Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Data Requirement: EPA DP Barcode D288775 EPA MRID 458677 11 EPA Guideline 70 1( Special Study)

Test material: Purity: not reported Common name: Atrazine Chemical name: IUPAC CAS name 6 chloro N ethyl N'( 1 methylethyl) 1,3,5 triazine 2,4 diamine CAS No. 1912 24 9 Synonyms EPA PC Code: 80803 Primary Reviewer: Thomas M. Steeger, Ph. D., Senior Biologist Date: April 6, 2003 Environmental Fate and Effects Division, ERB 4, U. S. Environmental Protection Agency Secondary Reviewer( s): Joseph E. Tietge, M. S., Research Aquatic Biologist Date: Mid Continent Ecology Division, National Health and Environmental Effects Research Laboratory ( Duluth), U. S. Environmental Protection Agency Stephanie Irene, Ph. D., Senior Advisor Date: Environmental Fate and Effects Division, ERB 3, U. S. Environmental Protection Agency Mary J. Frankenberry, Senior Statistician Date: Environmental Fate and Effects Division, ERB 3, U. S. Environmental Protection Agency EPA PC Code 080803 Date Evaluation Completed: 06/ 01/ 2003 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 2 of 70 CITATION: Du Preez, L., K. R. Solomon and A. M. 2003. Exposure of Xenopus laevis larvae to different concentrations of atrazine in semi natural microcosms. School of Environmental Sciences and Development, Zoology Department, Potchefstroom University of CHE, Potchefstroom 2520, South Africa. Sponsor: Syngenta Crop Protection, Inc., Laboratory Study ID ECORISK Number SA 01 D. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 3 of 70 EXECUTIVE SUMMARY:

This is an interim report to assess the effects of atrazine on gonadal abnormalities in Xenopus laevis metamorphs through the use of microcosm studies. X. laevis adults were collected from an earthen pond north ( pond C6) of Potchefstroom, South Africa, based on the reported absence of atrazine use and corn production in its watershed and the presence of X. laevis. Adults were induced to spawn in the laboratory and the 96 hr old progeny were divided among 4 treatments ( 0, 1, 10 and 25

g/ L atrazine) with three replicates per treatment ( 800 tadpoles per replicate). Microcosms consisted of 1600 L cement tanks, lined with polyethylene with a 3 cm sand bottom, and filled with 1,100 L of tap water. Larvae were exposed until they reached NF stage 66 ( forelimb emergence and complete tail resorption); the study was terminated after 133 days of exposure. A total of 150 stage 66 frogs were sampled per treatment. Snout vent length, weight, days to stage 66 ( metamorphosis) were recorded and animals were examined for gross gonadal morphology.

Mean measured concentrations of atrazine ranged from 0.9 to 1.82, 10 to 15.9 and 23.8 to 39.7

g/ L for the 1, 10 and 25

g/ L nominal treatments, respectively. Atrazine in reference tanks ranged from 0 to 0.1

g/ L. Although some animals reached stage 66 by 70 days, the majority did not reach metamorphosis until between 126 to 133 days. The study authors attributed low water temperatures to the delayed development of the tadpoles. Based on gross morphology, the incidence of gonadal deformities in 1, 10 and 25

g/ L atrazine groups was 1.3, 0.7 and 3.3% of the total frogs examined ( 150), respectively; reference frogs exhibited a 4% incidence of gonadal deformities. Discontinuous testis was the only gonadal abnormality identified; no abnormalities were observed in the ovaries except for one ovary that was reduced in size. Males comprised 48%, 39% and 47% of the 1.0, 10 and 25

g/ L atrazine treated samples, respectively, while the reference samples was 45% male.

This interim microcosm study represents a reasonable step forward in testing for effects observed in the laboratory and may provide useful information when all the analyses are completed. The present study, though, showed that developmental rates across all treatments, in terms of time to metamorphosis, were delayed. Fluctuating water quality in the microcosm units may have impacted the developmental rate. Additionally, the study was designed with the assumption that phytoplankton growth would serve as a source of food for developing tadpoles. Although the authors note that phytoplankton " flourished," no measure ( e. g., chlorophyl a concentration) of phytoplankton growth was reported and supplemental food ( rabbit pellets) had to be provided to the developing tadpoles. However, phytoplankton growth could have been significantly limited by atrazine treatment, thereby resulting in indirect effects of atrazine on development. In support of this possibility, atrazine treated male and female frogs were significantly smaller in terms of length and weight than their " untreated" counterparts, further complicating interpretation of the study results. This assessment was based on gross morphology alone, and the results may change when histology is completed. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 4 of 70 I. MATERIALS AND METHODS GUIDELINE FOLLOWED: Nonguideline Study COMPLIANCE: Not conducted in accordance with the Good Laboratory Practices as outlined in 40 CFR Part 160, August 19, 1989. However, GLP elements were incorporated into study conduct including:

° This study was conducted in accordance with a written protocol, signed by the Study Director and Principal Investigator ° Standard Operating Procedures were developed/ available for official activities. ° Protocol amendments/ deviations were written and signed by the Study Director.

A. MATERIALS:

1. Test Material Atrazine Description: Not reported Lot No./ Batch No. : Not reported Purity: Not reported Stability of compound uder Test Conditions: Not reported Storage conditions of test chemicals: Not reported 2. Test organism:

Species: African clawed frog ( Xenopus laevis) . Age at test initiation : 96 hr larvae Weight at study initiation ( mean and range): Not reported Length at study initiation ( mean and range): Not reported Source: Adult X. laevis obtained from earthen pond ( C6) 16 km north of the city of Potchefstroom, South Africa. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 5 of 70 B. STUDY DESIGN:

Objective: 1. To assess the effects of exposure to atrazine on gonadal abnormalities in Xenopus laevis metamorphs and sub adults under semi natural conditions in microcosms.

1. Experimental Conditions a) Range finding Study:

b) Definitive Study Table 1 . Experimental Parameters Parameter Details Acclimation: period: Conditions: ( same as test or not) Feeding: Health: ( any mortality observed) Microcosms stabilized for 5 months Adults paired and injected with chorionic gonadotropin ( Pregnyl ® ) to induce spawning and placed in 300 x 240 x 240 mm) spawning tanks at 30oC. After spawning, adults were removed from tanks and water was oxygenated.

Duration of the test NF Stage 66 ( forelimb emergence and tail resorbtion) or 19 weeks total exposure whichever came first.

Test condition static/ flow through Type of dilution system for flowthrough method.

Renewal rate for static renewal static water maintained at 1,100 L mark by addition of tap water when necessary Aeration, if any not reported Test vessel Material: ( glass/ stainless steel) Size: Fill volume: Cement with polyethylene liner 2.25 m x 0.6 m x 1.2 m = 1.62 m3 ( 1,620 L) 1,100 L plastic liner covered with 3 cm layer of sandy soil Source of dilution water Quality: Tap water Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Parameter Details Page 6 of 70 Water parameters: Hardness pH Dissolved oxygen Total Organic carbon Particulate Matter Ammonia Nitrite Metals Pesticides Chlorine Temperature { Salinity for marine or estuarine species}

Intervals of water quality measurement Measured every Thursday mid morning.

Number of replicates/ groups: negative control: water treated ones: 3 Number of organisms per replicate / groups: 800 ( from two separate spawns: 600 from on Sept. 5th and 200 from Sept. 12th )

Biomass loading rate 800 tadpoles per 1,100 L Test concentrations: nominal: measured: 1, 10 and 25

g/ L 0.91  1.82

g/ L; 10  15.9

g/ L; 23.8  39.7

g/ L Solvent ( type, percentage, if used) none Lighting natural sunlight Feeding supplemental rabbit food ( Complete Rabbit Pellets); 20 grams homogenized in 500 ml of tap water provided once per week but later twice per week. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Parameter Details Page 7 of 70 Recovery of chemical Level of Quantitation Level of Detection GC/ MS used to measure atrazine 0.1

g/ L Positive control { if used, indicate the chemical and concentrations} None Other parameters, if any 2. Observations:

Table 2: Observations Criteria Details Parameters measured including the sublethal effects/ toxicity symptoms body weight, snout vent length, time ( days) to metamorphosis; gross morphology and histology of gonads.

Observation intervals Every 14 days 1 L water samples collected for atrazine measurement; water chemistry weekly. From 11/ 5/ 02 until mid January 2003, metamorphs removed from microcosms every 2nd day.

Were raw data included? Yes Other observations, if any Report notes that 800 tadpoles per microcosm were well below the effective density of 1 tadpole/ L for a frog to reach metamorphosis successfully ( Weldon 1999). Xenopus bucket traps baited with marrow bones used to catch metamorphs.

The report notes that 150 metamorphs were collected from each treatment. The researchers attempted to collect 50 from each of the three replicates; however, they were apparently unable to do this in all cases and therefore had to sample more from one replicate than the others. According to the protocol, no more than 75 metamorphs would be collected from a single tank.

II. RESULTS and DISCUSSION: [ All results discussed in this section and the next are those reported by the study authors. Although supplemental data are typically used in a qualitative manner only, EFED verified spreadsheet data and ran basic statistical analyses on the major study parameters. See attached appendix. If results differed in any substantive way, the difference was reported in the text below.] Atrazine concentrations were relatively consistent with nominal levels. On one occasion, atrazine was detected in one of the reference microcosms at a concentration equal to the method detection limit ( 0.1

g/ L). Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 8 of 70 Microcosms were colder than natural water bodies ( no data provided). Extensive early mortality was observed in microcosm 1 ( 1

g/ L), while extensive mortality and slow development was observed in microcosm 8( 10

g/ L); both were excluded from the analysis.

First metamorphs reached development stage 66 in the 10th week. Collection of metamorphs terminated in week 19 when the target number ( 150/ treatment) was reached. The majority of frogs reaching stage 66 appears to have occurred between 126 and 133 days of exposure.

All 600 specimens ( 150 frogs/ treatment * 4 treatments) were examined visually and under a dissecting microscope for gonadal deformities. Of the total number of frogs exposed to 1, 10 and 25

g/ L of atrazine, 1.33%, 0.67% and 3.33% exhibited gonadal deformities, respectively, compared to 4% in reference microcosms. No intersex gonads were observed. Discontinuous testis was the most frequently observed anomaly and ranged from 1.7% to 7.1% in atrazine  treated animals and 8.8% in controls. No gonadal abnormalities were detected in females except for one female with a reduced size ovary. The study concludes that based on gross morphology, atrazine had no adverse effect on gonadal development at mean concentrations ranging from 1.4 to 30.8

g/ L under semi field conditions.

Table 3. Total number of frogs examined and percentage of gonadal deformities.

Nominal Concentration ( measured)

g/ L Total Number Percent Gonadal Abnormalities Frogs Collected Males Females Gonadal Abnormalitie s Total Frogs Males 0 ( 0  0.1) 150 68 82 6 4.0 8.8 1 ( 0.91  1.82) 150 72 78 2 1.3 2.8 10 ( 10  15.9) 150 59 91 1 0.7 1.7 25 ( 23.8  39.7) 150 70 80 5 3.3 7.1 C. REPORTED STATISTICS: Little to no statistics are reported in this interim report.

D. VERIFICATION OF STATISTICAL RESULTS: Statistical analyses run using SAS ® ( Statistical Analysis System, Release 8.01, Cary, North Carolina); see attached output.

E. STUDY DEFICIENCIES: The purity of atrazine used to make up exposure solutions was not reported. The length of exposure for test animals differed; one group consisting of 600 were exposed for approximately one week longer than the complement of 200 additional animals added later. Tap water was used as the Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 9 of 70 source of dilution water. Dissolved oxygen in several of the microcosms dropped below 3 mg/ L. Exposure tanks were not all at the same temperature, but were dependent on the amount of shade from a nearby tree. The number of animals sampled from each replicate may have been different; a total of 150 were sampled from each treatment, but up to 75 may have been sampled from a replicate rather than 50 from each replicate. Atrazine was detected at 0.1

g/ L ( detection limit of method) in a reference tank ( microcosm 3)

F. REVIEWER'S COMMENTS:

Frogs were collected from site C6 because of the absence of atrazine use in the watershed; however, this site was characterized in previous studies ( Smith et al. 2003) as having measurable atrazine concentrations averaging 0.15

g/ L. Although this concentration is relatively low, it does represent a level that has been reported to cause gonadal deformities in males ( Hayes et al. 2002). The potential for gonadal deformities in the control limits the usefulness of this study in testing the hypothesis. .

The conditions under which the tests were conducted probably resulted in poor water quality, which contributed to the relatively retarded development, although low temperatures in the initial few weeks of the study were also a factor. However, the exposure media was never changed and the total volume was not replenished to make up for evaporative losses. This approach would lead to the accumulation of waste products, especially since the study was run for 19 weeks. Unfortunately, mortality was not evaluated throughout the study, so it is difficult to assess the appropriateness of the exposure protocol. All that one can tell is that a maximum of 75 organisms were sampled from any one tank, representing less than 10% of the initial set. It is unclear why the study started out with so many organisms in each tank.

Microcosms 1 and 8 reportedly exhibited extensive mortality early in the study and the authors alluded to possible temperature shock because microcosm 1 was shaded; however, several of the microcosms were shaded by trees and it is unclear why only one would have had such excessively low temperature. Also, temperature shock usually results from too high a temperature, not too low a temperature. Microcosm 8 exhibited slow metamorphosis and the frogs were not at stage 66 by 133 days. Although microcosms were excluded from the stud, the cause of mortality and delayed development was not adequately characterized.

The method detection limit ( 0.1

g/ L) seems unusually high for GC/ MS given that ELISA assays have a detection limit of 0.025

g/ L.

Frogs were relatively slow to reach metamorphosis stage 66. According to the authors, under controlled laboratory conditions of 20  25oC, X. laevis tadpoles take 58 days to complete metamorphosis ( Nieuwkoop and Faber 1967) and under natural conditions where water temperatures may fluctuate widely, metamorphosis may take from 56 to 63 days, still a shorter period of time than the minimal 70 days required in the current study.

Because the test animals were added in two stages, exposure times differed. Animals ( 600) added to the tank first were exposed for roughly a week longer than the second complement ( 200) of animals added to bring the total number of tadpoles per microcosm to 800.

Although the first metamorphs reached stage 66 by 70 days, the majority of study animals did not reach stage 66 until 126 to 133 days, which implies that the study conditions delayed development and may have rendered the animals less sensitive to any developmental effects of atrazine should they exist. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 10 of 70 Although raw water quality data were not provided, a series of figures depicted water temperature, pH, dissolved oxygen ( DO) and conductivity over the study period. It appears from these graphs that something ( perhaps a high rain event) happened around Day 90 to 100 that caused conductivity, temperature and pH to drop substantially. Conductivity in all but three of the exposure tanks tended to steadily increase from days 0 to 90; however, tanks 2,4, 6 and 9 had very erratic conductivity with steep declines. In tanks 4 and 6, conductivity plummeted Day 40 and in tank 2 conductivity plummeted by day 60.

By Day 100, dissolved oxygen in some replicates of the reference, 10 and 25

g/ L treatments dropped below 2 mg/ L. Dissolved oxygen and conductivity in the 25

g/ L treatment were unusually variable relative to the other treatments. It is possible that the supplemental feeding resulted in food residues that increased conductivity during days 0 through 90 and that a high rain event may have resulted in a rapid influx of water that lowered conductivity and dissolved oxygen. The low oxygen content of the water is difficult to explain because water samples were collected mid day when photosynthesis by phytoplankton should supplement dissolved oxygen levels. Heavy rain charged with carbon dioxide could have lowered DO; however, it should have also altered pH which does not appear to have occurred. However, given that the report states that very little rain fell during the study, it is uncertain why water quality changed so dramatically.

The study was intentionally designed to keep the loading rate below 1 tadpole per liter. Because the reference was from 1999, it suggests that the authors were aware that higher loading rates would have compromised the development of tadpoles.

Total time to metamorphosis cannot be determined from this study because an unknown number of animals were sacrificed by Day 133. It is likely that tadpoles could have died and not been accounted for during the course of the study. What is apparent, though, is that metamorphosis was considerably delayed for the majority of animals in this study.

Although no intersex gonads were observed, the current analysis is based on gross morphology alone and does not represent an analysis based on histology.

Although the stated objectives were to analyze snout vent length ( SVL) and mass, the report did not present an analysis of these data. In the table below, the mean SVL and mean mass for each treatment replicate were extracted from the appendix. No statistical analysis was done on this data set, but it appears as if there may be significant reductions in SVL and mass associated with atrazine exposure. EFED's analysis ( see attached output) revealed that on average, male and female frogs not exposed to atrazine were longer 21.17 mm and 22.07 mm, respectively, than frogs from atrazine treated groups ( Table 4); non exposed males and females also tended to weigh more, i. e., 1.08 and 1.20 g, respectively, than frogs treated with atrazine. Analysis of variance ( see attached printout) revealed that both males and females were significantly larger in terms of weight ( p < 0.02) and length ( p< 0.03) than atrazine treated males and females. It is unclear whether the difference in size is due to the direct effects of atrazine on growth or whether it is due to reduced phytoplankton caused by atrazine treatment. At each treatment though, there was no difference in weight or length of males versus females.

Table 4. Summary of average male and female Xenopus laevis lengths and weights with associated standard deviations by atrazine concentration. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 11 of 70 Atrazine Concentration Sex Length Standard Deviation Weight Standard Deviation 0 Female 21.17 3.80 1.08 0.75 0 Male 22.07 3.97 1.20 0.78 1 Female 18.55 2.70 0.70 0.32 1 Male 18.78 2.36 0.71 0.28 10 Female 18.82 3.36 0.73 0.51 10 Male 18.84 3.17 0.75 0.61 25 Female 17.50 3.11 0.59 0.40 25 Male 18.17 2.79 0.64 0.34 Similarly, there was no analysis on time to metamorphosis, as originally stated in the objective section of the study. However, given the method by which metamorphs were removed, and given the confounding effects of the differential exposure during the first week of the study, this analysis may not be possible. Individual treatment data for each tank is needed for Figure 2. Clearly, the data were collected for each tank, but for some reason they were not presented in this study. Also, some very large organisms were collected that were up to 4 to 6 times the mean weight of the entire sample, suggesting that these organisms were not collected at completion of metamorphosis, but some time much later.

In terms of the gonadal analysis, only the gonads of the first 150 stage 66 organisms in each treatment were evaluated. This sampling would bias the evaluation toward the faster developing organisms, which is not representative of the population being tested. Only 150 organisms were sampled, representing 6.25% of the original organisms in a treatment.

H. REFERENCES:

Hayes, T. B., A. Collins, M. Lee, M. Mendoza, N. Noriega, A. S. Stuart, and A. Vonk. 2002a. Hermaphroditic, demasculinized frogs after exposure to the herbicide atrazine at low ecologically relevant doses. Proceedings of the National Academy of Sciences of the United States of America 99( 8): 5476  5480 Nieuwkoop, P. D. and J. Faber. 1967. Normal table of Xenopus laevis ( Daudin). North Holland Publishing Company, Amsterdam.

Smith, E. E., L. Du Preez, and K. Solomon. 2003. Gonadal and laryngeal responses to field exposure of Xenopus laevis to atrazine in areas of corn production in South Africa. The Institute of Environmental and Human Health, Texas Tech University, Lubbock, TX and School of Environmental Sciences and Development, Potchefstroom University for CHE, Potchefstroom, South Africa. Sponsor: Syngenta Crop Protection, Inc., Laboratory Study ID: ECORISK Number SA 01C. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 12 of 70 Weldon, C. 1999. The sustainable utilization of the African clawed frog Xenopus laevis ( Daudin). M. Sc. Thesis. Department of Zoology and Entomology, University of the Orange Free State, Bloemfontein, South Africa. Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 13 of 70 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 14 of 70 MEAN LENGTH AND WEIGHT FOR ALL FROGS COLLECTED AT EACH SITE 621 Obs ATRAZINE SITE _ TYPE_ _ FREQ_ LENGTH WEIGHT L_ SD W_ SD 1 0 12 0 61 21.3213 1.09951 4.09761 0.76949 2 0 3 0 67 21.5955 1.10149 3.49729 0.67835 3 0 7 0 22 22.2409 1.34364 4.50816 0.96341 4 1 11 0 75 17.5907 0.57760 1.82380 0.18347 5 1 4 0 75 19.7347 0.83400 2.70368 0.34261 6 10 10 0 75 18.1227 0.61533 1.63381 0.20565 7 10 5 0 75 19.5400 0.86227 4.22665 0.73003 8 25 2 0 75 18.4040 0.66387 2.59393 0.37961 9 25 6 0 47 17.5574 0.60936 3.52807 0.39261 10 25 9 0 28 16.5679 0.48536 2.52866 0.28908 MEAN LENGTH AND WEIGHT FOR MALE AND FEMALE FROGS COLLECTED AT EACH SITE 622 Obs ATRAZINE SITE Sex _ TYPE_ _ FREQ_ LENGTH WEIGHT L_ SD W_ SD 1 0 12 F 0 29 20.5931 1.03276 4.17312 0.81263 2 0 12 M 0 32 21.9813 1.16000 3.97772 0.73595 3 0 3 F 0 41 21.5585 1.07951 3.24700 0.59480 4 0 3 M 0 26 21.6538 1.13615 3.92624 0.80419 5 0 7 F 0 12 21.2417 1.19250 4.70483 1.07726 6 0 7 M 0 10 23.4400 1.52500 4.17591 0.82481 7 1 11 F 0 39 17.4103 0.56821 1.99272 0.18953 8 1 11 M 0 36 17.7861 0.58778 1.62659 0.17879 9 1 4 F 0 39 19.6974 0.84026 2.85671 0.36757 10 1 4 M 0 36 19.7750 0.82722 2.56742 0.31844 11 10 10 F 0 47 18.0426 0.61362 1.66417 0.21999 12 10 10 M 0 28 18.2571 0.61821 1.60241 0.18286 13 10 5 F 0 43 19.6791 0.86488 4.40317 0.68274 14 10 5 M 0 32 19.3531 0.85875 4.03908 0.80039 15 25 2 F 0 41 18.4561 0.68610 2.88037 0.44866 16 25 2 M 0 34 18.3412 0.63706 2.24150 0.27891 17 25 6 F 0 21 17.1619 0.57143 3.59256 0.39082 18 25 6 M 0 26 17.8769 0.64000 3.51287 0.39905 19 25 9 F 0 19 15.8316 0.41368 2.22862 0.18860 20 25 9 M 0 9 18.1222 0.63667 2.52971 0.40503 MEAN LENGTH AND WEIGHT FOR MALE AND FEMALE FROGS BY ATRAZINE CONC 623 Obs ATRAZINE Sex _ TYPE_ _ FREQ_ LENGTH WEIGHT L_ SD W_ SD 1 0 F 0 82 21.1707 1.07951 3.79852 0.75050 2 0 M 0 68 22.0706 1.20456 3.97068 0.77549 3 1 F 0 78 18.5538 0.70423 2.70405 0.32116 4 1 M 0 72 18.7806 0.70750 2.35723 0.28334 5 10 F 0 90 18.8244 0.73367 3.35507 0.51080 6 10 M 0 60 18.8417 0.74650 3.17032 0.60543 7 25 F 0 81 17.5049 0.59247 3.10676 0.39879 8 25 M 0 69 18.1377 0.63812 2.78832 0.34000 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 15 of 70 PLOT OF FROG WEIGHT OVER ATRAZINE CONCENTRATION FOR MALE AND FEMALE FROGS 624  Sex= F

Plot of WEIGHT* ATRAZINE. Symbol used is '*'.

1.1 ê é * é é é é é 1.0 ê é é é é é é 0.9 ê é é WEIGHT é é é é 0.8 ê é é é é é * é 0.7 ê * é é é é é é 0.6 ê é * é é é é é 0.5 ê èââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââ 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 ATRAZINE Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 16 of 70 PLOT OF FROG WEIGHT OVER ATRAZINE CONCENTRATION FOR MALE AND FEMALE FROGS 625  Sex= M

Plot of WEIGHT* ATRAZINE. Symbol used is '*'.

1.2 ê * é é é é é é 1.1 ê é é é é é é 1.0 ê é é WEIGHT é é é é 0.9 ê é é é é é é 0.8 ê é é é é * é é * 0.7 ê é é é é * é é 0.6 ê èââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââ 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 ATRAZINE Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 17 of 70 ANALYSIS OF VARIANCE FOR FROG WEIGHT BETWEEN TREATMENT GROUPS BY SEX 626  Sex= F

The GLM Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 331 Dependent Variable: MASS Sum of Source DF Squares Mean Square F Value Pr > F Model 3 10.8440965 3.6146988 13.21 <. 0001 Error 327 89.5108805 0.2737336 Corrected Total 330 100.3549770 R Square Coeff Var Root MSE MASS Mean 0.108057 67.26132 0.523196 0.777855 Source DF Type I SS Mean Square F Value Pr > F ATRAZINE 3 10.84409653 3.61469884 13.21 <. 0001 Source DF Type III SS Mean Square F Value Pr > F ATRAZINE 3 10.84409653 3.61469884 13.21 <. 0001 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 18 of 70 ANALYSIS OF VARIANCE FOR FROG WEIGHT BETWEEN TREATMENT GROUPS BY SEX 628  Sex= M

The GLM Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 269 Dependent Variable: MASS Sum of Source DF Squares Mean Square F Value Pr > F Model 3 13.56982643 4.52327548 15.88 <. 0001 Error 265 75.47985684 0.28482965 Corrected Total 268 89.04968327 R Square Coeff Var Root MSE MASS Mean 0.152385 64.76464 0.533694 0.824052 Source DF Type I SS Mean Square F Value Pr > F ATRAZINE 3 13.56982643 4.52327548 15.88 <. 0001 Source DF Type III SS Mean Square F Value Pr > F ATRAZINE 3 13.56982643 4.52327548 15.88 <. 0001 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 19 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG WEIGHT BY SEX 630  Sex= F

The UNIVARIATE Procedure Variable: Resid Moments N 331 Sum Weights 331 Mean 0 Sum Observations 0 Std Deviation 0.52081195 Variance 0.27124509 Skewness 3.47817499 Kurtosis 16.8585986 Uncorrected SS 89.5108805 Corrected SS 89.5108805 Coeff Variation . Std Error Mean 0.02862641 Basic Statistical Measures Location Variability Mean 0.00000 Std Deviation 0.52081 Median  0.12423 Variance 0.27125 Mode  0.24951 Range 4.39000 Interquartile Range 0.29944 Tests for Location: Mu0= 0 Test  Statistic  p Value

Student's t t 0 Pr > | t| 1.0000 Sign M  60.5 Pr >= | M| <. 0001 Signed Rank S  9276 Pr >= | S| <. 0001 Tests for Normality Test  Statistic  p Value

Shapiro Wilk W 0.670492 Pr < W < 0.0001 Kolmogorov Smirnov D 0.218809 Pr > D < 0.0100 Cramer von Mises W Sq 5.537051 Pr > W Sq < 0.0050 Anderson Darling A Sq 29.21036 Pr > A Sq < 0.0050 Quantiles ( Definition 5)

Quantile Estimate 100% Max 3.6004878 99% 2.4504878 95% 0.9657692 90% 0.4175309 75% Q3 0.0557692 50% Median  0.1242308 25% Q1  0.2436667 10%  0.3495122 5%  0.4436667 1%  0.7195122 0% Min  0.7895122 Extreme Observations  Lowest  Highest

Value Obs Value Obs  0.789512 1 2.18753 279 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 20 of 70  0.739512 38 2.45049 31  0.739512 6 2.92633 250  0.719512 42 3.38049 71  0.669512 7 3.60049 13 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG WEIGHT BY SEX 632  Sex= F

The UNIVARIATE Procedure Variable: Resid Histogram # Boxplot 3.7+* 1 * . .* 1 * . .* 1 * . .* 1 * . .* 1 * . .* 1 * 1.5+* 3 * .* 2 * .** 4 * .*** 9 0 .** 4 0 .*** 7 0 .****** 18 | .****************** 52 +++ .************************************** 114 ** .******************************* 92 ++ .***** 13 |  0.7+*** 7 0 +++++++ * may represent up to 3 counts The UNIVARIATE Procedure Variable: Resid Normal Probability Plot 3.7+ * | | * | | * | | * | | * | | * 1.5+ *** | * ++ | ** ++++ | ***++ | ++*+ | ++++ ** | ++++ *** | ++++ ****** | ********** | ************ | ***** ++++  0.7+* *** ++++ +++++++++++ Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 21 of 70  2  1 0 + 1 + 2 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 22 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG WEIGHT BY SEX 634  Sex= M

The UNIVARIATE Procedure Variable: Resid Moments N 269 Sum Weights 269 Mean 0 Sum Observations 0 Std Deviation 0.53069884 Variance 0.28164126 Skewness 3.56627975 Kurtosis 19.3195663 Uncorrected SS 75.4798568 Corrected SS 75.4798568 Coeff Variation . Std Error Mean 0.03235728 Basic Statistical Measures Location Variability Mean 0.00000 Std Deviation 0.53070 Median  0.12650 Variance 0.28164 Mode  0.18650 Range 4.69806 Interquartile Range 0.36900 NOTE: The mode displayed is the smallest of 4 modes with a count of 4.

Tests for Location: Mu0= 0 Test  Statistic  p Value

Student's t t 0 Pr > | t| 1.0000 Sign M  40.5 Pr >= | M| <. 0001 Signed Rank S  4884.5 Pr >= | S| 0.0001 Tests for Normality Test  Statistic  p Value

Shapiro Wilk W 0.698635 Pr < W < 0.0001 Kolmogorov Smirnov D 0.177585 Pr > D < 0.0100 Cramer von Mises W Sq 3.346828 Pr > W Sq < 0.0050 Anderson Darling A Sq 18.12 Pr > A Sq < 0.0050 Quantiles ( Definition 5)

Quantile Estimate 100% Max 3.863500 99% 3.115441 95% 0.885441 90% 0.491884 75% Q3 0.112500 50% Median  0.126500 25% Q1  0.256500 10%  0.384559 5%  0.474559 1%  0.784559 0% Min  0.834559 Extreme Observations  Lowest  Highest

Value Obs Value Obs  0.834559 39 1.34544 8  0.784559 27 2.14544 62 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 23 of 70  0.784559 24 3.11544 44  0.774559 63 3.19544 11  0.764559 5 3.86350 196 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG WEIGHT BY SEX 636  Sex= M

The UNIVARIATE Procedure Variable: Resid Histogram # Boxplot 3.9+* 1 * . . . .* 2 * . . . . .* 1 * . . 1.5+ .* 2 * .*** 5 0 .** 4 0 .**** 8 0 .**** 8 | .*********** 21 | .********************* 42 +++ .***************************************** 82 ** .************************************* 73 ++ .****** 12 | .**** 7 |  0.9+* 1 0 ++++++++ * may represent up to 2 counts The UNIVARIATE Procedure Variable: Resid Normal Probability Plot 3.9+ * | | | | * * | | | | | * | | 1.5+ | ** +++ | ***++ | **+ | ++*** | ++++*** | ++++ **** | ++++****** | ********* | *********** | ***** ++++ Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 24 of 70 |* ***** ++++  0.9+* +++ +++++++++++  2  1 0 + 1 + 2 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 25 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG WEIGHT BY SEX 638 The UNIVARIATE Procedure Variable: Resid Schematic Plots | 4 + | * | | * 3.5 + | * | * | * 3 + | * | | 2.5 + * | | * | * 2 + | | | * 1.5 + * | * * | * * | * 0 1 + 0 0 | 0 0 | 0 0 | 0 0 0.5 + | | | | | | | | | | ++ 0 + +++ | + | | ** ** | ++ ++ | | |  0.5 + | | | | | | 0 | | 0  1 + ++ Sex F M Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 26 of 70 REGRESSION ANALYSIS OF WEIGHT OVER ATRAZINE CONCENTRATION 639  Sex= F

The REG Procedure Model: MODEL1 Dependent Variable: WEIGHT Analysis of Variance Sum of Mean Source DF Squares Square F Value Pr > F Model 1 0.06563 0.06563 1.96 0.2969 Error 2 0.06711 0.03356 Corrected Total 3 0.13274 Root MSE 0.18318 R Square 0.4944 Dependent Mean 0.77747 Adj R Sq 0.2416 Coeff Var 23.56145 Parameter Estimates Parameter Standard Variable DF Estimate Error t Value Pr > | t|

Intercept 1 0.89246 0.12309 7.25 0.0185 ATRAZINE 1  0.01278 0.00914  1.40 0.2969 REGRESSION ANALYSIS OF WEIGHT OVER ATRAZINE CONCENTRATION 640  Sex= M

The REG Procedure Model: MODEL1 Dependent Variable: WEIGHT Analysis of Variance Sum of Mean Source DF Squares Square F Value Pr > F Model 1 0.07648 0.07648 1.25 0.3800 Error 2 0.12248 0.06124 Corrected Total 3 0.19896 Root MSE 0.24747 R Square 0.3844 Dependent Mean 0.82417 Adj R Sq 0.0766 Coeff Var 30.02629 Parameter Estimates Parameter Standard Variable DF Estimate Error t Value Pr > | t|

Intercept 1 0.94830 0.16628 5.70 0.0294 ATRAZINE 1  0.01379 0.01234  1.12 0.3800 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 27 of 70 PLOT OF FROG LENGTH OVER ATRAZINE CONCENTRATION FOR MALE AND FEMALE FROGS 641  Sex= F

Plot of LENGTH* ATRAZINE. Symbol used is '*'.

LENGTH é 21.5 ê é é é * é 21.0 ê é é é é 20.5 ê é é é é 20.0 ê é é é é 19.5 ê é é é é 19.0 ê é é * é é * 18.5 ê é é é é 18.0 ê é é é é 17.5 ê * é èââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââ 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 ATRAZINE Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 28 of 70 PLOT OF FROG LENGTH OVER ATRAZINE CONCENTRATION FOR MALE AND FEMALE FROGS 642  Sex= M

Plot of LENGTH* ATRAZINE. Symbol used is '*'.

LENGTH é é 22.5 ê é é é * 22.0 ê é é é 21.5 ê é é é 21.0 ê é é é 20.5 ê é é é 20.0 ê é é é 19.5 ê é é é 19.0 ê é * é * é 18.5 ê é é é * 18.0 ê é èââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââêââ 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 ATRAZINE Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 29 of 70 ANALYSIS OF VARIANCE FOR FROG LENGTH BETWEEN TREATMENT GROUPS BY SEX 643  Sex= F

The GLM Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 331 Dependent Variable: SVL Sum of Source DF Squares Mean Square F Value Pr > F Model 3 585.622241 195.207414 18.21 <. 0001 Error 327 3505.727849 10.720880 Corrected Total 330 4091.350091 R Square Coeff Var Root MSE SVL Mean 0.143137 17.21578 3.274276 19.01903 Source DF Type I SS Mean Square F Value Pr > F ATRAZINE 3 585.6222415 195.2074138 18.21 <. 0001 Source DF Type III SS Mean Square F Value Pr > F ATRAZINE 3 585.6222415 195.2074138 18.21 <. 0001 ANALYSIS OF VARIANCE FOR FROG LENGTH BETWEEN TREATMENT GROUPS BY SEX 645  Sex= M

The GLM Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 269 Dependent Variable: SVL Sum of Source DF Squares Mean Square F Value Pr > F Model 3 640.258332 213.419444 21.98 <. 0001 Error 265 2572.541817 9.707705 Corrected Total 268 3212.800149 R Square Coeff Var Root MSE SVL Mean 0.199284 16.01009 3.115719 19.46097 Source DF Type I SS Mean Square F Value Pr > F ATRAZINE 3 640.2583321 213.4194440 21.98 <. 0001 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 30 of 70 Source DF Type III SS Mean Square F Value Pr > F ATRAZINE 3 640.2583321 213.4194440 21.98 <. 0001 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 31 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG LENGTH BY SEX 647  Sex= F

The UNIVARIATE Procedure Variable: Resid Moments N 331 Sum Weights 331 Mean 0 Sum Observations 0 Std Deviation 3.25935848 Variance 10.6234177 Skewness 1.6487435 Kurtosis 4.17321286 Uncorrected SS 3505.72785 Corrected SS 3505.72785 Coeff Variation . Std Error Mean 0.17915054 Basic Statistical Measures Location Variability Mean 0.00000 Std Deviation 3.25936 Median  0.62444 Variance 10.62342 Mode  1.42444 Range 22.72940 Interquartile Range 2.77060 NOTE: The mode displayed is the smallest of 2 modes with a count of 6.

Tests for Location: Mu0= 0 Test  Statistic  p Value

Student's t t 0 Pr > | t| 1.0000 Sign M  36.5 Pr >= | M| <. 0001 Signed Rank S  5490 Pr >= | S| 0.0015 Tests for Normality Test  Statistic  p Value

Shapiro Wilk W 0.871872 Pr < W < 0.0001 Kolmogorov Smirnov D 0.1515 Pr > D < 0.0100 Cramer von Mises W Sq 2.170587 Pr > W Sq < 0.0050 Anderson Darling A Sq 12.12968 Pr > A Sq < 0.0050 Quantiles ( Definition 5)

Quantile Estimate 100% Max 15.775556 99% 11.395062 95% 6.729268 90% 3.646154 75% Q3 0.946154 50% Median  0.624444 25% Q1  1.824444 10%  3.004938 5%  4.104938 1%  5.670732 0% Min  6.953846 Extreme Observations  Lowest  Highest

Value Obs Value Obs  6.95385 83 10.4756 248 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 32 of 70  6.17073 1 11.3951 279  5.87073 6 13.5293 71  5.67073 38 13.9293 13  5.57073 5 15.7756 250 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 33 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG LENGTH BY SEX 649  Sex= F

The UNIVARIATE Procedure Variable: Resid Histogram # Boxplot 15.5+* 1 * . .* 2 * . .* 1 * .* 2 * .** 3 * .* 2 0 .*** 5 0 .*** 6 0 .*** 6 0 4.5+** 3 | .***** 10 | .******** 15 | .************ 23 | .************************* 50 +++ .****************************** 59 ** .************************************ 71 ++ .******************* 38 | .******** 16 | .****** 11 | .*** 5 |  6.5+* 2 0 +++++++ * may represent up to 2 counts The UNIVARIATE Procedure Variable: Resid Normal Probability Plot 15.5+ * | | ** | | * | ** | *** | * + | ** +++ | ** +++ | **+++ 4.5+ **+ | ++** | +++** | +++**** | +++***** | +****** | ******* | ******+ | ****+++ | *****+++ | **** +++  6.5+* +++ +++++++++++  2  1 0 + 1 + 2 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 34 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG LENGTH BY SEX 651  Sex= M

The UNIVARIATE Procedure Variable: Resid Moments N 269 Sum Weights 269 Mean 0 Sum Observations 0 Std Deviation 3.09823121 Variance 9.59903663 Skewness 1.47756279 Kurtosis 4.18236889 Uncorrected SS 2572.54182 Corrected SS 2572.54182 Coeff Variation . Std Error Mean 0.18890249 Basic Statistical Measures Location Variability Mean 0.00000 Std Deviation 3.09823 Median  0.57059 Variance 9.59904 Mode  1.68056 Range 22.72892 Interquartile Range 2.85713 Tests for Location: Mu0= 0 Test  Statistic  p Value

Student's t t 0 Pr > | t| 1.0000 Sign M  29.5 Pr >= | M| 0.0004 Signed Rank S  2839.5 Pr >= | S| 0.0259 Tests for Normality Test  Statistic  p Value

Shapiro Wilk W 0.903548 Pr < W < 0.0001 Kolmogorov Smirnov D 0.137053 Pr > D < 0.0100 Cramer von Mises W Sq 1.231739 Pr > W Sq < 0.0050 Anderson Darling A Sq 6.644115 Pr > A Sq < 0.0050 Quantiles ( Definition 5)

Quantile Estimate 100% Max 15.858333 99% 12.229412 95% 6.029412 90% 4.162319 75% Q3 1.119444 50% Median  0.570588 25% Q1  1.737681 10%  3.080556 5%  3.880556 1%  5.870588 0% Min  6.870588 Extreme Observations  Lowest  Highest

Value Obs Value Obs  6.87059 27 8.45833 183 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 35 of 70  6.27059 63 8.52941 62  5.87059 24 12.22941 44  5.77059 39 13.32941 11  5.37059 5 15.85833 196 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 36 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG LENGTH BY SEX 653  Sex= M

The UNIVARIATE Procedure Variable: Resid Histogram # Boxplot 15.5+* 1 * . .* 1 * .* 1 * . . . .** 4 0 .* 2 0 .*** 5 0 .** 4 | 4.5+***** 10 | .****** 11 | .****** 12 | .*********** 21 ++ .***************** 33 | + | .************************** 51 ** .**************************** 56 ++ .*************** 30 | .******** 16 | .*** 6 | .** 3 |  6.5+* 2 0 +++++ * may represent up to 2 counts The UNIVARIATE Procedure Variable: Resid Normal Probability Plot 15.5+ * | | * | * | | | | *** | * +++ | *** +++ | **+++ 4.5+ ***+ | +** | ++** | ++**** | +++**** | +****** | ******* | ******+ | *****+++ | *** ++++ | *** +++  6.5+* +++ +++++++++++  2  1 0 + 1 + 2 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 37 of 70 PROC UNIVARIATE OUTPUT FOR RESIDUALS FROM GLM PROCEDURE USING FROG LENGTH BY SEX 655 The UNIVARIATE Procedure Variable: Resid Schematic Plots | 17.5 + | | | * * 15 + | | * | * 12.5 + * | | * | * 10 + * | * | 0 0 | 0 0 7.5 + 0 0 | 0 | 0 0 | 0 | 5 + 0 | | | | | | | | | | 2.5 + | | | | | | ++ ++ | | | | | 0 + | + | | + | | ** ** | | | | | | ++ ++  2.5 + | | | | | | | | | | |  5 + | | | | | | 0 0 | 0 0  7.5 + ++ Sex F M Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 38 of 70 REGRESSION ANALYSIS OF LENGTH OVER ATRAZINE CONCENTRATION 656  Sex= F

The REG Procedure Model: MODEL1 Dependent Variable: LENGTH Analysis of Variance Sum of Mean Source DF Squares Square F Value Pr > F Model 1 3.99283 3.99283 2.51 0.2541 Error 2 3.18361 1.59180 Corrected Total 3 7.17643 Root MSE 1.26167 R Square 0.5564 Dependent Mean 19.01349 Adj R Sq 0.3346 Coeff Var 6.63564 Parameter Estimates Parameter Standard Variable DF Estimate Error t Value Pr > | t|

Intercept 1 19.91044 0.84775 23.49 0.0018 ATRAZINE 1  0.09966 0.06293  1.58 0.2541 REGRESSION ANALYSIS OF LENGTH OVER ATRAZINE CONCENTRATION 657  Sex= M

The REG Procedure Model: MODEL1 Dependent Variable: LENGTH Analysis of Variance Sum of Mean Source DF Squares Square F Value Pr > F Model 1 3.94737 3.94737 1.45 0.3522 Error 2 5.46029 2.73015 Corrected Total 3 9.40766 Root MSE 1.65232 R Square 0.4196 Dependent Mean 19.45762 Adj R Sq 0.1294 Coeff Var 8.49187 Parameter Estimates Parameter Standard Variable DF Estimate Error t Value Pr > | t|

Intercept 1 20.34946 1.11024 18.33 0.0030 ATRAZINE 1  0.09909 0.08241  1.20 0.3522 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 39 of 70 ANOVA FOR FROG LENGTH BETWEEN TREATMENTS 658  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 10 Dependent Variable: LENGTH Sum of Source DF Squares Mean Square F Value Pr > F Model 3 24.31994787 8.10664929 6.17 0.0290 Error 6 7.88332214 1.31388702 Corrected Total 9 32.20327001 R Square Coeff Var Root MSE LENGTH Mean 0.755201 6.043316 1.146249 18.96722 Source DF Anova SS Mean Square F Value Pr > F ATRAZINE 3 24.31994787 8.10664929 6.17 0.0290 Levene's Test for Homogeneity of LENGTH Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F ATRAZINE 1 1.4602 1.4602 2.90 0.1640 Error 4 2.0163 0.5041 Bartlett's Test for Homogeneity of LENGTH Variance Source DF Chi Square Pr > ChiSq ATRAZINE 3 1.6226 0.6543 Dunnett's t Tests for LENGTH NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 6 Error Mean Square 1.313887 Critical Value of Dunnett's t 3.12383 Comparisons significant at the 0.05 level are indicated by ***.

Difference ATRAZINE Between Simultaneous 95% Comparison Means Confidence Limits 10  0  2.2703  5.5390 0.9984 1  0  2.5773  5.8460 0.6915 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 40 of 70 25  0  3.9812  6.9049  1.0576 *** Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 41 of 70 ANOVA FOR FROG LENGTH BETWEEN TREATMENTS 662  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 10 Dependent Variable: LENGTH Sum of Source DF Squares Mean Square F Value Pr > F Model 3 32.02748490 10.67582830 14.25 0.0039 Error 6 4.49480602 0.74913434 Corrected Total 9 36.52229092 R Square Coeff Var Root MSE LENGTH Mean 0.876930 4.402765 0.865525 19.65868 Source DF Anova SS Mean Square F Value Pr > F ATRAZINE 3 32.02748490 10.67582830 14.25 0.0039 Levene's Test for Homogeneity of LENGTH Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F ATRAZINE 1 0.4820 0.4820 3.52 0.1337 Error 4 0.5470 0.1368 Bartlett's Test for Homogeneity of LENGTH Variance Source DF Chi Square Pr > ChiSq ATRAZINE 3 3.1456 0.3697 Dunnett's t Tests for LENGTH NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 6 Error Mean Square 0.749134 Critical Value of Dunnett's t 3.12383 Comparisons significant at the 0.05 level are indicated by ***.

Difference ATRAZINE Between Simultaneous 95% Comparison Means Confidence Limits 10  0  3.5532  6.0214  1.0851 *** Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 42 of 70 1  0  3.5778  6.0460  1.1096 *** 25  0  4.2449  6.4525  2.0373 *** Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 43 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH ACROSS TREATMENTS 666  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable LENGTH Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 27.0 16.50 4.387482 9.000000 1 2 10.0 11.00 3.829708 5.000000 10 2 10.0 11.00 3.829708 5.000000 25 3 8.0 16.50 4.387482 2.666667 Kruskal Wallis Test Chi Square 6.7455 DF 3 Pr > Chi Square 0.0805 Median Scores ( Number of Points Above Median) for Variable LENGTH Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 3.0 1.50 0.763763 1.00 1 2 1.0 1.00 0.666667 0.50 10 2 1.0 1.00 0.666667 0.50 25 3 0.0 1.50 0.763763 0.00 Median One Way Analysis Chi Square 5.4000 DF 3 Pr > Chi Square 0.1447 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 44 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH ACROSS TREATMENTS 668  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable LENGTH Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 27.0 16.50 4.387482 9.000000 1 2 8.0 11.00 3.829708 4.000000 10 2 10.0 11.00 3.829708 5.000000 25 3 10.0 16.50 4.387482 3.333333 Kruskal Wallis Test Chi Square 6.0909 DF 3 Pr > Chi Square 0.1073 Median Scores ( Number of Points Above Median) for Variable LENGTH Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 3.0 1.50 0.763763 1.00 1 2 1.0 1.00 0.666667 0.50 10 2 1.0 1.00 0.666667 0.50 25 3 0.0 1.50 0.763763 0.00 Median One Way Analysis Chi Square 5.4000 DF 3 Pr > Chi Square 0.1447 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 45 of 70 ANOVA FOR FROG WEIGHT BETWEEN TREATMENTS 670  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 10 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 3 0.47376094 0.15792031 7.93 0.0165 Error 6 0.11947701 0.01991283 Corrected Total 9 0.59323795 R Square Coeff Var Root MSE WEIGHT Mean 0.798602 17.94657 0.141113 0.786294 Source DF Anova SS Mean Square F Value Pr > F ATRAZINE 3 0.47376094 0.15792031 7.93 0.0165 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F ATRAZINE 1 0.000095 0.000095 1.45 0.2953 Error 4 0.000264 0.000066 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq ATRAZINE 3 0.9200 0.8206 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 6 Error Mean Square 0.019913 Critical Value of Dunnett's t 3.12383 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous ATRAZINE Between 95% Confidence Comparison Means Limits 10  0  0.3623  0.7647 0.0401 1  0  0.3974  0.7998 0.0050 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 46 of 70 25  0  0.5445  0.9044  0.1846 *** Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 47 of 70 ANOVA FOR FROG WEIGHT BETWEEN TREATMENTS 674  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values ATRAZINE 4 0 1 10 25 Number of observations 10 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 3 0.73743515 0.24581172 9.66 0.0103 Error 6 0.15260050 0.02543342 Corrected Total 9 0.89003565 R Square Coeff Var Root MSE WEIGHT Mean 0.828546 18.48632 0.159479 0.862684 Source DF Anova SS Mean Square F Value Pr > F ATRAZINE 3 0.73743515 0.24581172 9.66 0.0103 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F ATRAZINE 1 0.00150 0.00150 4.00 0.1161 Error 4 0.00150 0.000376 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq ATRAZINE 3 12.4660 0.0059 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 6 Error Mean Square 0.025433 Critical Value of Dunnett's t 3.12383 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous ATRAZINE Between 95% Confidence Comparison Means Limits Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 48 of 70 10  0  0.5352  0.9900  0.0805 *** 1  0  0.5662  1.0210  0.1114 *** 25  0  0.6358  1.0426  0.2290 *** Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 49 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT ACROSS TREATMENTS BY SEX 678  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 27.0 16.50 4.387482 9.00 1 2 8.0 11.00 3.829708 4.00 10 2 11.0 11.00 3.829708 5.50 25 3 9.0 16.50 4.387482 3.00 Kruskal Wallis Test Chi Square 6.5455 DF 3 Pr > Chi Square 0.0879 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 3.0 1.50 0.763763 1.00 1 2 1.0 1.00 0.666667 0.50 10 2 1.0 1.00 0.666667 0.50 25 3 0.0 1.50 0.763763 0.00 Median One Way Analysis Chi Square 5.4000 DF 3 Pr > Chi Square 0.1447 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 50 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT ACROSS TREATMENTS BY SEX 680  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 27.0 16.50 4.387482 9.00 1 2 7.0 11.00 3.829708 3.50 10 2 9.0 11.00 3.829708 4.50 25 3 12.0 16.50 4.387482 4.00 Kruskal Wallis Test Chi Square 5.8364 DF 3 Pr > Chi Square 0.1198 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable ATRAZINE Sum of Expected Std Dev Mean ATRAZINE N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ 0 3 3.0 1.50 0.763763 1.00 1 2 1.0 1.00 0.666667 0.50 10 2 1.0 1.00 0.666667 0.50 25 3 0.0 1.50 0.763763 0.00 Median One Way Analysis Chi Square 5.4000 DF 3 Pr > Chi Square 0.1447 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 51 of 70 ANOVA FOR WEIGHT BETWEEN SEXES 682  ATRAZINE= 0

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 6 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.04444191 0.04444191 1.64 0.2697 Error 4 0.10848814 0.02712203 Corrected Total 5 0.15293005 R Square Coeff Var Root MSE WEIGHT Mean 0.290603 13.86664 0.164688 1.187654 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.04444191 0.04444191 1.64 0.2697 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F Sex 1 0.00111 0.00111 2.89 0.1646 Error 4 0.00153 0.000384 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq Sex 1 1.3299 0.2488 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 4 Error Mean Square 0.027122 Critical Value of Dunnett's t 2.77630 Minimum Significant Difference 0.3733 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous Sex Between 95% Confidence Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 52 of 70 Comparison Means Limits M  F 0.1721  0.2012 0.5454 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 53 of 70 ANOVA FOR WEIGHT BETWEEN SEXES 686  ATRAZINE= 1

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 4 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00001069 0.00001069 0.00 0.9872 Error 2 0.06567277 0.03283639 Corrected Total 3 0.06568346 R Square Coeff Var Root MSE WEIGHT Mean 0.000163 25.67177 0.181208 0.705865 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.00001069 0.00001069 0.00 0.9872 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F Sex 0 0 . . . Error 0 0 .

Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq Sex 1 0.0108 0.9171 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 2 Error Mean Square 0.032836 Critical Value of Dunnett's t 4.30263 Minimum Significant Difference 0.7797 Comparisons significant at the 0.05 level are indicated by ***.

Difference Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 54 of 70 Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F 0.003269  0.776402 0.782940 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 55 of 70 ANOVA FOR WEIGHT BETWEEN SEXES 690  ATRAZINE= 10

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 4 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00000059 0.00000059 0.00 0.9969 Error 2 0.06049619 0.03024810 Corrected Total 3 0.06049678 R Square Coeff Var Root MSE WEIGHT Mean 0.000010 23.53874 0.173920 0.738866 Source DF Anova SS Mean Square F Value Pr > F Sex 1 5.9017464E 7 5.9017464E 7 0.00 0.9969 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F Sex 0 0 . . . Error 0 0 .

Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq Sex 1 0.00127 0.9716 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 2 Error Mean Square 0.030248 Critical Value of Dunnett's t 4.30263 Minimum Significant Difference 0.7483 Comparisons significant at the 0.05 level are indicated by ***.

Difference Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 56 of 70 Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F  0.0007682  0.7490802 0.7475438 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 57 of 70 ANOVA FOR WEIGHT BETWEEN SEXES 694  ATRAZINE= 25

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 6 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00980227 0.00980227 1.05 0.3639 Error 4 0.03742040 0.00935510 Corrected Total 5 0.04722267 R Square Coeff Var Root MSE WEIGHT Mean 0.207575 16.18803 0.096722 0.597489 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.00980227 0.00980227 1.05 0.3639 Levene's Test for Homogeneity of WEIGHT Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F Sex 1 0.000233 0.000233 4.00 0.1162 Error 4 0.000233 0.000058 Bartlett's Test for Homogeneity of WEIGHT Variance Source DF Chi Square Pr > ChiSq Sex 1 11.6016 0.0007 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 4 Error Mean Square 0.009355 Critical Value of Dunnett's t 2.77630 Minimum Significant Difference 0.2193 Comparisons significant at the 0.05 level are indicated by ***.

Difference Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 58 of 70 Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F 0.08084  0.13841 0.30009 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 59 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT BETWEEN SEXES FOR EACH EXPOSURE GROUP 698  ATRAZINE= 0

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 8.0 10.50 2.291288 2.666667 M 3 13.0 10.50 2.291288 4.333333 Wilcoxon Two Sample Test Statistic 8.0000 Normal Approximation Z  0.8729 One Sided Pr < Z 0.1914 Two Sided Pr > | Z| 0.3827 t Approximation One Sided Pr < Z 0.2113 Two Sided Pr > | Z| 0.4227 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.1905 DF 1 Pr > Chi Square 0.2752 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 1.0 1.50 0.670820 0.333333 M 3 2.0 1.50 0.670820 0.666667 Median Two Sample Test Statistic 1.0000 Z  0.7454 One Sided Pr < Z 0.2280 Two Sided Pr > | Z| 0.4561 Median One Way Analysis Chi Square 0.5556 DF 1 Pr > Chi Square 0.4561 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 60 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT BETWEEN SEXES FOR EACH EXPOSURE GROUP 700  ATRAZINE= 1

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 5.0 5.0 1.290994 2.50 M 2 5.0 5.0 1.290994 2.50 Wilcoxon Two Sample Test Statistic 5.0000 Normal Approximation Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 t Approximation One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 1.0 1.0 0.577350 0.50 M 2 1.0 1.0 0.577350 0.50 Median Two Sample Test Statistic 1.0000 Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Median One Way Analysis Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 61 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT BETWEEN SEXES FOR EACH EXPOSURE GROUP 702  ATRAZINE= 10

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 5.0 5.0 1.290994 2.50 M 2 5.0 5.0 1.290994 2.50 Wilcoxon Two Sample Test Statistic 5.0000 Normal Approximation Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 t Approximation One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 The NPAR1WAY Procedure Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 1.0 1.0 0.577350 0.50 M 2 1.0 1.0 0.577350 0.50 Median Two Sample Test Statistic 1.0000 Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Median One Way Analysis Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 62 of 70 NONPARAMETRIC COMPARISON OF FROG WEIGHT BETWEEN SEXES FOR EACH EXPOSURE GROUP 704  ATRAZINE= 25

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 9.0 10.50 2.291288 3.0 M 3 12.0 10.50 2.291288 4.0 Wilcoxon Two Sample Test Statistic 9.0000 Normal Approximation Z  0.4364 One Sided Pr < Z 0.3313 Two Sided Pr > | Z| 0.6625 t Approximation One Sided Pr < Z 0.3404 Two Sided Pr > | Z| 0.6807 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.4286 DF 1 Pr > Chi Square 0.5127 The NPAR1WAY Procedure Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 1.0 1.50 0.670820 0.333333 M 3 2.0 1.50 0.670820 0.666667 Median Two Sample Test Statistic 1.0000 Z  0.7454 One Sided Pr < Z 0.2280 Two Sided Pr > | Z| 0.4561 Median One Way Analysis Chi Square 0.5556 DF 1 Pr > Chi Square 0.4561 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 63 of 70 ANOVA FOR LENGTH BETWEEN SEXES 706  ATRAZINE= 0

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 6 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.04444191 0.04444191 1.64 0.2697 Error 4 0.10848814 0.02712203 Corrected Total 5 0.15293005 R Square Coeff Var Root MSE WEIGHT Mean 0.290603 13.86664 0.164688 1.187654 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.04444191 0.04444191 1.64 0.2697 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 4 Error Mean Square 0.027122 Critical Value of Dunnett's t 2.77630 Minimum Significant Difference 0.3733 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous Sex Between 95% Confidence Comparison Means Limits M  F 0.1721  0.2012 0.5454 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 64 of 70 ANOVA FOR LENGTH BETWEEN SEXES 709  ATRAZINE= 1

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 4 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00001069 0.00001069 0.00 0.9872 Error 2 0.06567277 0.03283639 Corrected Total 3 0.06568346 R Square Coeff Var Root MSE WEIGHT Mean 0.000163 25.67177 0.181208 0.705865 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.00001069 0.00001069 0.00 0.9872 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 2 Error Mean Square 0.032836 Critical Value of Dunnett's t 4.30263 Minimum Significant Difference 0.7797 Comparisons significant at the 0.05 level are indicated by ***.

Difference Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F 0.003269  0.776402 0.782940 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 65 of 70 ANOVA FOR LENGTH BETWEEN SEXES 712  ATRAZINE= 10

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 4 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00000059 0.00000059 0.00 0.9969 Error 2 0.06049619 0.03024810 Corrected Total 3 0.06049678 R Square Coeff Var Root MSE WEIGHT Mean 0.000010 23.53874 0.173920 0.738866 Source DF Anova SS Mean Square F Value Pr > F Sex 1 5.9017464E 7 5.9017464E 7 0.00 0.9969 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 2 Error Mean Square 0.030248 Critical Value of Dunnett's t 4.30263 Minimum Significant Difference 0.7483 Comparisons significant at the 0.05 level are indicated by ***.

Difference Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F  0.0007682  0.7490802 0.7475438 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 66 of 70 ANOVA FOR LENGTH BETWEEN SEXES 715  ATRAZINE= 25

The ANOVA Procedure Class Level Information Class Levels Values Sex 2 F M Number of observations 6 Dependent Variable: WEIGHT Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00980227 0.00980227 1.05 0.3639 Error 4 0.03742040 0.00935510 Corrected Total 5 0.04722267 R Square Coeff Var Root MSE WEIGHT Mean 0.207575 16.18803 0.096722 0.597489 Source DF Anova SS Mean Square F Value Pr > F Sex 1 0.00980227 0.00980227 1.05 0.3639 Dunnett's t Tests for WEIGHT NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 4 Error Mean Square 0.009355 Critical Value of Dunnett's t 2.77630 Minimum Significant Difference 0.2193 Comparisons significant at the 0.05 level are indicated by ***.

Difference Sex Between Simultaneous 95% Comparison Means Confidence Limits M  F 0.08084  0.13841 0.30009 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 67 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH BETWEEN SEXES FOR EACH EXPOSURE GROUP 718  ATRAZINE= 0

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 8.0 10.50 2.291288 2.666667 M 3 13.0 10.50 2.291288 4.333333 Wilcoxon Two Sample Test Statistic 8.0000 Normal Approximation Z  0.8729 One Sided Pr < Z 0.1914 Two Sided Pr > | Z| 0.3827 t Approximation One Sided Pr < Z 0.2113 Two Sided Pr > | Z| 0.4227 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.1905 DF 1 Pr > Chi Square 0.2752 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 1.0 1.50 0.670820 0.333333 M 3 2.0 1.50 0.670820 0.666667 Median Two Sample Test Statistic 1.0000 Z  0.7454 One Sided Pr < Z 0.2280 Two Sided Pr > | Z| 0.4561 Median One Way Analysis Chi Square 0.5556 DF 1 Pr > Chi Square 0.4561 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 68 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH BETWEEN SEXES FOR EACH EXPOSURE GROUP 720  ATRAZINE= 1

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 5.0 5.0 1.290994 2.50 M 2 5.0 5.0 1.290994 2.50 Wilcoxon Two Sample Test Statistic 5.0000 Normal Approximation Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 t Approximation One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 1.0 1.0 0.577350 0.50 M 2 1.0 1.0 0.577350 0.50 Median Two Sample Test Statistic 1.0000 Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Median One Way Analysis Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 69 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH BETWEEN SEXES FOR EACH EXPOSURE GROUP 722  ATRAZINE= 10

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 5.0 5.0 1.290994 2.50 M 2 5.0 5.0 1.290994 2.50 Wilcoxon Two Sample Test Statistic 5.0000 Normal Approximation Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 t Approximation One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 2 1.0 1.0 0.577350 0.50 M 2 1.0 1.0 0.577350 0.50 Median Two Sample Test Statistic 1.0000 Z 0.0000 One Sided Pr < Z 0.5000 Two Sided Pr > | Z| 1.0000 Median One Way Analysis Chi Square 0.0000 DF 1 Pr > Chi Square 1.0000 Data Evaluation Report on Exposure of Xenopus laevis larvae to Different Concentrations of Atrazine in Semi Natural Microcosms EPA MRID Number 458677 11 Page 70 of 70 NONPARAMETRIC COMPARISON OF FROG LENGTH BETWEEN SEXES FOR EACH EXPOSURE GROUP 724  ATRAZINE= 25

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 9.0 10.50 2.291288 3.0 M 3 12.0 10.50 2.291288 4.0 Wilcoxon Two Sample Test Statistic 9.0000 Normal Approximation Z  0.4364 One Sided Pr < Z 0.3313 Two Sided Pr > | Z| 0.6625 t Approximation One Sided Pr < Z 0.3404 Two Sided Pr > | Z| 0.6807 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.4286 DF 1 Pr > Chi Square 0.5127 Median Scores ( Number of Points Above Median) for Variable WEIGHT Classified by Variable Sex Sum of Expected Std Dev Mean Sex N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ F 3 1.0 1.50 0.670820 0.333333 M 3 2.0 1.50 0.670820 0.666667 Median Two Sample Test Statistic 1.0000 Z  0.7454 One Sided Pr < Z 0.2280 Two Sided Pr > | Z| 0.4561 Median One Way Analysis Chi Square 0.5556 DF 1 Pr > Chi Square 0.4561