overall Quality Determination^ Low 2.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Yoshioka, Y.,Ose, Y. ,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Metric 2: Test Substance Identity Test Substance Source Metric 3: Test Substance Purity High Low Low x 2 x 1 x 1 Source of CC14 was not reported, but it was noted that analytical grade CC14 was used. Purity not reported Domain 2: Test Design Metric 4: Metric 5: Metric 6: Negative Controls Negative Control Response Randomized Allocation Unacceptable x 2 N/A Low x 1 N/A 3 The study does not mention a control anywhere. The study refers to a blank for Dugesia japonica (planarian) but doesn't say what's in the blank, and doesn't mention a blank for M. macrocopa (water flea) No control reported Study does not report whether animals were ran- domly allocated. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Low tion Metric 8: Consistency of Exposure Administration Low Metric 9: Measurement of Test Substance Concentra- Low tion Metric 10: Exposure Duration and Frequency Low x 2 x 1 x 1 x 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Unacceptable x 1 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 It is not reported whether the container was closed or open, and CCL4 is a volatile chemical. Exposure methods were not reported for each study group It was not reported whether nominal or measured cone were used. Exposure occurred over 3 hours, and OECD recom- mends 48 hours for invertebrate acute tests. Number of exposure groups and spacing of exposure levels not reported Domain 4: Test Organism Metric 13: Test Organism Characteristics Low X 2 6 Test species is a saltwater invertebrate, and were used at 5 days old, but the source of the species is not reported. Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y. ,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Study did not report acclimating water fleas. 10 organisms per exposure group. For freshwater in- vertebrates, OECD recommends at least 20. Num- ber of replicates not reported. "Ten M. macrocopa in 100 ml of test solution were put in a 250-ml vial vessel at 20 " 1"C and the sur- vivors were counted after 3 hr in order to determine LC50." Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Metric 18: Consistency of Outcome Assessment Low X 1 3 Details of outcome assessment were not reported. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and Unacceptable X 2 8 The study did not provide enough information to Procedures allow a comparison of environmental conditions or other non treatment related factors across study groups. Metric 20: Outcomes Unrelated to Exposure Low X 1 3 Data on health and attrition were not reported for each study group. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Medium X 1 2 Methods used to calculate LC50 were not described Metric 22: Reporting of Data Low X 2 6 Data for exposure related findings were not reported for each study group Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination"'" Unacceptable 4.0 Metric mean score**: 2.7. Extracted No Metric 14: Acclimitization and Pretreatment Conditions Low x 1 3 Metric 15: Number of Organisms and Replicates per Low x 1 3 Group Metric 16: Adequacy of Test Conditions Medium x 1 2 Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ ** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4), EPA will determine the study to be unacceptable. In this case, three of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score is presented solely to increase transparency. * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = J]. (Metric Score; x MWF;) / J] . MWFj (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Yoshioka, Y.,Ose, Y. ,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Metric 2: Test Substance Identity Test Substance Source Metric 3: Test Substance Purity High Low Medium x 2 x 1 x 1 Source of CC14 was not reported, but it was noted that analytical grade CC14 was used. Analytical grade CC14 was used. Domain 2: Test Design Metric 4: Negative Controls Unacceptable x 2 Metric 5: Negative Control Response Metric 6: Randomized Allocation N/A Low x 1 N/A 3 The study does not mention a control anywhere. The study refers to a blank for Dugesia japonica (planarian), and Figure 1 indicates the blank for nitrobenzene is a concentration of 0 mg/L. Study doesn't mention a blank for the O. latipes (red kil- lifish) LC50 test.. No control reported Study does not report how test organisms were allo- cated Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Low tion Metric 8: Consistency of Exposure Administration Low Metric 9: Measurement of Test Substance Concentra- Low tion Metric 10: Exposure Duration and Frequency Low X 2 6 LC50 test methods do not describe measures taken to minimize loss of test substance and concentra- tions of test substance not reported as being mea- sured during study. For the oxygen uptake test, test was completed in a closed container (sealed with an electrode), but there were uncertainties about how much air space there was in the flask. X 1 3 Exposure methods were not reported for each study group X 1 3 It was not reported whether nominal or measured cone were used. X 2 6 Exposure occurred over 48 hours, and it sounds like a static test but it is not clear. OECD recommends 96 hours for fish acute tests. Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y. ,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Metric 11: Number of Exposure Groups/Spacing of Ex- Low posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 x 1 For CC14, it is unclear how many exposure groups were used for the LC50 determination. (For the oxy- gen uptake it looks like 5 exposure groups according to figure 2 but that was a different test. ) Domain 4: Test Organism Metric 13: Test Organism Characteristics Metric 14: Acclimitization and Pretreatment Conditions Medium Metric 15: Number of Organisms and Replicates per Medium Group Metric 16: Adequacy of Test Conditions Medium X 2 4 Minor uncertainties about the quality of the test or- ganisms given they were collected from the market. Study reports, "Orizias latipes (ca. 3 cm, 0. 3 g) was obtained from the market and acclimated for at least 1 week in dechlorinated water at 20" C (total hardness was about 80 mg/liter). X 1 2 Fish were acclimatized for at least 1 week and OECD recommends 12 days before they are used for testing. X 1 2 10 organisms per exposure group. OECD recom- mends at least 7. Number of replicates was not re- ported Medium x 1 2 10 fish in 2 liters of water which is a little more than what OECD would recommend. At 0.3 g each and 10 fish per container, it should be a 3 liter flask. Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High Low x 2 x 1 Details of outcome assessment were not reported. Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and Low Procedures Metric 20: Outcomes Unrelated to Exposure Low x 2 x 1 Study did not provide enough information to allow a comparison of environmental conditions or other non-treatment-related factors across study groups, and the omitted information is likely to have a sub- stantial impact on study results. Data on health and attrition were not reported for each study group. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Low x 1 Methods used to calculate LC50 were not described Continued on next page ------- . continued from previous page Study Citation: Yoshioka, Y.,Ose, Y.,Sato, T.. 1986. Correlation of the Five Test Methods to Assess Chemical Toxicity and Relation to Physical Properties. 12:15-21 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3617749 Domain Metric Rating^ MWF* Score Comments^ Metric 22: Reporting of Data Metric 23: Explanation of Unexpected Outcomes Low x 2 High x 1 6 1 Data for exposure related findings not reported for each study group Overall Quality Determination"'" Unacceptable 4.0 Metric mean score**: 2.5. Extracted No ** Consistent with our Application of Systematic Review in TSCARisk Evaluations document, if a metric for a data source receives a score of Unacceptable (score = 4), EPA will determine the study to be unacceptable. In this case, one of the metrics were rated as unacceptable. As such, the study is considered unacceptable and the score is presented solely to increase transparency. * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / J] . MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by a Closed-System Technique. Environmental Toxicology and Chemistry 26:1931-1939 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 3617867 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Metric 2 Metric 3 Test Substance Identity Test Substance Source Test Substance Purity High x 2 Medium x 1 Medium x 1 Source was not provided Purity was not provided. Authors described the chemical purity as "reagent grade" Domain 2: Test Design Metric 4: Negative Controls Medium x 2 Metric 5: Negative Control Response Metric 6: Randomized Allocation Low Low x 1 x 1 Authors referred to a control when discussing how they calculated their EC50 value, but additional de- tails were not reported. The authors indicated that the details of the test setup can be found at the fol- lowing source: Lin JH, Kao WC, Tsai KP, Chen CY. 2005. A novel algal toxicity testing technique for assessing the toxicity of both metallic and organic toxicants. Water Res 39:1869" 1877. Negative Control response was not specifically re- ported in the study, but was incorporated into the calculation of the percent inhibition. Researchers did not report how organisms were al- located to study groups Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion Metric 8: Consistency of Exposure Administration Metric 9: Measurement of Test Substance Concentra- tion Metric 10: Exposure Duration and Frequency High High Medium Metric 11: Number of Exposure Groups/Spacing of Ex- Low posure Levels x 2 x 1 x 1 Medium x 2 x 1 Test concentrations were reported in terms of nom- inal concentrations, but analytical confirmation of the test concentrations was performed at the begin- ning and end of the test by HPLC. This was intended to quantify any potential degradation. The test was 48 hours, but should be 72/96 hrs in duration. The study report indicated that both a range finding and definitive test were conducted but did not report the test concentrations. Continued on next page ------- . continued from previous page Study Citation: Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by a Closed-System Technique. Environmental Toxicology and Chemistry 26:1931-1939 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 3617867 Domain Metric Rating^ MWF* Score Comments^ Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High x 2 2 Metric 14: Acclimitization and Pretreatment Conditions High x 1 1 Metric 15: Number of Organisms and Replicates per High x 1 1 Group Metric 16: Adequacy of Test Conditions High x 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure Medium x 1 2 Data on attrition was not reported for each study group, but is unlikely to have a substantial impact on results. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data Medium X 2 4 Quantitative results were not provided. Metric 23: Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.5 Extracted Yes Continued on next page . . . ------- . continued from previous page Study Citation: Tsai, K. P.,Chen, C. Y.. 2007. An Algal Toxicity Database of Organic Toxicants Derived by a Closed-System Technique. Environmental Toxicology and Chemistry 26:1931-1939 Data Type: Acute (0-96 hour); Aquatic; Plants Hero ID: 3617867 Domain Metric Rating^ MWF* Score Comments^ * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = ^ (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. ^ Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. -I o ------- Study Citation: Schell, J. D. J.. 1987. Interactions of Halogenated Hydrocarbon Mixtures in the Embryo of the Japanese Medaka (Oryzias latipes). Data Type: Other; Aquatic; Fish Hero ID: 3625489 Domain Metric Rating^ MWF* Score Comments"^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Study did not report whether allocation to study groups was random. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- tion Metric 8: Consistency of Exposure Administration Metric 9: Measurement of Test Substance Concentra- tion Metric 10: Exposure Duration and Frequency Metric 11: Number of Exposure Groups/Spacing of Ex- posure Levels Metric 12: Testing at or Below Solubility Limit High High Low High High High x 2 x 1 x 1 x 2 x 1 x 1 Nominal concentrations were used. An experiment was conducted to evaluate rate of loss of CC14 from the exposure vials. After 24 hours, the solution CC14 concentration was 46 percent of the initial nominal concentration Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per Low X 1 3 10 embryos per dose group, Group many replicates. Metric 16: Adequacy of Test Conditions High X 1 1 Continued on next page ------- . continued from previous page Study Citation: Schell, J. D. J.. 1987. Interactions of Halogenated Hydrocarbon Mixtures in the Embryo of the Japanese Medaka (Oryzias latipes). Data Type: Other; Aquatic; Fish Hero ID: 3625489 Domain Metric Ratingt MWF* Score Commentstt Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology Metric 18: Consistency of Outcome Assessment High High x 2 x 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure Medium x 1 2 Data on attrition was reported in each exposure group. Other health outcomes were not reported. Adults were periodically treated with a chemical regime to prevent disease. Eggs were not collected from females of a breeding group that had been chemically treated for disease until at least one week following the treatment. Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods Metric 22: Reporting of Data High x 1 Medium x 2 Metric 23: Explanation of Unexpected Outcomes High x 1 Most but not all outcomes were reported; only minor uncertainties. Overall Quality Determination^ High 1.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Comparisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3634436 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High x 2 2 tion Metric 8: Consistency of Exposure Administration High x 1 1 Metric 9: Measurement of Test Substance Concentra- High x 1 1 tion Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Allocation not reported; does state that procedures in ASTM. 1980. Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. E729-80, were followed Continued on next page ------- . continued from previous page Study Citation: Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Comparisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3634436 Domain Metric Rating^ MWF* Score Comments^ Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21 Metric 22 Metric 23 Statistical Methods High x 1 1 Reporting of Data High x 2 2 Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. ^ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. 4 if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating. U Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Comparisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3634436 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source High X 1 1 Metric 3 Test Substance Purity High X 1 1 Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported; does state that procedures in ASTM. 1980. Standard practice for conducting acute toxicity tests with fishes, macroinvertebrates, and amphibians. E729-80, were followed Report states "all test chambers were open to the atmosphere" but water samples were collected for analysis at 0, 48 and 96 hours., and at 24 or 72 hours in odd- or even-numbered tanks. Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Medium x 2 4 tion Metric 8: Consistency of Exposure Administration High x 1 1 Metric 9: Measurement of Test Substance Concentra- High x 1 1 tion Metric 10: Exposure Duration and Frequency High x 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High x 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High x 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics Medium x 2 Metric 14: Acclimitization and Pretreatment Conditions High x 1 1 Metric 15: Number of Organisms and Replicates per Medium x 1 2 Group Adult amphipods were collected from the Eau Claire River. Douglas County, WI. The number of organisms in each test chamber was five or ten for amphipods. Number used in the CC14 test not specified Continued on next page ------- . continued from previous page Study Citation: Brooke, L.. 1987. Report of the Flow-Through and Static Acute Test Comparisons with Fathead Minnows and Acute Tests with an Amphipod and a Cladoceran. Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3634436 Domain Metric Rating^ MWF* Score Comments^ Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.2 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Geiger, D. L.,Brooke, L. T.,Call, D. J.. 1990. Acute toxicities of organic chemicals to fathead minnows (Pimephales promelas): Volume V. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3660853 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source High X 1 1 Metric 3 Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation High X 1 1 Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Continued on next page ------- . continued from previous page Study Citation: Geiger, D. L.,Brooke, L. T.,Call, D. J.. 1990. Acute toxicities of organic chemicals to fathead minnows (Pimephales promelas): Volume V. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3660853 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.0 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Weber, L. J.,Gingerich, W. H.,Pfeifer, K. F.. 1979. Alterations in Rainbow Trout Liver Function and Body Fluids Following Treatment with Carbon Tetrachloride or Monochlorobenzene. 99:401-413 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3662132 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Medium X 1 2 Only source listed, no other details Metric 3 Test Substance Purity Low X 1 3 Purity/Grade not reported Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- Medium X 2 4 Injection dosing described but test chambers and tion set-up not described Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- Low X 1 3 Not measured tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 1 study only has 1 concentration posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High x 2 2 Continued on next page ------- . continued from previous page Study Citation: Weber, L. J.,Gingerich, W. H.,Pfeifer, K. F.. 1979. Alterations in Rainbow Trout Liver Function and Body Fluids Following Treatment with Carbon Tetrachloride or Monochlorobenzene. 99:401-413 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3662132 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Richie, J. P., Jr.,Mills, B. J.,Lang, C. A.. 1984. The Verification of a Mammalian Toxicant Classification Using a Mosquito Screening Method. 4:1029-1035 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3673049 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source High X 1 1 Metric 3: Test Substance Purity Low X 1 3 The info was not provided Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation method not reported Domain 3: Exposure Characterization x 2 Metric 7: Experimental System/Test Media Prepara- High tion Metric 8: Consistency of Exposure Administration High Metric 9: Measurement of Test Substance Concentra- Low tion Metric 10: Exposure Duration and Frequency High Metric 11: Number of Exposure Groups/Spacing of Ex- High posure Levels Metric 12: Testing at or Below Solubility Limit Medium x 1 Exposure concentrations were not reported, though their determination was described Solubility of some of the test chemicals and solvents used were described, but not pertaining to CC14 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Continued on next page ------- . continued from previous page Study Citation: Richie, J. P., Jr.,Mills, B. J.,Lang, C. A.. 1984. The Verification of a Mammalian Toxicant Classification Using a Mosquito Screening Method. 4:1029-1035 Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3673049 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data High x 2 2 Metric 23: Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.3 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Koskinen, H.,Pehkonen, P.,Vehniainen, E.,Krasnov, A.,Rexroad, C.,Afanasyev, S.,Molsa, H.,Oikari, A.. 2004. Response of Rainbow Trout Transcriptome to Model Chemical Contaminants. 320:745-753 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3684136 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 The info was not provided Metric 3: Test Substance Purity Low X 1 3 The info was not provided Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation Low X 1 3 Allocation not described Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- Medium x 2 tion Metric 8: Consistency of Exposure Administration High x 1 Metric 9: Measurement of Test Substance Concentra- Low x 1 tion Metric 10: Exposure Duration and Frequency Metric 11: Number of Exposure Groups/Spacing of Ex- Low posure Levels Metric 12: Testing at or Below Solubility Limit Medium x 2 x 1 Low x 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Test system described but not in great detail Only nominal concentrations were reported Justification for exposure duration and frequency not provided Details about exposure groups and concentration levels not provided Solvents were discussed, but not for CC14 Continued on next page ------- . continued from previous page Study Citation: Koskinen, H.,Pehkonen, P.,Vehniainen, E.,Krasnov, A.,Rexroad, C.,Afanasyev, S.,Molsa, H.,Oikari, A.. 2004. Response of Rainbow Trout Transcriptome to Model Chemical Contaminants. 320:745-753 Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3684136 Domain Metric Rating^ MWF* Score Comments^ Metric 17: Outcome Assessment Methodology High x 2 2 Metric 18: Consistency of Outcome Assessment High x 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High x 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High x 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High x 1 1 Metric 22: Reporting of Data High x 2 2 Metric 23: Explanation of Unexpected Outcomes High x 1 1 Overall Quality Determination"'" High 1.5 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. $ The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. if any metric is Unacceptable Overall rating = (Metric Score; X MWF;) / J] . MWF, (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, tt Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/Information not reported Metric 3 Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Acute (0-96 hour); Aquatic; Fish Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/Information not reported Metric 3 Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Acute (0-96 hour); Aquatic; Invertebrates Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/information not reported Metric 3 Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Chronic (>21 days); Aquatic; Invertebrates Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1 Test Substance Identity High X 2 2 Metric 2 Test Substance Source Low X 1 3 Source/Information not reported Metric 3 Test Substance Purity High X 1 1 Domain 2: Test Design Metric 4 Negative Controls High X 2 2 Metric 5 Negative Control Response High X 1 1 Metric 6 Randomized Allocation Low X 1 3 Allocation not reported Domain 3: Exposure Characterization Metric 7 Experimental System/Test Media Prepara- High X 2 2 tion Metric 8 Consistency of Exposure Administration High X 1 1 Metric 9 Measurement of Test Substance Concentra- High X 1 1 tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- High X 1 1 posure Levels Metric 12: Testing at or Below Solubility Limit High X 1 1 Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Kimball, G.. 1978. The Effects of Lesser Known Metals and One Organic to Fathead Minnows (Pimephales promelas) and Daphnia magna. Data Type: Chronic (>21 days); Aquatic; Invertebrates Hero ID: 3684293 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.1 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- Study Citation: Kotsanis, N.,Metcalfe, C. D.. 1988. Accelerating an in vivo trout carcinogenesis assay with carbon tetrachloride and partial hepatec- tomy. 15th Annual Aquatic Toxicity Workshop Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 4338225 Domain Metric Rating^ MWF* Score Comments^ Domain 1: Test Substance Metric 1: Test Substance Identity High X 2 2 Metric 2: Test Substance Source Low X 1 3 Not reported Metric 3: Test Substance Purity Low X 1 3 Not reported Domain 2: Test Design Metric 4: Negative Controls High X 2 2 Metric 5: Negative Control Response High X 1 1 Metric 6: Randomized Allocation High X 1 1 Domain 3: Exposure Characterization Metric 7: Experimental System/Test Media Prepara- High X 2 2 tion Metric 8: Consistency of Exposure Administration High X 1 1 Metric 9: Measurement of Test Substance Concentra- Low X 1 3 nominal injection tion Metric 10: Exposure Duration and Frequency High X 2 2 Metric 11: Number of Exposure Groups/Spacing of Ex- Low X 1 3 There was only a single injection dose. posure Levels Metric 12: Testing at or Below Solubility Limit Low X 1 3 This was not discussed. Domain 4: Test Organism Metric 13: Test Organism Characteristics High X 2 2 Metric 14: Acclimitization and Pretreatment Conditions High X 1 1 Metric 15: Number of Organisms and Replicates per High X 1 1 Group Metric 16: Adequacy of Test Conditions High X 1 1 Domain 5: Outcome Assessment Metric 17: Outcome Assessment Methodology High X 2 2 Continued on next page . . . ------- . continued from previous page Study Citation: Kotsanis, N.,Metcalfe, C. D.. 1988. Accelerating an in vivo trout carcinogenesis assay with carbon tetrachloride and partial hepatec- tomy. 15th Annual Aquatic Toxicity Workshop Data Type: Chronic (>21 days); Aquatic; Fish Hero ID: 4338225 Domain Metric Rating^ MWF* Score Comments^ Metric 18: Consistency of Outcome Assessment High X 1 1 Domain 6: Confounding / Variable Control Metric 19: Confounding Variables in Test Design and High X 2 2 Procedures Metric 20: Outcomes Unrelated to Exposure High X 1 1 Domain 7: Data Presentation and Analysis Metric 21: Statistical Methods High X 1 1 Metric 22: Reporting of Data High X 2 2 Metric 23: Explanation of Unexpected Outcomes High X 1 1 Overall Quality Determination^ High 1.4 Extracted Yes * MWF = Metric Weighting Factor t High = 1; Medium = 2; Low = 3; Unacceptable = 4; N/A has no value. * The overall rating is calculated as necessary. EPA may not always provide a comment for a metric that has been categorized as High. Overall rating = (Metric Score; x MWF;) / ^ MWFj if any metric is Unacceptable (round to the nearest tenth) otherwise where High: > 1 to < 1.7; Medium: > 1.7 to < 2.3; Low: > 2.3 to < 3. If the reviewer determines that the overall rating needs adjustment, the original rating is crossed out and an arrow points to the new rating, ft Metrics that are rated 'High' met the criteria for high confidence as expected for this type of study, and may not require additional comments. ------- |