Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Data Requirement: EPA DP Barcode D288775 EPA MRID 458677 08 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: March 31, 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: 05/ 31/ 2003 CITATION: Villeneuve, D. L., K. Coady, M. Hecker, M. B. Murphy, P. D. Jones and J. P. Giesy. 2003. Methods development for the study of mechanism of action of atrazine in adult and metamorphosing Xenopus laevis and Rana clamitans: aromatase induction. Aquatic Toxicology Laboratory, Michigan State University, National Food Safety and Toxicology Center, E. Lansing, MI. Sponsor: Syngenta Crop Protection, Inc., Laboratory Study ID ECORISK Number MSU 01. Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 2 of 28 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 3 of 28 EXECUTIVE SUMMARY:

In three separate studies, two involving adult male African clawed frogs ( Xenopus laevis) and one with adult female Xenopus, frogs were exposed to either atrazine at 25

g/ L or to freshwater under static renewal conditions with 50% exposure solution changes every 72 hours. In the first study, 15 males were exposed for 26 days ( single tank per treatment); in the second study 15 males were exposed for 43 days ( single tank per treatment); and in the third study 13 females ( 6 in one replicate and 7 in the second replicate) were exposed for 47 days. Overall mortality was 3%, 7% and 19% in the 26, 43 and 47 day exposures, respectively; mortality was primarily associated with disease ( fungal/ bacterial) and was positively correlated ( r= 0.77) with the number of exposure days. Homogenates from a single testes and from brain were used to measure aromatase activity ( CYP19 gene expression) in males. Aromatase activity in the testes was at or near the level of detection ( LOD = 0.025 fmol/ h/ mg protein). Mean brain aromatase activity from atrazine treated males ( 8.4 ± 4.2 fmol/ h/ mg protein) was not statistically different ( p = 0.678) from controls ( 7.1 ± 4.2 fmol/ h/ mg protein) after 26 days of exposure. Following 43 days of exposure, mean brain aromatase activity in atrazine treated males ( 5.8 ± 3.4 fmol/ h/ mg protein) was not statistically different ( p = 0.199) from controls ( 10.4 ± 7.1 fmol/ h/ mg protein); however, in the second study, atrazine contaminated the controls at levels as high 0.25

g/ L. In the third exposure using female frogs, ovarian aromatase activity in atrazine treated females averaged 4.5 ± 1.7 fmol/ h/ mg protein and did not differ statistically ( p= 0.447) from controls ( 5.4 ± 2.1 fmol/ h/ mg protein). Mean aromatase activity of brain homogenates from atrazine treated females was 7.3 ± 5.0 fmol/ h/ mg protein did not differ significantly ( p= 0.582) from control females ( 8.9 ± 4.2 fmol/ h/ mg protein).

With a single tank per treatment in the first two studies, it is not possible to document tank effects. Hhowever, the authors note that there was considerable variability in aromatase activity between frogs receiving the same treatment with coefficients of variability at or exceeding 100%. The third exposure where treatments were replicated confirmed that tank effects were high enough to potentially confound the ability of the study design to detect treatment effects. Additionally, with only one atrazine concentration tested, the ability of the study to discriminate potential " low dose" effects is limited. Atrazine contamination of controls in the second study further confounded the test's ability to discriminate subtle effects.

The study recommends that future testing use replication, higher sample sizes and a broader range of atrazine levels to test for potentially subtle effects. Although not discussed in the study, water quality may have also compromised the study given the correlation of mortality with the number of days the frogs were confined and the references to the diseased state of the animals that had succumbed. Apparently, Xenopus are susceptible to bacterial septicemia ( red leg disease) if poor water quality persists.

Although it is indicated in the introduction that plasma sex steroids were also analyzed, these data and associated methods were not presented in this report. The only mention of sex steroid measurements was an indication that blood samples were collected by cardiac puncture. Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 4 of 28 I. MATERIALS AND METHODS GUIDELINE FOLLOWED: Nonguideline Study COMPLIANCE: Not conducted under full GLP; however, most practices as defined by 40 CFR Part 160, August 19, 1989 were established for this study, including but not limited to:

° Written, authorized protocol ° Written, authorized Standard Operating Procedures for all key procedures. ° Organization and Personnel were sufficient in terms of number, education, training and experience. ° Facilities were of suitable size and construction ° Equipment used was of appropriate design and adequate capacity. ° Independent A Inspection was conducted of raw data. ° Final Report was written ° Raw data, documentation, records, protocols, and final report was archieved.

A. MATERIALS:

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

Species: African clawed frog ( Xenopus laevis) . Age at test initiation Adults Weight at study initiation: ( mean and range) Not reported Length at study initiation: ( mean and range) Not reported Source: Adult X. laevis obtained from Xenopus Express ® ( Plant City, FL) Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 5 of 28 B. STUDY DESIGN:

Objective: 1. To test hypothesis that waterborne exposure to 25

g/ L atrazine could up regulate aromatase activity in sexually mature X. laevis; male and female X. laevis were tested.

2. Test hypothesis that exposure to atrazine causes decreases in plasma testosterone and increases in estradiol ( consistent with an increase in aromatase activity).

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) several weeks freshwater ( treated well water) not reported mortality observed during study; frog with leision Duration of the test Exposure 1: 26 day Exposure 2: 43 day Exposure 3: 47 day Test condition static/ flow through Type of dilution system for flowthrough method.

Renewal rate for static renewal static renewal NA 50% test solution change every 72 hours Aeration, if any not reported Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Parameter Details Page 6 of 28 Test vessel Material: ( glass/ stainless steel) Size: Fill volume: fiberglass 600 L 120 L Source of dilution water Quality: Treated well water ( MSU University Research Containment Facility;

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 no water quality data provided at all No mention of whether water quality measurements were taken at all Number of replicates/ groups: negative control: water treated ones: atrazine at 25

g/ L Exposure 1: single tank for each treatment ( 25

g atrazine/ L plus control) Exposure 2: single tank for each treatment ( 25

g atrazine/ L plus control) Exposure 3: 2 replicates per treatment Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Parameter Details Page 7 of 28 Number of organisms per replicate / groups: Exposure 1: 15 adult frogs/ treatment Exposure 2: 15 adult frogs/ treatment Exposure 3: 6 adults in Rep 1 and 7 adults in Rep 2 Biomass loading rate Exposure 1 and 2: 15 adults/ 120 L = 0.13 frogs/ L Exposure 3: 6 to 7 adults/ 120 L = 0.05  0.06 frogs/ L Test concentrations: nominal: measured: 25

g/ L Exposure 1: 30.6 ± 2.1

g/ L Exposure 2: 34 ± 8

g/ L Exposure 3: 32 ± 16

g/ L Solvent ( type, percentage, if used) none Lighting not reported Feeding not reported Recovery of chemical Level of Quantitation Level of Detection ELISA ( Envirogard Triazine ® ; Strategic Diagnostics Newark, DE)

LOD 0.025

g/ L ( Envirogard); 0.05

g/ L ( Beacon)

Positive control { if used, indicate the chemical and concentrations} none Other parameters, if any NA 2. Observations:

Table 2: Observations Criteria Details Parameters measured including the sublethal effects/ toxicity symptoms Catalytic activity of aromatase ( CYP19) measured in gonad and brain homogenates. Blood analyzed for steroid hormone levels Observation intervals daily Were raw data included? Yes Other observations, if any Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 8 of 28 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 appeared to differ in any substantive way, the difference was reported in the text below.]

Although blood was collected for steroid hormone analysis, these data were not reported in this study. Catalytic activity of aromatase ( CYP19) was measured using the tritiated water release assay based on method of Lephart and Simpson ( 1991); protein concentration was determined using fluorescamine assay based on method of Kennedy and Jones ( 1994). Aromatase level was expressed as fmol/ h/ mg protein.

Atrazine concentrations in exposure tanks are based on samples collected following solution renewal after 72 hrs static exposure Only 50% of the exposure solutions were changed at renewal though.

Exposure 1 ( 26 days): one atrazine treated frog died apparently from disease ( open lesions on its legs). Single testes were taken for analysis; none of the testes homogenates had measurable aromatase activity; mean aromatase activity to the testicular homogenates was not statistically different among treatments ( p= 0.586). Positive control ( Xenopus ovarian tissue).

Immature Xenopus ovarian exhibited roughly similar aromatase levels as 2 mg/ mL bovine serum albumin; whereas X. laevis adult ovary contained roughly 26 fmol/ h/ mg protein. Mean aromatase activity from atrazinetreated males ( 8.4 ± 4.2 fmol/ h/ mg protein) was not statistically different ( p = 0.678) from controls ( 7.1 ± 4.2 fmol/ h/ mg protein).

Atrazine concentrations averaged 30.6 ± 2.1

g/ L, while the control tank contained less than 0.025

g/ L ( LOD for ELISA).

Exposure 2 ( 43 days): one animal from each treatment died of unknown causes. Animals were reported to appear healthy. Testicular homogenates had low to non detectable levels of aromatase and again, there was no statistical difference ( p = 0.764) between treatments. Positive control X. laevis ovarian homogenates yielded measurable aromatase activity, and activity was inhibited by addition of 7.5

L of 559

M 4 androstein 4 ol 3, 17 dione ( a specific inhibitor of aromatase).

Atrzine treated mean brain aromatase activity ( 5.8 ± 3.4 fmol/ h/ mg protein) was not statistically different ( p = 0.199) from controls ( 10.4 ± 7.1 fmol/ h/ mg protein). A negative control ( Rana pipiens ovarian homogenate) did not yield a significant response in the aromatase assay. There were significant differences among mean aromatase activity in replicate tanks ( tank effects) within each treatment group.

Atrazine concentrations over the 43 day exposure period averaged 34 ± 8

g/ L but ranged as high as 50

g/ L. Atrazine in control tanks ranged up to 0.26

g/ L but average 0.07 ± 0.09

g/ L. Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 9 of 28 Exposure 3 ( 47 days): two atrazine treated and three control females died; overall mortality was 19%. Fungal or bacterial infections were considered the probable cause of the mortalities although the surviving animals are reported as appearing healthy.

Ovarian aromatase activity in atrazine treated females averaged 4.5 ± 1.7 fmol/ h/ mg protein and did not differ statistically ( p= 0.447) from controls ( 5.4 ± 2.1 fmol/ h/ mg protein). The report notes that overall ovarian aromatase activity was approximately 33 times greater than that observed for testicular homogenates from Exposure II.

The mean aromatase activity of brain homogenates from atrazine treated females was 7.3 ± 5.0 fmol/ h/ mg; protein did not differ significantly ( p= 0.582) from control females ( 8.9 ± 4.2 fmol/ h/ mg protein) .

Atrazine concentrations in each of the atrazine groups averaged 32 ± 16

g/ L and 28 ± 15

g/ L; however concentrations ranged as high as 70

g/ L. Atrazine concentrations in control tanks were generally less than the limit of detection and averaged 0.018 ± 0.02

g/ L and 0.016 ± 0.019

g/ L in the two replicates.

The report concludes that there were no statistically significant differences in mean brain homogenate aromatase activity among treatment groups in all three exposures ( p= 0.75). For ovarian homogenates from exposure 3, there were no statistically significant differences between atrazine and control females. As a whole " the results of the three exposures do no support the hypothesis that waterborne exposure to approximately 25

g/ L atrazine causes changes in brain or gonadal aromatase activity in adult X. laevis relative to that in X. laevis exposed to laboratory water."

C. REPORTED STATISTICS:

D. VERIFICATION OF STATISTICAL RESULTS: Basic analyses in attached SAS © [ Statistical Analysis System, Release 8.01, Cary, North Carolina] output)

E. STUDY DEFICIENCIES: Water quality data were not provided; the purity of atrazine was not provided; and atrazine was detected in the negative control in Exposure 2. In Exposures 1 and 3, mortality was attributed to disease. The feeding rate and lighting cycle were not discussed in the study.

F. REVIEWER'S COMMENTS:

It is unclear why a negative control ( Rana pipiens ovarian homogenate) did not yield a significant response in the aromatase assay. In other words, why didn't aromatase from a different species convert testosterone to estrogen since the substrates are presumably the same?

In Exposure 2, atrazine in control tanks ranged up to 0.26

g/ L, but averaged 0.07 ± 0.09

g/ L. While the report states that this suggests " atrazine concentrations in control water was, on average, at least 485 times lower than that in atrazine treated water," the levels detected were well above the detection limit of the ELISA assay ( LOD = 0.025). Atrazine in controls was at 0.1

g/ L on March 18, 0.25

g/ L on March 21, and 0.24

g/ L on March 30. Thus, exposure to atrazine in the controls was not an isolated event and may have confounded the ability of the study to detect treatment related effects. Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 10 of 28 The level of total mortality was positively correlated ( r= 0.77) with the length of exposure and in most cases was associated with either bacterial and/ or fungal disease.

In the second exposure, there were significant differences among mean aromatase activity in replicate tanks ( tank effects) within each treatment group.

Although the report concluded that " the results of the three exposures do no support the hypothesis that waterborne exposure to approximately 25

g/ L atrazine causes changes in brain or gonadal aromatase activity in adult X. laevis relative to that in X. laevis exposed to laboratory water," the authors acknowledged that the magnitude of variability in gonadal and brain aromatase activity among individual frogs from the same treatment limited the power of the experiments to detect subtle differences even if they occurred. Large sample sizes would be required to test for subtle effects. Additionally, with only one tank per treatment in Exposures 1 and 2, the ability to account for tank effects is very limited and as the authors noted could potentially yield a false negative or false positive response.

Furthermore, the studies only examined a single atrazine exposure concentration, i. e., 25

g/ L. The authors note that one hypothesis that has been raised is that high concentrations of atrazine may up regulate aromatase to the point where the estrogen produced causes negative feedback on the hypothalmic pituitary axis resulting in down regulation of aromatase expression. They conclude that future testing should use a wider range of concentrations.

Although it is indicated in the introduction that plasma sex steroids were also analyzed, these data and associated methods were not presented in this report. The only mention of sex steroid measurements was an indication that blood samples were collected by cardiac puncture.

G. CONCLUSIONS:

Aromatase activity in testes was too low to reliably quantify. While atrazine treatment of adult male X. laevis did not significantly affect brain aromatase activity relative to controls, the ability of the current studies to detect treatment effects appears limited. Since mortality over the three studies was positively correlated with the length of the study and because disease appeared to be an issue in two out of three studies, water quality may have been a problem; however, water quality data were not reported for the studies. Additionally, the second study ( Exposure 2) was compromised by the discovery of atrazine in the control exposure. With a single tank per treatment ( no replicates) and a single atrazine treatment ( 25

g/ L) per study, variability within treatments made it difficult to clearly differentiate treatment effects and to test the hypothesis.

H. REFERENCES:

Kennedy, S. W. and S. P. Jones. 1994. Simultaneous measurement of cytochrome P4501A catalyitc activity and total protein concentration with fluorescence plate reader. Analytical Biochemistry 222( 1): 217  223.

Laphart, E. D. and E. R. Simpson. 1991. Assay of aromatase activity. Methods in Enzymology 206: 477  483. Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 11 of 28 NONPARAMETRIC COMPARISON OF GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED 264 Obs SAMPLE _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 0 2 11.1919 5.11863 45.735 2 A10 0 2 2.1999 0.02387 1.085 3 A11 0 2 1.8184 0.09016 4.958 4 A12 0 2 0.6189 0.14642 23.658 5 A13 0 2 18.8785 0.89090 4.719 6 A14 0 2  0.3838 0.34889  90.914 7 A15 0 2  1.7780 0.36618  20.595 8 A2 0 2 19.5097 0.08537 0.438 9 A3 0 2 1.0145 0.35979 35.465 10 A4 0 2 13.9604 0.21978 1.574 11 A7 0 2 17.5941 0.01244 0.071 12 A8 0 2 6.4782 1.36797 21.116 13 A9 0 2 17.6115 0.57149 3.245 14 C1 0 2  0.8158 0.25213  30.907 15 C10 0 2 12.0858 0.74018 6.124 16 C11 0 2 14.7327 0.07916 0.537 17 C12 0 2 3.1996 0.31515 9.850 18 C13 0 2 7.2257 0.47387 6.558 19 C14 0 2 0.5245 0.04568 8.709 20 C15 0 2 12.5170 0.11766 0.940 21 C2 0 2  0.0897 0.25792  287.405 22 C3 0 2 14.6997 5.50738 37.466 23 C4 0 2 1.7527 0.36438 20.790 24 C5 0 2 1.2051 1.46082 121.218 25 C6 0 2 13.9926 3.28426 23.471 26 C8 0 2 18.9569 0.13915 0.734 27 C9 0 2  0.2912 0.07412  25.456 MEAN BRAIN AROMATASE ACTIVITY IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 1) 265 Obs GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 ATRAZIN 0 13 8.36265 8.27945 99.0051 2 CONTROL 0 14 7.12111 7.06613 99.2279 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 12 of 28 ANOVA FOR BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( EXP 1) 266 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 ATRAZIN CONTROL Number of observations 27 Dependent Variable: MEAN Sum of Source DF Squares Mean Square F Value Pr > F Model 1 10.390314 10.390314 0.18 0.6780 Error 25 1471.684807 58.867392 Corrected Total 26 1482.075121 R Square Coeff Var Root MSE MEAN Mean 0.007011 99.39913 7.672509 7.718889 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 10.39031393 10.39031393 0.18 0.6780 Levene's Test for Homogeneity of MEAN Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 1928.1 1928.1 1.54 0.2255 Error 25 31216.8 1248.7 Bartlett's Test for Homogeneity of MEAN Variance Source DF Chi Square Pr > ChiSq GROUP 1 0.3013 0.5831 Dunnett's t Tests for MEAN NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 25 Error Mean Square 58.86739 Critical Value of Dunnett's t 2.05959 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous GROUP Between 95% Confidence Comparison Means Limits CONTROL  ATRAZIN  1.242  7.328 4.845 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 13 of 28 NONPARAMETRIC COMPARISON OF BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FR 270 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 13 190.0 182.0 20.607442 14.615385 CONTROL 14 188.0 196.0 20.607442 13.428571 Wilcoxon Two Sample Test Statistic 190.0000 Normal Approximation Z 0.3639 One Sided Pr > Z 0.3579 Two Sided Pr > | Z| 0.7159 t Approximation One Sided Pr > Z 0.3594 Two Sided Pr > | Z| 0.7188 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.1507 DF 1 Pr > Chi Square 0.6979 Median Scores ( Number of Points Above Median) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 13 6.0 6.259259 1.321968 0.461538 CONTROL 14 7.0 6.740741 1.321968 0.500000 Median Two Sample Test Statistic 6.0000 Z  0.1961 One Sided Pr < Z 0.4223 Two Sided Pr > | Z| 0.8445 Median One Way Analysis Chi Square 0.0385 DF 1 Pr > Chi Square 0.8445 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 14 of 28 MEAN GONADAL AROMATASE ACTIVITY IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 1) 273 Obs SAMPLE _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 0 2 0.40032 1.40426 350.79 2 A10 0 2  0.06537 0.04967  75.99 3 A11 0 2 0.00952 0.22021 2314.17 4 A12 0 2  0.33007 0.17387  52.68 5 A13 0 2  0.04765 0.14865  311.96 6 A14 0 2  0.41950 0.00438  1.04 7 A15 0 2  0.14911 0.14151  94.90 8 A2 0 2  0.52470 0.90598  172.67 9 A3 0 2  0.39147 0.30402  77.66 10 A4 0 2  0.47445 0.00000 0.00 11 A5 0 2 0.11161 0.20003 179.21 12 A7 0 2  0.21175 0.02073  9.79 13 A8 0 2  0.34509 0.02312  6.70 14 A9 0 2  0.08582 0.10868  126.65 15 C1 0 2  0.59029 0.29282  49.61 16 C10 0 2  0.84140 0.11866  14.10 17 C11 0 2  0.01878 0.11230  597.83 18 C12 0 2  0.19693 0.05968  30.30 19 C13 0 2  0.05506 0.02920  53.03 20 C14 0 2  0.03542 0.36070  1018.23 21 C15 0 2 0.40039 0.08395 20.97 22 C2 0 2  0.46908 0.05875  12.52 23 C3 0 2  0.27553 0.13632  49.48 24 C4 0 2  0.38774 0.48604  125.35 25 C5 0 2  0.03013 0.00000 0.00 26 C6 0 2  0.27841 0.00000 0.00 27 C8 0 2  0.33768 0.12295  36.41 28 C9 0 2  0.21742 0.07923  36.44 MEAN GONADAL AROMATASE ACTIVITY IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 1) 273 Obs GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 ATRAZIN 0 14  0.18025 0.25669  142.408 2 CONTROL 0 14  0.23811 0.29774  125.044 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 15 of 28 ANOVA FOR GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( EXP 1) 274 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 ATRAZIN CONTROL Number of observations 28 Dependent Variable: MEAN Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.02342954 0.02342954 0.30 0.5866 Error 26 2.00900245 0.07726932 Corrected Total 27 2.03243199 R Square Coeff Var Root MSE MEAN Mean 0.011528  132.8879 0.277974  0.209179 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 0.02342954 0.02342954 0.30 0.5866 Levene's Test for Homogeneity of MEAN Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 0.00313 0.00313 0.25 0.6229 Error 26 0.3280 0.0126 Bartlett's Test for Homogeneity of MEAN Variance Source DF Chi Square Pr > ChiSq GROUP 1 0.2744 0.6004 Dunnett's t Tests for MEAN NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 26 Error Mean Square 0.077269 Critical Value of Dunnett's t 2.05558 Minimum Significant Difference 0.216 Comparisons significant at the 0.05 level are indicated by ***.

Difference GROUP Between Simultaneous 95% Comparison Means Confidence Limits CONTROL  ATRAZIN  0.05785  0.27382 0.15811 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 16 of 28 NONPARAMETRIC COMPARISON OF GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED 278 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 14 208.0 203.0 21.763884 14.857143 CONTROL 14 198.0 203.0 21.763884 14.142857 Wilcoxon Two Sample Test Statistic 208.0000 Normal Approximation Z 0.2068 One Sided Pr > Z 0.4181 Two Sided Pr > | Z| 0.8362 t Approximation One Sided Pr > Z 0.4189 Two Sided Pr > | Z| 0.8377 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0528 DF 1 Pr > Chi Square 0.8183 Median Scores ( Number of Points Above Median) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 14 8.0 7.0 1.347151 0.571429 CONTROL 14 6.0 7.0 1.347151 0.428571 Median Two Sample Test Statistic 8.0000 Z 0.7423 One Sided Pr > Z 0.2290 Two Sided Pr > | Z| 0.4579 Median One Way Analysis Chi Square 0.5510 DF 1 Pr > Chi Square 0.4579 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 17 of 28 AVERAGE ATRAZINE CONCENTRATIONS IN CONTROL EXPOSURE TANKS BY DATE 280 Obs MONTH DAY _ TYPE_ _ FREQ_ MEAN 1 3 3 0 2 0.03820 2 3 6 0 2 0.06553 3 3 9 0 2 0.01873 4 3 12 0 2 0.00515 5 3 15 0 2 0.03088 6 3 18 0 2 0.11453 7 3 21 0 2 0.26317 8 3 24 0 2 0.01652 9 3 30 0 2 0.22902 10 4 5 0 2 0.00473 11 4 8 0 2 0.00725 AVERAGE ATRAZINE CONCENTRATION IN CONTROL EXPOSURE TANK ACROSS ENTIRE STUDY ( EXP 1) 281 Obs _ TYPE_ _ FREQ_ MEAN STD CV 1 0 11 0.072156 0.092187 127.761 AVERAGE ATRAZINE CONCENTRATIONS IN 25 UG/ L EXPOSURE 1 TANKS BY DATE 282 Obs MONTH DAY _ TYPE_ _ FREQ_ MEAN 1 3 3 0 2 22.5051 2 3 6 0 2 32.4627 3 3 9 0 2 25.4194 4 3 12 0 2 25.8225 5 3 15 0 2 38.2852 6 3 18 0 2 34.8589 7 3 21 0 2 44.3471 8 3 24 0 2 49.5251 9 3 30 0 2 35.0879 10 4 5 0 2 33.9842 11 4 8 0 2 32.9775 AVERAGE ATRAZINE CONCENTRATION IN 25 UG/ L EXPOSURE TANK ACROSS ENTIRE STUDY 283 Obs _ TYPE_ _ FREQ_ MEAN STD CV 1 0 11 34.1160 8.02663 23.5275 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 18 of 28 MEAN BRAIN AROMATASE ACTIVITY FOLLOWING EXPOSURE 2 284 Obs _ TYPE_ _ FREQ_ MEAN STD CV 1 0 28 8.07322 9.41514 116.622 ANOVA FOR BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( EXPOSURE 2 285 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 EXP REF Number of observations 28 Dependent Variable: FMOL Sum of Source DF Squares Mean Square F Value Pr > F Model 1 150.032040 150.032040 1.74 0.1988 Error 26 2243.380177 86.283853 Corrected Total 27 2393.412216 R Square Coeff Var Root MSE FMOL Mean 0.062685 115.0583 9.288910 8.073222 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 150.0320396 150.0320396 1.74 0.1988 Levene's Test for Homogeneity of FMOL Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 70229.1 70229.1 3.77 0.0632 Error 26 484906 18650.2 Bartlett's Test for Homogeneity of FMOL Variance Source DF Chi Square Pr > ChiSq GROUP 1 6.2027 0.0128 Dunnett's t Tests for FMOL NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 26 Error Mean Square 86.28385 Critical Value of Dunnett's t 2.05558 Minimum Significant Difference 7.2169 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous GROUP Between 95% Confidence Comparison Means Limits REF  EXP 4.630  2.587 11.846 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 19 of 28 NONPARAMETRIC COMPARISON OF BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FR 289 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable FMOL Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ EXP 14 186.0 203.0 21.763884 13.285714 REF 14 220.0 203.0 21.763884 15.714286 Wilcoxon Two Sample Test Statistic 186.0000 Normal Approximation Z  0.7581 One Sided Pr < Z 0.2242 Two Sided Pr > | Z| 0.4484 t Approximation One Sided Pr < Z 0.2275 Two Sided Pr > | Z| 0.4549 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.6101 DF 1 Pr > Chi Square 0.4347 Median Scores ( Number of Points Above Median) for Variable FMOL Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ EXP 14 6.0 7.0 1.347151 0.428571 REF 14 8.0 7.0 1.347151 0.571429 Median Two Sample Test Statistic 6.0000 Z  0.7423 One Sided Pr < Z 0.2290 Two Sided Pr > | Z| 0.4579 Median One Way Analysis Chi Square 0.5510 DF 1 Pr > Chi Square 0.4579 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 20 of 28 MEAN ADULT MALE GONADAL AROMATASE ACTIVITY IN ATRAZINE TREATED AND CONTROL FROGS ( EXP 2) 291 Obs GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 ATRAZIN 0 14 0.13818 0.09607 69.525 2 CONTROL 0 14 0.15601 0.19786 126.827 ANOVA FOR ADULT MALE GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( 292 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 ATRAZIN CONTROL Number of observations 28 Dependent Variable: FMOL Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.00222571 0.00222571 0.09 0.7640 Error 26 0.62891899 0.02418919 Corrected Total 27 0.63114470 R Square Coeff Var Root MSE FMOL Mean 0.003526 105.7346 0.155529 0.147094 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 0.00222571 0.00222571 0.09 0.7640 Levene's Test for Homogeneity of FMOL Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 0.00540 0.00540 1.08 0.3074 Error 26 0.1296 0.00499 Bartlett's Test for Homogeneity of FMOL Variance Source DF Chi Square Pr > ChiSq GROUP 1 6.0347 0.0140 Dunnett's t Tests for FMOL NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control. Alpha 0.05 Error Degrees of Freedom 26 Error Mean Square 0.024189 Critical Value of Dunnett's t 2.05558 Minimum Significant Difference 0.1208 Comparisons significant at the 0.05 level are indicated by ***.

Difference GROUP Between Simultaneous 95% Comparison Means Confidence Limits CONTROL  ATRAZIN 0.01783  0.10300 0.13867 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 21 of 28 NONPARAMETRIC COMPARISON OF ADULT MALE GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZI 296 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable FMOL Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 14 224.0 203.0 21.763884 16.0 CONTROL 14 182.0 203.0 21.763884 13.0 Wilcoxon Two Sample Test Statistic 224.0000 Normal Approximation Z 0.9419 One Sided Pr > Z 0.1731 Two Sided Pr > | Z| 0.3462 t Approximation One Sided Pr > Z 0.1773 Two Sided Pr > | Z| 0.3546 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.9310 DF 1 Pr > Chi Square 0.3346 Median Scores ( Number of Points Above Median) for Variable FMOL Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score âââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZIN 14 9.0 7.0 1.347151 0.642857 CONTROL 14 5.0 7.0 1.347151 0.357143 Median Two Sample Test Statistic 9.0000 Z 1.4846 One Sided Pr > Z 0.0688 Two Sided Pr > | Z| 0.1376 Median One Way Analysis Chi Square 2.2041 DF 1 Pr > Chi Square 0.1376 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 22 of 28 MEAN ATRAZINE LEVEL BY SAMPLING DATE FOR CONTROL ( C) AND ATRAZINE ( A) TREATED TANKS 298 Obs MONTH DAY SAMPLE GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 4 17 A1 ATRAZINE 0 2 21.3668 0.25186 1.1788 2 4 17 A2 ATRAZINE 0 2 6.8501 0.20184 2.9465 3 4 17 C1 CONTROL 0 2 0.0072 0.00365 50.6899 4 4 17 C2 CONTROL 0 2 0.0041 0.00070 17.0096 5 4 20 A1 ATRAZINE 0 2 22.4385 2.50752 11.1751 6 4 20 A2 ATRAZINE 0 2 4.5276 1.69332 37.3999 7 4 20 C1 CONTROL 0 2 0.0070 0.00132 18.7495 8 4 20 C2 CONTROL 0 2 0.0254 0.00179 7.0668 9 4 23 A1 ATRAZINE 0 2 18.2464 3.42110 18.7495 10 4 23 A2 ATRAZINE 0 2 32.0647 4.89450 15.2644 11 4 23 C1 CONTROL 0 2 0.0769 0.00724 9.4163 12 4 23 C2 CONTROL 0 2 0.0771 0.00045 0.5894 13 5 2 A1 ATRAZINE 0 2 25.6397 2.86526 11.1751 14 5 2 A2 ATRAZINE 0 2 17.2724 6.55444 37.9475 15 5 2 C1 CONTROL 0 2 0.0234 0.00963 41.2067 16 5 2 C2 CONTROL 0 2 0.0186 0.00865 46.5324 17 5 5 A1 ATRAZINE 0 2 17.7872 4.56176 25.6463 18 5 5 A2 ATRAZINE 0 2 17.4941 5.47574 31.3004 19 5 5 C1 CONTROL 0 2 0.0057 0.00027 4.7134 20 5 5 C2 CONTROL 0 2 0.0044 0.00008 1.7681 21 5 8 A1 ATRAZINE 0 2 21.1917 0.49956 2.3573 22 5 8 C1 CONTROL 0 2 0.0068 0.00052 7.6546 23 5 8 C2 CONTROL 0 2 0.0048 0.00057 11.7606 24 5 11 C1 CONTROL 0 2 0.0636 0.01413 22.2114 25 5 11 C2 CONTROL 0 2 0.0121 0.00445 36.8512 26 5 14 C1 CONTROL 0 2 0.0202 0.00107 5.3020 27 5 14 C2 CONTROL 0 2 0.0140 0.00530 37.9475 28 5 17 C1 CONTROL 0 2 0.0064 0.00300 46.5324 29 5 17 C2 CONTROL 0 2 0.0072 0.00025 3.5356 30 5 20 C1 CONTROL 0 2 0.0058 0.00219 37.9475 31 5 20 C2 CONTROL 0 2 0.0040 0.00073 18.1701 32 5 23 C1 CONTROL 0 2 0.0110 0.00672 61.1231 33 5 23 C2 CONTROL 0 2 0.0203 0.00892 43.8868 34 5 26 C1 CONTROL 0 2 0.0094 0.00094 10.0029 35 5 26 C2 CONTROL 0 2 0.0177 0.00239 13.5146 36 5 29 C1 CONTROL 0 2 0.0055 0.00197 35.7501 37 5 29 C2 CONTROL 0 2 0.0055 0.00065 11.7606 38 6 1 C1 CONTROL 0 2 0.0064 0.00101 15.8467 39 6 1 C2 CONTROL 0 2 0.0047 0.00087 18.7494 MEAN ATRAZINE CONCENTRATION ACROSS SAMPLING DATES FOR EACH OF THE TWO CONTROL ( C) AND ATRAZI 299 Obs SAMPLE GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 ATRAZINE 0 6 21.1117 2.8844 13.662 2 A2 ATRAZINE 0 5 15.6418 10.9158 69.786 3 C1 CONTROL 0 14 0.0182 0.0229 125.436 4 C2 CONTROL 0 14 0.0157 0.0191 121.373 MEAN ATRAZINE CONCENTRATION IN CONTROL AND ATRAZINE TREATED TANKS OVER ENTIRE STUDY ( EXP 3 300 Obs GROUP _ TYPE_ _ FREQ_ MEAN MIN MAX STD CV 1 ATRAZINE 0 11 18.6254 4.52761 32.0647 7.74480 41.582 2 CONTROL 0 28 0.0170 0.00403 0.0771 0.02070 121.980 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 23 of 28 MEAN BRAIN AROMATASE ACTIVITY BY SAMPLE AND NUMBER IN CONTROL ( C) AND ATRAZINE TREATETED ( A) 301 Obs SAMPLE NUMBER GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 1 ATRAZINE 0 2 20.9131 0.52971 2.5329 2 A1 2 ATRAZINE 0 2 1.9509 0.02533 1.2982 3 A1 3 ATRAZINE 0 2 1.3884 0.00843 0.6070 4 A1 4 ATRAZINE 0 2 3.8467 0.18493 4.8075 5 A1 5 ATRAZINE 0 2 5.6692 0.02706 0.4774 6 A1 6 ATRAZINE 0 2 0.9863 0.02995 3.0367 7 A2 1 ATRAZINE 0 2 7.5578 0.14937 1.9764 8 A2 2 ATRAZINE 0 2 8.7285 1.14801 13.1524 9 A2 3 ATRAZINE 0 2 7.6883 0.05418 0.7047 10 A2 4 ATRAZINE 0 2  0.0514 0.00540  10.5056 11 A2 5 ATRAZINE 0 2 21.1885 0.02985 0.1409 12 C1 1 CONTROL 0 2 11.8105 0.01860 0.1575 13 C1 2 CONTROL 0 2 2.1175 0.20755 9.8015 14 C1 3 CONTROL 0 2 6.2637 0.10634 1.6977 15 C1 4 CONTROL 0 2 21.4478 0.23323 1.0874 16 C1 5 CONTROL 0 2 4.1599 0.02262 0.5438 17 C2 1 CONTROL 0 2 15.0054 2.98537 19.8953 18 C2 2 CONTROL 0 2 1.0907 0.04346 3.9843 19 C2 3 CONTROL 0 2 7.0063 0.10293 1.4692 20 C2 4 CONTROL 0 2 8.3976 0.10458 1.2454 21 C2 5 CONTROL 0 2 12.2114 0.15241 1.2481 MEAN BRAIN AROMATASE ACTIVITY BY SAMPLE IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 302 Obs SAMPLE _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 0 6 5.79244 7.61168 131.407 2 A2 0 5 9.02234 7.65441 84.838 3 C1 0 5 9.15987 7.76212 84.741 4 C2 0 5 8.74230 5.31368 60.781 MEAN BRAIN AROMATASE ACTIVITY IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 303 Obs GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 ATRAZINE 0 11 7.26057 7.43303 102.375 2 CONTROL 0 10 8.95109 6.27499 70.103 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 24 of 28 ANOVA FOR BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 304 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 ATRAZINE CONTROL Number of observations 21 Dependent Variable: MEAN Sum of Source DF Squares Mean Square F Value Pr > F Model 1 14.9695821 14.9695821 0.31 0.5820 Error 19 906.8789562 47.7304714 Corrected Total 20 921.8485383 R Square Coeff Var Root MSE MEAN Mean 0.016239 85.65689 6.908724 8.065579 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 14.96958206 14.96958206 0.31 0.5820 Levene's Test for Homogeneity of MEAN Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 1145.7 1145.7 0.31 0.5865 Error 19 71107.0 3742.5 Bartlett's Test for Homogeneity of MEAN Variance Source DF Chi Square Pr > ChiSq GROUP 1 0.2554 0.6133 Dunnett's t Tests for MEAN NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 19 Error Mean Square 47.73047 Critical Value of Dunnett's t 2.09309 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous GROUP Between 95% Confidence Comparison Means Limits CONTROL  ATRAZINE 1.691  4.628 8.009 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 25 of 28 NONPARAMETRIC COMPARISON OF BRAIN AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FR 308 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZINE 11 107.0 121.0 14.200939 9.727273 CONTROL 10 124.0 110.0 14.200939 12.400000 Wilcoxon Two Sample Test Statistic 124.0000 Normal Approximation Z 0.9506 One Sided Pr > Z 0.1709 Two Sided Pr > | Z| 0.3418 t Approximation One Sided Pr > Z 0.1766 Two Sided Pr > | Z| 0.3531 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.9719 DF 1 Pr > Chi Square 0.3242 Median Scores ( Number of Points Above Median) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZINE 11 5.0 5.238095 1.171274 0.454545 CONTROL 10 5.0 4.761905 1.171274 0.500000 Median Two Sample Test Statistic 5.0000 Z 0.2033 One Sided Pr > Z 0.4195 Two Sided Pr > | Z| 0.8389 Median One Way Analysis Chi Square 0.0413 DF 1 Pr > Chi Square 0.8389 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 26 of 28 MEAN GONADAL AROMATASE ACTIVITY BY SAMPLE AND NUMBER IN CONTROL ( C) AND ATRAZINE TREATETED ( 310 Obs SAMPLE NUMBER GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 1 ATRAZINE 0 2 4.3309 0.13618 3.1445 2 A1 2 ATRAZINE 0 2 3.8744 0.02522 0.6510 3 A1 3 ATRAZINE 0 2 7.3398 0.02316 0.3155 4 A1 4 ATRAZINE 0 2 5.1150 0.06204 1.2129 5 A1 5 ATRAZINE 0 2 8.3364 0.01106 0.1327 6 A1 6 ATRAZINE 0 2 7.3282 0.02467 0.3366 7 A2 1 ATRAZINE 0 2 1.7495 0.02643 1.5105 8 A2 2 ATRAZINE 0 2 4.9783 0.12473 2.5055 9 A2 3 ATRAZINE 0 2 3.3030 0.07831 2.3710 10 A2 4 ATRAZINE 0 2 0.2169 0.02263 10.4327 11 A2 5 ATRAZINE 0 2 2.8070 0.34990 12.4655 12 C1 1 CONTROL 0 2 3.5497 0.02948 0.8304 13 C1 2 CONTROL 0 2 1.6790 0.04290 2.5551 14 C1 3 CONTROL 0 2 4.5738 0.50058 10.9444 15 C1 4 CONTROL 0 2 3.9795 0.02188 0.5497 16 C1 5 CONTROL 0 2 3.1596 0.03938 1.2463 17 C2 1 CONTROL 0 2 10.6332 0.14104 1.3264 18 C2 2 CONTROL 0 2 9.6644 0.09341 0.9665 19 C2 3 CONTROL 0 2 3.3400 1.00685 30.1450 20 C2 4 CONTROL 0 2 6.0585 0.00074 0.0122 21 C2 5 CONTROL 0 2 7.5248 0.31761 4.2209 MEAN GONADAL AROMATASE ACTIVITY BY SAMPLE IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 311 Obs SAMPLE _ TYPE_ _ FREQ_ MEAN STD CV 1 A1 0 6 6.05412 1.84866 30.5355 2 A2 0 5 2.61093 1.77449 67.9639 3 C1 0 5 3.38835 1.09028 32.1774 4 C2 0 5 7.44418 2.90987 39.0892 MEAN GONADAL AROMATASE ACTIVITY IN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 312 Obs GROUP _ TYPE_ _ FREQ_ MEAN STD CV 1 ATRAZINE 0 11 4.48904 2.49031 55.4753 2 CONTROL 0 10 5.41627 2.97674 54.9592 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 27 of 28 ANOVA FOR GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED FROGS ( EXP 3) 313 The ANOVA Procedure Class Level Information Class Levels Values GROUP 2 ATRAZINE CONTROL Number of observations 21 Dependent Variable: MEAN Sum of Source DF Squares Mean Square F Value Pr > F Model 1 4.5034675 4.5034675 0.60 0.4468 Error 19 141.7650045 7.4613160 Corrected Total 20 146.2684720 R Square Coeff Var Root MSE MEAN Mean 0.030789 55.40006 2.731541 4.930574 Source DF Anova SS Mean Square F Value Pr > F GROUP 1 4.50346755 4.50346755 0.60 0.4468 Levene's Test for Homogeneity of MEAN Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F GROUP 1 28.6086 28.6086 0.49 0.4920 Error 19 1107.1 58.2672 Bartlett's Test for Homogeneity of MEAN Variance Source DF Chi Square Pr > ChiSq GROUP 1 0.2867 0.5923 Dunnett's t Tests for MEAN NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 19 Error Mean Square 7.461316 Critical Value of Dunnett's t 2.09309 Comparisons significant at the 0.05 level are indicated by ***.

Difference GROUP Between Simultaneous 95% Comparison Means Confidence Limits CONTROL  ATRAZINE 0.9272  1.5709 3.4253 Data Evaluation Report on Methods Development for the Study of Mechanism of Action of Atrazine in Adult and Metamorphosing Xenopus laevis and Rana clamitans: Aromatase Induction EPA MRID Number 458677 08 Page 28 of 28 NONPARAMETRIC COMPARISON OF GONADAL AROMATASE ACTIVITY BETWEEN CONTROL AND ATRAZINE TREATED 317 The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZINE 11 113.0 121.0 14.200939 10.272727 CONTROL 10 118.0 110.0 14.200939 11.800000 Wilcoxon Two Sample Test Statistic 118.0000 Normal Approximation Z 0.5281 One Sided Pr > Z 0.2987 Two Sided Pr > | Z| 0.5974 t Approximation One Sided Pr > Z 0.3016 Two Sided Pr > | Z| 0.6032 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.3174 DF 1 Pr > Chi Square 0.5732 Median Scores ( Number of Points Above Median) for Variable MEAN Classified by Variable GROUP Sum of Expected Std Dev Mean GROUP N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ ATRAZINE 11 5.0 5.238095 1.171274 0.454545 CONTROL 10 5.0 4.761905 1.171274 0.500000 Median Two Sample Test Statistic 5.0000 Z 0.2033 One Sided Pr > Z 0.4195 Two Sided Pr > | Z| 0.8389 Median One Way Analysis Chi Square 0.0413 DF 1 Pr > Chi Square 0.8389


59173 Federal Register / Vol. 68, No. 198 / Tuesday, October 14, 2003 / Notices Comments: Comments presented during the meeting should be submitted to Edocket. In addition, please submit one unbound original set of comments with pages numbered consecutively at the meeting. For attachments, provide an index and number pages consecutively with the comments. Because all comments received will be placed in Edocket, commentors should not submit personal information ( such as medical data or home address), Confidential Business Information, or information protected by copyright.

SUPPLEMENTARY INFORMATION: During their September 2002 review of the Agency's draft action plan for the development of a metals framework, the EPA Science Advisory Board emphasized the importance of engaging the outside community so as to contribute to the knowledge base the Agency would draw from in developing guidance on the risk assessment of metals. As part of the effort to engage stakeholders and the scientific community and to build on existing experience, the Agency commissioned ERG to organize external experts to lead the development of scientific papers on issues and state of the art approaches to metals risk assessment. ( Some individual EPA experts contributed specific discussions on topic( s) for which he or she has scientific expertise or knowledge of current Agency practice.) On September 22, 2003, EPA published a Federal Register Notice announcing the availability of, and opportunity to comment on, five draft scientific white papers titled: ( 1) Issue Paper on the Environmental Chemistry of Metals, ( 2) Issue Paper on Metal Exposure Assessment, ( 3) Issue Paper on the Ecological Effects of Metals, ( 4) Issue Paper on the Human Health Effects of Metals, and ( 5) Issue Paper on the Bioavailability and Bioaccumulation of Metals. The draft issue papers are available primarily via the Internet on the Risk Assessment Forum's web page at http:// cfpub. epa. gov/ ncea/ raf/ recordisplay. cfm? deid= 59052. Comments will be received on the draft papers until November 7, 2003. The public meeting will provide further opportunity for the Agency to receive stakeholder input. EPA is particularly interested in the public's perspectives on the following: 1. For the purpose of deriving general principles that can be applied in the assessment of metals, do the issue papers provide an appropriate level of detail? 2. Are there additional chemical, biological and physical processes that should be considered for metals assessment? If so, please describe and provide references. 3. Are you aware of any models, approaches or methods not considered in the reports that, if implemented, would substantially reduce uncertainty in the Agency's metal assessments? If so, which ones are ready for application now ( or in the next few years), and which types of assessments would benefit most from their application ( e. g., hazard ranking/ characterization, national, or site specific assessments)? 4. What other suggestions do you have to improve the utility of these papers as the Agency develops a metals assessment framework? The purpose of this meeting is for EPA to receive additional scientific comment from the public on the issue papers. Therefore, although EPA staff will be present, EPA will not evaluate or respond to comments at the meeting. Likewise, EPA is not soliciting comments on past Agency actions. The views expressed in the draft papers are those of the authors and do not necessarily reflect the views or policies of the EPA. For this reason, disposition of scientific comments received on these papers will be made at the author's or author team's discretion. The material contained in the papers may be used in total, or in part, as source material for the Agency's framework for metals risk assessment and EPA's evaluation of this material will therefore include consideration of the Assessment Factors recently published by EPA for use in evaluating the quality of scientific and technical information.

Dated: October 7, 2003. Peter W. Preuss, Director, National Center for Environmental Assessment. [ FR Doc. 03   25935 Filed 10   10   03; 8: 45 am]

BILLING CODE 6560   50   P ENVIRONMENTAL PROTECTION AGENCY [ OPP   2003   0342; FRL   7331   4]

Pesticide Program Dialogue Committee and PPDC Registration Review Work Group; Notice of Public Meetings AGENCY: Environmental Protection Agency ( EPA). ACTION: Notice.

SUMMARY: EPA's Pesticide Program Dialogue Committee ( PPDC) will hold a public meeting on October 29 and 30, 2003. An agenda is being developed and will be posted on EPA's website. Agenda topics will include a report from the PPDC's Registration Review Work Group; endangered species; nongovernmental stewardship activities; pesticide budget/ fees and output efficiencies; certification and training; and follow up to issues discussed at the April 2003 PPDC meeting. In addition, the PPDC Registration Review Work Group will hold two separate public meetings on the following dates: October 17 and October 28, 2003. Agendas for the PPDC Registration Review Work Group meetings will also be posted on EPA's website. DATES: The PPDC meeting will be held on Wednesday, October 29, 2003, from 9 a. m. to 5: 30 p. m., and Thursday, October 30, 2003, from 9 a. m. to 1 p. m. The PPDC Registration Review Work Group will hold a public meeting on Friday, October 17, 2003, from 8: 30 a. m. to 5 p. m., and on Tuesday, October 28, 2003, from 9 a. m. to 5 p. m. ADDRESSES: The PPDC meeting will be held at the National Rural Electric Cooperative Association ( NRECA) Conference Center, 4301 Wilson Boulevard, Arlington, VA. The telephone number to the NRECA Conference Center is ( 703) 907   5500. The NRECA Conference Center is located adjacent to the Ballston Metro Station and about a 15 minute taxi ride from Ronald Reagan Washington National Airport. The PPDC Registration Review Work Group meeting on October 17, 2003, will be held at EPA's Offices at 1921 Jefferson Davis Highway, Crystal Mall # 2, Room 1110 ( the Fishbowl), Arlington, VA. The PPDC Registration Review Work Group meeting on October 28, 2003, will be held at the Hyatt Regency Crystal City, 2799 Jefferson Davis Highway, Arlington, VA. The telephone number to the Hyatt Regency Crystal City is ( 703) 413   6873.

FOR FURTHER INFORMATION CONTACT: Margie Fehrenbach, Office of Pesticide Programs, Mail code 7501C, Environmental Protection Agency, 1200 Pennsylvania Avenue, NW., Washington, DC 20460; telephone number: 703   308   4775; fax number: 703   308   4776; e mail address: Fehrenbach. Margie@ epa. gov.

SUPPLEMENTARY INFORMATION:

I. General Information A. Does this Action Apply to Me?

This action is directed to the public in general; however, persons may be interested who work in agricultural settings or persons who are concerned about implementation of the Federal VerDate jul< 14> 2003 15: 29 Oct 10, 2003 Jkt 203001 PO 00000 Frm 00020 Fmt 4703 Sfmt 4703 E:\ FR\ FM\ 14OCN1. SGM 14OCN1 59174 Federal Register / Vol. 68, No. 198 / Tuesday, October 14, 2003 / Notices Insecticide, Fungicide, and Rodenticide Act ( FIFRA); the Federal Food, Drug, and Cosmetic Act ( FFDCA); and the amendments to both of these major pesticide laws by the Food Quality Protection Act ( FQPA) of 1996. Since other entities may also be interested, the Agency has not attempted to describe all the specific entities that may be affected by this action. If you have any questions regarding the applicability of this action to a particular entity, consult the person listed under FOR FURTHER INFORMATION CONTACT. Potentially affected entities may include but are not limited to: Agricultural workers and farmers; pesticide industry and trade associations; environmental, consumer and farmworker groups; pesticide users and growers; pest consultants; State, local and Tribal governments; academia; public health organizations; food processors; and the public. If you have any questions regarding the applicability of this action to a particular entity, consult the person listed under FOR FURTHER INFORMATION CONTACT.

B. How Can I Get Copies of this Document and Other Related Information?

1. Docket. EPA has established an official public docket for this action under docket identification ( ID) number OPP   2003   0342. The official public docket consists of the documents specifically referenced in this action, any public comments received, and other information related to this action. Although a part of the official docket, the public docket does not include Confidential Business Information ( CBI) or other information whose disclosure is restricted by statute. The official public docket is the collection of materials that is available for public viewing at the Public Information and Records Integrity Branch ( PIRIB), Rm. 119, Crystal Mall # 2, 1921 Jefferson Davis Hwy., Arlington, VA. This docket facility is open from 8: 30 a. m. to 4 p. m., Monday through Friday, excluding legal holidays. The docket telephone number is ( 703) 305   5805. 2. Electronic access. You may access this Federal Register document electronically through the EPA Internet under the `` Federal Register'' listings at http:// www. epa. gov/ fedrgstr/. An agenda for the PPDC meeting is being developed and will be posted on EPA's website at www. epa. gov/ pesticides/ ppdc. Agendas for the PPDC Registration Review Work Group meetings will also be posted on EPA's website at www. epa. gov/ pesticides/ ppdc. An electronic version of the public docket is available through EPA's electronic public docket and comment system, EPA Dockets. You may use EPA Dockets at http:// www. epa. gov/ edocket/ to view public comments, access the index listing of the contents of the official public docket, and to access those documents in the public docket that are available electronically. Although not all docket materials may be available electronically, you may still access any of the publicly available docket materials through the docket facility identified in Unit I. B. 1. Once in the system, select `` search,'' then key in the appropriate docket ID number.

II. Background The Office of Pesticide Programs is entrusted with the responsibility of ensuring the safety of the American food supply, the protection and education of those who apply or are exposed to pesticides occupationally or through use of products from unreasonable risk, and general protection of the environment and special ecosystems from potential risks posed by pesticides. PPDC was established under the Federal Advisory Committee Act ( FACA), Public Law 92   463 in September 1995, for a 2   year term and renewed every 2 years since that time. EPA is seeking to renew the current PPDC Charter, which expires in November 2003, for another 2   year term. PPDC provides advice and recommendations to the Office of Pesticide Programs on a broad range of pesticide regulatory, policy and program implementation issues that are associated with evaluating and reducing risks from use of pesticides. The following sectors are represented on the PPDC: Pesticide industry and trade associations; environmental/ public interest and consumer groups; farm worker organizations; pesticide user, grower, and commodity groups; Federal and State/ local/ Tribal governments; the general public; academia; and public health organizations. Copies of the PPDC Charter are filed with appropriate committees of Congress and the Library of Congress and are available upon request.

List of Subjects Environmental protection, Agricultural workers, Agriculture, Chemicals, Foods, Pesticides, Pests, Public health, Risk assessment, Tolerance reassessment. Dated: October 9, 2003. James Jones, Director, Office of Pesticide Programs.

[ FR Doc. 03   25984 Filed 10   9   03; 10: 10 am]

BILLING CODE 6560   50   S ENVIRONMENTAL PROTECTION AGENCY [ FRL   7574   2]

Integrated Risk Information System ( IRIS); Administrative Changes AGENCY: U. S. Environmental Protection Agency. ACTION: Notice; administrative changes in the Integrated Risk Information System hotline and records center.

SUMMARY: IRIS is an EPA data base that contains EPA scientific consensus positions on human health effects that may result from chronic exposure to chemical substances in the environment. In this action, EPA is announcing an upcoming change to the location and phone number of EPA's contractor operated IRIS hotline and publically accessible records center. DATES: On or about October 17, 2003. ADDRESSES: The new address of the IRIS Hotline and records center will be IRIS Hotline, ( Mail Code 28221T) EPA West Building, 1301 Constitution Avenue, NW, Washington DC 20005. FOR FURTHER INFORMATION CONTACT: For information on this administrative change, contact Rick Johnson, IRIS staff, National Center for Environmental Assessment ( mail code 8601D), Office of Research and Development, U. S. Environmental Protection Agency, Washington, DC 20460, or call ( 202) 564   3291, or send electronic mail inquiries to johnson. rickc@ epa. gov. For information on the IRIS program, contact Amy Mills, Program Director, National Center for Environmental Assessment ( mail code 8601D), Office of Research and Development, U. S. Environmental Protection Agency, Washington, DC 20460, or call ( 202) 564   3204, or send electronic mail inquiries to mills. amy@ epa. gov. SUPPLEMENTARY INFORMATION:

Background IRIS is an EPA data base containing Agency consensus scientific positions on potential adverse human health effects that may result from chronic ( or lifetime) exposure to chemical substances found in the environment. IRIS currently provides information on health effects associated with more than 500 chemical substances. EPA has provided outreach to facilitate the use VerDate jul< 14> 2003 15: 29 Oct 10, 2003 Jkt 203001 PO 00000 Frm 00021 Fmt 4703 Sfmt 4703 E:\ FR\ FM\ 14OCN1. SGM 14OCN1


[Federal Register: March 20, 2007 (Volume 72, Number 53)]
[Notices]
[Page 13156]
[DOCID:fr20mr07-103]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-55447; File No. SR-NYSEArca-2006-50]

Self-Regulatory Organizations; NYSE Arca, Inc.; Order Granting
Approval of Proposed Rule Change Relating to Amendments to Registration
Rules of NYSE Arca Equities, Inc.

March 12, 2007.

I. Introduction

    On November 14, 2006, NYSE Arca, Inc. (``NYSE Arca'' or
``Exchange''), through its wholly owned subsidiary NYSE Arca Equities,
Inc. (``NYSE Arca Equities'' or ``Corporation''), filed with the
Securities and Exchange Commission (``Commission'') pursuant to Section
19(b)(1) of the Securities Exchange Act of 1934 (``Act'') \1\ and Rule
19b-4 thereunder,\2\ a proposed rule change relating to amendments to
registration rules of the Corporation. NYSE Arca filed Amendment No. 1
to the proposed rule change on January 12, 2007. The proposed rule
change, as amended, was published for comment in the Federal Register
on February 7, 2007.\3\ The Commission received no comments on the
proposal. This order approves the proposed rule change, as amended.
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    \1\ 15 U.S.C. 78s(b)(l).
    \2\ 17 CFR 240.19b-4.
    \3\ See Securities Exchange Act Release No. 55214 (January 31,
2007), 72 FR 5780 (February 7, 2007).
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II. Description of the Proposal

    The Exchange, through its wholly owned subsidiary NYSE Arca
Equities, proposed to amend certain NYSE Arca Equities Rules governing
registration procedures and ongoing compliance obligations for Equity
Trading Permit (``ETP'') Holders \4\ and their registered persons in
order to clarify registration procedures and make them consistent with
the procedures of other self-regulatory organizations (``SROs'') and
with the operation of the Central Registration Depository (``CRD'')
system maintained by the National Association of Securities Dealers,
Inc. (``NASD'').
---------------------------------------------------------------------------

    \4\ See NYSE Arca Equities Rule 1.1(n).
---------------------------------------------------------------------------

    Specifically, the Exchange proposed to amend Rule 2.4(c), its
waiver standards, so that the Exchange's practices are generally
consistent with the criterion in NASD Rule 1070(d) and Supplementary
Material .15(1)(b) to NYSE Rule 345. The Exchange also proposed to
amend Rule 2.21 to provide manual registration procedures for
registration categories (e.g., floor clerk) for which CRD does not
provide electronic registration. In addition, the Exchange is
consolidating its continuing education requirements in Rule 2.21(d) and
deleting the continuing education requirements in Rule 9.27(c) and (d)
to avoid needless repetition and risk of inconsistencies.

III. Discussion and Commission Findings

    The Commission has reviewed carefully the proposed rule change and
finds that it is consistent with the requirements of the Act and the
rules and regulations thereunder applicable to a national securities
exchange.\5\ In particular, the Commission finds that the proposed rule
change is consistent with Section 6(b)(5) of the Act,\6\ which, among
other things, requires that the rules of a national securities exchange
be designed to promote just and equitable principles of trade, to
foster cooperation and coordination with persons engaged in regulating
transactions in securities, to remove impediments to and perfect the
mechanism of a free and open market and a national market system and,
in general, to protect investors and the public interest.
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    \5\ In approving this proposed rule change, the Commission has
considered the proposed rule's impact on efficiency, competition,
and capital formation. See 15 U.S.C. 78c(f).
    \6\ 15 U.S.C. 78f(b)(5).
---------------------------------------------------------------------------

    The Commission believes that clarifying the registration procedures
and ongoing compliance obligations and making the registration
procedures consistent with the procedures of the other SROs will
benefit ETP Holders and their registered persons by making the
registration process easier and more efficient. Furthermore, amending
Exchange rules to be generally consistent with other SROs' rules,
market practices, and the operation of the CRD should help simplify
such procedures and administrative matters for ETP Holders and their
registered persons.

IV. Conclusion

    It is therefore ordered, pursuant to Section 19(b)(2) of the
Act,\7\ that the proposed rule change (SR-NYSEArca-2006-50), as
modified by Amendment No. 1, be, and hereby is, approved.
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    \7\ 15 U.S.C. 78s(b)(2).
    \8\ 17 CFR 200.30-3(a)(12).

    For the Commission, by the Division of Market Regulation,
pursuant to delegated authority.\8\
Florence E. Harmon,
Deputy Secretary.
[FR Doc. E7-4975 Filed 3-19-07; 8:45 am]

BILLING CODE 8010-01-P


[Federal Register Volume 77, Number 78 (Monday, April 23, 2012)]
[Notices]
[Pages 24189-24190]
[FR Doc No: 2012-9659]

=======================================================================
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DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

[Docket No. IC12-11-000]

Commission Information Collection Activities (FERC-725); Comment
Request; Extension

AGENCY: Federal Energy Regulatory Commission, DOE.

ACTION: Notice of information collection and request for comments.

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SUMMARY: In compliance with the requirements of the Paperwork Reduction
Act of 1995, 44 U.S.C. 3506(c)(2)(A), the Federal Energy Regulatory
Commission (Commission or FERC) is soliciting public comment on the
currently approved information collection, FERC-725, Certification of
Electric Reliability Organization; Procedures for Electric Reliability
Standards.

DATES: Comments on the collection of information are due June 22, 2012.

ADDRESSES: You may submit comments (identified by Docket No. IC12-11-
000) by either of the following methods:
     eFiling at Commission's Web Site: http://www.ferc.gov/docs-filing/efiling.asp
     Mail/Hand Delivery/Courier: Federal Energy Regulatory
Commission, Secretary of the Commission, 888 First Street NE.,
Washington, DC 20426.
    Instructions: All submissions must be formatted and filed in
accordance with submission guidelines at: http://www.ferc.gov/help/submission-guide.asp. For user assistance contact FERC Online Support
by email at<EMAIL_ADDRESS>or by phone at:<PHONE_NUMBER>
(toll-free), or<PHONE_NUMBER> for TTY.
    Docket: Users interested in receiving automatic notification of
activity in this docket or in viewing/downloading comments and
issuances in this docket may do so at http://www.ferc.gov/docs-filing/docs-filing.asp.

FOR FURTHER INFORMATION CONTACT: Ellen Brown may be reached by email at
<EMAIL_ADDRESS>telephone at<PHONE_NUMBER>, and fax at (202)
273-0873.

SUPPLEMENTARY INFORMATION:
    Title: FERC-725 and Certification of Electric Reliability
Organization; Procedures for Electric Reliability Standards.
    OMB Control No.: 1902-0225.
    Type of Request: Three-year extension of the FERC-725 information
collection requirements with no changes to the current reporting
requirements. However, FERC is making adjustments to the burden
estimates based on current information.
    Abstract: The Commission uses the information collected under the
requirements of FERC-725 \1\ to implement the statutory provisions of
Section 215 of the Federal Power Act (FPA).\2\
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    \1\ ``Electric Reliability Organization'' or ``ERO'' means the
organization certified by the Commission the purpose of which is to
establish and enforce Reliability Standards for the Bulk-Power
System, subject to Commission review.
    \2\ Section 215 was added by the Energy Policy Act of 2005,
Public Law 109-58, 119 Stat. 594 (2005) (codified at 42 U.S.C.
16451, et seq.)
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    Section 215 of the FPA aids the Commission's efforts to strengthen
the reliability of the interstate grid through the granting of new
authority to provide for a system of mandatory Reliability Standards
developed by the Electric Reliability Organization (ERO) and reviewed
and approved by FERC.
    On February 3, 2006, the Commission issued Order No.672 \3\
certifying a single ERO [the North American Electric Reliability
Corporation (NERC)], to oversee the reliability of the United States'
portion of the interconnected North American Bulk-Power System, subject
to Commission oversight. The ERO is responsible for developing and
enforcing the mandatory Reliability Standards. The Reliability
Standards apply to all users, owners and operators of the Bulk-Power
System. The Commission has the authority to approve all ERO actions, to
order the ERO to carry out its responsibilities under these statutory
provisions, and (as appropriate) to enforce Reliability Standards.
---------------------------------------------------------------------------

    \3\ Rules Concerning Certification of the Electric Reliability
Organization; and Procedures for the Establishment, Approval, and
Enforcement of Electric Reliability Standards ] 31,204 71 FR 8662
(2006) Order on rehearing, 71 FR 19,814 (2006), FERC Statutes and
Regulations ] 31,212 (2006).
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    The ERO can delegate its enforcement responsibilities to a Regional
Entity. Delegation is effective only after the Commission approves the
delegation agreement. A Regional Entity can also propose a Reliability
Standard to the ERO for submission to the Commission for approval.
    The FERC-725 contains the following information collection
elements:
    Self Assessment and ERO Application: The Commission requires the
ERO to submit to FERC a performance assessment report every five years.
Each of regional entity submits a performance assessment report to the
ERO. Submitting an application to become an ERO is also part of this
collection.\4\
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    \4\ The Commission does not expect any new ERO applications to
be submitted in the next three years and is not including any burden
for this requirement in the burden estimate.
---------------------------------------------------------------------------

    Reliability Assessments: 18 CFR 39.11 requires the ERO to assess
the reliability and adequacy of the Bulk-Power System in North America.
Subsequently, the ERO must report to the Commission on its findings.
Regional entities perform similar assessments within individual
regions.
    Reliability Compliance: Reliability Standards are mandatory and
enforceable. In addition to the specific information collection
requirements contained in each standard, there are general compliance,
monitoring and enforcement information collection requirements imposed
on applicable entities. Audits, spot checks, self-certifications,
exception data submittals, violation reporting, and mitigation plan
confirmation are included in this area.
    Stakeholder Survey: The ERO used a stakeholder survey to solicit
feedback from registered entities in preparation for its three year
performance assessment. The Commission assumes that the ERO will
perform another survey prior to the 2014 performance assessment.
    Other Reporting: This category refers to all other reporting
requirements imposed on the ERO or regional entities in order to comply
with the Commission's regulations.
    The Commission implements its responsibilities through the Code of
Federal Regulations in 18 CFR part 39.
    Type of Respondents: Electric reliability organization, regional
entities, and registered entities.

    Estimate of Annual Burden \5\: The Commission estimates the total
public reporting burden for this information collection as:
---------------------------------------------------------------------------

    \5\ Burden is defined as the total time, effort, or financial
resources expended by persons to generate, maintain, retain, or
disclose or provide information to or for a Federal agency. For
further explanation of what is included in the information
collection burden, reference 5 Code of Federal Regulations 1320.3.

                                  FERC-725--Certification of the ERO \4\; Procedures for Electric Reliability Standards
--------------------------------------------------------------------------------------------------------------------------------------------------------
                                                                                Number of                    Average burden     Estimated
            Type of respondent                    Type of reporting          Number of     responses per   Total number      hours per     total annual
                                                   requirement \4\          respondents     respondent     of responses      response         burden
                                            ............................             (A)         (B) \6\     (A) x (B) =             (D)       (C) x (D)
                                                                                (C)
--------------------------------------------------------------------------------------------------------------------------------------------------------
Electric Reliability Organization (ERO)...  Self-Assessment.............  ..............            0.33            0.33          10,400           3,432
                                            Reliability Assessments.....  ..............              11              11           3,120          34,320
                                            Reliability Compliance......  ..............               1               1          76,837          76,837
                                            Standards Development.......  ..............               1               1          51,834          51,834
                                            Other Reporting.............               1               1               1           2,080           2,080
Regional Entities.........................  Self-Assessment.............  ..............            0.33            2.64          16,640          43,930
                                            Reliability Assessments.....  ..............               1               8          16,679         133,432
                                            Reliability Compliance......  ..............               1               8          46,788         374,304
                                            Standards Development.......  ..............               1               8         * 4,142          33,134
                                            Other Reporting.............               8               1               8           1,040           8,320
Registered Entities.......................  Stakeholder Survey..........  ..............            0.33             537               4           2,148
                                            Reliability Compliance......           1,627               1           1,627           * 483         786,342
                                           -------------------------------------------------------------------------------------------------------------
Subtotals:
    ERO...................................                                                                                168,503
    Regional..............................                                             \7\ N/A                                                   593,120
    Registered............................                                                                                788,490
                                           -------------------------------------------------------------------------------------------------------------
        Total.............................  ............................           1,636         \7\ N/A         \7\ N/A         \7\ N/A       1,550,113
--------------------------------------------------------------------------------------------------------------------------------------------------------
* (Rounded)

     The Commission derived the figures above using NERC's Business
Plan and Budget Submissions, NERC's Compliance, Enforcement and
Monitoring Plans, NERC's Performance Assessments, other information on
NERC's Web site (http://www.nerc.com/), and internal FERC staff
estimates. See the appendix for more details regarding the burden
estimates.\8\
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    \6\ In all instances below where the number of responses per
respondent is ``1'' the Commission acknowledges that actual number
of responses varies and cannot be estimated clearly.
    \7\ N/A = not applicable.
    \8\ The appendix will not be published in the Federal Register.
The appendix is available in FERC's eLibrary system under the notice
issuance in Docket No. IC12-11-000.
---------------------------------------------------------------------------

    The total estimated annual cost burden to respondents is
$115,655,020 ($15,128,199 + $46,121,011 + $54,405,810).
    ERO Cost: 168,503 hours @ $89.78/hr = $15,128,199.
    Regional Entity Cost: 593,120 hours @ $77.76/hr = $46,121,011.
    Registered Entity Cost: 788,490 hours @ $69/hr = $54,405,810 .
    The hourly cost figures are loaded (i.e. includes salary and other
personnel costs). The Commission used NERC's 2012 Business Plan and
internal FERC salary estimates for these cost figures.
    Comments: Comments are invited on: (1) Whether the collection of
information is necessary for the proper performance of the functions of
the Commission, including whether the information will have practical
utility; (2) the accuracy of the agency's estimate of the burden and
cost of the collection of information, including the validity of the
methodology and assumptions used; (3) ways to enhance the quality,
utility and clarity of the information collection; and (4) ways to
minimize the burden of the collection of information on those who are
to respond, including the use of automated collection techniques or
other forms of information technology.

    Dated: April 16, 2012.
Kimberly D. Bose,
Secretary.
[FR Doc. 2012-9659 Filed 4-20-12; 8:45 am]
BILLING CODE 6717-01-P