Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Data Requirement: : EPA DP Barcode D288775 EPA MRID 458677 06 EPA Guideline 70 1( Special Study)

Test material: Purity: {............} 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 24, 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, Statistician Date: Environmental Fate and Effects Division, ERB 3, U. S. Environmental Protection Agency EPA PC Code 080803 Date Evaluation Completed: 05/ 31/ 2003 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 2 of 54 CITATION: Sepulveda, M. S. and T. S. Gross. 2003. Characterization of Atrazine Exposures and Potential Effects in Florida Ecosystems Dominated by Sugarcane Agriculture: A Reconnaissance Survey of Amphibians in South Florida for the Assessment of Potential Atrazine Effects. Department of Physiological Sciences, University of Florida, Caribbean Science Center, Gainesville, Florida. Sponsor: Syngenta Crop Protection, Inc. Study ID: ECORISK Number UFL 02. Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 3 of 54 EXECUTIVE SUMMARY:

A reconnaissance survey for cane toads ( Bufo marinus) in south Florida sugarcane dominated agricultural sites ( Belle Glade and Canal Point) and nonagricultural sites ( University of Miami) indicated increased incidence of intersex ( ovarian tissue in Bidder's organ) in frogs identified as having testes. B. marinus is typically a sexually dimorphic amphibian; however, 100% of the cane toads collected at Belle Glade and roughly 55% of the male/ intersex frogs collected at Canal Point exhibited female coloration. Additionally, males typically exhibit nuptual pads; however 71% and 0% of the of the intersex frogs collected from Canal Point and Belle Glade, respectively, had nuptual pads. Vitellogenin, a female specific protein, is not typically expressed in males; however, intersex frogs had vitellogenin levels ( 774 ± 29 PO4/ mg protein) similar to the females ( 853 ± 34 PO4/ mg protein) and was roughly double that of male toads ( 375 ± 34 PO4/ mg protein). Plasma sex steroids ( 17   estradiol and testosterone) were relatively gender specific; however, testosterone levels in intersex males exhibited roughly twice the amount of variability as similar estimates for males. Although agricultural sites had atrazine concentrations ranging from < 0.01 to 24.45

g/ L over the six month sampling period, no atrazine levels were measured at the University of Miami ( nonagricultural) site.

The study authors clearly recognized the potential for other chemicals to confound the interpretation of this study, but they failed to monitor for any other chemicals. The study authors also failed to characterize atrazine levels in nonagricultural sites and water quality parameters that could have impacted the development of frogs.

In this study, the southern toad ( B. terrestris) was also examined and found to have an increased incidence of intersex ( Bidder's organ containing ovarian tissue) in both agricultural and nonagricultural sites. The study authors speculate that the presence of a Bidder's organ may have rendered the animals more sensitive to developmental effects. Bidder's organ is characterized as a nonfunctional, rudimentary ovary; however, no information is available on whether the organ has an endocrine function at any time during the development of the animal. Additionally, while bufonids typically exhibit Bidder's organs at the anterior of each testis, oogenesis is normally abortive because the oocytes fail to reach vitellogenesis and undergo a degenerative process. However, the one slide depicting an intersex male in this study suggested that the oocytes possessed yolk and were therefore vetellogenic.

This study was useful in identifying the incidence of hermaphroditism in field collected toads. As with the previous studies, toads with testes also appeared to have ovarian tissue, but unlike previous studies the ovarian tissue was associated with the Bidder's organ rather than the testes. While toads collected in agricultural sites may have been exposed to atrazine during development, it is unknown whether atrazine was present at the nonagricultural sites. Other problems with the the study design include exposure to other agrochemicals and environmental conditions relevant to development of toads, e. g., water quality characteristics. These factors limit an interpretation of the findings and make it difficult to establish causality. Based on these data, it is difficult to conclude that atrazine exposure was associated with an increased incidence of intersex. Also, the study does not provide insights on the ecological relevance of the data. If toads depend on coloration to attract mates, though, toads from agricultural sites may have an impaired ability to attract mates because of their distinctly female appearance. Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 4 of 54 I. MATERIALS AND METHODS GUIDELINE FOLLOWED: Nonguideline Study COMPLIANCE: Not conducted under GLP; however, most practices as defined by 40CFR Part 160, August 19, 1989, were established for this study, including but not limited to: 1. Written, authorized preliminary protocol 2. Written, authorized Standard Operating Procedures for all key procedures. 3. Organization and personnel were sufficient in terms of number, education, training and experience. 4. Independent QA inspections were conducted 5. Final Report was written 6. Raw data, documentation, records, protocols, and final report were archived A. MATERIALS:

1. Test Material [ Complete this subsection using the information provided in the methodology section of the study report. Name of test material as cited in the study report.]

Description: Atrazine Lot No./ Batch No. :

Purity: NA Stability of compound under test conditions:

Storage conditions of NA test chemicals:

2. Test organism:

Species: Cane toad ( Bufo marinus); Southern toad ( B. terrestris); Green Treefrog ( Hyla cinera); Cuban Treefrog ( Osteopilus. septentrionalis) . Age at test initiation: Field collected animals; age not determined Weight at study initiation: ( mean and range) Male: 125.1 ± 8.3 g Female: 133.9 ± 10.3 g Intersex: 123.0 ± 14.5 g Length at study initiation: ( mean and range) Male: 100.6 ± 2.5 mm Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 5 of 54 Female: 104.9 ± 2.4 mm Intersex: 99.9 ± 6.7 mm Source: Field collected at Belle Glade and Canal Point ( sugar cane sites); University of Miami ( nonagricultural site)

B. STUDY DESIGN:

Objective:

1) To determine whether exposure of frogs to sugarcane agricultural areas in South Florida would result in a higher incidence of intersex and/ or other gonadal/ developmental anomalies 2) to examine amphibian populations exposed to a complex mixture of several pesticides and assorted agrichemicals.

1. Experimental Conditions a) Range finding Study: Preliminary b) Definitive Study Table 1 . Experimental Parameters Parameter Details Acclimation: period: Conditions: ( same as test or not) Feeding: Health: ( any mortality observed) NA Duration of the test NA Test condition static/ flow through Type of dilution system for flow through method.

Renewal rate for static renewal NA Aeration, if any NA Test vessel NA Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Parameter Details Page 6 of 54 Source of dilution water Quality: NA Water parameters: Hardness pH Dissolved oxygen Total Organic carbon Particulate Matter Metals Pesticides Chlorine Temperature { Salinity for marine or estuarine species}

Intervals of water quality measurement NA Number of replicates/ groups: control: solvent control: treated ones: NA Number of organisms per replicate / groups: control: solvent control: treated ones: NA Biomass loading rate NA Test concentrations: nominal: measured: NA Solvent ( type, percentage, if used) NA Lighting NA Feeding NA Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Parameter Details Page 7 of 54 Recovery of chemical Level of Quantitation Level of Detection NA Positive control { if used, indicate the chemical and concentrations} NA Other parameters, if any NA Reconnaissance survey was conducted in the field:

Initially, distribution and concentration of atrazine at south Florida sites ( Belle Glade and Canal Point) was measured by collecting multiple water samples from several canals/ ditches every 2 weeks from February through June with an additional sample in mid July. A scoping survey of atrazine did not include University of Miami site. No chemicals other than atrazine were screened.

A scoping survey of amphibians for Bufo marinus was conducted; however, only the University of Miami site was identified as nonagricultural and having B. marinus.

Although B. terrestris was located at Belle Glade, it was not located at Canal Point. Two nonagricultural sites ( Archibald Biological Station and Fisheater Creek) did have the Southern Toad. Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 8 of 54 2. Observations: Table 2: Observations Criteria Details Parameters measured including the sublethal effects/ toxicity symptoms Observation intervals Were raw data included?

Other observations, if any 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.]

Canals and ditches were sampled for atrazine at both of the agricultural sites. Atrazine levels ranged from < 0.10 to 24.45 ppb at Belle Glade and from < 0.10 to 19.54 ppb at Canal Point. Atrazine concentration at Belle Glade ( consisting of 8 sampling sites) peaked in early February ( February 10) and then rapidly declined to a more or less steady state for the remainder of the sampling period. At Canal Point ( consisting of 8 sampling sites), atrazine concentrations peaked on March 16 and again on April 14, but remained sporadic.

Distribution of Amphibians Bufo marinus was located in two sugarcane areas ( Belle Glade and Canal Point) and only one nonagricultural area, i. e., University of Miami. Bufo terrestris was located in one sugarcane area ( Belle Glade) and two nonagricultural sites, i. e., Archibald Biological Station and Fisheater Creek. Hyla cinera was present at Belle Glade and Archibald. O. septentrionalis was present at Belle Glade and Fisheating Creek.

Body weight, length and coloration were recorded; blood was collected by cardiac puncture.

Gender defined by: female: present of ovarian tissue and absence of testicular tissue male: presence of testes and absence of developing eggs and/ or ovarian tissue intersex: presence of testes and developing eggs and/ or ovarian tissue.

Gonadal Deformities:

segmented testes: testes with clear demarcations or splits highly segmented: testes with demarcations making testes appear multiple abnormal shaped: twisted or curled multiple testes: not defined in text Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 9 of 54 intersex: not defined in text but likely refers to mix of ovarian and testicular tissue in same organism Only testes were examined histologically; sections were fixed in formalin and then embedded in parafin. Approximately 29% of the males collected from Belle Glade and 39% of the males collected from Canal Point were intersexed based on ovarian tissue located in the Bidder's organ of males.

For B. terristris, 6 of 18 ( 33%) were intersex from Archibaold ( nonagricultural site); 3 of 21 ( 14%) were intersex from Belle Glade; and 5 of 23 ( 22%) were intersex from Fisheater Creek ( cattle/ citrus site).

Plasma was analyzed for 17   estradiol and testosterone using radioimmunoassay and for alkali labile phosphate as in indirect measure of vitellogenin.

Table 3 . Distribution of amphibians in both agricultural ( sugarcane) and nonagricultural sites.

Species Agricultural ( Sugarcane Site) Nonagricultural Site Belle Glade Canal Point Univ. of Miami Archibald Fisheater Cane Toad ( B. marinus) X X X Southern Toad ( B. terrestris) X X X Green Treefrog ( Hyla cinera) X X Cuban Treefrog ( Ostropilus septentrionalis) X X C. VERIFICATION OF STATISTICAL RESULTS: Basic statistics run using SAS ® ( Statistical Analysis System, Release 8.01, Cary, North Carolina). ( See attached printout).

D. STUDY DEFICIENCIES: The study failed to measure atrazine in nonagricultural sites and to collect any information on other chemicals present at the sites. The study also failed to characterize water quality at any of the collection sites.

The University of Miami nonagricultural site was added late in the study and was therefore not included in any Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 10 of 54 of the atrazine sampling efforts.

According to the protocol, animals collected will include: tadpoles at early, mid, and late metamorphosis (> 30 per site), metamorphs (> 30 per site), and adults ( 10 males and 10 females per site) as available for each species. Each site will contain 3 replicates/ location. Blood will be collected for subsequent endocrine biomarker analysis ( E, T, P and DHT). Gonadal tissue from adults will be fixed for subsequent evaluation of gonadal tissues and laryngeal development and subsequent atrazine analysis. The protocol was amended to show that only adults were collected because tadpoles were " difficult to collect." Does this mean that tadpoles were too few in number to collect?

No effort was made to characterize the larynxes because the technique for doing so had not been validated for B. marinus.

No P or DHT analysis was performed for B. marinus because validated procedures were not available for this species.

Proposed experimental start date on UFL 02 protocol ( April 1, 2001) does not agree with proposed experimental start and termination date, i. e., March 1, 2002, and October 1, 2003, respectively also reported in the protocol E. REVIEWER'S COMMENTS:

Although the study objective states that a range of chemicals are used on sugarcane and implies a recognition that other chemicals would potentially confound any effort to link anomalies to atrazine, the study failed to measure any chemicals in the nonagricultural ( University of Miami) site and only measured atrazine in each of the agricultural sites ( Belle Glade and Canal Point). Atrazine concentrations in agricultural sites ranged over three orders of magnitude. While some sampling sites exhibited relatively high atrazine concentrations during a sampling date, others samples collected from different areas of the same site exhibited low atrazine concentrations. Although no intersex animals were detected in the nonagricultural site, the authors suggested that the incidence of intersex in agricultural sites may be reflective of " background". The authors also suggested that Bidder's organ may potentially render Bufonidae more susceptible to gonad effects. It is unclear whether the characterization of Bidder's organ as being a " nonfunctional, rudimentary ovary" is entirely correct because literature suggests that if normal males are castrated, the Bidder's organ will differentiate into an ovary. If the theory that atrazine up regulates is correct and testosterone is transformed into estrogen, is there sufficiently low testosterone or sufficiently high estrogen to result in differentiation of the Bidder's organ?

Across treatment sites, estradiol in intersex animals was only slightly ( 302 ± 33 pg/ mL) above levels found in normal males ( 293 ± 23 pg/ mL); serum testosterone in intersex males was slightly lower ( 503 ± 307 pg/ mL) than normal males ( 552 ± 174 pg/ mL). It is clear, though, that the variability in testosterone levels for intersex males ( CV= 61%) was roughly double that of normal males ( 31.5%). Additionally, labile phosphate as an indirect measure of plasma vitellogenin levels in intersex males ( 774 ± 29 PO4/ mg protein) was roughly double the level in normal males ( 375 ± 34 PO4/ mg protein) and was similar to the levels contained in females ( 853 ± 34 PO4/ mg protein). If the labile phosphate ( ALP) is indicative of vitellogenin, then vitellogenesis in intersex animals has been up regulated by exposure to either an endogenous and/ or exogenous estrogen. Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 1Petrini, S. and F. Zaccanti. 1998. The effects of Aromatize and 5   reductase inhibitors, antiandrogen, and sex steroids on Bidder's organs development and gonadal differentiation in Bufo bufo tadpoles. The Journal of Experimental Zoology 280: 245  259.

Page 11 of 54 It is unclear why the bidder's organ makes the toad a generally more sensitive species to the effects of atrazine or other endocrine disrupting chemicals. It could be argued that this natural hermaphroditic condition is a confounding characteristic since the ovarian tissue of bidder's organ is presumably capable of estrogen synthesis and therefore capable of inducing vitellogenin synthesis in the absence of an exogenous estrogen. This study seems to indicate that vitellogenin levels in the intersex organisms approximates the normal female condition in the absence of elevated plasma estrogen. But, the lack of a strong vitellogenic response of the normal males from the same site suggests that they have not been exposed to an estrogenic chemical. This presumes that the ALP measurement is a good surrogate for vitellogenin and that the male levels at the reference site represents baseline vitellogenin concentrations. This issue clearly needs more research.

Furthermore, although Bufo marinus is typically a sexually dimorphic species with gender specific coloration being detected in the nonagricultural site collected animals, female coloration pattern was detected in 100% of the frogs ( male, female and intersex) collected at Belle Glade and 57%, 54% and 100% of the intersex, male and female frogs collected from Canal Point, respectively.

Nuptual pads were not identified for females from any of the sites. In males collected from nonagricultural sites, 90% had nuptual pads, while 82% and 32% of the males had nuptual pads from Canal Point and Belle Glade, respectively. For males identified as intersex from Canal Point and Belle Glade, 71% and 0% had nuptual pads, respectively.

Testicular abnormalities other than intersex were somewhat similar across collection sites with 44% for Canal Point, 61% for Belle Glade and 45% for University of Miami.

F. CONCLUSIONS: It is not clear why the authors concluded that Bufo marinus may be more sensitive to factors that influence gonadal development and sexual differentiation because it possesses a nonfunctional rudimentary ovary ( Bidder's organ). For that reason, the species may be a useful sentinel for factors influencing gonadal development. Additionally, while bufonids typically exhibit Bidder's organs at the anterior of each testis, oogenesis is normally abortive because the oocytes fail to reach vitellogenesis and undergo a degenerative process ( Petrini and Zaccanti 1998) 1. However, the one slide ( Figure 3F) depicting an intersex male in this study suggests that the oocytes possessed yolk and were therefore vitellogenic. Although the authors state that previous studies have hypothesized a potential developmental mechanism of action for atrazine and that the current study was indirectly designed to evaluate a similar mechanism of action in amphibians, the current study's focus on adults does not permit direct evaluation of this effect. Also, it is difficult to test the hypothesis that atrazine exposure resulted in developmental effects in amphibians because atrazine was not measured in nonagricultural sites, other chemicals were not measured at any of the sites, and no water quality data were collected to characterize the study sites. While it may have been possible to derive some correlation between the incidence of intersex in males and atrazine patterns within particular areas of the sample sites, these data are not provided.

Since intersex, i. e., ovarian tissue in the Bidder's organ, did not appear to involve the testes per se, it is unclear Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 12 of 54 whether the reproductive capacity of the adult frogs was diminished due to this effect. However, the intersex frogs appear to be responding to an environmental estrogen given their elevated vitellogenin levels and a distinctly female appearance. If these frogs rely heavily of coloration to attract mate, their ability to reproduce may be impaired The incidence of intersex in Bufo terrestris also appeared to be elevated; however this was true for both agricultural and nonagricultural sites. The authors speculate that this increased incidence may be indicative of the variability in the background occurrence of intersex among amphibians with a Bidder's organ. Since atrazine levels at the nonagricultural sites were not characterized, it is difficult to speculate on the relevancy of the proposition of " background". Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 13 of 54 MEAN LENGTH WEIGHT GONAD AND GONADOSOMATIC INDEX BY SITE AND SEX 1 Obs site Sex _ TYPE_ _ FREQ_ LENGTH WEIGHT GONAD GSE SD_ LT SD_ WT SD_ GONAD SD_ GSI 1 BG F 0 19 92.979 94.005 2.22374 1.11278 19.0811 67.773 4.3402 1.16438 2 BG I 0 9 96.332 94.639 0.21622 0.23778 10.6652 40.275 0.0927 0.08913 3 BG M 0 22 90.763 79.609 0.23955 0.31182 7.4531 21.577 0.2006 0.29333 4 CP F 0 37 109.067 143.762 6.29178 3.35892 15.8198 65.904 10.8004 5.11631 5 CP I 0 7 104.639 159.486 0.53771 0.37286 39.9476 59.198 0.1515 0.18901 6 CP M 0 11 106.042 128.105 0.41200 0.32727 10.1743 47.890 0.1693 0.11934 7 UM F 0 4 122.835 232.685 9.56100 4.33500 13.9482 136.862 5.1713 1.41630 8 UM M 0 20 108.400 174.345 0.40530 0.23050 24.0066 57.599 0.1495 0.05375 ANOVA FOR LENGTH BETWEEN SITES 2  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 Dependent Variable: SVL Sum of Source DF Squares Mean Square F Value Pr > F Model 2 4629.32548 2314.66274 8.17 0.0008 Error 57 16146.86161 283.27827 Corrected Total 59 20776.18709 R Square Coeff Var Root MSE SVL Mean 0.222819 16.04614 16.83087 104.8905 Source DF Anova SS Mean Square F Value Pr > F site 2 4629.325476 2314.662738 8.17 0.0008 Levene's Test for Homogeneity of SVL Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 194186 97093.2 0.56 0.5751 Error 57 9906646 173801 Bartlett's Test for Homogeneity of SVL Variance Source DF Chi Square Pr > ChiSq site 2 1.0035 0.6055 Dunnett's t Tests for SVL NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 57 Error Mean Square 283.2783 Critical Value of Dunnett's t 2.28365 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits UM  BG 29.856 8.712 51.000 *** CP  BG 16.088 5.240 26.936 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 14 of 54 ANOVA FOR LENGTH BETWEEN SITES 6  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 Dependent Variable: SVL Sum of Source DF Squares Mean Square F Value Pr > F Model 1 271.66953 271.66953 0.36 0.5566 Error 14 10484.85064 748.91790 Corrected Total 15 10756.52018 R Square Coeff Var Root MSE SVL Mean 0.025256 27.37560 27.36636 99.96625 Source DF Anova SS Mean Square F Value Pr > F site 1 271.6695337 271.6695337 0.36 0.5566 Levene's Test for Homogeneity of SVL Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 6318154 6318154 1.83 0.1976 Error 14 48349862 3453562 Bartlett's Test for Homogeneity of SVL Variance Source DF Chi Square Pr > ChiSq site 1 9.8175 0.0017 Dunnett's t Tests for SVL NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 14 Error Mean Square 748.9179 Critical Value of Dunnett's t 2.14485 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 8.306  21.274 37.887 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 15 of 54 ANOVA FOR LENGTH BETWEEN SITES 10  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 Dependent Variable: SVL Sum of Source DF Squares Mean Square F Value Pr > F Model 2 3671.52388 1835.76194 6.98 0.0021 Error 50 13151.65680 263.03314 Corrected Total 52 16823.18068 R Square Coeff Var Root MSE SVL Mean 0.218242 16.12326 16.21830 100.5894 Source DF Anova SS Mean Square F Value Pr > F site 2 3671.523883 1835.761942 6.98 0.0021 ANOVA FOR LENGTH BETWEEN SITES 12 Levene's Test for Homogeneity of SVL Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 2890854 1445427 0.89 0.4168 Error 50 81136179 1622724 Bartlett's Test for Homogeneity of SVL Variance Source DF Chi Square Pr > ChiSq site 2 26.2881 <. 0001 Dunnett's t Tests for SVL NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 50 Error Mean Square 263.0331 Critical Value of Dunnett's t 2.28707 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits UM  BG 17.637 6.177 29.097 *** CP  BG 15.279 1.582 28.976 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 16 of 54 NONPARAMETRIC COMPARISON OF LENGTH BETWEEN SITES 14  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 363.0 579.50 62.927869 19.105263 CP 37 1276.0 1128.50 65.771701 34.486486 UM 4 191.0 122.00 33.744135 47.750000 Kruskal Wallis Test Chi Square 13.9187 DF 2 Pr > Chi Square 0.0009 Median Scores ( Number of Points Above Median) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 4.0 9.50 1.816823 0.210526 CP 37 22.0 18.50 1.898929 0.594595 UM 4 4.0 2.00 0.974245 1.000000 Median One Way Analysis Chi Square 11.4979 DF 2 Pr > Chi Square 0.0032 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 17 of 54 NONPARAMETRIC COMPARISON OF LENGTH BETWEEN SITES 16  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 56.0 76.50 9.447222 6.222222 CP 7 80.0 59.50 9.447222 11.428571 Wilcoxon Two Sample Test Statistic 80.0000 Normal Approximation Z 2.1170 One Sided Pr > Z 0.0171 Two Sided Pr > | Z| 0.0343 t Approximation One Sided Pr > Z 0.0257 Two Sided Pr > | Z| 0.0514 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 4.7087 DF 1 Pr > Chi Square 0.0300 The NPAR1WAY Procedure Median Scores ( Number of Points Above Median) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 2.0 4.50 1.024695 0.222222 CP 7 6.0 3.50 1.024695 0.857143 Median Two Sample Test Statistic 6.0000 Z 2.4398 One Sided Pr > Z 0.0073 Two Sided Pr > | Z| 0.0147 Median One Way Analysis Chi Square 5.9524 DF 1 Pr > Chi Square 0.0147 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 18 of 54 NONPARAMETRIC COMPARISON OF LENGTH BETWEEN SITES 18  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 313.0 594.0 55.398556 14.227273 CP 11 348.0 297.0 45.596052 31.636364 UM 20 770.0 540.0 54.497706 38.500000 Kruskal Wallis Test Chi Square 27.1304 DF 2 Pr > Chi Square <. 0001 The NPAR1WAY Procedure Median Scores ( Number of Points Above Median) for Variable SVL Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 1.0 10.792453 1.810436 0.045455 CP 11 7.0 5.396226 1.490088 0.636364 UM 20 18.0 9.811321 1.780996 0.900000 Median One Way Analysis Chi Square 31.1926 DF 2 Pr > Chi Square <. 0001 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 19 of 54 ANOVA FOR WEIGHT BETWEEN SITES 20  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 Dependent Variable: wt Sum of Source DF Squares Mean Square F Value Pr > F Model 2 72873.3117 36436.6559 7.03 0.0019 Error 57 295228.9890 5179.4559 Corrected Total 59 368102.3007 R Square Coeff Var Root MSE wt Mean 0.197970 53.73440 71.96844 133.9336 Source DF Anova SS Mean Square F Value Pr > F site 2 72873.31174 36436.65587 7.03 0.0019 Levene's Test for Homogeneity of wt Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 3.5729E8 1.7864E8 2.91 0.0624 Error 57 3.4941E9 61300449 Bartlett's Test for Homogeneity of wt Variance Source DF Chi Square Pr > ChiSq site 2 4.3547 0.1133 Dunnett's t Tests for wt NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 57 Error Mean Square 5179.456 Critical Value of Dunnett's t 2.28365 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits UM  BG 138.68 48.27 229.09 *** CP  BG 49.76 3.37 96.14 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 20 of 54 ANOVA FOR WEIGHT BETWEEN SITES 24  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 Dependent Variable: wt Sum of Source DF Squares Mean Square F Value Pr > F Model 1 16557.62363 16557.62363 6.82 0.0205 Error 14 34003.16446 2428.79746 Corrected Total 15 50560.78809 R Square Coeff Var Root MSE wt Mean 0.327480 40.06429 49.28283 123.0094 Source DF Anova SS Mean Square F Value Pr > F site 1 16557.62363 16557.62363 6.82 0.0205 Levene's Test for Homogeneity of wt Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 9605650 9605650 1.11 0.3100 Error 14 1.2117E8 8654926 Bartlett's Test for Homogeneity of wt Variance Source DF Chi Square Pr > ChiSq site 1 0.9592 0.3274 Dunnett's t Tests for wt NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 14 Error Mean Square 2428.797 Critical Value of Dunnett's t 2.14485 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits CP  BG 64.85 11.58 118.12 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 21 of 54 ANOVA FOR WEIGHT BETWEEN SITES 28  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 Dependent Variable: wt Sum of Source DF Squares Mean Square F Value Pr > F Model 2 94122.4446 47061.2223 24.58 <. 0001 Error 50 95747.7804 1914.9556 Corrected Total 52 189870.2250 R Square Coeff Var Root MSE wt Mean 0.495720 34.88999 43.76021 125.4234 Source DF Anova SS Mean Square F Value Pr > F site 2 94122.44456 47061.22228 24.58 <. 0001 Levene's Test for Homogeneity of wt Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 77865494 38932747 3.38 0.0420 Error 50 5.7584E8 11516787 Bartlett's Test for Homogeneity of wt Variance Source DF Chi Square Pr > ChiSq site 2 16.9429 0.0002 Dunnett's t Tests for wt NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 50 Error Mean Square 1914.956 Critical Value of Dunnett's t 2.28707 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits UM  BG 94.74 63.81 125.66 *** CP  BG 48.50 11.54 85.45 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 22 of 54 NONPARAMETRIC COMPARISON OF WEIGHT BETWEEN SITES 32  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 371.0 579.50 62.926995 19.526316 CP 37 1270.0 1128.50 65.770787 34.324324 UM 4 189.0 122.00 33.743666 47.250000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 12.9558 DF 2 Pr > Chi Square 0.0015 Median Scores ( Number of Points Above Median) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 3.0 9.50 1.816823 0.157895 CP 37 23.0 18.50 1.898929 0.621622 UM 4 4.0 2.00 0.974245 1.000000 Average scores were used for ties.

Median One Way Analysis Chi Square 14.8325 DF 2 Pr > Chi Square 0.0006 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 23 of 54 NONPARAMETRIC COMPARISON OF WEIGHT BETWEEN SITES 34  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 59.0 76.50 9.447222 6.555556 CP 7 77.0 59.50 9.447222 11.000000 Wilcoxon Two Sample Test Statistic 77.0000 Normal Approximation Z 1.7995 One Sided Pr > Z 0.0360 Two Sided Pr > | Z| 0.0719 t Approximation One Sided Pr > Z 0.0460 Two Sided Pr > | Z| 0.0921 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 3.4314 DF 1 Pr > Chi Square 0.0640 Median Scores ( Number of Points Above Median) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 3.0 4.50 1.024695 0.333333 CP 7 5.0 3.50 1.024695 0.714286 Median Two Sample Test Statistic 5.0000 Z 1.4639 One Sided Pr > Z 0.0716 Two Sided Pr > | Z| 0.1432 Median One Way Analysis Chi Square 2.1429 DF 1 Pr > Chi Square 0.1432 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 24 of 54 NONPARAMETRIC COMPARISON OF WEIGHT BETWEEN SITES 36  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 331.0 594.0 55.397439 15.045455 CP 11 313.0 297.0 45.595133 28.454545 UM 20 787.0 540.0 54.496608 39.350000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 26.0714 DF 2 Pr > Chi Square <. 0001 Median Scores ( Number of Points Above Median) for Variable wt Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 1.0 10.792453 1.810436 0.045455 CP 11 7.0 5.396226 1.490088 0.636364 UM 20 18.0 9.811321 1.780996 0.900000 Average scores were used for ties.

Median One Way Analysis Chi Square 31.1926 DF 2 Pr > Chi Square <. 0001 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 25 of 54 ANOVA FOR GONAD WEIGHT BETWEEN SITES 38  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 Dependent Variable: gonad Sum of Source DF Squares Mean Square F Value Pr > F Model 2 288.453909 144.226955 1.78 0.1779 Error 57 4618.623706 81.028486 Corrected Total 59 4907.077615 R Square Coeff Var Root MSE gonad Mean 0.058783 172.3940 9.001582 5.221517 Source DF Anova SS Mean Square F Value Pr > F site 2 288.4539090 144.2269545 1.78 0.1779 Levene's Test for Homogeneity of gonad Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 128737 64368.3 1.67 0.1977 Error 57 2199727 38591.7 Bartlett's Test for Homogeneity of gonad Variance Source DF Chi Square Pr > ChiSq site 2 15.4430 0.0004 Dunnett's t Tests for gonad NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 57 Error Mean Square 81.02849 Critical Value of Dunnett's t 2.28365 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits UM  BG 7.337  3.971 18.646 CP  BG 4.068  1.734 9.870 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 26 of 54 ANOVA FOR GONAD WEIGHT BETWEEN SITES 42  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 Dependent Variable: gonad Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.40696877 0.40696877 27.59 0.0001 Error 14 0.20647098 0.01474793 Corrected Total 15 0.61343975 R Square Coeff Var Root MSE gonad Mean 0.663421 34.02901 0.121441 0.356875 Source DF Anova SS Mean Square F Value Pr > F site 1 0.40696877 0.40696877 27.59 0.0001 Levene's Test for Homogeneity of gonad Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 0.000571 0.000571 0.92 0.3534 Error 14 0.00868 0.000620 Bartlett's Test for Homogeneity of gonad Variance Source DF Chi Square Pr > ChiSq site 1 1.5542 0.2125 Dunnett's t Tests for gonad NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 14 Error Mean Square 0.014748 Critical Value of Dunnett's t 2.14485 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 0.32149 0.19023 0.45276 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 27 of 54 ANOVA FOR GONAD WEIGHT BETWEEN SITES 46  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 Dependent Variable: gonad Sum of Source DF Squares Mean Square F Value Pr > F Model 2 0.36407367 0.18203683 5.85 0.0052 Error 50 1.55651365 0.03113027 Corrected Total 52 1.92058732 R Square Coeff Var Root MSE gonad Mean 0.189564 52.21800 0.176438 0.337887 Source DF Anova SS Mean Square F Value Pr > F site 2 0.36407367 0.18203683 5.85 0.0052 Levene's Test for Homogeneity of gonad Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 0.00324 0.00162 0.27 0.7670 Error 50 0.3041 0.00608 Bartlett's Test for Homogeneity of gonad Variance Source DF Chi Square Pr > ChiSq site 2 1.6748 0.4328 Dunnett's t Tests for gonad NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 50 Error Mean Square 0.03113 Critical Value of Dunnett's t 2.28707 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 0.17245 0.02344 0.32147 *** UM  BG 0.16575 0.04108 0.29043 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 28 of 54 NONPARAMETRIC COMPARISON OF GONAD WEIGHT BETWEEN SITES 50  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 346.0 579.50 62.926995 18.210526 CP 37 1282.0 1128.50 65.770787 34.648649 UM 4 202.0 122.00 33.743666 50.500000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 16.7428 DF 2 Pr > Chi Square 0.0002 Median Scores ( Number of Points Above Median) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 4.0 9.50 1.816823 0.210526 CP 37 22.0 18.50 1.898929 0.594595 UM 4 4.0 2.00 0.974245 1.000000 Average scores were used for ties.

Median One Way Analysis Chi Square 11.4979 DF 2 Pr > Chi Square 0.0032 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 29 of 54 NONPARAMETRIC COMPARISON OF GONAD WEIGHT BETWEEN SITES 52  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 48.0 76.50 9.433319 5.333333 CP 7 88.0 59.50 9.433319 12.571429 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 88.0000 Normal Approximation Z 2.9682 One Sided Pr > Z 0.0015 Two Sided Pr > | Z| 0.0030 t Approximation One Sided Pr > Z 0.0048 Two Sided Pr > | Z| 0.0096 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 9.1277 DF 1 Pr > Chi Square 0.0025 Median Scores ( Number of Points Above Median) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 2.0 4.50 1.024695 0.222222 CP 7 6.0 3.50 1.024695 0.857143 Average scores were used for ties.

Median Two Sample Test Statistic 6.0000 Z 2.4398 One Sided Pr > Z 0.0073 Two Sided Pr > | Z| 0.0147 Median One Way Analysis Chi Square 5.9524 DF 1 Pr > Chi Square 0.0147 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 30 of 54 NONPARAMETRIC COMPARISON OF GONAD WEIGHT BETWEEN SITES 54  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 382.50 594.0 55.386271 17.386364 CP 11 377.50 297.0 45.585941 34.318182 UM 20 671.00 540.0 54.485621 33.550000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 14.5995 DF 2 Pr > Chi Square 0.0007 Median Scores ( Number of Points Above Median) for Variable gonad Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 4.0 10.792453 1.810436 0.181818 CP 11 8.0 5.396226 1.490088 0.727273 UM 20 14.0 9.811321 1.780996 0.700000 Average scores were used for ties.

Median One Way Analysis Chi Square 14.0970 DF 2 Pr > Chi Square 0.0009 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 31 of 54 ANOVA FOR GONADOSOMATIC INDEX BETWEEN SITES 56  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 NOTE: Due to missing values, only 59 observations can be used in this analysis.

Dependent Variable: GSI Sum of Source DF Squares Mean Square F Value Pr > F Model 2 72.010680 36.005340 2.08 0.1350 Error 56 971.423418 17.346847 Corrected Total 58 1043.434098 R Square Coeff Var Root MSE GSI Mean 0.069013 152.0150 4.164955 2.739831 Source DF Anova SS Mean Square F Value Pr > F site 2 72.01068044 36.00534022 2.08 0.1350 Levene's Test for Homogeneity of GSI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 8045.3 4022.6 1.28 0.2865 Error 56 176236 3147.1 Bartlett's Test for Homogeneity of GSI Variance Source DF Chi Square Pr > ChiSq site 2 32.7955 <. 0001 Dunnett's t Tests for GSI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 56 Error Mean Square 17.34685 Critical Value of Dunnett's t 2.28391 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits UM  BG 3.2222  2.0359 8.4804 CP  BG 2.2461  0.4875 4.9797 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 32 of 54 ANOVA FOR GONADOSOMATIC INDEX BETWEEN SITES 60  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 Dependent Variable: GSI Sum of Source DF Squares Mean Square F Value Pr > F Model 1 0.07184534 0.07184534 3.62 0.0779 Error 14 0.27789841 0.01984989 Corrected Total 15 0.34974375 R Square Coeff Var Root MSE GSI Mean 0.205423 47.45756 0.140890 0.296875 Source DF Anova SS Mean Square F Value Pr > F site 1 0.07184534 0.07184534 3.62 0.0779 Levene's Test for Homogeneity of GSI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 0.00219 0.00219 1.21 0.2896 Error 14 0.0253 0.00180 Bartlett's Test for Homogeneity of GSI Variance Source DF Chi Square Pr > ChiSq site 1 3.5402 0.0599 Dunnett's t Tests for GSI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 14 Error Mean Square 0.01985 Critical Value of Dunnett's t 2.14485 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 0.13508  0.01721 0.28737 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 33 of 54 ANOVA FOR GONADOSOMATIC INDEX BETWEEN SITES 64  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 Dependent Variable: GSI Sum of Source DF Squares Mean Square F Value Pr > F Model 2 0.09486143 0.04743072 1.18 0.3147 Error 50 2.00424045 0.04008481 Corrected Total 52 2.09910189 R Square Coeff Var Root MSE GSI Mean 0.045191 70.41295 0.200212 0.284340 Source DF Anova SS Mean Square F Value Pr > F site 2 0.09486143 0.04743072 1.18 0.3147 Levene's Test for Homogeneity of GSI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 0.0746 0.0373 1.60 0.2121 Error 50 1.1661 0.0233 Bartlett's Test for Homogeneity of GSI Variance Source DF Chi Square Pr > ChiSq site 2 42.9859 <. 0001 Dunnett's t Tests for GSI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 50 Error Mean Square 0.040085 Critical Value of Dunnett's t 2.28707 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 0.01545  0.15364 0.18454 UM  BG  0.08132  0.22279 0.06015 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 34 of 54 NONPARAMETRIC COMPARISON OF GONADOSOMATIC INDEX ( GSI) BETWEEN SITES 68  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 18 306.50 540.0 60.741820 17.027778 CP 37 1265.50 1110.0 63.792823 34.202703 UM 4 198.00 120.0 33.164309 49.500000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 17.6412 DF 2 Pr > Chi Square 0.0001 Median Scores ( Number of Points Above Median) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 18 4.0 8.847458 1.783289 0.222222 CP 37 21.0 18.186441 1.872862 0.567568 UM 4 4.0 1.966102 0.973655 1.000000 Average scores were used for ties.

Median One Way Analysis Chi Square 10.0440 DF 2 Pr > Chi Square 0.0066 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 35 of 54 NONPARAMETRIC COMPARISON OF GONADOSOMATIC INDEX ( GSI) BETWEEN SITES 70  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 56.50 76.50 9.440273 6.277778 CP 7 79.50 59.50 9.440273 11.357143 Average scores were used for ties.

Wilcoxon Two Sample Test Statistic 79.5000 Normal Approximation Z 2.0656 One Sided Pr > Z 0.0194 Two Sided Pr > | Z| 0.0389 t Approximation One Sided Pr > Z 0.0283 Two Sided Pr > | Z| 0.0566 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 4.4884 DF 1 Pr > Chi Square 0.0341 Median Scores ( Number of Points Above Median) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 2.0 4.50 1.024695 0.222222 CP 7 6.0 3.50 1.024695 0.857143 Average scores were used for ties.

Median Two Sample Test Statistic 6.0000 Z 2.4398 One Sided Pr > Z 0.0073 Two Sided Pr > | Z| 0.0147 Median One Way Analysis Chi Square 5.9524 DF 1 Pr > Chi Square 0.0147 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 36 of 54 NONPARAMETRIC COMPARISON OF GONADOSOMATIC INDEX ( GSI) BETWEEN SITES 72  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 524.50 594.0 55.289008 23.840909 CP 11 423.00 297.0 45.505889 38.454545 UM 20 483.50 540.0 54.389940 24.175000 Average scores were used for ties.

Kruskal Wallis Test Chi Square 7.6716 DF 2 Pr > Chi Square 0.0216 Median Scores ( Number of Points Above Median) for Variable GSI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 10.00 10.792453 1.758432 0.454545 CP 11 8.50 5.396226 1.447286 0.772727 UM 20 7.50 9.811321 1.729838 0.375000 Average scores were used for ties.

Median One Way Analysis Chi Square 4.8749 DF 2 Pr > Chi Square 0.0874 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 37 of 54 ANOVA FOR CONDITION INDEX BETWEEN SITES 74  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 Dependent Variable: CI Sum of Source DF Squares Mean Square F Value Pr > F Model 2 2.87929138 1.43964569 6.71 0.0024 Error 57 12.22812021 0.21452842 Corrected Total 59 15.10741159 R Square Coeff Var Root MSE CI Mean 0.190588 38.52713 0.463172 1.202197 Source DF Anova SS Mean Square F Value Pr > F site 2 2.87929138 1.43964569 6.71 0.0024 Levene's Test for Homogeneity of CI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 0.6135 0.3067 3.66 0.0320 Error 57 4.7813 0.0839 Bartlett's Test for Homogeneity of CI Variance Source DF Chi Square Pr > ChiSq site 2 4.3397 0.1142 Dunnett's t Tests for CI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 57 Error Mean Square 0.214528 Critical Value of Dunnett's t 2.28365 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits UM  BG 0.8712 0.2893 1.4531 *** CP  BG 0.3133 0.0148 0.6119 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 38 of 54 ANOVA FOR CONDITION INDEX BETWEEN SITES 78  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 Dependent Variable: CI Sum of Source DF Squares Mean Square F Value Pr > F Model 1 8.11930253 8.11930253 2.32 0.1499 Error 14 48.97043484 3.49788820 Corrected Total 15 57.08973737 R Square Coeff Var Root MSE CI Mean 0.142220 118.0374 1.870264 1.584467 Source DF Anova SS Mean Square F Value Pr > F site 1 8.11930253 8.11930253 2.32 0.1499 Levene's Test for Homogeneity of CI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 183.9 183.9 1.95 0.1844 Error 14 1320.7 94.3380 Bartlett's Test for Homogeneity of CI Variance Source DF Chi Square Pr > ChiSq site 1 23.5316 <. 0001 Dunnett's t Tests for CI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 14 Error Mean Square 3.497888 Critical Value of Dunnett's t 2.14485 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 1.4360  0.5856 3.4576 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 39 of 54 ANOVA FOR CONDITION INDEX BETWEEN SITES 82  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 Dependent Variable: CI Sum of Source DF Squares Mean Square F Value Pr > F Model 2 22.6025084 11.3012542 2.26 0.1149 Error 50 250.0105867 5.0002117 Corrected Total 52 272.6130951 R Square Coeff Var Root MSE CI Mean 0.082911 151.8343 2.236115 1.472734 Source DF Anova SS Mean Square F Value Pr > F site 2 22.60250839 11.30125420 2.26 0.1149 Levene's Test for Homogeneity of CI Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 1898.9 949.5 0.93 0.4022 Error 50 51177.9 1023.6 Bartlett's Test for Homogeneity of CI Variance Source DF Chi Square Pr > ChiSq site 2 122.7 <. 0001 Dunnett's t Tests for CI NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 50 Error Mean Square 5.000212 Critical Value of Dunnett's t 2.28707 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits UM  BG 1.4305  0.1496 3.0105 CP  BG 0.3156  1.5730 2.2041 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 40 of 54 NONPARAMETRIC COMPARISON OF CONDITION INDEX ( CI) BETWEEN SITES 86  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 393.0 579.50 62.927869 20.684211 CP 37 1253.0 1128.50 65.771701 33.864865 UM 4 184.0 122.00 33.744135 46.000000 Kruskal Wallis Test Chi Square 10.5265 DF 2 Pr > Chi Square 0.0052 Median Scores ( Number of Points Above Median) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 19 4.0 9.50 1.816823 0.210526 CP 37 23.0 18.50 1.898929 0.621622 UM 4 3.0 2.00 0.974245 0.750000 Median One Way Analysis Chi Square 9.3983 DF 2 Pr > Chi Square 0.0091 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 41 of 54 NONPARAMETRIC COMPARISON OF CONDITION INDEX ( CI) BETWEEN SITES 88  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 60.0 76.50 9.447222 6.666667 CP 7 76.0 59.50 9.447222 10.857143 Wilcoxon Two Sample Test Statistic 76.0000 Normal Approximation Z 1.6936 One Sided Pr > Z 0.0452 Two Sided Pr > | Z| 0.0903 t Approximation One Sided Pr > Z 0.0555 Two Sided Pr > | Z| 0.1110 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 3.0504 DF 1 Pr > Chi Square 0.0807 Median Scores ( Number of Points Above Median) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 3.0 4.50 1.024695 0.333333 CP 7 5.0 3.50 1.024695 0.714286 Median Two Sample Test Statistic 5.0000 Z 1.4639 One Sided Pr > Z 0.0716 Two Sided Pr > | Z| 0.1432 Median One Way Analysis Chi Square 2.1429 DF 1 Pr > Chi Square 0.1432 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 42 of 54 NONPARAMETRIC COMPARISON OF CONDITION INDEX ( CI) BETWEEN SITES 90  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 349.0 594.0 55.398556 15.863636 CP 11 305.0 297.0 45.596052 27.727273 UM 20 777.0 540.0 54.497706 38.850000 Kruskal Wallis Test Chi Square 23.2397 DF 2 Pr > Chi Square <. 0001 Median Scores ( Number of Points Above Median) for Variable CI Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 1.0 10.792453 1.810436 0.045455 CP 11 7.0 5.396226 1.490088 0.636364 UM 20 18.0 9.811321 1.780996 0.900000 Median One Way Analysis Chi Square 31.1926 DF 2 Pr > Chi Square <. 0001 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 43 of 54 ANOVA FOR E2/ T RATIO BETWEEN SITES 92  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 NOTE: Due to missing values, only 42 observations can be used in this analysis.

Dependent Variable: RATIO Sum of Source DF Squares Mean Square F Value Pr > F Model 1 21.055872 21.055872 0.15 0.6999 Error 40 5587.442993 139.686075 Corrected Total 41 5608.498865 R Square Coeff Var Root MSE RATIO Mean 0.003754 161.1670 11.81889 7.333315 Source DF Anova SS Mean Square F Value Pr > F site 1 21.05587231 21.05587231 0.15 0.6999 Levene's Test for Homogeneity of RATIO Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 53436.8 53436.8 0.32 0.5745 Error 40 6670870 166772 Bartlett's Test for Homogeneity of RATIO Variance Source DF Chi Square Pr > ChiSq site 1 1.4679 0.2257 Dunnett's t Tests for RATIO NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 40 Error Mean Square 139.6861 Critical Value of Dunnett's t 2.02111 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits CP  BG 1.478  6.215 9.170 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 44 of 54 ANOVA FOR E2/ T RATIO BETWEEN SITES 96  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 NOTE: Due to missing values, only 15 observations can be used in this analysis.

Dependent Variable: RATIO Sum of Source DF Squares Mean Square F Value Pr > F Model 1 3.54538874 3.54538874 0.57 0.4636 Error 13 80.81750386 6.21673107 Corrected Total 14 84.36289260 R Square Coeff Var Root MSE RATIO Mean 0.042025 100.0048 2.493337 2.493217 Source DF Anova SS Mean Square F Value Pr > F site 1 3.54538874 3.54538874 0.57 0.4636 Levene's Test for Homogeneity of RATIO Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 7.8877 7.8877 0.13 0.7221 Error 13 776.1 59.6964 Bartlett's Test for Homogeneity of RATIO Variance Source DF Chi Square Pr > ChiSq site 1 0.0674 0.7951 Dunnett's t Tests for RATIO NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 13 Error Mean Square 6.216731 Critical Value of Dunnett's t 2.16042 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG  0.9924  3.8314 1.8466 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 45 of 54 ANOVA FOR E2/ T RATIO BETWEEN SITES 100  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 NOTE: Due to missing values, only 42 observations can be used in this analysis.

Dependent Variable: RATIO Sum of Source DF Squares Mean Square F Value Pr > F Model 2 6.0121986 3.0060993 0.88 0.4222 Error 39 132.9691436 3.4094652 Corrected Total 41 138.9813422 R Square Coeff Var Root MSE RATIO Mean 0.043259 132.2498 1.846474 1.396201 Source DF Anova SS Mean Square F Value Pr > F site 2 6.01219864 3.00609932 0.88 0.4222 Levene's Test for Homogeneity of RATIO Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 148.8 74.3982 0.47 0.6311 Error 39 6230.2 159.7 Bartlett's Test for Homogeneity of RATIO Variance Source DF Chi Square Pr > ChiSq site 2 9.7484 0.0076 Dunnett's t Tests for RATIO NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control. Alpha 0.05 Error Degrees of Freedom 39 Error Mean Square 3.409465 Critical Value of Dunnett's t 2.31338 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits CP  BG  0.3223  2.0859 1.4413 UM  BG  0.8799  2.4128 0.6531 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 46 of 54 NONPARAMETRIC COMPARISON OF E2/ T RATIO BETWEEN SITES 104  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 15 269.0 322.50 38.095275 17.933333 CP 27 634.0 580.50 38.095275 23.481481 Wilcoxon Two Sample Test Statistic 269.0000 Normal Approximation Z  1.3912 One Sided Pr < Z 0.0821 Two Sided Pr > | Z| 0.1642 t Approximation One Sided Pr < Z 0.0858 Two Sided Pr > | Z| 0.1717 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.9723 DF 1 Pr > Chi Square 0.1602 Median Scores ( Number of Points Above Median) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 15 5.0 7.50 1.571468 0.333333 CP 27 16.0 13.50 1.571468 0.592593 Median Two Sample Test Statistic 5.0000 Z  1.5909 One Sided Pr < Z 0.0558 Two Sided Pr > | Z| 0.1116 Median One Way Analysis Chi Square 2.5309 DF 1 Pr > Chi Square 0.1116 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 47 of 54 NONPARAMETRIC COMPARISON OF E2/ T RATIO BETWEEN SITES 106  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 83.0 72.0 8.485281 9.222222 CP 6 37.0 48.0 8.485281 6.166667 Wilcoxon Two Sample Test Statistic 37.0000 Normal Approximation Z  1.2374 One Sided Pr < Z 0.1080 Two Sided Pr > | Z| 0.2159 t Approximation One Sided Pr < Z 0.1181 Two Sided Pr > | Z| 0.2363 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 1.6806 DF 1 Pr > Chi Square 0.1949 Median Scores ( Number of Points Above Median) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 5.0 4.20 0.979796 0.555556 CP 6 2.0 2.80 0.979796 0.333333 Median Two Sample Test Statistic 2.0000 Z  0.8165 One Sided Pr < Z 0.2071 Two Sided Pr > | Z| 0.4142 Median One Way Analysis Chi Square 0.6667 DF 1 Pr > Chi Square 0.4142 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 48 of 54 NONPARAMETRIC COMPARISON OF E2/ T RATIO BETWEEN SITES 108  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 542.0 473.0 39.707262 24.636364 CP 8 157.0 172.0 31.219652 19.625000 UM 12 204.0 258.0 35.916570 17.000000 Kruskal Wallis Test Chi Square 3.2394 DF 2 Pr > Chi Square 0.1980 Median Scores ( Number of Points Above Median) for Variable RATIO Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 22 14.0 11.0 1.637964 0.636364 CP 8 4.0 4.0 1.287842 0.500000 UM 12 3.0 6.0 1.481594 0.250000 Median One Way Analysis Chi Square 4.5260 DF 2 Pr > Chi Square 0.1040 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 49 of 54 ANOVA FOR VITELLOGENIN ( P_ mg) BETWEEN SITES 110  Sex= F

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 60 NOTE: Due to missing values, only 20 observations can be used in this analysis.

Dependent Variable: P_ mg Sum of Source DF Squares Mean Square F Value Pr > F Model 2 11649.5373 5824.7687 0.32 0.7294 Error 17 308087.4127 18122.7890 Corrected Total 19 319736.9500 R Square Coeff Var Root MSE P_ mg Mean 0.036435 15.97021 134.6209 842.9500 Source DF Anova SS Mean Square F Value Pr > F site 2 11649.53730 5824.76865 0.32 0.7294 Levene's Test for Homogeneity of P_ mg Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 33916350 16958175 0.05 0.9474 Error 17 5.3185E9 3.1285E8 Bartlett's Test for Homogeneity of P_ mg Variance Source DF Chi Square Pr > ChiSq site 2 0.0751 0.9631 Dunnett's t Tests for P_ mg NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 17 Error Mean Square 18122.79 Critical Value of Dunnett's t 2.42696 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits CP  BG 54.37  110.29 219.02 UM  BG 25.72  170.61 222.06 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 50 of 54 ANOVA FOR VITELLOGENIN ( P_ mg) BETWEEN SITES 114  Sex= I

The ANOVA Procedure Class Level Information Class Levels Values site 2 BG CP Number of observations 16 NOTE: Due to missing values, only 14 observations can be used in this analysis. Dependent Variable: P_ mg Sum of Source DF Squares Mean Square F Value Pr > F Model 1 532.1488 532.1488 0.04 0.8535 Error 12 179378.2083 14948.1840 Corrected Total 13 179910.3571 R Square Coeff Var Root MSE P_ mg Mean 0.002958 15.80060 122.2628 773.7857 Source DF Anova SS Mean Square F Value Pr > F site 1 532.1488095 532.1488095 0.04 0.8535 Levene's Test for Homogeneity of P_ mg Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 1 75518806 75518806 0.30 0.5920 Error 12 2.9897E9 2.4914E8 Bartlett's Test for Homogeneity of P_ mg Variance Source DF Chi Square Pr > ChiSq site 1 0.1445 0.7038 Dunnett's t Tests for P_ mg NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 12 Error Mean Square 14948.18 Critical Value of Dunnett's t 2.17886 Comparisons significant at the 0.05 level are indicated by ***.

Difference Simultaneous site Between 95% Confidence Comparison Means Limits CP  BG  12.46  156.33 131.41 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 51 of 54 ANOVA FOR VITELLOGENIN ( P_ mg) BETWEEN SITES 118  Sex= M

The ANOVA Procedure Class Level Information Class Levels Values site 3 BG CP UM Number of observations 53 NOTE: Due to missing values, only 24 observations can be used in this analysis.

Dependent Variable: P_ mg Sum of Source DF Squares Mean Square F Value Pr > F Model 2 182143.0833 91071.5417 8.25 0.0023 Error 21 231848.7500 11040.4167 Corrected Total 23 413991.8333 R Square Coeff Var Root MSE P_ mg Mean 0.439968 27.98847 105.0734 375.4167 Source DF Anova SS Mean Square F Value Pr > F site 2 182143.0833 91071.5417 8.25 0.0023 Levene's Test for Homogeneity of P_ mg Variance ANOVA of Squared Deviations from Group Means Sum of Mean Source DF Squares Square F Value Pr > F site 2 5.3083E8 2.6541E8 0.75 0.4831 Error 21 7.399E9 3.5233E8 Bartlett's Test for Homogeneity of P_ mg Variance Source DF Chi Square Pr > ChiSq site 2 2.8489 0.2406 Dunnett's t Tests for P_ mg NOTE: This test controls the Type I experimentwise error for comparisons of all treatments against a control.

Alpha 0.05 Error Degrees of Freedom 21 Error Mean Square 11040.42 Critical Value of Dunnett's t 2.37033 Minimum Significant Difference 124.53 Comparisons significant at the 0.05 level are indicated by ***.

Difference site Between Simultaneous 95% Comparison Means Confidence Limits CP  BG 51.63  72.90 176.15 UM  BG  153.50  278.03  28.97 *** Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 52 of 54 NONPARAMETRIC COMPARISON OF VITELLOGENIN ( P_ mg) BETWEEN SITES 122  Sex= F

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 84.0 94.50 13.162447 9.333333 CP 7 81.0 73.50 12.619429 11.571429 UM 4 45.0 42.00 10.583005 11.250000 Kruskal Wallis Test Chi Square 0.6439 DF 2 Pr > Chi Square 0.7247 Median Scores ( Number of Points Above Median) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 9 4.0 4.50 1.141329 0.444444 CP 7 4.0 3.50 1.094243 0.571429 UM 4 2.0 2.00 0.917663 0.500000 Median One Way Analysis Chi Square 0.2413 DF 2 Pr > Chi Square 0.8864 Data Evaluation Report on the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 53 of 54 NONPARAMETRIC COMPARISON OF VITELLOGENIN ( P_ mg) BETWEEN SITES 124  Sex= I

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 8 61.0 60.0 7.745967 7.625000 CP 6 44.0 45.0 7.745967 7.333333 Wilcoxon Two Sample Test Statistic 44.0000 Normal Approximation Z  0.0645 One Sided Pr < Z 0.4743 Two Sided Pr > | Z| 0.9485 t Approximation One Sided Pr < Z 0.4748 Two Sided Pr > | Z| 0.9495 Z includes a continuity correction of 0.5.

Kruskal Wallis Test Chi Square 0.0167 DF 1 Pr > Chi Square 0.8973 Median Scores ( Number of Points Above Median) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 8 4.0 4.0 0.960769 0.50 CP 6 3.0 3.0 0.960769 0.50 Median Two Sample Test Statistic 3.0000 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 the Reconnaissance Survey of South Florida Amphibians for the Assessment of Potential Atrazine Effects EPA MRID Number 458677 06 Page 54 of 54 NONPARAMETRIC COMPARISON OF VITELLOGENIN ( P_ mg) BETWEEN SITES 126  Sex= M

The NPAR1WAY Procedure Wilcoxon Scores ( Rank Sums) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 8 117.0 100.0 16.329932 14.6250 CP 8 138.0 100.0 16.329932 17.2500 UM 8 45.0 100.0 16.329932 5.6250 Kruskal Wallis Test Chi Square 11.8950 DF 2 Pr > Chi Square 0.0026 The NPAR1WAY Procedure Median Scores ( Number of Points Above Median) for Variable P_ mg Classified by Variable site Sum of Expected Std Dev Mean site N Scores Under H0 Under H0 Score ââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââââ BG 8 5.0 4.0 1.179536 0.6250 CP 8 7.0 4.0 1.179536 0.8750 UM 8 0.0 4.0 1.179536 0.0000 Median One Way Analysis Chi Square 12.4583 DF 2 Pr > Chi Square 0.0020


[Federal Register: December 28, 2007 (Volume 72, Number 248)]
[Notices]
[Page 73937-73939]
[DOCID:fr28de07-194]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-57019; File No. SR-ISE-2007-120]

Self-Regulatory Organizations; International Securities Exchange,
LLC; Notice of Filing and Immediate Effectiveness of Proposed Rule
Change Relating to Rule 710, Minimum Trading Increments

December 20, 2007.
    Pursuant to Section 19(b)(1) of the Securities Exchange Act of 1934
(``Act'')\1\ and Rule 19b-4 thereunder,\2\ notice is hereby given that
on December 17, 2007, the International Securities Exchange, LLC
(``ISE'' or ``Exchange'') filed with the Securities and Exchange
Commission (``Commission'') the proposed rule change as described in
Items I and II below, which Items have been substantially prepared by
the Exchange. The Exchange filed the proposal pursuant to Section
19(b)(3)(A) of the Act\3\ and Rule 19b-4(f)(6) thereunder,\4\ which
renders the proposal effective upon filing with the Commission. The
Commission is publishing this notice to solicit comments on the
proposed rule change from interested persons.
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    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
    \3\ 15 U.S.C. 78s(b)(3)(A).
    \4\ 17 CFR 240.19b-4(f)(6).
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I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    ISE proposes to amend Rule 710, Minimum Trading Increments, to
decrease the size of the minimum quoting and trading increments
applicable to the Exchange's foreign currency options (``FX options'').
The text of the proposed rule change is available at ISE, the
Commission's Public Reference Room, and http://www.ise.com.

II. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

    In its filing with the Commission, the Exchange included statements
concerning the purpose of, and basis for, the proposed rule change and
discussed any comments it received on the proposed rule change. The
text of these statements may be examined at the places specified in
Item IV below. ISE has prepared summaries, set forth in Sections A, B,
and C below, of the most significant aspects of such statements.

A. Self-Regulatory Organization's Statement of the Purpose of, and the
Statutory Basis for, the Proposed Rule Change

1. Purpose
    ISE proposes to amend its Rule 710, Minimum Trading Increments, to
decrease the size of the minimum quoting and trading increments
applicable to the Exchange's FX options.\5\ The Exchange believes that
by reducing the minimum trading increments applicable to ISE's FX
options, the proposed rule change will provide market participants with
additional trading opportunities in this product. Further, quoting and
trading in smaller increments will enable market participants to trade
FX options with greater precision as to price.
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    \5\ The Exchange began trading FX options on the Euro, the
British pound, the Japanese yen and the Canadian dollar on April 17,
2007. See Securities Exchange Act Release No. 55575 (April 3, 2007),
72 FR 17963 (April 10, 2007) (approving SR-ISE-2006-59).
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    Currently, FX options traded on the Exchange have minimum
increments of $0.05 or $0.10 depending on the price at which an FX
option is quoting. Specifically, under the Exchange's current rules,
the minimum trading increment for an FX options contract trading at
less than $3.00 is $0.05, and for an FX options contract trading at
$3.00 or higher, the minimum trading increment is $0.10. The proposed
amendment to Rule 710 would set the minimum increment for all FX
options at $0.01 regardless of the price at which the option is
quoting. Although FX options would be trading in these narrower
increments, they would not actually be trading in pennies\6\ and would
not be considered part of the Exchange's pilot program currently
applicable to certain equity options.\7\
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    \6\ The Exchange notes that ISE's FX options have underlying
values that modify the magnitude of traditionally quoted exchange
rates that appear in the underlying foreign currency markets. As a
result, the ``rate-modified'' FX options traded on the Exchange are
quoted to reflect the sub-penny movements in the actual exchange
rate of any underlying currency. Since all premiums in ISE's FX
options are quoted in U.S. Dollars, customers will be able to trade
this product in one-cent increments. Thus, while the Exchange's
proposal seeks to set the minimum increment for all FX options at
$0.01, the quoted values reflect much smaller currency increments
with respect to the exchange rate of the underlying currency.
    \7\ The penny pilot, which permits certain options series to be
quoted and traded in increments of $0.01, began on January 26, 2007.
See Securities Exchange Act Release No. 55161 (January 24, 2007), 72
FR 4754 (February 1, 2007) (approving SR-ISE-2006-62). The penny
pilot was extended through September 27, 2007. See Securities
Exchange Act Release No. 56151 (July 26, 2007), 72 FR 42452 (August
2, 2007) (approving SR-ISE-2007-68). The penny pilot has been
extended again through March 27, 2009. See Securities Exchange Act
Release No. 56564 (September 27, 2007), 72 FR 56412 (October 3,
2007) (approving SR-ISE-2007-74). With one exception, all series in
options included in the penny pilot trading at a price of less than
$3.00 are currently quoted and traded in minimum increments of
$0.01, and those with a price of $3.00 or higher are currently
quoted and traded in minimum increments of $0.05. A list of the
options to be included in the penny pilot was communicated to the
Exchange's members via a Regulatory Information Circular.
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    Currently, options on currency futures trade in these smaller
increments on the Chicago Mercantile Exchange. Also, currencies trade
on the cash market in these smaller increments. Further, the Commission
recently approved a proposed rule change by the Philadelphia Stock
Exchange (``Phlx'') permitting that exchange to trade its U.S. dollar-
settled foreign currency options in $0.01 increments.\8\ As a
competitive matter, ISE seeks the opportunity to offer market
participants those same, more refined increments. The Exchange notes
that providing these more refined increments will permit the Exchange's
market makers the opportunity to provide better fills (meaning less
spread than the current wider minimum increments rules allow) to
customers.
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    \8\ See Securities Exchange Act Release No. 56933 (December 7,
2007), 72 FR 71185 (December 14, 2007) (approving SR-Phlx-2007-70).
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2. Statutory Basis
    The Exchange believes that the proposed rule change is consistent
with Section 6(b) of the Act in general and furthers the objectives of
Section 6(b)(5) in particular, in that it is designed to prevent
fraudulent and manipulative acts and practices, to promote just and
equitable principles of trade, 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.

B. Self-Regulatory Organization's Statement on Burden on Competition

    The Exchange does not believe that the proposed rule change will
impose any burden on competition that is not necessary or appropriate
in furtherance of the purposes of the Act.

C. Self-Regulatory Organization's Statement on Comments on the Proposed
Rule Change Received from Members, Participants or Others

    No written comments were solicited or received with respect to the
proposed rule change.

III. Date of Effectiveness of the Proposed Rule Change and Timing for
Commission Action

    Because the foregoing proposed rule change does not:
    (i) Significantly affect the protection of investors or the public
interest;
    (ii) Impose any significant burden on competition; and
    (iii) Become operative for 30 days from the date on which it was
filed, or such shorter time as the Commission may designate, if
consistent with the protection of investors and the public interest, it
has become effective pursuant to Section 19(b)(3)(A) of the Act and
Rule 19b-4(f)(6) thereunder. As required under Rule 19b-4(f)(6)(iii)
under the Act, the Exchange provided the Commission with written notice
of its intent to file the proposed rule change, along with a brief
description and text of the proposed rule change, at least five
business days prior to the date of the filing of the proposed rule
change.
    A proposed rule change filed under 19b-4(f)(6) normally may not
become operative prior to 30 days after the date of filing. However,
Rule 19b-4(f)(6)(iii) permits the Commission to designate a shorter
time if such action is consistent with the protection of investors and
the public interest. The Exchange has requested that the Commission
waive the 30-day operative delay and render the proposed rule change to
become operative on January 2, 2008.\9\ The Commission believes that
waiving the 30-day operative delay is consistent with the protection of
investors and the public interest. Waiver of the 30-day operative delay
would enable the Exchange to start trading FX options in the same
increments and at the same time as Phlx. For the reasons stated above,
the Commission therefore designates the proposal to become operative
immediately.
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    \9\ The Exchange also may decide to start using these smaller
trading increments later than January 2, 2007. Telephone
conversation between Samir M. Patel, Assistant General Counsel, ISE,
and Natasha Cowen, Special Counsel, Division of Trading and Markets,
Commission, dated December 19, 2007.
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    At any time within 60 days of the filing of the proposed rule
change, the Commission may summarily abrogate

such rule change if it appears to the Commission that such action is
necessary or appropriate in the public interest, for the protection of
investors, or otherwise in furtherance of the purposes of the Act.

IV. Solicitation of Comments

    Interested persons are invited to submit written data, views, and
arguments concerning the foregoing, including whether the proposed rule
change is consistent with the Act. Comments may be submitted by any of
the following methods:

Electronic Comments

     Use the Commission's Internet comment form (http://www.sec.gov/rules/sro.shtml.
 ); or     Send an e-mail to<EMAIL_ADDRESS>Please include

File Number SR-ISE-2007-120 on the subject line.

Paper Comments

     Send paper comments in triplicate to Nancy M. Morris,
Secretary, Securities and Exchange Commission, 100 F Street, NE.,
Washington, DC 20549-1090.

All submissions should refer to File Number SR-ISE-2007-120. This file
number should be included on the subject line if e-mail is used. To
help the Commission process and review your comments more efficiently,
please use only one method. The Commission will post all comments on
the Commission's Internet Web site (http://www.sec.gov/rules/sro.shtml
). Copies of the submission, all subsequent amendments, all

written statements with respect to the proposed rule change that are
filed with the Commission, and all written communications relating to
the proposed rule change between the Commission and any person, other
than those that may be withheld from the public in accordance with the
provisions of 5 U.S.C. 552, will be available for inspection and
copying in the Commission's Public Reference Room, 100 F Street, NE.,
Washington, DC 20549, on official business days between the hours of 10
a.m. and 3 p.m. Copies of such filing also will be available for
inspection and copying at the principal office of ISE. All comments
received will be posted without change; the Commission does not edit
personal identifying information from submissions. You should submit
only information that you wish to make available publicly. All
submissions should refer to File Number SR-ISE-2007-120 and should be
submitted on or before January 18, 2008.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\10\
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    \10\ 17 CFR 200.30-3(a)(12).
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Florence E. Harmon,
Deputy Secretary.
[FR Doc. E7-25208 Filed 12-27-07; 8:45 am]

BILLING CODE 8011-01-P